http://link.springer.com/journal/volumesAndIssues/13765

Sungryung Kim, Hongseok Oh, Woon-Seok Yeo. Analysis of alkanethiolates on gold with matrix-assisted laser desorption/ionization time-of-fl ight mass spectrometry. J. Korean Soc. Appl. Biol. Chem. 58(1):1-8Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been effectively utilized in the analysis of self-assembled monolayers (SAMs) on gold, which comprise a broad range of research activities at interfaces, including chip-based sensors, various assay and drug delivery systems, and basic cell research. This review provides an overview of the most recent literatures regarding the use of MALDI-TOF MS as a tool for the surface analysis of SAMs on gold, with a particular focus on bioanalytical applications. Several seminal papers are also briefly discussed.

Ji-Yeon Yang, Sang-Min Lee, Hoi-Seon Lee. Phototactic behavior 6: behavioral responses of Myzus persicae (Hemiptera: Aphididae) to light-emitting diodes. J. Korean Soc. Appl. Biol. Chem. 58(1):9-12The behavioral responses of Myzus persicae adults to light-emitting diodes (LEDs) were evaluated under optimal conditions (luminance intensities, light exposure times, and specific wavelengths). Based on the attraction rate (%) of M. persicae adults under optimal conditions, the green LED (520 ± 5 nm) showed the highest potential attraction rate (85.3 %), followed by the blue (470 ± 10 nm, 75.0 %), yellow (590 ± 5 nm, 73.7 %), and red LED (625 ± 10 nm, 69.7 %). The green LED (520 ± 5 nm) was approximately 1.4 times (61.7 %) more attractive than that of a black light bulb, which served as the positive control. In conclusion, the green LED (520 ± 5 nm) was suitable for attracting M. persicae adults under optimal conditions (40 lx and 120 min).

Yeonjoong Yong, Soon Young Shin, Yearam Jung, Hyeryoung Jung, Seunghyun Ahn, Youhoon Chong, Yoongho Lim. Flavonoids activating adenosine monophosphate-activated protein kinase. J. Korean Soc. Appl. Biol. Chem. 58(1):13-19Eighteen flavonoids were tested as adenosine monophosphate-activated protein kinase (AMPK) activators. The AMPK activation induced by kaempferide was 1.75 times better than the value obtained at 100 % AMPK phosphorylation. Western blotting analysis of kaempferide demonstrated that it increased the phosphorylation of AMPK in vitro. In silico docking results carried out to elucidate the binding mode between kaempferide and AMPK showed the docking pose of kaempferide in the binding site of AMPK was similar to that of the substrate contained in the crystallographic structure of AMPK. Flavonoids, including kaempferide, are promising potential AMPK activators.

Da Yeon Kim, So-Hyun Kim, Hye Min Ahn, Sa Rang Lim, Junsang Oh, Seulgi Choi,Hong-Jin Le, Joong-Hyuck Auh, Hyung-Kyoon Choi. Differentiation of highbush blueberry (Vaccinium corymbosum L.) fruit cultivars by GC–MS-based metabolic profiling. J. Korean Soc. Appl. Biol. Chem. 58(1):21-28Highbush blueberries (Vaccinium corymbosum L.) are cultivated worldwide for their fruit with unique taste and potential health benefits. Blueray, Bluecrop, and Spartan are prominent among the various blueberry cultivars. We performed gas chromatography–mass spectrometry (GC–MS)-based metabolic profiling to differentiate the fruits of these three cultivars, and built an optimal partial least squares-discriminant analysis (PLS-DA) model to separate them. Amino acids, fatty acids, organic acids, phenolic compounds, and sugars were identified in the fruits. Theoptimized PLS-DA model for different cultivars of the fruits was obtained by selecting variables based on a variable importance in the projection (VIP) cut-off value of 1.0. Caffeic acid, aspartic acid, acetic acid, threonolactone, inositol, xylose, glucoside, linolenic acid, mannose, altrose, glycine alanine, and valine were found to be relevant and contributing compounds for differentiating cultivars. In addition, a hierarchical cluster analyses dendrogram pattern was correlated with the PLS-DA. This study

suggested that GC–Msbased metabolic profiling coupled with multivariate statistical analysis could be used to differentiate the fruits of three major highbush blueberry cultivars.

Dong Uk Kim, Hee Jung Kim, Yu Seok Jeong, Han Beur Na, Young-Lok Cha, Bon-Cheol Koo, Jungho Kim, Han Dae Yun, Jung-Kul Lee, Hoon Kim. Enhanced saccharification of reed and rice straws by the addition of β-1,3-1,4-glucanase with broad substrate specificity and calcium Ion. J. Korean Soc. Appl. Biol. Chem. 58(1):29-33The possibility of using additive enzymes to improve the saccharification of lignocellulosic substrates with commercial cellulolytic enzymes was studied. Reed (Phragmites communis) and rice (Oryza sativa) straw powders were pretreated with NaOH/steam via a hightemperatureexplosion system. The saccharification of untreated reed and rice straw powders by commercialenzymes (Celluclast 1.5 L + Novozym 188) was not significantly increased by the addition of xylanases (Xyn10J, XynX), a cellulase (Cel6H), and a β-1,3-1,4-glucanase (BGlc8H) with broad substrate specificity. The saccharification of the pretreated reed and rice straw powders by thecommercial enzymes was increased by 10.4 and 4.8 %, respectively, by the addition of BGlc8H. In the presence of Ca2+ and BGlc8H, the saccharification of the pretreated reed and rice straw powders by the commercial enzymes was increased by 18.5 and 11.7 %, respectively. No sucheffect of Ca2+ was observed with Xyn10J, XynX, or Cel6H. The results suggest that the enzymatic conversion of lignocellulosic biomass to reducing sugars could be enhanced by certain additive enzymes such as b-1,3-1,4-glucanase, and that the enhancement could further be increased by Ca2+.

Kee Sung Kyung , Ki Chang Ahn, Jeong Wook Kwon, Yong Pil Lee, Eun Young Lee, Yun Jeong Kim, Fritz Führ, Jae Koo Lee. Long-term fate of the herbicide mefenacet in a rice-grown lysimeter over a period of 6 consecutive years. J. Korean Soc. Appl. Biol. Chem. 58(1):35-43To show the long-term fate of the herbicide mefenacet in rice paddies, [aniline-14C]mefenacet was applied to a lysimeter of loamy soil with a depth of 1 m, and rice plants were cultivated for 6 consecutive years according to the conventional methods in Korea. Mineralization of [14C] mefenacet to 14CO2 and volatilization from the soil surface were 12.01 and 0.02 %, respectively, of the originally applied amount during the first 23 weeks following application. Throughout the 6-year period, the total proportion of 14C-radioactivity that leached through the lysimeter soil was 0.778 % of the original radioactivity. The total 14C-radioactivity absorbed and translocated by rice plants throughout the 6 years was 2.46 % of the applied 14C. Measurement of the 14C-radioactivity distributed in each part of the rice plant (the straw, the ears without rice grain, the chaff, and the brown rice grain) indicated that the amount of 14C in straw was 25.87 times higher than that in brown rice grain over the 6 years. The 14C-radioactivity remaining in the soil layer after 6 years was 44.58 % of that applied, 91.45 % of which was distributed in the upper 0–20-cm layer. These results strongly indicate that mefenacet moved downward very slowly and more than half of the herbicide applied was released into the air, mainly through mineralization to CO2 in soil during the experimental period.

J. K. Kim, E. C. Shin, H. G. Park. Fructooligosaccharides decreased the ability of probiotic Escherichia coli Nissle 1917 to adhere to co-cultures of human intestinal cell lines. J. Korean Soc. Appl. Biol. Chem. 58(1):45-52In the present study, we investigated effects of prebiotic fructooligosaccharides (FOS) on the ability of Escherichia coli Nissle 1917 (EcN), a probiotic bacterial strain, to adhere to co-cultures of human intestinal cells (Caco-2 and HT29-MTX). For the bacterial adhesion assay, the co-cultures were seeded together at a ratio of 3:1 and then incubated EcN in the presence of various prebiotics including FOS. To eliminate possible bias, both manual

and automated cell counting methods were utilized. In results, FOS significantly suppressed the adhesion abilities of probiotic EcN to the co-cultures. Taken altogether, we carefully suggest that an unbalanced supplementation of prebiotics may adversely impact on gut microflora; hence, further investigations are warranted in regards to such unexpected adverse effects and responsible mechanisms on colonization of probiotics in the gastrointestinal tract utilizing in vivo models.

Soyoung Lee, Han-Seung Shin. Effect of lotus seed and seedpod extracts on oxidative stability against lard during storage. J. Korean Soc. Appl. Biol. Chem. 58(1):53-60The total phenolic and total proanthocyanidin contents of six kinds of seed and seedpod extracts of lotus (Nelumbo nucifera Gaertner) were investigated. The inhibitory effects of the edible extracts including 80 % ethanol and water from lotus seeds (LS) and lotus seedpods (LSP) against lard were evaluated by determining acid value, peroxide value, and TBARS during storage at 10 °C for 14 days. Total phenolic contents of the LS water, LSP water, and LS 80 % ethanol extracts were 469.175, 313.842, and 359.008 mg CE/g dry weight, and the proanthocyanidin contents of the LSP water, LSP 80 % ethanol, and LS 80 % ethanol were 76.526, 68.526, and 40.896 mg CE/g dry weight at a concentration of 1.0 mg/mL, respectively. In a storage stability test, the LSP water fraction provided the antioxidant effect against lard. The high lipid oxidation inhibition effect of the LSP water fraction might be useful as a natural antioxidant in food.

Young Hwa Kim, Jae Woong Lee, Sungwook Chae, Sang Ho Moon, Eui Jeong Do, Seung Eun Oh, Gui Jun Zhang, Mi Young Lee. Development of a PCR-based assay to differentiate Cervus elaphus sibiricus from Cervus antlers. J. Korean Soc. Appl. Biol. Chem. 58(1):61-66Deer antlers (Cervi Parvum Cornu) are reported to possess various pharmacological properties, including anti-infective, anti-arthritic, anti-allergic, and anti-endometriotic properties, and are credited with reversing memory impairment. To determine the global distribution of Cervus subspecies by analyzing antlers, 166 samples of Cervus subspecies antlers were collected from Russia, Canada, China, New Zealand, and Korea. The respective deer subspecies were identified by amplifying the D-loop region of mitochondrial DNA isolated from the antler samples. On the basis of the mitochondrial DNA sequence, a C. elaphus sibiricus-specific primer was developed that differentiates C. e. sibiricus from four C. e. subspecies (C. e. manitobensis, C. e. nelsoni, C. e. canadensis, and C. e. bactrianus), C. elaphus, and C. nippon. This was confirmed using agarose gel electrophoresis. This primer set produced a specific 396-bp fragment that amplified only C. e. sibiricus in antler samples. A 468-bp fragment derived from the cytochrome b gene was used as the internal control to verify the success of amplification in all samples. This method may assist in rapid and effective differentiation between products from Cervus species and other deer products.

In Hee Cho. Volatile compounds of ginseng (Panaxsp.): a review. J. Korean Soc. Appl. Biol. Chem. 58(1):67-75The compositions of ginseng volatiles and their differences therein based on the species, cultivation age, and cultivation method are reviewed in this paper. Some sesquiterpene hydrocarbons (e.g., β-panasinsene, α-panasinsene, α-neoclovene, β-neoclovene, bicyclogermacrene, β-farnesene, aromadendrene, and (E)-caryophyllene) and sesquiterpene alcohols (e.g., (+)-spathulenol, ginsenol, panasinsenol A, and panasinsenol B) were reportedly the main volatile compounds of ginseng. The differences between ginseng species were mainly associated with sesquiterpene hydrocarbons and monoterpenes, such as α-selinene, α-terpinolene, β-bisabolene, β-phellandrene, β-sesquiphellandrene, zingiberene, germacrene D, limonene, α-gurjunene, (E)-caryophyllene, δ-cadinene, (E)-β-farnesene, α-humulene, bicyclogermacrene, longiborn-8-ene, β-neoclovene, and (+)-spathulenol. Also, the amounts of α-cadinol, α-bisabolol, thujopsene, and n-hexadecanoic acid significantly increased with the cultivation ages. Moreover, aromadendrene, calarene, β-panasinsene, (E)-caryophyllene, α-neoclovene, and α-farnensene contributed to the discrimination among ginsengs cultivated using

different methods.

Soon Young Shin , Yeonjoong Yong , Dae Sun Hong, Da Hyun Lee, Da Young Lee, Young Han Lee. Identification of flavonoids from Eriodictyon californicum and their cytotoxicity against HCT116 colon cancer cells. J. Korean Soc. Appl. Biol. Chem. 58(1):77-81The extract of Eriodictyon californicum has been widely used as an essential emollient, and as an anti-inflammatory, antibacterial, and antifungal agent. In this study, antitumor effects of E. californicum were evaluated against HCT116 human colon cancer cells. Treatment with the ethyl acetate extract of E. californicum (EEC) inhibited the short-term viability and long-term clonogenicity of HCT116 cells. We identified several flavonoids, including rosmarinic acid and luteolin, from the EEC by using high-performance liquid chromatography and nuclear magnetic resonance. Luteolin inhibited clonogenicity, activated caspase-7, and induced apoptosis of HCT116 cells. These findings suggest that EEC contains bioactive flavonoids that exhibit antitumor activities.

Nak-Bum Song, Ji-Hyun Lee, Kyung Bin Song. Preparation of perilla seed meal protein composite films containing various essential oils and their application in sausage packaging. J. Korean Soc. Appl. Biol. Chem. 58(1):83-90Perilla seed meal protein (PSMP) was extracted from perilla seed oil residue, and its composite films were developed using different amounts of red algae (RA) to prepare edible films. The PSMP composite film that contained 3 % RA had the highest tensile strength (11.52 MPa) and the lowest water vapor permeability (1.82 × 10−9 g m/m2 s Pa). The incorporation of clove oil into the PSMP composite film had the highest antimicrobial activity against Listeria monocytogenes. Pork sausages packaged with the composite films containing clove oil showed antimicrobial and antioxidative activities during storage at 4 °C. The composite film containing 1.2 % clove oil reduced the microbial growth by 1.24 log CFU/g compared to the control, and thiobarbituric acid reactive substances and peroxide values were reduced by 40 and 37 %, respectively. These results suggest that the PSMP/RA composite film containing 1.2 % clove oil can be utilized in sausage packaging.

Chinaka O. Nwaehujor , Rita I. Udegbunam, Julius O. Ode, Sunday O. Udegbunam. Analgesic anti-inflammatory anti-pyretic activities of Garcinia hydroxybiflavanonol (GB1) from Garcinia kola. J. Korean Soc. Appl. Biol. Chem. 58(1):91-96This study was conducted to evaluate the analgesic, anti-inflammatory, and anti-pyretic effects of GB1, a hydroxybiflavanonol of Garcinia kola. The analgesic effect was studied using acetic acid-induced writhing and tail withdrawal tests. Its anti-inflammatory effect was studied using carrageenan-induced paw edema and adjuvant-induced arthritis tests. Brewer’s yeast-induced pyrexia test was used to evaluate the anti-pyretic effect of GB1. GB1 reduced writhing induced by 0.6 % acetic acid. The number of writhing in 40- and 80-mg/kg GB1-treated groups were significantly lower than writhing in mice treated with distilled water. After 15-min latency period following oral GB1 (40 and 80 mg/kg), there was significant increase in tail withdrawal time compared to control rats. The tail withdrawal time at 30, 45, and 60 min were significantly prolonged in GB1 (20, 40, and 80 mg/kg) treated mice. GB1 (20, 40, and 80 mg/kg) groups had significantly lower paw volumes at 2 and 3 h post carrageenan injection. Also paw volume were significantly lower in GB1 (40 and 80 mg/kg) groups at 18 h post arthritis induction. By day 30 paw volumes of all GB1 groups were significantly lower than paw volume of control group. Rectal temperatures of GB1 (80 mg/kg)-treated group were significantly lower at 1, 2, 3, 4, and 5 h post brewer’s yeast injection compared to rectal temperatures of control and 20-mg/kg GB1 groups. These finding shows that GB1 may serve as an alternative to NSAIDs in clinical practice.

Hyun Ho Noh, Dae Kyu Kim, Eun Young Lee, Moon Ik Chang, Moo Hyeog Im, Young Deuk Lee, Kee Sung Kyung. Effects of oven drying on pesticide residues in field-grown chili peppers. J. Korean Soc. Appl. Biol. Chem. 58(1):97-104

The effects of drying on nine pesticides (chlorfenapyr, clothianidin, diethofencarb, folpet, imidacloprid, indoxacarb, methomyl, methoxyfenozide, and tetraconazole) in chili peppers were studied. The mean concentration factor calculated from weight reduction after drying by water loss was 5.28 for field 1 and 5.41 for field 2 (n = 10). Processing factors for pesticide residues in/on chili peppers after drying were 2.45–5.14 for field 1 and 1.71–4.53 for field 2, indicating that pesticide residues remaining in/on chili peppers were concentrated by usual drying process for pepper powder. The reduction factors reflecting net loss of pesticide residues during drying were 0.50–0.98 for field 1 and 0.44–0.98 for field 2. Drying caused a large reduction (37–49 %) in clothianidin, diethofencarb, imidacloprid, and tetraconazole; moderate reductions (16 and 22 %) in methomyl and methoxyfenozide; while did not affect the levels of chlorfenapyr, folpet, and indoxacarb (using conservative higher reduction factors from the two field trials).

Soon Young Shin, Mi So Lee, Da Hyun Lee, Da Young Lee, Dongsoo Koh, Young Han Lee. The synthetic compound 2′-hydroxy-2,4,6′-trimethoxychalcone overcomes P-glycoprotein-mediated multi-drug resistance in drug-resistant uterine sarcoma MES-SA/DX5 cells. J. Korean Soc. Appl. Biol. Chem. 58(1):105-109The antitumor activity of a novel synthetic chalcone derivative, 2′-hydroxy-2,4,6′-trimethoxychalcone (1H3MC), was evaluated using the multi-drug-resistant human uterine sarcoma MES-SA/Dx5 cells. Treatment with 1H3MC reduced P-glycoprotein expression in a time-dependent manner and inhibited MES-SA/Dx5 cell proliferation. Cisplatin alone had no effect on cell viability, but combined treatment with cisplatin and 1H3MC exhibited synergistic cytotoxicity. Furthermore, the combination of cisplatin and 1H3MC synergistically cleaved both caspase-3 and its substrate protein, poly(ADP-ribose) polymerase, which resulted in the fragmentation of genomic DNA, a hallmark of apoptosis. These results suggest that 1H3MC is a promising adjuvant agent for overcoming P-glycoprotein-mediated multi-drug resistance in cancer cells.

Chao Chen , Yumei Wang , Chun Su , Xinqing Zhao , Ming Li, Xiaowei Meng, Yingyu Jin, Seung-Hwan Yang, Yushu Ma, Dongzhi Wei, Joo-Won Suh. Antifungal activity of Streptomycesalbidoflavus L131 against the leaf mold pathogen Passalora fulva involves membrane leakage and oxidative damage. J. Korean Soc. Appl. Biol. Chem. 58(1):111-119Passalora fulva (or Fulvia fulva) is the causal microorganism of tomato leaf mold, the outbreak of which occurs worldwide in greenhouse especially when humidity is high. However, studies on antifungal agents of P. fulva are still very limited. In this study, a marine-derived Streptomycesalbidoflavus strain L131 showing potent inhibitory activities against P. fulva was identified and characterized. The active antifungal components were obtained, and studies on the antifungal mechanisms of the crude extract showed that the antifungal metabolites of L131 caused damage of hyphae and spore development, as well as plasma membrane of P. fulva. In addition, accumulation of endogenous reactive oxygen species of the leaf pathogen was also observed after treatment by culture extracts of L131. To our knowledge, this is the first report on the studies of the antifungal mechanisms against P. fulva, which benefit further development of biocontrol agent against tomato leaf mold disease.

Jun-Hwan Park, Hoi-Seon Lee. Antimicrobial activities of 1,2-dihydroxyanthraquinone derivatives against food-borne bacteria. J. Korean Soc. Appl. Biol. Chem. 58(1):121-125The antimicrobial activities of materials derived from Cassia obtusifolia seeds were evaluated against the seven food-borne bacteria. The active constituent of C. obtusifolia seeds was isolated using various chromatographic techniques and characterized as 1,2-dihydroxyanthraquinone (alizarin). The purified alizarin exhibited weak activity against Bacillus cereus (clean zone diameter, 11.0 mm), and moderate activity against Staphylococcus epidermidis (17.5 mm) and Salmonella enterica (16.2 mm) at 2.0 mg/disc. When compared with alizarin analogues, alizarin-3-methyliminodiacetic acid exhibited the strongest antimicrobial activity against B.

cereus, S. intermedius, S. epidermidis, and S. enterica at 2.0 mg/disc, whereas the other analogues exhibited no antimicrobial activity against the seven food-borne bacteria. Taken together, these results indicate that alizarin isolated from C. obtusifolia seeds and its structural analogues may be useful as natural preservatives.

Bou-Hee Kang, Kenneth Racicot, Sarah J. Pilkenton, Young-In Kwon, Emmanouil Apostolidis. Blueberry extract inhibits carbohydrate-hydrolyzing enzymes and these inhibitory activities are not proanthocyanidin dependent. J. Korean Soc. Appl. Biol. Chem. 58(1):127-136This study investigates the carbohydrate-hydrolyzing inhibitory potential of blueberry extract on carbohydrate-hydrolyzing enzymes and evaluates if the inhibitory activity is proanthocyanidin (PAC) or lower molecular weight phenolic dependent. Freeze-dried blueberry powder was extracted using acetone and subjected to C18 extraction (BAE). Low-molecular weight phenolics (BAE-LMW) and PACs (BAE-PAC) were separated from BAE with gel filtration chromatography using LH-20 column. Total phenolic content, PAC content, and phenolic profiles using HPLC, as well as rat α-glucosidase, sucrase, and maltase inhibitory activities, were determined for all samples. The rat alpha-glucosidase inhibitory activity of BAE (IC50 0.390 mg/mL TP basis) was enhanced in BAE-LMW (IC500.242 mg/mL TP basis) and reduced in BAE-PAC (IC50 0.915 mg/mL TP basis). Similar trends were observed with maltase and sucrase inhibitory activities. Our findings suggest that blueberry acetone extract has inhibitory activity on carbohydrate-hydrolyzing enzymes and this effect is dependent on LMWs rather than PAC.

Jung Yeon Kim, Ji Yeon Choi, Chang Yong Yoon, Sooyeul Cho, Woo Seong Kim, Jung Ah Do. LC–MS/MS monitoring of 22 illegal antihistamine compounds in health food products from the Korean market. J. Korean Soc. Appl. Biol. Chem. 58(1):137-147With the increasing popularity of dietary supplements, a critical analysis of safety issues concerning their use has become imperative. Despite several regulations and laws being in place, there have been several instances of adulterated health food-induced accidents. Therefore, in light of the growing seriousness of this problem, we attempted to detect the presence of 22 antihistamines in health foods sold in South Korea. 117 samples, representative of the various types of health foods, were screened by liquid chromatography with electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS). The limit of detections (LODs) and limit of quantifications (LOQs) of the instrument ranged from 0.0003 to 0.3 μg/mL and from 0.0009 to 0.6 μg/mL, respectively. The LODs and LOQs of the method ranged from 0.006 to 6.0 μg/mL and from 0.018 to 12.0 μg/mL, respectively. The calibration curve was linear with R 2 between 0.997 and 0.999. The mean recovery efficiency ranged from 89.7 to 111.8 %. On applying our method to screen 117 commercially available samples, we found that 116 samples were not detected targeted compounds and one sample contained diphenhydramine component.

Hyun-Jung Kim, Yong Jae Lee, Jong-Bang Eun. Effects of ultraviolet radiation on the physicochemical characteristics of Korean native cattle (Hanwoo) beef. J. Korean Soc. Appl. Biol. Chem. 58(1):149-156The effects of ultraviolet (UV) radiation on the physicochemical characteristics of Korean native cattle (Hanwoo) beef with regards to shelf life were investigated in this research. The Hanwoo beef was exposed to UV radiation (4.5 mW s/cm2) for 0, 5, 10, 15, and 20 min. The results showed no differences in physicochemical parameters such as pH, drip loss, and shear force for all of the beef samples that were treated during storage. Lightness (L* value) and yellowness (b* value) did increase during storage. The redness (a* value) of all of the Hanwoo beef tended to increase until day 6 of storage, but then decreased afterward. The non-radiated beef had higher L*, a*, and b* values than UV-radiated Hanwoo beef. Furthermore, the UV-radiated beef showed significantly higher thiobarbituric acid values during storage as compared to those of non-radiated beef. No differences in the volatile basic nitrogen values in any of the beef samples were observed during storage. The sensory evaluation also showed no significant differences between

the UV radiation treatments. The results verified that the physicochemical characteristics of beef were not significantly affected by UV radiation treatments.

Joungsu Joo, Sang Ik Song. Rice ASR1 regulates sugar levels and participates in sugar signaling in roots. J. Korean Soc. Appl. Biol. Chem. 58(2):161-167The ASR gene is found in various plant species. Although ASR is also expressed in sink tissues, most functional studies on this gene have been performed in source leaves. Here, we report that OsASR1 is involved in the abscisic acid (ABA) response via regulation of sugar levels in the roots. Overexpression of OsASR1 altered root sugar content by increasing fructose levels and decreasing sucrose levels. Moreover, the transcript levels of sugar transporters and sucrose metabolism-related genes were altered in the roots of OsASR1-OX lines. Upon ABA treatment, an additional increase in the glucose and fructose levels was observed in the roots of OsASR1-OX lines compared with ABA-treated NT controls and untreated OsASR1-OX lines. Compared with NT control, the root growth of OsASR1-OX lines was more strongly inhibited by 10 % glucose. Our results suggest that OsASR1 plays an important role in the ABA response and sugar signaling in the roots.

Sangyong Lim, Jihae Choi, Minjeong Kim, Hyunjin Yoon. Temporal regulation of Salmonellapathogenicity Island 1 (SPI-1) hilA by Hfq in Salmonella enterica serovar typhimurium. J. Korean Soc. Appl. Biol. Chem. 58(2):169-172Hfq is a global regulatory RNA-binding protein abundant in many bacteria. In terms of virulence regulation in Salmonella, Hfq binds to and stabilizes the mRNA of hilD, a master regulator ofSalmonella pathogenicity island 1 (SPI-1), and Hfq activates SPI-1 gene transcription during the late exponential-growth phase. In contrast to the positive roles of Hfq in SPI-1 regulation, we observed that the absence of Hfq increased the expression of SPI-1 regulators, including HilA and InvF, during the late stationary phase. This result suggests that Hfq has pleiotropic functions, controlling SPI-1 positively or negatively depending on the growth conditions.

M. I. R. Mamun, A. M. Abd El-Aty , Md. Musfiqur Rahman, Jeong-Heui Choi, Kyeong Won Yun, Ho-Chul Shin, Jae-Han Shim. Characterization of secondary metabolite compounds correlated with the seasons in Artemisia princeps var. orientalis (Pamp.) H. Hara leaves using direct sample injection and gas chromatography–mass spectrometry: contribution to phytotoxicity. J. Korean Soc. Appl. Biol. Chem. 58(2):173-183Leaves from a natural population of Artemisia princeps var. orientalis (Pamp.) H. Hara were collected monthly from April through October and characterized for composition of secondary metabolite compounds and their phytotoxic effects on seed germination and seedling growth ofAchyranthes japonica and Lactuca sativa. The compounds were identified using gas chromatography/mass spectrometry (GC/MS) coupled with a solvent-free solid injector (SFSI). GC/MS analyses of all samples revealed qualitative variability in the composition of secondary metabolites. The greatest number of compounds was identified in July (56) followed by September (30) and April (24), and the lowest number was found in June (2) and August (2). Among 92 compounds, the major compounds were various terpenes (23) (mono-, sesqui, di-, and tri-terpenes) followed by heterocyclic compounds (18) and hydrocarbons (14). The higher the concentration of the secondary metabolites, the lower the seed germination and seedling growth of A. japonica and L. sativa. Plant samples collected in July and August were most detrimental. Taken together, variability in the secondary metabolites compounds of A. princeps var. orientalis was verified during different seasons, and the compounds were successfully identified by a combination of SFSI and GC/MS. Notably, the antimicrobial and antioxidative effects were inconsistent throughout the various seasons.

Sundramurthy Venkatesa Prabhu, Rajoo Baskar. Kinetics of heavy metal biosolubilization from electroplating sludge: effects of sulfur concentration. J. Korean Soc. Appl. Biol. Chem. 58(2):185-194The removal of heavy metals from industrial sludge through biosolubilization using sulfur-

oxidizing bacteria has been shown to be a promising technology, but the process with surplus concentration of sulfur causes re-acidification of the treated residues and creates environmental issues. Thus, the study for investigating the effect of sulfur concentration on the heavy metal biosolubilization system, with an emphasis on optimizing the sulfur concentration, is of immense importance. In this study, the experiments to investigate the effect of sulfur concentration on the performance of biosolubilization were carried out using 2–10 g/L elemental sulfur on heavy metal-laden electroplating sludge (50 g/L). The sludge-acclimatized, sulfur-grown Acidithiobacillus ferrooxidans isolate was used as sulfur-oxidizing bacteria. For the type of sludge used in this study, high pH reduction, short lag phase, and high heavy metal solubilization efficiencies were obtained in the treatment with 6 g/L sulfur. The kinetic study showed that the rate constant values of heavy metal solubilization were relatively high while using sulfur concentration of 6 g/L. The analysis using shrinking core model of fluid–particle reaction kinetics explicated that chemical reaction step controls the rate of heavy metal biosolubilization. The study provides an optimized strategy to design an efficient biosolubilization system for anticipated energy source.

Griselda Herman Natadiputri, Antonius Suwanto, Hyung Kwoun Kim. One-step transesterification reaction using methanol-stable lipase for omega-3 fatty acid ethyl ester production. J. Korean Soc. Appl. Biol. Chem. 58(2):195-202We investigated transesterification activity of methanol-stable lipase from Staphylococcus haemolyticus L62. Lipase L62 transesterified olive oil and menhaden oil using various alcohols including methanol, ethanol, 1-propanol, and 1-butanol. Based on the high stability of lipase L62 toward alcohols, high molar ratios (3:1 or 6:1) of alcohol to oil were employed in the reaction systems. Lipase L62 produced fatty acid esters efficiently by one-step transesterification. Reaction conditions for production of fatty acid methyl esters (FAMEs) and omega-3 fatty acid ethyl esters were further optimized. Lipase L62 produced those esters efficiently when 30–40 % water content was present in the reaction medium. The lipase L62 conversion yield was as high as 92 % for FAME and omega-3 ethyl ester production. These results indicate that lipase L62 is a suitable catalyst for producing an omega-3 ethyl ester concentrate in the pharmaceutical industry as well as for producing biodiesel in the bio-refinery industry.

Liang Dong, Feng Li, Yongzhe Piao, Dong Sun, Rui Zhao, Cheng Li, Lina Cong, Changxin Zhao. Characterization of proteinase A excretion from Saccharomyces cerevisiaein high sugar stress conditions. J. Korean Soc. Appl. Biol. Chem. 58(2):203-208High sugar concentration culturing was commonly used in modern fermentation industry. However, it leads to the reduction of the foam stability in beer brewing due to the excess secretion of proteinase A. To better understand the characterization of proteinase A excretion from Saccharomyces cerevisiae in high sugar stress conditions, the cultures were grown by using YNB medium with a high concentration of glucose. Pro-PrA isolated from the medium was purified by gel exclusion chromatography, and the PrA activity was detected using fluorescent substrate analysis. The relative molecular weight of the purified PrA and pro-PrA was estimated at 42 and 54 kDa, respectively, by SDS-PAGE. It indicated that the metabolic behavior of PrA in the high glucose culturing was quite different from the normal conditions, and glucose concentration may have a big influence on its secreted process. Further study showed that PrA was released at the logarithmic growth phase of the culturing, and the amount of PrA was 11 times higher compared with the normal culturing. PrA was considered to be activated by itself under acidic conditions. And it was also confirmed in this work that the step-wise pathway for the autoactivation known as a pseudo-PrA has a major contribution to the autoactivation process of PrA zymogen outside the cell.

Euiyoung Bae , Sojin Moon, George N. PhillipsJr.. Molecular dynamics simulation of a psychrophilic adenylate kinase. J. Korean Soc. Appl. Biol. Chem. 58(2):209-212Proteins from psychrophilic organisms can function at low temperatures and have shown

potential for industrial applications. In the present study, thermal denaturation of a psychrophilic adenylate kinase (AKpsychro) from Bacillus globisporus was measured using circular dichroism spectroscopy and its molecular dynamics (MD) simulations were performed. The results from the unfolding experiment and MD simulations of AKpsychro were compared with those of its mesophilic and thermophilic homologues to study the relationship between dynamic motion and the cold adaptation of proteins.

Ah-Young Ko , A. M. Abd El-Aty , Jin Jang , Jeong-Heui Choi, Md. Musfiqur Rahman, Sung-Woo Kim, Ho-Chul Shin, Jae-Han Shim. Detecting fludioxonil residues in brown rice and rice straw using gas chromatography-nitrogen phosphorus detector. J. Korean Soc. Appl. Biol. Chem. 58(2):213-217In the present study, brown rice was steeped in 20 % wettable powder fludioxonil for 24 h, subsequently germinated, and transplanted in paddy fields. The harvested rice was tested at 156 days to detect residue levels using gas chromatography-nitrogen phosphorus detector. Validation was carried out to assess the following parameters: linearity, limit of detection and limit of quantitation (LOQ), recovery, and storage stability. Using matrix-matched calibrations, the determination coefficients were >0.999 in both matrices. Mean recoveries were 73.5–101.0 % with relative standard deviations <10 % in both matrices. The LOQ (0.006 mg/kg) was lower than the maximum residue limit (MRL = 0.02 mg/kg) set by the Ministry of Food and Drug Safety, Republic of Korea. The developed method was applied successfully, and no residues were detected in field-incurred rice and/or rice straw samples.

Munira Taj Muhammad, Lubna, Nida Fayyaz, Saima Tauseef, Ummarah Razaq, Muhammad Ali Versiani, Aqeel Ahmad, Shaheen Faizi, Munawwer Rasheed. Antibacterial activity of flower of Melia azedarach Linn. and identification of its metabolites. J. Korean Soc. Appl. Biol. Chem. 58(2):219-227Constituents of eight different extracts and fractions, obtained from flowers of Melia azedarach, were identified by gas chromatography–mass spectrometry and mass spectral library search. Altogether, 38 phytochemicals were identified, all for the first time from the flowers. Only 14 of these were known from parts of M. azedarach other than flowers, while 24 are reported for the first time from any part of the plant. These metabolites included branched and n-hydrocarbons, aromatics, a polyisoprenoid, fatty acids, fatty acid methyl esters, and fatty alcohols, which were found to be different from its essential oil constituents. One major constituent 4-hydroxybenzoic acid (4) was not only identified but also isolated as a pure compound from ethyl acetate extract. Extracts, compound 4 and its derivatives gallic acid and methyl gallate were tested for antimicrobial potential. Gallic acid and methyl gallate exhibited antibacterial activity against Staphylococcus saprophyticus. Methyl gallate also showed some activity against Corynebacterium diphtheriae and Corynebacterium. hofmannii. Among the extracts, only methanol extract exhibited activity against Pseudomonas sp. The extracts only inhibited the growth of dermatophytic fungi.

Eun-Seon Lee, Shin Young Park, Young-Gil Jeong, Byung-Chul Jo, Minhui Kim, Sang-Do Ha. Quality evaluation and estimation of shelf life of retort-pouched tomato-based and Korean traditional fermented food-based sauces. J. Korean Soc. Appl. Biol. Chem. 58(2):229-236

The shelf life of four different sauces in retort pouches, Gochujang chicken, Kimchi, ham beef, and beef salsa, was examined. Samples of each sauce pouch were stored at 25, 35, and 45 °C for 6 months; changes in the chemical were measured, and sensory evaluation was done. Finally, this study predicted the shelf life of products. Concentrations of thiobarbituric acid (TBA) inside the retort-pouched sauces were found to increase over the course of storage. Likewise, the overall pH of the sauces was found to decrease generally. Finally, sensory evaluation, ranked on a 7-point scale, was found to be greater than 5 (“like moderately”) when processed by one of 15 panel. The TBA value was used to estimate the shelf life of the four sauces, which was found to be greater than 3 years for each. Therefore, the use of retort pouches is an effective packaging method for the

long-time storage of sauces at temperatures lower than 25 °C.

Md. Ashaduzzaman Siddikee, Subbiah Sundaram, Murugesan Chandrasekaran, Kiyoon Kim, Gopal Selvakumar, Tongmin Sa. Halotolerant bacteria with ACC deaminase activity alleviate salt stress effect in canola seed germination. J. Korean Soc. Appl. Biol. Chem. 58(2):237-241Amelioration of salt stress effect on canola seed germination was investigated using 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing plant growth promoting halotolerant bacteria. NaCl at 120 mM concentration reduced canola seed germination by 50 %. The inoculation of ACC deaminase-producing halotolerant Brevibacterium epidermidis RS15 and Bacillus aryabhattai RS341 at 120 mM NaCl significantly increased the seed germination with decreased seed ACC content. Notably, the hydrolytic enzymes activities like amylase, invertase, and protease also increased due to inoculation of RS15 and RS341 compared to uninoculated salt stress imposed germinating canola seeds. Ethylene emission of salt stress exposed eight-day-old canola seedlings was reduced by 35.4 and 41.1 % compared to uninoculated salt stressed control due to respective inoculation of RS341 and RS15. The amelioration of salt stress inhibitory effect on the canola seed germination was attributed to the inoculation of ACC deaminase-producing halotolerant bacteria modulating ethylene emission and inducing hydrolytic enzymes.

Woong Kim, Bo Ram Kang, Hye Yun Kim, Soo Min Cho, Yong-Deok Lee, Sehoon Kim, Jung Young Kim, Dong Jin Kim, YoungSoo Kim. Real-time imaging of glioblastoma using bioluminescence in a U-87 MG xenograft model mouse. J. Korean Soc. Appl. Biol. Chem. 58(2):243-248Glioblastoma multiforme (GBM), the most common malignant brain tumor, is characterized by aggressive proliferation and invasive potential. Xenograft animal models of GBM have critically contributed to evaluation of novel therapeutic agents, drug delivery system, and diagnostic tools. To mimic intrinsic behavior of GBM, orthotopic transplantation of cancer cells and continuous observation of cell growth should be conducted in animal study. Here, we generated xenograft model mouse of GBM in which U-87 MG human glioblastoma cells were intracranially implanted for live imaging. Introducing luciferase gene into U-87 MG cell line enabled real-time observation and quantification of tumor survival and propagation by detecting photon emission derived from luciferase. Our GBM model mouse has potentials to bring great advantages in pharmacological and mechanistic investigation on brain tumors.

Jia-Le Song , Kai Zhu , Xia Feng, Xin Zhao. Protective effect Malus pumila Mill leaf polyphenols in reserpine-induced gastric ulcer in mice. J. Korean Soc. Appl. Biol. Chem. 58(2):249-256The protective effect of polyphenols from Malus pumila leaf (MALP) on reserpine (10 mg/kg)-induced gastric ulcer in Kunming (KM) mice was investigated. Gastric juice secretion, pH of gastric juice, serum neuropeptides, including motilin (MOT), substance P (SP), vasoactive intestinal peptide (VIP), and somatostatin (SS) levels, as well as serum levels of inflammatory cytokines (TNF-α, IL-6, IL-12, and IFN-γ) were measured. Gastric occludin and p38MAPK phosphorylation was analyzed by RT-PCR or Western blotting. MALP reduced reserpine-induced gastric juice secretion and increased the pH of gastric juice. MALP increased the serum levels of VIP and SS and reduced MOT, SP, and inflammatory cytokines in serum. It also modulated the gastric expression of occludin and p38MAPK phosphorylation in mice. These results suggest that MALP showed a protective effect against reserpine-induced ulcer in mice by reducing gastric juice secretion, modulating serum neuropeptide levels, attenuating serum inflammatory cytokines, and regulating gastric levels of occludin and p38MAPK phosphorylation.

Yeon Woo Song, Sabina Shrestha, Rajendra Gyawali, Dong-Sun Lee, Somi Kim Cho. Citrus unshiu leaf extract containing phytol as a major compound induces autophagic cell death in human gastric adenocarcinoma AGS cells. J. Korean Soc. Appl. Biol. Chem. 58(2):257-265

The pharmaceutical potential of the methanolic extract of Citrus unshiu leaves (MECL) was assessed through analysis of its inhibitory effect on cancer cells. The antiproliferative activities of the leaves were evaluated using several cancer cell lines and considerable cytotoxicity was observed in human gastric adenocarcinoma AGS cells. Inhibition of AGS cell viability was both time- and dose-dependent, and MECL induced non-apoptotic cell death. AGS cells treated with MECL increased the formation of acidic vesicular organelles and GFP-LC3 puncta. Pretreatment with an autophagy inhibitor, 3-methyladenine, inhibited MECL-induced cell death. These results indicated that the mechanism underlying the anticancer effects of MECL in AGS cells could be via the induction of autophagic cell death. The major compounds of MECL were identified as phytol, 4-ethenyl-2-methoxyphenol, hexadecanoic acid, 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one, and vitamin E using gas chromatography–mass spectrometry. These results indicate that C. unshiuleaves can be exploited for numerous pharmaceutical applications as a source of anticancer ingredients.

Mir Sadiq Shah, Mark Eppinger, Safia Ahmed, Aamer Ali Shah, Abdul Hameed, Fariha Hasan. Multidrug-resistant diarrheagenic E. colipathotypes are associated with ready-to-eat salad and vegetables in Pakistan. J. Korean Soc. Appl. Biol. Chem. 58(2):267-273Diarrheagenic E. coli pathotypes are major foodborne pathogens causing gastrointestinal tract infections leading to hemolytic uremic syndrome and hemorrhagic colitis. Consumption of raw vegetables is encouraged due to its nutrient content and antioxidant properties, although their ingestion is linked to a series of foodborne disease outbreaks. The aim of this study was to evaluate the prevalence of Enterotoxigenic, Enteropathogenic, and Shiga toxin-producing E. coli pathotypes in raw vegetables and ready-to-eat salad for the development of better risk management. A total of 260 vegetable and salad mix (cucumber, lettuce, spinach, and carrot) samples were collected from commercial food markets in Southern districts of Khyber Pakhtunkhawa, Pakistan. About 34 % vegetable samples were contaminated with E. coli strains. 32.4 % E. coli strains from vegetable sample were identified as diarrheagenic E. coli pathotypes. Similarly, 26.7 % of salad samples were contaminated with E. coli pathotypes. 15 % [two Enterotoxigenic E. coli and one Enteropathogenic E. coli] were isolated from spinach salad samples. 25 % [three Enterotoxigenic E. coli, one Enteropathogenic E. coli and one Shiga toxin-producing E. coli] were isolated from mixed salad type A. 40 % [four Enterotoxigenic E. coli, two Enteropathogenic E. coli and two Shiga toxin-producing E. coli] were isolated from mixed salad type B. 92 % diarrheagenic E. coli pathotypes showed resistance against Tetracycline and 87 % to Ampicillin. This study showed that fresh vegetable and their products were contaminated with multidrug-resistant E. coli pathotypes.

Jae-Whan Lee, Soon-Ah Kang, Chul Cheong. Quality characteristics of distilled alcohols prepared with different fermenting agents. J. Korean Soc. Appl. Biol. Chem. 58(2):275-283This study was performed to investigate the quality characteristics of distilled spirits made with different fermenting agents in copper distiller. The highest distillate yield was observed in the mashing made with fermenting agent A. The distillate made with fermenting agent A showed also significantly the highest level of ester and higher-alcohol concentration. In addition, the distilled spirit made with fermenting agent A indicated significantly higher preferences in appearance, smell, taste, quality, and overall acceptability than those of other agents. However, the aging in bottle for 6 months had no significant differences in sensory improvement in distilled spirits regardless in kind of fermenting agents. This study emphasizes the importance of rice koji as fermenting agent in distilled spirit prepared with rice and using with multistage distiller made of copper to improve aroma substances and yield ratio of distillate.

Hee-Sun Kim, Kyung-Mi Kim, Gwi-Jung Han, Hyeon-Gyu Lee, Myung-Hwan Kim. Effect of wheat flour addition on retardation of retrogradation in waxy rice cake, Ingeolmi. J. Korean Soc. Appl. Biol. Chem. 58(2):285-291

Reduction of the retrogradation of Injeolmi (IM) during storage for 3 days at 4 °C due to the addition of 0.8 % (w/w) wheat flour was analyzed. The optimum reaction temperature for the hydrolytic enzyme activity of α-amylase in the wheat flour was 30 °C for gelatinized waxy rice. Activity sharply increased until 7 h and then leveled off. The amylose content of IM remained almost constant, while that of wheat flour-added Injeolmi (WFIM) increased by 24 % during storage. Average amylopectin chain lengths of the starches in IM and WFIM increased from 27.91 to 30.36 and from 29.57 to 57.98, respectively, during storage. In the X-ray diffraction pattern, IM starch showed the characteristic peak located at 17°, while no distinct diffraction peak was observed for WFIM starch after storage. Enthalpy changes (ΔH) indicated by differential scanning calorimetry of IM and WFIM were 1.1060 and 0.6159 J/g, respectively. The hardness of IM significantly increased from 149.27 to 2202.52 g f during storage while that of WFIM increased only from 112.14 to 128.08 g f which was 5.82 % compared to that of IM.

Sang Gon Kim, Jin-Seok Lee, Seong-Hyu Shin, Sung Cheol Koo, Jung Tae Kim, Hwan Hee Bae, Beom-Young Son, Yul-Ho Kim, Sun Lim Kim, Seong-Bum Baek, Young-Up Kwon. Profiling of differentially expressed proteins in mature kernels of Korean waxy corn cultivars using proteomic analysis. J. Korean Soc. Appl. Biol. Chem. 58(3):293-303Maize (Zea mays L.) is the most important crop such as food for humans and feed for animals. Although new varieties of maize have been extensively developed in Korea, little is known about differences in the proteomes of mature kernels among maize varieties. Three Korean waxy corn cultivars (Ilmichal, Eolrukchal 1, and Heukjinjuchal) have previously been developed. The total protein content is higher in Ilmichal than in Eolrukchal 1 or Heukjinjuchal, while the starch and fatty acid contents are similar among these three Korean waxy corns. To uncover the differences in proteomic profiles among these three Korean waxy corns, we performed proteomic analysis and compared their protein compositions. We detected 37 differentially expressed protein spots and identified the proteins using MALDI-TOF mass spectrometry. Of these proteins, 37.8 % were identified as storage proteins, 18.9 % as stress-related proteins, and 18.9 % as metabolism-related proteins. Storage proteins (globulin-2) and stress-related proteins (heat shock proteins and general stress proteins) were highly expressed in Ilmichal or Heukjinjuchal. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis of corresponding genes of five randomly selected proteins, including glyoxalase family protein (accession number, B6SGF3), globulin-2 (Q7M1Z8), heat shock protein 1 and 3 (B6TGQ2 and B6TDB5), and vicilin-like embryo storage protein (Q03865), showed that the expression levels of the tested genes were well-correlated with protein abundance, suggesting that these proteins are also differentially regulated at the transcriptional level. Taken together, these results provide a better understanding of proteomic differences among Korean waxy corn cultivars and may support further molecular breeding efforts.

Jeong Han Kang, Sung-Yong Yang, Jaeho Ha, Kwang-Won Lee. Perilla frutescens modulates CYP1A1/2 and HO-1 and activates Nrf2 in oxidative stress-induced hepatotoxicity. J. Korean Soc. Appl. Biol. Chem. 58(3):305-315We speculated that lipid peroxidation induced by tert-butyl hydroperoxide (t-BHP) in liver is closely linked with the metabolism mediated by CYPs. In this study, we have examined the effect of Perillaleaf extract (PLE) on CYPs using 7-ethoxyresorufin-O-deethylase (EROD, indicator of CYP1A1), 7-methoxyresorufin-O-demethylase (MROD, indicator of CYP1A2), erythromycin N-demethylase (ERDM, indicator of CYP3A), and p-nitrophenol hydroxylase (PNPH, indicator of CYP2E1) in rat liver. Rats orally pretreated with PLE (250, 500, and 1,000 mg/kg b.w.) for 5 days were administered with a single i.p. dose of t-BHP (0.5 mmol/kg). Kinetic analysis of CYP1A1/2 activities in t-BHP-treated liver demonstrated that PLE inhibits the enzyme activities by competitive and noncompetitive inhibitions. The pretreatment with PLE decreased the expression of CYP1A1/2 mRNA and protein compared with t-BHP treatment alone. A Phase II enzyme, heme oxygenase-1 (HO-1), is involved in hepatoprotection against oxidative damage, and we confirmed that PLE increases the levels of HO-1 mRNA and protein, as well as its activity in t-BHP-induced

liver damage. PLE administration resulted in enhanced nuclear translocation and ARE binding of NF-E2-related factor 2. These findings suggest that PLE protects against t-BHP-induced hepatotoxicity through modulated activity and expression of selective CYPs, and ARE-driven induction of HO-1 expression and its activity.

Kwang-Su Park, Hyungmi Kim, Mi Kyoung Kim, Kyungdo Kim, Youhoon Chong. Synthesis and biological evaluation of flavonol-glucose conjugates for cosmeceutical development. J. Korean Soc. Appl. Biol. Chem. 58(3):317-323Quercetin and kaempferol, two well-known flavonols, were chemically conjugated with glucose to produce the corresponding flavonol glucosides, and the following biological activities were evaluated for cosmeceutical development: antioxidant activity, ability to increase collagen synthesis, and moisturizing activity. Among the synthetic flavonol glucosides, quercetin-3-O-β-D-glucoside significantly enhanced collagen synthesis (60 %) compared to quercetin. Kaempferol-3,7-di-O-β-D-glucoside showed promising skin-moisturizing effects, inducing a sixfold increase in the expression of aquaporin-3. Thus, both quercetin-3-O-β-D-glucoside and kaempferol-3,7-di-O-β-D-glucoside were shown to possess interesting biological activities which warrant their further development as cosmetic ingredients.

Soyoung Lee, Yoo-Jung Lee, Jung-Suk Sung, Han-Seung Shin. Influence of roasting conditions on the chemical properties and antioxidant activity of perilla oils. J. Korean Soc. Appl. Biol. Chem. 58(3):325-334Chemical properties of perilla oils under a variety of roasting conditions were evaluated. Roasting was performed in a hot-air roaster, with inlet air temperatures of 150–240 °C. The major fatty acid contents in perilla oil were α-linolenic, oleic, and linoleic acid. Trans-fatty acid content was 0.10–0.31 % from perilla oil under different roasting conditions. Maillard reaction products increased significantly from perilla oils exposed to higher temperatures for longer times. In particular, 2-methylpyrazine and 2,5-dimethylpyrazine were highest in the P210/30 (roasting temp. 210 °C, roasting time 30 min) and P210/30 treatments, respectively. Total benzo[a]pyrene of 0.21 µg/kg was detected in perilla oil under different roasting conditions. Evaluation of radical scavenging activity revealed that antioxidant function of perilla oil was promoted by increasing the roasting temperature of perilla seeds. As roasting temperature and time increased, the oxidative stability of perilla oils increased. The average iodine value of perilla oil under different roasting conditions was 154.23. Oxidation stability increased in the P240/20 group.

Sang Won Lee, Yu Geon Lee, Jeong-Yong Cho, Young Chool Kim, Sang-Hyun Lee, Wol-Soo Kim, Jae-Hak Moon. Establishment of a simple method for purification of high purity chlorogenic acid from immature fruit of pear (Pyrus pyrifolia Nakai). J. Korean Soc. Appl. Biol. Chem. 58(3):335-341Chlorogenic acid [3-(3,4-dihydroxycinnamoyl)quinic acid, CGA] is one of the main compounds contained in immature pear fruits. In the present study, we established simple purification of CGA in high purity from the immature fruit of pear (Pyrus pyrifolia Nakai cv. Chuhwangbae) which contains CGA in high concentration. The 80 % MeOH extract (1.4 g) of immature pear fruits (20 g fresh wt.) was partitioned with ethyl acetate (EtOAc) under acidic condition (pH 2.65). Then, the EtOAc-soluble acidic layer (110.3 mg) was further purified using Diaion HP-20 column chromatography (H2O/MeOH, step wise). Then, the CGA-contained fractions were finally purified by preparative ODS-HPLC. The isolated CGA (24.3 mg) showed high purity (>99 %) and the total recovery was estimated to be 63.9 %.

Mihyang Kim, Youngrong Park, Sooyoung Cho, Supawadee Burapan, Jaehong Han. Synthesis of alkyl quercetin derivatives. J. Korean Soc. Appl. Biol. Chem. 58(3):343-348Series of alkyl quercetin derivatives were synthesized from the reactions between quercetin (1) and the corresponding alkyl halides. Three methyl quercetin derivatives, 5,3′-dihydroxy-3,7,4′-trimethoxyflavone (2), 5-hydroxy-3,7,3′,4′-tetramethoxyflavone (3), 3,5,7,3′,4′-pentamethoxyflavone (4), three ethyl quercetin derivatives, 5,3′-dihydroxy-

3,7,4′-triethoxyflavone (5), 5-hydroxy-3,7,3′,4′-tetraethoxyflavone (6), 3,5,7,3′,4′-pentaethoxyflavone (7), two propyl quercetin derivatives, 5,3′-dihydroxy-3,7,4′-tripropoxyflavone (8) and 5-hydroxy-3,7,3′,4′-tetrapropoxyflavone (9), two butyl quercetin derivatives, 5,3′-dihydroxy-3,7,4′-tributoxyflavone (10), 2-(3,4-dibutyoxybenzoyloxy)-4-butoxy-6-hydroxybenzoic acid (11), and two benzyl quercetin derivatives, 5,3′-dihydroxy-3,7,4′-tribenzoxyflavone (12), 5-hydroxy-3,7,3′,4′-tetrabenzoxyflavone (13), were purified by column chromatography. Among these, synthesis and physical properties of 5, 8, 9, 10, and 11 were reported for the first time. The chemical structures were determined by spectroscopy, including NMR, MS, and IR, and the physical properties of the quercetin derivatives were also characterized. Formation of 11 from the alkylation was of a great interest, and it was rationalized by the singlet oxygen-mediated product decomposition. Antioxidant and anti-bacterial activities of the synthesized alkyl quercetin derivatives were measured and found less active than 1.

Seon-A Yoo, Chang-Su Na, Seong-Eun Park, Seung-Ho Seo, Hong-Seok Son. Characterization of fermented sausages using Lactobacillus plantarum MLK 14-2 as starter culture. J. Korean Soc. Appl. Biol. Chem. 58(3):349-358The effect of adding starter culture (Lactobacillus plantarum MLK 14-2) isolated from Kimchi on the microbial counts, physicochemical parameters, and free amino acid composition of fermented sausage was determined during the ripening period. The MLK 14-2 populations in the sausage treatments increased to above 9 log cfu/g and remained at this level during ripening. The pH decreased sharply in the sausage fermented using MLK 14-2, in accordance with the dramatic increase of lactic acid during the early stages of fermentation. The concentrations of most free amino acids increased during fermentation, and the highest total free amino acids concentration of 16.3 mg/g was observed in the sausage inoculated with MLK 14-2 after 36 days of fermentation. Significantly higher amounts of arginine, methionine, isoleucine, and leucine were detected in the sausage inoculated with MLK 14-2. In conclusion, L. plantarum MLK 14-2 was considered to have high potential as a substitute of starter culture for making fermented sausage.

Ji-Yeon Yang, Bo-Kyung Sung, Hoi-Seon Lee. Phototactic behavior 8: phototactic behavioral responses of western flower thrips, Frankliniella occidentalisPergande (Thysanoptera: Thripidae), to light-emitting diodes. J. Korean Soc. Appl. Biol. Chem. 58(3):359-363The phototactic behavioral responses of western flower thrips to light-emitting diodes (LEDs) were evaluated under optimal conditions (light sources, luminance intensities, and light-exposure durations). Based on the attraction rate (%) of the western flower thrips under optimal conditions in the dark, the yellow LED (590 ± 5 nm) showed the greatest attraction rate (48.0 %), followed by green (520 ± 5 nm, 44.3 %), blue (470 ± 10 nm, 40.3 %), red (625 ± 10 nm, 25.0 %), white (450–620 nm, 16.0 %), and infrared LEDs (730 nm, 10.3 %). Compared with the black light bulb (BLB), which served as the positive control, the yellow LED was approximately 1.2 times more active than that of the BLB (39.0 %) toward western flower thrips. In this regard, the yellow LED should be suitable for managing western flower thrips.

Hye Min Kim, Dong Gu Lee, Sanghyun Lee. Plant-derived molecules from Saussurea grandifolia as inhibitors of aldose reductase. J. Korean Soc. Appl. Biol. Chem. 58(3):365-371Saussurea grandifolia was tested for aldose reductase (AR) inhibition as part of a search for potentially therapeutic natural compounds. Stepwise polarity fractions were tested for in vitro inhibition of rat lens AR. Of these, the ethyl acetate (EtOAc) and n-butanol (n-BuOH) fractions exhibited AR inhibitory activities. A chromatographic separation of the active EtOAc and n-BuOH fractions led to the isolation of nine compounds, which were known and identified as vitexin (1), genkwanin-5-O-glucoside (2), vitexin 2″-p-hydroxybenzoate (3), kaempferol-3-O-rutinoside (4), isovitexin (5), quercitrin (6), isoquercitrin (7), 1,5-di-O-caffeoylquinic acid (8), and vicenin 1 (9). Among these compounds, compounds 6, 7, and 8 showed very strong inhibitory activities against AR

(IC50’s = 0.34, 0.32, and 0.08 μM, respectively). The results suggest that S. grandifolia is clearly a potential source for therapeutic agent that might be useful for the treatment of diabetic complications.

Jun-Hwan Park, Bo-Kyung Sung, Hoi-Seon Lee. Phototactic behavior 7: phototactic response of the maize weevil, Sitotroga zeamais motsch (Coleopter: Curculionidae), to light-emitting diodes. J. Korean Soc. Appl. Biol. Chem. 58(3):373-376Phototactic responses of maize weevil, Sitotroga zeamais, adults to five light-emitting diodes (LEDs) were evaluated and compared with the commonly used luring lamp (BLB). Under optimal light conditions, the red LED (625 ± 10 nm) exhibited the highest attraction rate (59.8 %), followed by the yellow LED (590 ± 5 nm, 52.3 %), the infrared LED (730 nm, 51.9 %), the green LED (520 ± 5 nm, 46.7 %), the blue LED (470 ± 10 nm, 45.3 %), the ultraviolet LED (365 nm, 32.7 %), and the BLB (27.3 %). Moreover, the red LED was approximately 2.19 times more attractive than that of the BLB. These results indicate that a red LED trap may be useful to control S. zeamais adults.

Md. Azizul Haque, Weon Taek Seo, Chung Eun Hwang, Hee Yul Lee, Min Ju Ahn, Kye Man Cho. Culture-independent analysis of yeast diversity in Korean traditional fermented soybean foods (doenjang and kanjang) based on 26S rRNA sequence. J. Korean Soc. Appl. Biol. Chem. 58(3):377-385The yeast-26S rRNA libraries were constructed from two different fermented soybean foods, doenjang and kanjang. A total of 42 clones, containing the partial 26S rRNA sequences, 0.6 kb in length, were sequenced and subjected to an online similarity search. All doenjang yeast (DY) clones only appeared in the Saccharomycotina class. The 21 clones from the doenjang library were classified into five groups: Debaryomyces hansenii (DY I, 76.0 %), Zygosaccharomyces pseudorouxii (DY II, 9.6 %), Candida versatilis (DY III, 4.8 %), Candida etchellsii (DY IV, 4.8 %), andDebaryomyces castellii (DY V, 4.8 %). The 21 kanjang yeast (KY) clones were affiliated with theSaccharomycotina (52.4 %), Urediniomycetes (19.0 %), Ustilaginomycetes (23.8 %), andHymenomycetes (4.8 %) classes and divided into six groups: D. hansenii (KY I, 38.0 %),Sterigmatomyces halophilus (KY II, 19.0 %), Malassezia restricta (KY III, 23.8 %), Cryptococcus magnus (KY V, 4.8 %), and Pichia triangularis (KY VI, 9.6 %). Yeast belonging to theSaccharomycotina class was predominant (76.2 %) in fermented soybean foods, doenjang andkanjang. These findings are of fundamental value for understanding the complexity of two different fermented soybean foods.

Jiaying Xin , Kai Lin, Yan Wang, Chungu Xia. Improved biological synthesis of gold nanoparticles using methanobactin. J. Korean Soc. Appl. Biol. Chem. 58(3):387-395The present study reported a novel approach for the green synthesis of gold nanoparticles (AuNPs) by using methanobactin (Mb) from the spent medium of Methylosinus trichosporium 3011. This paper explored the synthesis of AuNPs under different Mb concentrations, temperatures, and reaction times. The biosynthesized AuNPs were characterized by using UV–Visible spectroscopy, fluorescence spectroscopy, transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). Comparing with the previous research using hydroquinone as an extra electron donor to synthesis of AuNPs, this research was the first to show that rising temperature improved the formation rate of AuNPs in a higher concentration of Mb itself. The formation of AuNPs was confirmed by surface plasmon resonance using UV–Visible spectrophotometry. Fluorescence spectroscopy experiments revealed that Mb sample showed three prominent peaks. In addition, the fluorescence exhibited by Mb was rapidly quenched upon addition of HAuCl4. According to the image analysis performed on a TEM micrograph, the obtained AuNPs were predominantly spherical with an average size of 19.3 ± 5.5 nm. XPS analysis demonstrated that Mb molecules reduced Au(III) to Au(0). FTIR spectrum confirmed the involvement of biomass groups in capping and reduction of AuNPs, which increased the stability of the synthesized AuNPs.

Sung Cheol Koo, Sang Gon Kim, Dong-Won Bae, Hyun Young Kim, Hyun Tae Kim, Young Hoon Lee, Beom Kyu Kang, Seong-Bum Baek, In Youl Baek, Hong Tai Yun, Man Soo Choi. Biochemical and proteomic analysis of soybean sprouts at different germination temperatures. J. Korean Soc. Appl. Biol. Chem. 58(3):397-407Soybean sprouts are available throughout the year and have gained popularity as a functional food owing to their high nutritional value. In the present study, soybean seeds were germinated at different temperatures and the effects on growth characteristics, nutrient composition, and secondary metabolites were investigated. Sprout qualities such as whole length and hypocotyl length were observed to increase at a higher temperature of germination (25 vs. 20 °C). The total protein content of the sprouts increased, whereas the total fatty acid content decreased upon germination at 25 °C. The total phenolic content was higher in soybean sprouts than in soybean seeds. Additionally, antioxidant activity increased in a temperature-dependent manner. Both DPPH and ABTS activity were higher at 25 °C than at 20 °C. Proteomic analysis was conducted to generate temperature responsive protein profiles of soybean sprouts. Using 2D gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, 33 differentially expressed spots were identified. Further analysis of these spots revealed potential function in protein storage and modification. Upon germination at 25 °C, 16 spots increased significantly, whereas 17 protein spots were observed to decrease. Interestingly, a trypsin inhibitor was highly expressed at 25 °C. Semi-quantitative RT-PCR analysis showed that mRNA expression level of most of genes encoding the identified proteins correlated well with their protein abundance, suggesting their temperature-dependent transcriptional regulation in soybean sprouts. In summary, our results clearly indicate an effect of temperature on growth of and secondary metabolite production in soybean sprouts.

Ju-Gyeong Lee, Sung-Ok Moon, Se-Yong Kim, Eun-Ju Yang, Ju-Sik Min, Ju-Hee An, Eun-A Choi, Kwang-Hyeon Liu, Eun Ji Park, Hwa-Dong Lee, Kyung-Sik Song. Rapid HPLC determination of gastrodin in Gastrodiae Rhizoma. J. Korean Soc. Appl. Biol. Chem. 58(3):409-413Gastrodin is a major biologically active ingredient in members of the genus Gastrodia. For this reason, there are many reports on the quantification of gastrodin in Gastrodiae Rhizoma (GR) and in GR-containing herbal preparations. HPLC, HPLC–MS, and TLC are the major approaches for gastrodin quantification; however, they usually require complicated pre-treatment, lengthy analysis, and expensive instruments. Therefore, a rapid and reliable method for determining gastrodin in GR is necessary. Optimal HPLC separation was achieved using a Chromolith Performance RP-18e (4.8 × 100 mm, 5 μm) stationary phase. The optimal mobile phase was a mixture of water (A) and acetonitrile (B) with a gradient of 1 % B at 0–8 min, 20 % B at 8–10 min, 80–100 % B at 10–12 min, and 100 % B at 12–13 min, followed by equilibration with 1 % B for 2 min at a flow rate of 1.5 mL/min. The detection wavelength was UV220 nm. Gastrodin appeared within 4 min under the above conditions. The calibration curve of gastrodin showed good linearity in the 0.25–10.0 range (r2 = 0.9998). The limit of detection (0.13 μg), limit of quantification (0.25 μg), and reproducibilities of area and retention time (0.04 and 3.24 %, respectively) were within acceptable ranges. In addition, the intra-day precision and accuracy of gastrodin were 0.74 and 100.63 ± 0.04 %, respectively, while the inter-day precision and accuracy were 0.06 and 99.25 ± 0.05 %, respectively. The range of mean gastrodin content in six GR samples, which were cultivated at different sites, was 0.37–0.79 %. This result may be a guideline for the quality control of GR and GR-containing medicinal preparations.

Do Huu Nghi, René Ullrich, Franco Moritz, Le Mai Huong, Vu Dinh Giap, Do Huu Chi, Martin Hofrichter, Christiane Liers. The ascomycete Xylaria polymorphaproduces an acetyl esterase that solubilises beech wood material to release water-soluble lignin fragments. J. Korean Soc. Appl. Biol. Chem. 58(3):415-421

Three ascomycetous soft-rot fungi belonging to the family Xylariaceae were found to produce high levels of a p-nitrophenyl acetate-hydrolyzing activity during growth on lignocellulosic materials, i.e., wheat straw and beech wood chips. Accordingly, Kretzschmaria deusta, Morchella elata, Xylaria polymorpha were seemingly most noteworthy acetyl esterase producers and, of which, X. polymorpha (strain A35) was chosen for further studies. Induction study indicated that raw carbohydrate sources, such as beech wood, rape straw, birch wood, and wheat straw, were extremely important for acetyl esterase production. Acetyl esterase of X. polymorpha was produced in solid-state culture on wheat straw and purified by different steps of anion-exchange and size-exclusion chromatography. This purified enzyme (M W = 44 kDa and pI values of 3.5–3.6) exhibited the capability to solubilise in vitro beech wood to release water-soluble lignin fragments with molecular masses of 1–3 kDa as analyzed by high performance size exclusion chromatography.

Hyun Chung, Mi Kyung Yoon, Jihye Han, Young-Suk Kim. Evaluation of volatile organic compounds in alcoholic beverages consumed in Korea. J. Korean Soc. Appl. Biol. Chem. 58(3):423-432Volatile organic compounds including acetaldehyde, methanol, and higher alcohols such as 1-propanol and 3-methyl-1-butanol in 75 domestic and imported alcoholic beverages consumed in Korea were investigated and evaluated using a gas chromatograph equipped with a mass spectrometer. The acetaldehyde contents in the studied samples ranged from 0.02 to 11.73 mg/L, and were highest in refined rice wines and fruit wines. Only the wine and fruit wine samples contained methanol, at concentrations in the range of 1.42–23.81 mg/L. The 1-propanol content was highest in whisky, while the 3-methyl-1-butanol content ranged from 4.45 to 280 mg/L in all samples other than Korean distilled liquor.

Byung Il Kim, Yong Hoon Joo, Pyo June Pak, Joong-Su Kim, Namhyun Chung. Different shapes of Al2O3 particles induce differential cytotoxicity via a mechanism involving lysosomal destabilization and reactive oxygen species generation. J. Korean Soc. Appl. Biol. Chem. 58(3):433-442The biological effects of nano- and micro-sized Al2O3 particles are hypothesized to differ according to the shapes as well as the sizes of the particles. Thus, the mechanisms of interleukin (IL)-1β production and the association between the shape of the Al2O3 particle and its cytotoxicity in macrophage-like THP-1 cells were investigated using particles with three different shapes [N-Al2O3(<30 nm), S-Al2O3 (2–4 nm × 100–800 nm), L-Al2O3 (2–4 nm × 2800 nm)]. Levels of IL-1β production and cytotoxicity were concentration-dependent and were in the following order: S-Al2O3 > N-Al2O3 > L-Al2O3. Stimulus of THP-1 cells by Al2O3 particles led to lysosomal destabilization, generation of intracellular reactive oxygen species (ROS), and subsequent release of cathepsin B. The magnitude of the stimulus was dependent on the shapes and aspect ratios of the particles. Additional results suggested that caspase-1 (NALP3 inflammasome) activation and IL-1β production followed cathepsin B release. In addition, the cell death induced by Al2O3 particles was closely related with cathepsin B leakage. The mode of cell death was necrotic as much as apoptotic. We conclude that Al2O3 particles induce different levels of IL-1β production and cytotoxicity depending on their particle shapes or aspect ratios. The current finding may support the development of safe forms of Al2O3 particles.

Farnoosh Zarinhadideh, Akram Amini, Mohammad-Ali Assarehzadegan, Seyed-Hamid Borsi, Najmeh Sepahi, Hosein Ali-Sadeghi. Immunochemical and molecular characterization of allergenic profilin (Koc s 2) from Kochia scoparia pollen. J. Korean Soc. Appl. Biol. Chem. 58(3):443-451Kochia scoparia pollen has been demonstrated as an important cause of pollinosis in tropical and sub-tropical regions of the world. The aim of this study was to characterize the IgE-binding protein of the Kochia pollen extract and production of recombinant form of allergenic profilin of this weed. To predict its allergenic cross-reactivity with profilins of common allergenic plants, nucleotide sequence homology of Kochia profilin was evaluated. Specific ELISA and immunoblotting assay were applied to determine the IgE-

binding reactivity of 28 sera collected from patients who were sensitized to Kochia pollen. In cloning procedure, the Kochia profilin-coding sequence was inserted into PTZ57R/T vector and expressed using pET-21b(+) vector. IgE-binding competence of purified recombinant Kochia profilin (rKoc s 2) was analyzed by in vitro assays. There were several protein bands in Kochia pollen extract with molecular weights approximately ranging from 14.5 to 85 kDa. Nucleotide sequencing revealed an open reading frame of 399 bp encoding for 133 amino acid residues which belonged to the profilin family; 15 patients (15/28, 53.1 %) had significant specific IgE levels for the rKoc s 2. Immunodetection and inhibition assays indicated that the purified rKoc s 2 might be the same as that in the crude extract. Koc s 2, the first allergen from the Kochia pollen was identified as a member of the family of profilins. High degree of homology was found among amino acid sequences of Kochia profilin and several profilin molecules from unrelated plant families.

Se-Hui Lee, Min-Ju Ahn, Ji-Sang Hong, Ju-Hoon Lee. Diversity and community analysis of fermenting bacteria isolated from eight major Korean fermented foods using arbitrary-primed PCR and 16S rRNA gene sequencing. J. Korean Soc. Appl. Biol. Chem. 58(3):453-461Korean fermented foods are known to be beneficial for human health. Bacterial community studies have been conducted to figure out what roles the bacteria used to ferment these foods play in food fermentation. The metagenomic approach identifies both culturable and unculturable bacterial compositions, but this technique is limited in its ability to accurately determine the bacterial species from short 16S rRNA PCR products. In this study, we revisited the culture-dependent method using a relatively large number of bacterial isolates in an attempt to overcome the problem of bacterial identification, accepting that the unculturable bacterial population in each fermented food would be undetectable. Eight Korean fermented foods including kimchi, jeotgal, and meju were collected, and 1589 fermenting bacterial strains were randomly isolated. Bacteria were grouped by banding pattern using arbitrary-primed (AP) PCR prior to bacterial identification and sorted into 219 groups; 351 strains were not grouped because there was no identical AP-PCR band pattern. 16S rRNA sequence analysis identified the bacterial compositions of the fermented foods. As dominant genera, Lactobacillus and Leuconostoc strains were detected in four kimchi samples, Staphylococcus in three jeotgal samples, and Enterococcus and Bacillus in the meju sample. Interestingly, S. Equorumwas most dominant in saeu-jeotgal, indicating that it is halophilic and may enhance the fermentation flavor. Further comparative analysis of this study with previous metagenomic results revealed that bacterial communities in fermented foods are highly similar at the genus level but often differ at the species level. This bacterial community study is useful for understanding the roles and functional properties of fermenting bacteria in the fermentation process of Korean fermented foods.

Sarah Louise Atulba, Jessie Gutierrez, Gil Won Kim, Sang Yoon Kim, Muhammad Israr Khan, Yong Bok Lee, Pil Joo Kim. Evaluation of rice root oxidizing potential using digital image analysis. J. Korean Soc. Appl. Biol. Chem. 58(3):463-471The aerenchymal transport of oxygen to rice roots has significantly influenced the anaerobic root zone of flooded paddy soils. Therefore, the visualization of redox dynamics may be useful to characterize rice root oxidation potentials and the dynamics of redox-influenced ions in the root zone of paddy soils. In order to investigate the interaction between root oxidation potential and Fe uptake of (a) six different rice cultivars (Oryza sativa L.; Chuchung, Dongjin, Ilmi, Junam, Nampyeong, andSamkwang) were monitored in a flooded paddy soil with the aid of rhizotron experiment throughout the vegetation period, (b) digital images of the root zone were taken at the important growing stages, and (c) rice Fe uptake was characterized simultaneously. The images were processed by image analysis to display the reduction and oxidation areas in the root zones, and the distinct areas which were colorized due to varying soil redox changes were localized and quantified. Oxidized areas were mainly observed in the surrounding active roots and in a distinct layer on the soil surface. The selected rice cultivars have shown significantly different root-oxidized areas at the same rice growing stage. Root-oxidized area was

significantly and positively correlated with total Fe content of rice root, but negatively correlated with the inner root Fe content. Rice cultivars having higher root oxidation potential precipitated more Fe on the outer root surface in the form of Fe plaques. In conclusion, digital image analysis is an effective tool for evaluating the oxidizing potential of rice root under anaerobic soil condition.

Mi Jeong Kim, Jae Hoon Kim, Jun Ho Kim, Young Jun Kim. Comparative studies on the antioxidant capacities and catechin profiles of conventional and organic green tea. J. Korean Soc. Appl. Biol. Chem. 58(4):475-480As one of the most consumed beverages worldwide, green tea has been extensively studied for its wide array of health benefits. In this study, the composition of active compounds and antioxidative activity was analyzed using various antioxidant assay systems and compared between organic green tea (OGT) and conventionally grown green tea (CGT). OGT extract was prepared in the same way as CGT with the exception of fertilizer and insecticide, which were used only for CGT preparation. There were no significant differences in total polyphenol content and antioxidant activity tested against 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radicals and 1,1-diphenyl-2-picrylhydrazyl radicals. In addition, the antioxidant activities shown with both assays were highly correlated with the polyphenol contents (r 2 > 0.99). No significant differences were observed in the cellular antioxidant activity assay using HepG2 cells (P > 0.05). However, substantially higher concentrations of catechins, especially epigallocatechin-3-gallate, were found in the OGT (7.64 vs 5.54/100 mg extract). Thus, OGT could be considered as a potent and safe antioxidative resource, being free of both fertilizer and herbicide uses. However, the advantages of OGT should be further optimized to provide a wide range of physiological effects, considering the price of production and parallel amounts of the active ingredients in CGT.

Ji Eun Moon, Dong-Min Kim, Ji Yeon Kim. Anti-inflammatory effect of Rhus verniciflua stokes extract in the murine macrophage cell line, Raw264.7. J. Korean Soc. Appl. Biol. Chem. 58(4):481-486Rhus verniciflua stokes (RVS) (Anacardiaceae), which contains the major flavonoids fustin, fisetin, and sulfuretin, is known to have anti-inflammatory effects. Using lipopolysaccharide (LPS)—induced RAW264.7 cells, we examined which flavonoids were the most active compounds in the RVS extract. RVS extract dose-dependently reduced the production of nitric oxide, prostaglandin E2, interleukin-6 (IL-6), and reactive oxygen species (ROS) induced by LPS. Only sulfuretin significantly suppressed IL-6 and ROS levels although the effects were smaller than those provided by the RVS extract at the equivalent concentration (0.25 µg/mL of pure sulfuretin and 100 µg/mL of RVS extract). Other flavonoids such as fisetin and fustin did not show anti-inflammatory effects at the levels typically contained in the RVS extract. Taken together, sulfuretin was postulated to be the major anti-inflammatory flavonoid in RVS extract.

Min Ah Kim, Seung Jun Choi, Tae Wha Moon. Digestibility of retrograded starches with A- and B-type crystalline structures. J. Korean Soc. Appl. Biol. Chem. 58(4):487-490This study investigated whether the crystalline structure of the retrograded starch affects the contents of slowly digestible starch (SDS) and resistant starch (RS). When the relative crystallinities were similar, there was no difference in RS content between the retrograded starches with A- and B-type crystallites. However, the SDS content in A-type crystalline structure was constantly higher than B-type, contradicting the observation that the granular starches with B-type crystallites were more enzyme resistant. Therefore, the primary factor that accounts for the digestibility of granular starch is not the crystalline structure, but the granular properties, including the surface pores and the size.

Manoharan Saravanan, Ji-Yoon Kim, Hea-Na Kim, Seong-Beom Kim, Dong-Hoon Ko, Jang-Hyun Hur. Ecotoxicological impacts of isoprothiolane on freshwater fishCyprinus carpio fingerlings: a multi-biomarker assessment. J. Korean Soc.

Appl. Biol. Chem. 58(4):491-499Extensive use of fungicides in agriculture has led to the detrimental effects on aquatic ecosystems and organisms even at very low concentrations. In this respect, the present study was aimed to investigate the impact of environmentally relevant concentrations of fungicide isoprothiolane (2.7 and 27 µg/L; reported concentration in Yung San River, Korea) on a freshwater fish Cyprinus carpio for a short-term period of 96 h using various biomarkers such as hematological, ionoregulatory, biochemical, and enzymological parameters. At both concentrations, hemoglobin, hematocrit, red blood cell, serum glucose, cholesterol, triglycerides, aspartate aminotransferase, and gill Na+/K+-ATPase levels were significantly (p < 0.05) decreased in fish treated with isoprothiolane. Contrastingly, white blood cell, serum sodium (Na+), chloride (Cl−), and albumin levels were increased (p < 0.05). However, the short-term exposure of isoprothiolane in carp resulted in slight alterations in mean cellular volume, mean cellular hemoglobin, mean cellular hemoglobin concentration, serum potassium (K+), protein, globulin, and alkaline phosphatase levels. The present study concludes that alterations of hematological, ionoregulatory, biochemical, and enzymological parameters can be used as efficient biomarkers in monitoring the toxicity of isoprothiolane as well as other pesticides in aquatic organisms. Further, more detailed studies on using of specific biomarkers to monitor the long-term effects of isoprothiolane are needed.

Dae Yoon Cho, Min Kyoung Lee, Eun Ae Kim, Sook Young Lee. Analysis of the isoflavone content, antioxidant activity, and SDS-PAGE of cheese analogs produced with different proteolysis and soymilk residue contents. J. Korean Soc. Appl. Biol. Chem. 58(4):501-509We investigated the heat coagulation, SDS-PAGE, isoflavone content, and hydrogen-donating activity of cheese analogs produced with different hydrolysis time (0.3 % Flavourzyme® for 0, 20, 25, and 30 min) and soymilk residue contents (0, 7, 14, and 21 %) to improve the protein functionality and to use the by-product of soymilk. SDS gel electrophoresis of samples treated with Flavourzyme confirmed that the Flavourzyme treatments not only decreased the molecular weight of the proteins but also removed lipoxygenase, which results in the beany flavor of soy cheese, and Kunitz inhibitor (KTI), the antinutritional factor. The results of SDS-PAGE in the soy flour group and the roasted soy flour group treated with Flavourzyme® for 25 min and the determined soy milk residue contents indicated the recovery of some fractions (50, 32, and 18 kDa) of 7S and 11S globulin. The heat coagulation ratio increased as the amount of added soymilk residue was increased up to 14 % (p < 0.001). The glycoside (daidzin and genistin) and total isoflavone content increased in both the soy flour group and the roasted soy flour group as the amount of added soymilk residue was increased, but the contents of non-glycosidic compounds and the anti-oxidative activity were the highest in soy flour samples to which 7 % soymilk residue was added(p < 0.001). The variation in the anti-oxidative activity was similar to the variation in the contents of non-glycosidic compounds.

Minkyu Moon, Kyu-Suk Kang, Il-Kwon Park, Taekyu Kim, Hyun Seok Kim. Effects of leaf nitrogen allocation on the photosynthetic nitrogen-use efficiency of seedlings of three tropical species in Indonesia. J. Korean Soc. Appl. Biol. Chem. 58(4):511-519The leaf nitrogen (N) content (N L) and its allocation are major factors that determine the photosynthetic characteristics and photosynthetic N-use efficiency (PNUE) of tree species. In this study, we investigated the allocation of N L to the photosynthetic apparatus (N P) and its effects on the photosynthetic characteristics and PNUE of three tropical tree species (Magnolia montanaBlume, Swietenia macrophylla King, and Acacia mangium Willd), which were grown in western Java, Indonesia. A. mangium (the only N-fixing species among three studied species) exhibited a greaterN L and N P than other two species, resulting in higher capacities of photosynthetic parameters such as the light-saturated photosynthetic rate (P max), maximum carboxylation rate (V Cmax), and maximum electronic transport rate (J max). However, S. macrophylla and A. mangium displayed a similar fraction of leaf N allocated to N P (N P/N L) and showed a higher N P/N L ratio

than M. montana. Accordingly, S. macrophylla presented a higher PNUE than M. montana. Although A. mangium exhibited a similar PNUE to S. macrophylla, A. mangium showed a higher N L than the other two species, which resulted in a lower PNUE at a given N P. In addition, stomatal conductance was positively correlated with PNUE, but the slopes of the correlations between species presented a negative relationship with the leaf mass per area (LMA), which caused a longer pathway to the photosynthetic apparatus. Our results suggested that the variation of PNUE between species was affected by various factors, including the fraction of N L allocated to N P and morphological characteristics such as LMA.

Sumi Oh, Bo Ra Yi, Jung Woo Park, Mi-Ja Kim, Jae Hwan Lee. Effects of relative humidity and neutral emulsifier on oxidative stability of corn oil during room temperature storage. J. Korean Soc. Appl. Biol. Chem. 58(4):521-526The effects of tween 20, one of neutral emulsifiers, on oxidative stability of corn oil were determined under a relative humidity (RH) range of 0–93 % at room temperature after 350 days of storage. The degree of oxidation was determined by headspace oxygen content and conjugated dienoic acids analyses. Moisture content and α-tocopherol were also analyzed. The degree of lipid oxidation was significantly influenced by the concentration of tween 20 and relative humidity. Samples without addition of tween 20 showed the highest oxidative stability in 11 % RH, whereas oxidative stability was lowest at 93 % RH. Adding 0.5 and 1.0 % tween 20 increased the rate of lipid oxidation in a concentration dependent manner. Samples containing tween 20 had higher moisture content than those without an amphiphilic compound. Stability of α-tocopherol was depending on the content of moisture and amphiphilic compounds in bulk oil.

Jae Kyeom Kim, Ho-Jeong Lim, Dong-Hoon Shin, Eui-Cheol Shin. Comparison of nutritional quality and thermal stability between peanut oil and common frying oils. J. Korean Soc. Appl. Biol. Chem. 58(4):527-532The aim of this work was to compare the performances and nutritional characteristics of peanut oil to common frying oils during repetitive frying. The peanut oil had the highest color stability, the lowest initial, and final total polar compounds (TPC) levels among all other oil samples throughout the frying of potato slices. The TPC levels increased in all oils over the course of frying, being impacted more by the initial quality rather than the type of oils. In tocopherol analysis, as a consequence of frying, γ-tocopherol decreased most among the other six tocopherol isomers. Owing to the lack of data on the repetitive frying of peanut oil, this work provides basic information for peanut oil to use for frying purposes.

Youngwook Lee, Ramasamy Krishnamoorthy, Gopal Selvakumar, Kiyoon Kim, Tongmin Sa. Alleviation of salt stress in maize plant by co-inoculation of arbuscular mycorrhizal fungi and Methylobacterium oryzae CBMB20. J. Korean Soc. Appl. Biol. Chem. 58(4):533-540Arbuscular mycorrhizal fungi (AMF) and plant growth-promoting bacteria (PGPB) can have multiple impacts on phytohormone production and nutrient uptake to improve plant growth. Co-inoculation of AMF and PGPB can mitigate the effects of salinity in plants. This study was conducted to evaluate the efficacy of co-inoculation of AMF (Glomus etunicatum) and Methylobacterium oryzae CBMB20 to alleviate salt stress and improve maize growth. Single and co-inoculation of AMF and M. oryzaeCBMB20 significantly increased dry biomass, AMF root colonization, and nutrient accumulation in maize plants under salt stress. The proline content and Na uptake were significantly reduced in co-inoculation of AMF and M. oryzae CBMB20. In addition, co-inoculation of AMF and M. oryzaeCBMB20 (soil) showed higher AMF density and AMF colonization compared to single AMF treatment. Among all treatments, co-inoculation of AMF and M. oryzae CBMB20 (soil) was more effective to ameliorate salt stress and improve plant growth. These results demonstrate that co-inoculation of AMF and M. oryzae CBMB20 (soil application) may be an effective alternative to enhance plant growth under salinity stress.

Jae-Woo Jung, Ji-Hae Park, Kyeong-Hwa Seo, Eun-Ji Oh, Dae-Young Lee, Dong-Wook Lim, Daeseok Han, Myoung-Chong Song, Nam-In Baek. New hydroxy fatty acid from the root bark of Morus alba L. J. Korean Soc. Appl. Biol. Chem. 58(4):541-543A new hydroxyl fatty acid, named sangbaimoric acid (1), was isolated from the root bark of Morus alba L. The chemical structure of compound 1 was established as 2-methoxyoctadeca-3Z, 5Z-dienoic acid on the basis of spectroscopic analyses including nuclear magnetic resonance, high-resolution electronic ionization mass spectroscopy, and infrared spectroscopy experiments.

Youngju Kim, Ro-Dong Park. Progress in bioextraction processes of chitin from crustacean biowastes. J. Korean Soc. Appl. Biol. Chem. 58(4):545-554The most common methods for the recovery of chitin from crustacean shells are chemical procedures using strong acids and bases. The chemical demineralization and deproteinization processes have several drawbacks such as depolymerization, anomerization, and deacetylation. Moreover, chemical treatments are hazardous, energy consuming, and environmentally unfriendly. As an alternative to chemical processes for extracting chitin, fermentation processes using microorganisms or proteolytic enzymes have been studied for decades for various crustacean shells. Bioextraction of chitin is emerging as a green and eco-friendly process, thus overcoming some of the shortcomings of chemical extraction. Microorganism-mediated fermentation processes are desirable due to easy handling, simplicity, controllability through optimization of process parameters, ambient temperature, and less environmental impact. The main drawbacks of the biotechnological process are the lower yield and quality of the products and thus a higher cost than the chemical processes. We briefly discuss the critical issues encountered in biotechnological processes for chitin recovery from crustacean shells, together with compiling the most advanced findings during the last decades.

Jagadeesh Sundaramoorthy, Gyu Tae Park, Jeong-Dong Lee, Jeong Hoe Kim, Hak Soo Seo, Jong Tae Song. Genetic and molecular regulation of flower pigmentation in soybean. J. Korean Soc. Appl. Biol. Chem. 58(4):555-562Flower color is one of the key traits, which has been widely considered for genetic studies on soybean. A variety of flower colors, such as dark purple, purple, purple blue, purple throat, magenta, pink, near white, and white, has been identified in cultivated soybean (Glycine max). Out of the 19,649 soybean accessions deposited in the United States Department of Agriculture-Germplasm Resources Information Network database, 67 % have purple flowers, 32 % have white flowers, and merely 1 % have flowers with different colors. In contrast, almost all accessions of wild soybean (Glycine soja) have only purple flowers. Flavonoids, mainly anthocyanins, are the most common pigments contributing to flower coloration in soybean. In the recent decades, the flavonoid biosynthesis pathway for anthocyanins has been well established, and some of the genes controlling flower color in soybean have been identified and characterized. Flower pigmentation of soybean is mainly controlled by six independent loci (W1, W2, W3, W4, Wm, and Wp) along with the combination of various other factors such as anthocyanin structure, vacuolar pH, and co-pigments. In this review, we summarize the current status of genetic and molecular regulation of flower pigmentation in cultivated and wild varieties of soybean.

Jeong-Ho Lee, Kicheol Lee, Da Hyun Lee, Soon Young Shin, Yeonjoong Yong, Young Han Lee. Anti-invasive effect of β-myrcene, a component of the essential oil from Pinus koraiensis cones, in metastatic MDA-MB-231 human breast cancer cells. J. Korean Soc. Appl. Biol. Chem. 58(4):563-569The invasive potential of malignant tumor cells is critical for their metastasis. This study was undertaken to evaluate the anti-invasive activity of β-myrcene, a natural compound found in the essential oil from Pinus koraiensis cones (EOPC), in metastatic MDA-MB-231 human breast cancer cells. Among four major constituents that included α-pinene, β-myrcene, 3-carene, and d-limonene, β-myrcene showed the most potent inhibition of tumor necrosis factor-α (TNFα)-induced nuclear factor κB (NF-κB) activity. Pretreatment with β-myrcene suppressed TNFα-induced phosphorylation of inhibitor of κB kinase and

NF-κB as well as matrix metalloproteinase-9 (MMP-9) gene expression in a dose-dependent fashion. Furthermore, β-myrcene inhibited TNFα-induced invasion of MDA-MB-231 cells as determined by three-dimensional spheroid invasion assays. These findings suggest that EOPC may promote anti-metastatic activity in breast cancer cells through its downregulation of NF-κB-mediated MMP-9 expression.

Kyung-Hye Seo, Ji-Eun Ra, Sung-Joon Lee, Ji Hae Lee, Sun Rim Kim, Jin Hwan Lee, Woo Duck Seo. Anti-hyperglycemic activity of polyphenols isolated from barnyard millet (Echinochloa utilis L.) and their role inhibiting α-glucosidase. J. Korean Soc. Appl. Biol. Chem. 58(4):571-579The extracts and chemical compounds isolated from barnyard millet (Echinochloa utilis) grains were investigated as part of a search for naturally derived anti-hyperglycemic medicinal plants. Among the five different solvent extracts, the ethyl acetate extract showed the lowest IC50 value againstSaccharomyces cerevisiae α-glucosidase (70.2 μg/mL). Chromatography yielded eight phenolic compounds that may have been responsible for this effect. Among them, N-p-coumaroyl serotonin (1, CS), feruloyl serotonin (2, FS), and luteolin (5) potently inhibited α-glucosidase with IC50 values of 1.3–17.8 μM compared with those of deoxynojirimycin (DNJ, IC50 = 2.5 ± 0.1 μM) and acarbose (IC50 = 255.1 ± 15.6 μM). Additionally, for the first time, we found that CS and FS were significantly inhibited mammalian rat intestinal sucrase (IC50 of 3.0 and 8.2 μM) and reduced glucose content (73 and 52 % at 0.1 mg/wells) in Caco-2 (human intestinal epithelial) cells. Furthermore, oral glucose tolerance test revealed improved glucose tolerance following treatment with the barnyard millet grains extract by retarding the postprandial rise in blood glucose in vivo. These results suggest that barnyard millet grain can be used as a natural functional medicine to prevent and alleviate type-2 diabetes.

Deepak Kumar, M. Poornima, R. N. Kushwaha, Tae-Jin Won, Chuljin Ahn, C. Ganesh Kumar, Kiwan Jang, Dong-Soo Shin. Antimicrobial and docking studies of (−)-catechin derivatives. J. Korean Soc. Appl. Biol. Chem. 58(4):581-585Antimicrobial activities of (−)-catechin derivatives were assayed for their antibacterial and antifungal activities against gram positive, gram negative bacteria, and fungi. Most of the compounds significantly active among which Compounds 1a, 1b, and 1c showed excellent antibacterial for both gram negative and gram positive bacteria, these compounds also exhibited excellent antifungal activity more than the standard drug. Molecular docking studies of Compounds 1a and 1bestablished good binding affinity with ATP-binding pocket of DNA gyrase and are in favor of the observed biological activity. These data collectively suggest that Compounds 1a and 1b could serve as a novel antimicrobial agent.

Young-Jun Kim, Youngjae Shin. Antioxidant profile, antioxidant activity, and physicochemical characteristics of strawberries from different cultivars and harvest locations. J. Korean Soc. Appl. Biol. Chem. 58(4):587-595Three major cultivars of strawberries (‘Yukbo,’ ‘Seolhyang,’ and ‘Janghee’) harvested from three different locations (Gyeongsan, Nonsan, and Daegu) in Korea were used for this study. The soluble solid content of ‘Yukbo’ harvested from Nonsan was the highest among the samples. The ranges of titratable acidity and pH from the samples were 0.48–0.65 and 3.70–4.01 %, respectively. Fructose, glucose, and citric acid contents of ‘Janghee’ harvested from Daegu were significantly higher than other strawberries (p < 0.05). The ascorbic acid contents of ‘Seolhyang’ and ‘Janghee’ grown in Nonsan were significantly higher than in other strawberries (p < 0.05). Total flavonoid and phenolic concentrations, and total antioxidant activities of ‘Janghee’ harvested from Daegu were significantly higher than other samples (p < 0.05). Total flavonoids and total phenolics were highly correlated with the antioxidant activities, and the relationships between total flavonoids and total phenolics were also strong. Cultivars and harvest locations may affect to the physicochemical quality and antioxidant activity of the fruit.

Hwa-Won Lee, Hoi-Seon Lee. Acaricidal potency of active constituent isolated

from Mentha piperita and its structural analogs against pyroglyphid mites. J. Korean Soc. Appl. Biol. Chem. 58(4):597-602This study was conducted to evaluate acaricidal toxicities of an active compound isolated fromMentha piperita leaves and its structural analogs against pyroglyphid mites, Dermatophagoides farinae, and D. pteronyssinus. Menthol was isolated through chromatographic techniques. Based on the LD50 values of M. piperita oil and menthol analogs against D. farinae and D. pteronyssinus,obtained using the impregnated cotton fabric disk method, the acaricidal activities of (−)-menthol (0.19 and 0.12 μg/cm3) were 194 and 164 times higher than those of DEET (36.89 and 19.72 μg/cm3), followed by that of (±)-menthol (0.23 and 0.21 μg/cm3), (+)-menthol (0.29 and 0.23 μg/cm3), (+)-neomenthol (0.32 and 0.26 μg/cm3), (+)-isomenthol (0.34 and 0.29 μg/cm3), M. piperita oil (1.50 and 1.00 μg/cm3), 4-methylcyclohexanol (5.45 and 4.76 μg/cm3), 2-methylcyclohexanol (7.64 and 5.67 μg/cm3), and 1-methylcyclohexanol (9.82 and 8.56 μg/cm3). Similarly, the acaricidal activities of M. piperita oil and menthol analogs in the filter paper method were similar to those in the impregnated cotton fabric disk method against Dermatophagoides spp. In the structure–activity relationships, the isopropyl (C3H7) functional group on the cyclohexane skeleton influenced acaricidal activity the most, followed by the hydroxyl (OH) and methyl (CH3) functional groups. This study indicates that M. piperita oil and menthol analogs have the potential for development as preventive agents for control of Dermatophagoides spp.

Eun-Kyung Seo, Hyeyoung Kwon, Su-Myeong Hong, Taek-Kyum Kim. Simultaneous determination of flonicamid, imidacloprid, and its metabolites in paprika by QuEChERS and tandem mass spectrometry. J. Korean Soc. Appl. Biol. Chem. 58(4):603-610An analytical method for the simultaneous determination of flonicamid, imidacloprid, and its metabolites is established. The method developed herein uses a rapid extraction and cleanup procedure based on quick, easy, cheap, effective, rugged, and safe followed by liquid chromatography–tandem mass spectrometry. The effects of various experimental parameters, including extraction, salting-out agents, and purification sorbents, were investigated to optimize the sample-preparation method; the optimized method was then validated using fortified samples. Flonicamid, imidacloprid, and its metabolites spiked at three concentration levels: 0.05, 0.1, and 0.5 mg kg−1; the means of their recovery were determined by replicate analysis at each level. Under the conditions described, the mean recoveries of flonicamid, imidacloprid, and its metabolites were in the acceptable range 70–120 % with a satisfactory precision (RSD ≤20 %). This modified method provides improved recoveries of flonicamid metabolites and may be a useful analytical technique for the simultaneous determination of flonicamid, imidacloprid, and its metabolites in fruits and vegetables.

Sangbuem Cho, Jeong-Mee Kim, Mi-Sang Yu, Su-Jung Yeon, Chi-Ho Lee, Soo-Ki Kim. Fermentation of hot pepper juice byBacillus licheniformis to reduce pungency. J. Korean Soc. Appl. Biol. Chem. 58(4):611-616Capsaicin, an active component of hot pepper, has considerable health benefits; however, the pungency of hot pepper has limited its usage. This study aimed to reduce the pungency of hot pepper by fermentation to enhance its application in the food industry. Hot pepper juice was fermented by Bacillus licheniformis SK1230, which was previously isolated as a capsaicin-degrading bacterium. B. licheniformis SK1230 was inoculated into the juice of red or green pepper, and the mixture was then fermented for 19 days to determine the degradation level of capsaicinoids. It was observed that with a gradual increase in pH, the growth of B. licheniformis SK1230 increased to 1.0 × 109 CFU/mL after day 1, and its viability persisted until the end of fermentation. The sugar content of green pepper drastically reduced at day 1 and that of red pepper reduced at day 5. The total polyphenol content of the medium containing red pepper was about 2-fold higher than that of the medium containing green pepper. The antioxidant activity of the medium containing red pepper was also higher than that of the medium containing green pepper; however, the activity gradually decreased during the course of the fermentation period. The capsaicinoid content of both types of peppers radically

decreased at day 5. Thus, we suggest that a fermentation period of 5 days would be optimal for the degradation of capsaicin with minimal loss of antioxidant activity and would provide higher polyphenol content.

Ting Shen , Xueqin Li , Weicheng Hu, Lijin Zhang, Xudong Xu, Haifeng Wu, Lilian Ji. Hepatoprotective effect of phenylethanoid glycosides fromIncarvillea compacta against CCl4- induced cytotoxicity in HepG2 cells. J. Korean Soc. Appl. Biol. Chem. 58(4):617-625The extraction and solvent partition of roots of Incarvillea compacta, a traditional Tibetan folk medicine, and repeated column chromatography and preparative high-performance liquid chromatography for n-butanol fraction yielded four phenylethanoid glycosides, crenatoside (1), 3′′′-O-methylcrenatoside (2), leucoseceptoside A (3), and martynoside (4). The chemical structures were identified on the basis of spectroscopic data analyses including NMR and MS. All compounds were isolated for the first time from the plant. Compound 1 exerted better 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity. In addition, compounds 1–4 were evaluated for their hepatoprotective activity on carbon tetrachloride (CCl4)-induced liver injury in HepG2 cells. Pretreatment of HepG2 cells with compound 1–4 significantly increased the viability on CCl4-induced cell death. Furthermore, compounds 1–4 also alleviated CCl4-induced hepatotoxicity by enhancement of the antioxidant enzyme activities of superoxide dismutase and reduction of the malondialdehyde content, intracellular ROS as well as NF-κB transactivation. Our results suggest that phenylethanoid glycosides ameliorate CCl4-induced cell injury, and this protection was likely due to antioxidative activity and down-regulation of NF-κB.

Jihye Seong. Lipid rafts signaling detected by FRET-based molecular biosensors. J. Korean Soc. Appl. Biol. Chem. 58(5):629-636Cells recognize and process various extracellular signals via transmembrane receptors and membrane-associated signaling molecules. Therefore, the precise regulation of these signaling events at the plasma membrane, which occur spatiotemporally, is necessary for proper cellular functions such as cell proliferation, migration, and survival. The plasma membrane may contain dynamic microdomains called lipid rafts, which are suggested to be crucial for the regulation of efficient and specific signaling pathways. However, because of the limitation in methodologies, the specific molecular mechanisms underlying dynamic signaling events at lipid rafts are largely unknown. This review discusses the traditional biochemical methods for the visualization of lipid rafts and the related signaling events at these microdomains. In addition, the review reports on fluorescence resonance energy transfer (FRET)-based molecular biosensors with lipid rafts targeting sequences as a powerful tool for live-cell imaging of spatiotemporal signaling events at lipid rafts. In particular, examples of dynamic lipid rafts signaling mechanisms visualized by FRET-based biosensors in live cells are covered in the last section.

Hyeon Ji Kim, Jeong-Ju Baek, Byung-Jung Min, Jong Hoon Kim, Sang-Ho Yoo, Suyong Lee. Utilization of brown rice flour as a functional ingredient in instant fried noodles. J. Korean Soc. Appl. Biol. Chem. 58(5):637-642Brown rice flour was utilized as a functional ingredient in instant fried noodles and their quality attributes were characterized from the standpoint of physicochemical characteristics. The pasting results showed that there was an increasing tendency in the peak, trough, and final viscosities of wheat flour with increasing amounts of brown rice flour. The thermal conductivity of the noodle dough with brown rice flour had a lower value of thermal conductivity. When the noodles were subjected to frying, the brown rice flour did not play a negative role in the oil absorption of the instant fried noodles. The use of brown rice flour contributed to retarding the oxidative deterioration of the instant fried noodles during storage. While the breaking stress of instant fried noodles before cooking increased with increasing amount of brown rice flour, the tensile properties of the noodles were lowered after cooking.

Hee-Sook Park, Soon-Mi Shim. Preventive responses to a standardized extract

of Houttuynia cordatasupplemented diet in obesity induced by a high-fat diet in mice. J. Korean Soc. Appl. Biol. Chem. 58(5):643-649The current study investigated preventive responses to Houttuynia cordata (HC) standardized extract in obesity induced by a high-fat diet (HFD) in mice. High-performance liquid chromatography was carried out to analyze bioactive components in HC. Mice were divided into HFD groups as a control and HFD groups fed with 1 and 4 % of HC supplementation diet. Changes of the body weight of mice, blood biological factors, and histopathological markers were measured. Bioactive components from HC were characterized to contain chlorogenic acid, caffeic acid, rutin, and quercitrin. HC supplementation reduced body weight, glucose, and total cholesterol levels, but it did not affect aspartic acid transaminase, alanine transaminase, or triglyceride amounts in plasma from mice induced by HFD. A 4 % HC supplementation showed a remarkable decrease in the number of large adipocyte in subcutaneous and abdominal adipose tissue. Histopathological evaluation of liver injury induced by HFD was observed to be improved by HC supplementation, leading to attenuated levels of cytoplasmic vacuolation of hepatocytes. From these results, we explored that standardized extract of HC could be a potential natural product to be directly associated with the control of weight.

Jun Yeon Lee, Kwi Hyung Cho, Jung Woo Kim, Sang Don Nam, Hee Yeon Kim, Jae Soo Shin, Dong Yeon Kim, Seung-Han Lee. Synthesis and biological screening of small molecule peptides based on the amino acid sequence of thymosin β4. J. Korean Soc. Appl. Biol. Chem. 58(5):651-658Small molecule peptides based on the amino sequence of thymosin β4 were prepared using solid-phase synthesis, and their bioactivities were investigated. Small molecule peptides are less than ten amino acids in size so can be produced on a large scale and, after modification, can easily penetrate blood vessels or tissues. We studied the ability of the small molecule peptides to promote wound healing, angiogenesis, and hair growth by examining their effects on the proliferation of human dermal fibroblasts, human umbilical vein endothelial cells, and human dermal papilla cells versus those of thymosin β4, vascular endothelial growth factor, and minoxidil, respectively. Several small molecule peptides promoted significantly more cell proliferation than the corresponding positive controls.

Bitnara Lee, Do-Won Jeong , Jong-Hoon Lee. Genetic diversity and antibiotic resistance of Staphylococcus saprophyticus isolates from fermented foods and clinical samples. J. Korean Soc. Appl. Biol. Chem. 58(5):659-668A multilocus sequence typing (MLST) scheme was developed to evaluate the genetic diversity and background of Staphylococcus saprophyticus strains of food and clinical origins. A total of 48 isolates from human urine samples (21 isolates), Doenjang (9 isolates), Myeolchi-jeotgal (6 isolates), Saeu-jeotgal (3 isolates), and sausages (9 isolates) were subjected to MLST using internal fragments of 7 housekeeping genes: aroE, dnaJ, glpF, gmk, hsp60, mutS, and pta. MLST analysis resulted in 26 sequence types (STs), and the eBURST algorithm clustered the STs into 4 clonal groups (CGs) and 6 singletons. The predominant STs, ST11 (16.7 %, 8/48), and ST7 (10.4 %, 5/48), belonged to the major CG, CG1 (12 STs, 60.4 %, 29/48), and comprised isolates fromDoenjang and urine. CG2 and CG3 comprised isolates from sausages, while CG4 included isolates from Myeolchi-jeotgal and Saeu-jeotgal. Twenty of the S. saprophyticus isolates exhibited resistance to erythromycin, lincomycin, or tetracycline. Lincomycin resistance was identified from the ST10–12 isolates, all of which are variants of ST08 and were isolated from Doenjang and human urine samples. This MLST scheme established the genetic diversity within S. saprophyticus, and clustering of the STs using eBURST revealed a correlation between the genetic backgrounds and the origins of isolates, and a link between genetic background and acquisition of lincomycin resistance.

Chi-Yeol Kim, Kieu Thi Xuan Vo, Gynheung An, Jong-Seong Jeon. A rice sucrose non-fermenting-1 related protein kinase 1, OSK35, plays an important role in fungal and bacterial disease resistance. J. Korean Soc. Appl. Biol. Chem.

58(5):669-675Rice blast, caused by the fungal pathogen Magnaporthe oryzae, and bacterial leaf blight, caused byXanthomonas oryzae pv. oryzae (Xoo), impose serious limitations on rice productivity worldwide. Sucrose non-fermenting-1 related protein kinases (SnRKs) have been suggested to play a role in global gene expression reprogramming in response to biotic stress. However, little is known in rice about the role of SnRKs in modulating disease resistance. Here, we characterized the function ofOSK35, a rice SnRK1b gene, in disease resistance by analyzing two T-DNA mutants, OSK35-D, an activation line, and osk35, a knockout line. Both mutants and their respective wild-type controls were challenged with two different rice pathogens, M. oryzae and Xoo. The OSK35-D activation mutant showed enhanced resistance against both rice pathogens. Conversely, the osk35 null mutant was more susceptible to these pathogens, compared with wild-type controls. These results clearly indicate that OSK35 is a positive regulator of rice disease resistance upon fungal and bacterial pathogen invasion.

Young-Jun Kim, Hyong Joo Lee, Youngjae Shin. Estimation of daily curcuminoid intake from commercial curry products. J. Korean Soc. Appl. Biol. Chem. 58(5):677-684Turmeric, the main spice in curry, contains curcuminoids, which have been closely linked to the prevention of various chronic diseases. However, the daily intake of curcuminoids from curry consumption remains unclear. This study optimized an analytical method for quantifying individual curcuminoids and then applied it to 50 commercial curry samples available in the retail market. The curcuminoids namely curcumin, demethoxycurcumin, and bisdemethoxycurcumin, which provide the yellow color present in turmeric, were analyzed, and a daily curcuminoid intake from a curry-based diet was estimated. Prior to data collection, the analytical method, incorporating high-performance liquid chromatography with a diode array detector, was validated for linearity, limit of detection (0.03–0.04 mg/kg), limit of quantification (0.10–0.14 mg/kg), and precision and accuracy (less than a relative standard deviation of 1.25 %). Total curcuminoid content in the curry products tested varied greatly depending on the product type, ranging from 24.59 to 161.02 mg/100 g (mean 75.92 mg/100 g) for powdered curry and from 1.04 to 10.78 mg/100 g (mean 4.12 mg/100 g) for retorted curry (precooked curry packaged in a retorted pouch). The daily intake of total curcuminoids from curry products, estimated from the data in this study and available domestic consumption data, was 0.339 mg/person/day for males and 0.299 mg/person/day for females. The distribution of total curcuminoids varied greatly by product owing to variation in the turmeric contents used by curry manufacturers in individual products. The results reveal the wide variation in the curcuminoid contents of commercial curry products and provide information on daily curcuminoid intake.

San-qing Lin, Zhong-liu Zhou, Hua-lin Zhang, Wen-qing Yin. Phenolic glycosides from the rhizomes ofCyperus rotundus and their antidepressant activity. J. Korean Soc. Appl. Biol. Chem. 58(5):685-691Depression is the most common psychiatric comorbidity in patients. Searching for antidepressant drugs from natural products is very important for the treatment of this disease. The ethanol extract and its fractions of Cyperus rotundus were evaluated for antidepressant activities in mice by our group. The aim of this study was to investigate the potential active components of C. rotundus. The extraction and solvent partition of the rhizomes of C. rotundus, and repeated column chromatography for n-BuOH fraction yielded three new phenolic glycosides, rotunduside D (1), rotunduside E (2), and rotunduside F (3). Their chemical structures were identified on the basis of spectroscopic data analyses including NMR, MS, UV, and IR. The potential antidepressant activity of all the isolated compounds was evaluated. In the despair mice models, rotunduside F (3) showed significant antidepressant activity at the dosage of 50 mg/kg, which was close to the positive control fluoxetine (20 mg/kg).

Jae-Hyun Yoon, Young-Min Bae, Sun-Young Lee. Predictive modeling on the growth ofStaphylococcus aureus inoculated on Korean style cooked foods

stored at various temperatures. J. Korean Soc. Appl. Biol. Chem. 58(5):693-701This study was conducted to develop predictive models for the growth of Staphylococcus aureus on Korean style cooked foods (potato salad, stir-fried boiled fish paste, seasoned baby aster, and stir-fried anchovies) as a function of various storage temperatures (from 15 to 35 °C). The modified Gompertz equation was applied to fit the growth curves. Potato salad, stir-fried boiled fish paste, and seasoned baby aster showed the greatest specific growth rate by 1.219, 0.782, and 1.282 log10CFU/h and the shortest lag time by 0.283, 0.407, and 0.374, respectively, when stored at 35 °C. No growth of S. aureus was observed in stir-fried anchovies due to its low water activity. Subsequently, these results were employed to establish the secondary models, which were further used for validation using accuracy factor and bias factor. Overall, accuracy factor of less than 1.461 and bias factor of less than 1.093 were observed depending on different storage temperatures and a sort of contaminated food samples. These results will be used to develop the manual representing the safe storage period of Korean foods contaminated with pathogenic bacteria.

Eun-Seo Lim. Purification and characterization of two bacteriocins from Lactobacillus brevisBK11 and Enterococcus faecalis BK61 showing anti-Helicobacter pylori activity. J. Korean Soc. Appl. Biol. Chem. 58(5):703-714The aim of this study was to investigate the antimicrobial effects of purified bacteriocins isolated from Lactobacillus brevis BK11 and Enterococcus faecalis BK61 on Helicobacter pylori. After the final purification step, the molecular weights of the purified bacteriocins from L. brevis BK11 and E. faecalis BK61 were estimated to be approximately 6.5 and 4.5 kDa, respectively. Significant decrease in the antimicrobial activity of these bacteriocins was observed when they were treated with several proteolytic enzymes. However, their antimicrobial activity was stable over long periods of storage and a wide range of pH and was relatively heat resistant. The inhibitory spectrum of the bacteriocin produced by L. brevis BK11 strain was quite narrow, whereas the bacteriocin produced from E. faecalis BK61 strain exhibited a broad antimicrobial spectrum against various foodborne pathogens such as Listeria monocytogenes, Escherichia coli O157, and Salmonella enteritidis. Treatment of H. pylori with these bacteriocins significantly reduced the number of cells of the pathogen found adhering to the monolayers of cultured human gastric adenocarcinoma epithelial cell line (p < 0.05). Furthermore, the urease activity of adherent H. pylori after treatment with the bacteriocins was lower than that for the control group, but there appeared to be no significant difference between the competition and displacement groups (p > 0.05).

Ho Bong Hyun, Sabina Shrestha, Kyung Hwan Boo, Somi Kim Cho. Evaluation of antioxidant potential of ethyl acetate fraction of Rosmarinus officinalis L. and its major components. J. Korean Soc. Appl. Biol. Chem. 58(5):715-722The solvent fractions of rosemary methanol extract were obtained by successive extraction with n-hexane, chloroform, ethyl acetate, butanol, and water. The ethyl acetate fraction (EAF) contained a remarkable amount of polyphenol and flavonoid as well as high levels of alkyl and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity. The activity guided fractionation and repeated chromatographic separations over silica gel, RP C18, and Sephadex LH-20 led to isolation of six compounds from the EAF. 1H NMR, 13C NMR, 2D NMR, MS, and IR spectroscopies determined the compounds to be caffeic acid (1), rosmarinic acid (2), rosmarinic acid methyl ester (3), luteolin (4), apigenin (5), and hispidulin (6), and high-performance liquid chromatography quantification was used to determine concentrations in EAF. Among the six isolated compounds, rosmarinic acid methyl ester showed the highest scavenging activities against di(phenyl)-(2,4,6-trinitrophenyl) iminoazanium, alkyl and ABTS radicals. The EAF mixture, but not individual isolated compounds, shielded dermal fibroblast cells from H2O2-induced cytotoxicity at concentrations that encompass the SC50 of alkyl and ABTS radical. Therefore, our findings suggested for the first time that antioxidant capacity of the EAF mixture result in a synergistic effect on the antioxidant action.

Yeon-Ki Kim, Baek Hie Nahm, Nam Iee Oh, Jong-Joo Cheong. Determination of the consensus sequence for FUS3-specific binding by protein binding microarray analysis. J. Korean Soc. Appl. Biol. Chem. 58(5):723-728A protein binding microarray (PBM) analysis was performed to determine the nucleotide sequence responsible for the specific binding of FUS3, a transcription factor in Arabidopsis that is considered to be a master regulator of seed development. Full-length FUS3 cDNA was fused in-frame with sequences encoding the fluorescent protein DsRed and a polyhistidine tag, and expressed in Escherichia coli. The partially purified fusion protein was incubated with the quadruple 9-mer PBM. An analysis of fluorescent images revealed that the fusion protein recognized and bound to the consensus sequence CATGCA. This result suggests that, among the nucleotides contained within the RY motif (CATGCA[TG]), six nucleotides (CATGCA) are sufficient for FUS3-specific binding. It appears that an additional ‘TG’ at the 3′ end of the RY motif found in seed-specific gene promoters was not necessary for binding specificity. Notably, FUS3 may also bind to the palindromic RY motif in its dimeric form.

Kang Hee Kho, Jung Woo Kim, Soo Cheol Kim, Myeong Rak Choi, Kyeong Ho Han, Won Kyo Lee, Kap Seong Choi. Identification and intracellular localization of carbonic anhydrase I in gills, heart, muscle, and intestine of rainbow trout, Oncorhynchus mykiss. J. Korean Soc. Appl. Biol. Chem. 58(5):729-733Carbonic anhydrases (CAs) play a key role in physiological functions such as pH homeostasis, calcification, photosynthesis, and ionic regulations. Among CA isozymes, CA I is a specific marker for the cytoplasm or apical cell membranes of colonic epithelial cells, and it is related to enterocyte proliferation in mammals. The major aim of the present study was to investigate the regional, cellular, and subcellular localization of CA I in a fish, rainbow trout, Oncorhynchus mykiss. The CA I was identified in the gills, heart, and intestine of the rainbow trout. CA I protein detected in the tissues of the rainbow trout was similar to a CA III with a molecular weight of 29 kDa. Immunohistochemical results demonstrated that CA I was localized in the various cells of gills, heart, muscle, and intestine of the rainbow trout.

Haechang Yi, Keum Taek Hwang, Heedon Choi, Hak-Tae Lim. Physicochemical and organoleptic characteristics of deep-fat fried and microwaved potato chips. J. Korean Soc. Appl. Biol. Chem. 58(5):735-740This study was carried out to examine the effects of deep-fat frying and microwaving on various physicochemical parameters and sensory characteristics of potato chips. The moisture content of deep-fat fried potato chips was found to decrease faster than that of microwaved chips during the cooking process. The fat content of the deep-fat fried potato chips increased with increasing frying time, while that of microwaved chips changed little. Microwaving was found to increase the hardness and lower the acrylamide content of the chips. Microwaving also resulted in chips with a lower browning index and a lighter color than deep-fat fried chips. Although deep-fat fried chips were found to be superior in terms of appearance, color, and overall acceptability, no significant differences were determined in the taste and crispiness between deep-fat fried and microwaved potato chips. Our findings indicate that microwaving as a cooking method has the potential to yield potato chips with low acrylamide levels, low fat content, and a desirable texture. Microwaving can therefore be considered a suitable method of preparing potato chips for modern people who consume excessive fats and oils and are interested in healthier and high-quality products.

Dong Ho Shin, Tae-Lim Kim, Man-Ho Cho, Sang-Won Lee , Seong Hee Bhoo. Characterization of Arabidopsis mutants insensitive to high sugar concentrations. J. Korean Soc. Appl. Biol. Chem. 58(5):741-744Sugar is appreciated to function as a signaling molecule by controlling gene expression, thereby regulating various developmental processes in plants. We screened sucrose-insensitive (sin) mutants in Arabidopsis that showed normal germination and seedling development under high concentration (438 mM) of sucrose. Compared to wild-type and other sugar mutants, sin mutants seedlings exhibited resistance to high glucose, high

salt, and paclobutrazol, inhibitor of gibberellin biosynthesis. We discuss the unique characteristics of these sin mutants in this report.

Seung Min Kim, Taeyoung Oh, Hyun Jung Kim. Antimicrobial resistance, molecular, and phenotypic diversity of Escherichia coliisolates from fresh vegetable products in Korea. J. Korean Soc. Appl. Biol. Chem. 58(5):745-750

Escherichia coli is generally considered as an indicator of antimicrobial resistance due to its genetic flexibility and adaptability. In the present study, antimicrobial resistance patterns, biofilm formation ability, and genotypes were assessed in E. coli isolates from fresh vegetable products in Korea. Among 120 isolates, 22 isolates (18.3 %) were resistant to one or more antimicrobials and 11 isolates were concurrently multidrug resistant against up to six antimicrobial agents. The highest resistance rate was detected on ampicillin (14.2 %), followed by piperacillin (11.7 %) and cefalotin (10.0 %). Rep-Polymerase chain reaction (PCR) revealed that five out of the 26 isolates were assigned to one cluster with a similarity >95 %, and strains carrying the same antimicrobial resistance profile had discrete rep-PCR patterns. A biofilm formation assay demonstrated that significantly larger amounts of biofilm were formed by an isolate, which showed no antimicrobial resistance. However, 60 % of the five intermediated biofilm-forming E. coli exhibited multidrug resistance against three or four antimicrobial agents. These results suggest that fresh vegetable products contaminated with E. coli are sources of antimicrobial resistance and other diverse virulence determinants. Therefore, the information provided in this study can be useful for improving food safety and public health.

Jong Hwan Park, Yong Sik Ok, Seong Heon Kim, Se Won Kang, Ju Sik Cho , Jong Soo Heo, Ronald D. Delaune, Dong Cheol Seo. Characteristics of biochars derived from fruit tree pruning wastes and their effects on lead adsorption. J. Korean Soc. Appl. Biol. Chem. 58(5):751-760The aim of this study was to evaluate the biochar characteristics derived from fruit tree pruning wastes (FTPW) and their effects on lead (Pb) adsorption. Based on results from Pb adsorption, surface area, and phosphorus content, the optimum pyrolysis temperature was 600 °C for Pb adsorption capacity. Using the Freundlich isotherm, the Pb adsorption capacity (K) of biochar obtained from various FTPW decreased in the order of pear (3.8001) ≫ persimmon (2.3977) ≥ apple (2.1968). Based on the Langmuir adsorption isotherm, the maximum Pb adsorption capacities (a; mg g−1) of biochar obtained from different FTPW were in the following order: pear (26.2) ≫ persimmon (19.9) ≥ apple (17.7). The maximum Pb adsorption capacity of the pruned pear tree waste biochar was greater than the other FTPW biochars. Pruned apple tree waste biochar had the lowest Pb adsorption capacity among the tested FTPW biochars. The positive correlation between the Langmuir maximum adsorption capacity (L M) values of the biochars and their phosphorus content and surface area indicated difference in adsorption capacity. However, adsorption capacity of the biochar from all FTPW studied could be used for removing Pb and other metal from wastewater.

Mi-Jung Park, Sook-Young Lee. Quality characteristics of soy yogurt produced using proteases and mixed microbial consortia. J. Korean Soc. Appl. Biol. Chem. 58(5):761-769In this study, we examined the quality characteristics of soy yogurts produced using two proteases(Flavourzyme® and Neutrase®) and the mixed microbial consortia of Lactobacillus bulgaricus and Streptococcus thermophilus (two-mixed) and Lactobacillus acidophilus,Bifidobacterium lactis, and S. thermophilus (three-mixed). The viscosity was the highest in the untreated yogurt. Among the proteolyzed samples, those treated with Flavourzyme® for 20 min showed the highest viscosity, whereas the samples treated with Neutrase® for 30 min and cultured with three-mixed microbial consortia showed the lowest viscosity. The numbers of viable bacteria in the Flavourzyme®-treated samples were slightly greater than those in the Neutrase®-treated samples. The number of viable bacteria increased as hydrolysis time increased. The

organic acid content was higher in the three-mixed group than in the two-mixed group, and it was higher in the Flavourzyme®-treated samples than in the Neutrase®-treated samples. Among the Flavourzyme®-treated samples, the three-mixed group showed higher total free amino acid content than the two-mixed group. In contrast, among the Neutrase®-treated samples, those cultured with both mixed microbial consortia produced the similar amounts of free amino acids. The above results showed that proteolytic treatment promoted the growth of lactic acid-producing bacteria. Soy yogurts produced using Flavourzyme® and three-mixed microbial consortia were thought to be more desirable because of more organic acids and free amino acids than those produced using Neutrase®and two-mixed microbial consortia.

Yu-Ra Son, Soon-Mi Shim. Various domestic heating processes changed content, digestibility, and radical scavenging capacities of Su Ri Chwi. J. Korean Soc. Appl. Biol. Chem. 58(5):771-778The hypothesis of the current study was that various domestic heating processes such as blanching and microwaving could change content, digestibility, and radical scavenging capacities of caffeoylquinic acids (CQAs) from Su Ri Chwi, Korean wild vegetable. The greatest amount of the total CQAs (300.18 μg/g fresh weight) was observed in blanching for 1 min. Blanching for 5 min showed the highest digestibility of total CQAs (44.72 %). The highest DPPH, ABTS, and ORAC values were found from Su Ri Chwi microwaved for 30 s, blanched for 3 min, and fresh, respectively. The total CQAs in fresh and microwaving for 1 min Su Ri Chwi were significantly correlated to ORAC assay (R = 0.99 and R = 0.97, respectively.). The significant correlations with DPPH and ORAC were measured in digestibility of microwaving for 2 min (R = 0.98) and microwaving for 30 s (R = 0.94), respectively. Our results suggest useful information to consumer for choice of processing procedures to get CQAs, improving the absorption and antioxidant capacity.

Jun Cui, Sang Yeol Lee, Ho Hee Jang. Yeast 2-Cys peroxiredoxin Tsa1 protects cells from DNA damage-induced reactive oxygen species through peroxidase activity. J. Korean Soc. Appl. Biol. Chem. 58(6):779-785Peroxiredoxins (Prxs) belong to a family of thiol-specific peroxidases that scavenge reactive oxygen species (ROS). Yeast Prx thiol-specific antioxidant (Tsa1) protects cells from oxidative stress and maintains genome stability. In this study, we investigated the role of Tsa1 in the response to DNA damage. We observed that Tsa1 among all five Prxs of yeast is the most responsive in cell viability and mutation rate assays. The Δtsa1 mutant is hypersensitive to DNA-damaging agents such as hydroxyurea, methyl methane sulfonate, and camptothecin. The Δtsa1 cells lead to reduced cell viability and cell growth after DNA damage. The Δtsa1-expressing wild-type Tsa1 rescues the sensitivity of Δtsa1 cells to DNA damage, but not in Δtsa1 cells carrying the active-site mutant Tsa1-C47S. We found that the level of ROS in Δtsa1 cells was increased more than in wild-type cells by DNA damage. Tsa1 completely restored the production of DNA damage-induced ROS in Δtsa1 cells, but not Tsa1-C47S. Finally, DNA damage-induced ROS resulted in a switch of Tsa1 from a low-molecular-weight to a high-molecular-weight complex. Taken together, these findings suggest that Tsa1 protects yeast cells from DNA damage by regulating the homeostasis of ROS through peroxidase activity.

Yoon-Su Baek, Na-Young Song, Tae-Gyu Nam, Dae-Ok Kim, Hee-Cheol Kang, Oh-Keun Kwon, Nam-In Baek. Flavonoids from Fragaria ananassacalyx and their antioxidant capacities. J. Korean Soc. Appl. Biol. Chem. 58(6):787-793The calyx of Fragaria ananassa (Rosaceae) has not been investigated for its chemical constituents and biological activity. This study focused on the isolation of active metabolites from the calyx of this plant. Compounds from F. ananassa calyx were extracted in 80 % aqueous MeOH and the concentrated extract was successively partitioned into EtOAc, n-BuOH, and H2O fractions. Repeated silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies afforded six purified flavonoids. On the basis of spectroscopic methods, including 1H, 13C nuclear magnetic resonance spectrometry (NMR), DEPT, and 2D NMR (gCOSY, gHSQC, gHMBC), the flavonoids were identified as kaempferol (1), astragalin (2), quercetin (3), isoquercitrin (4),

sexangularetin (5), and sinensin (6). All the flavonoids were isolated for the first time from the F. ananassa calyx in this study. The flavonoids were evaluated for radical-scavenging capacity 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid ammonium salt), 2,2-diphenyl-1-picrylhydrazyl, and oxygen radical absorbance capacity methods. In conclusion, quercetin (3) consistently showed the highest ABTS, DPPH, and ORAC radical-scavenging activities compared with the other isolated flavonoids.

Soo-In Sohn, Young-Ju Oh, Byung-Yong Kim, Soon-Jong Kweon, Hyun-Suk Cho, Tae-Hoon Ryu. Effect of genetically modified rice producing resveratrol on the soil microbial communities. J. Korean Soc. Appl. Biol. Chem. 58(6):795-805Cultivation of genetically modified (GM) crops has rapidly increased in the global agricultural area. However, cultivation of GM crops in the field evoked the concern of the possibility of unintentional consequences from transgenic plant into environment. In our present study, we have assessed the effects of RS526, GM rice producing resveratrol on the surrounding soil microbial community. The effects of RS526 on the soil microbial community in its field of growth were assessed using a conventional culture technique and culture-independent molecular methods. Three replicate field plots were planted with single GM rice and a non-GM counterpart, Dongjin. The soil microbial communities around these plants were compared using colony counting, denaturing gradient gel electrophoresis (DGGE), pyrosequencing analysis, and community-level physiological profiling during the growing periods. The bacterial, fungal, and actinomycetes population densities of the RS526 soils were found to be within the range of those of the non-GM rice cultivar. The DGGE banding patterns of the GM and non-GM soils were also similar, suggesting that the bacterial community structures were stable within a given month and were unaffected by the presence of a GM plant. The data obtained from pyrosequencing analysis showed that the bacterial community distribution at the phylum level were highly similar to DGGE patterns between two tested groups. The substrate utilization pattern of RS526 and Dongjin rice soil was quite similar between each sampling time. These results indicate that soil microbial communities are not significantly affected by the cultivation of RS526 within the experimental time frame.

Ying Shan, Ying Dong, Dong Jiang. Recombinant expression of a novel antimicrobial peptide consisting of human α-defensin 5 and Mytilus coruscus mytilin-1 in Escherichia coli. J. Korean Soc. Appl. Biol. Chem. 58(6):807-812Antibiotic peptides are a battery of broad-spectrum antibacterial cationic polypeptides widely distributed in the plant and animal kingdoms. Among them, the human defensins are the first line of defense against pathogens and Mytilin, which is isolated from mussel serum, plays a key role in the mussel defense system. The antibacterial activity of these two peptides is generally ascribed to their overall positive charge, which enables them to disrupt bacterial membrane integrity and function. The aim of this study was to develop an effective method for the biosynthesis of a fusion peptide containing human α-defensin 5 (HD5) and Mytilin-1 in Escherichia coli to improve the antimicrobial activity. The individual HD5 and Mytilin-1 peptides were also synthesized for comparison with the fusion peptide. All the peptides, expressed as soluble fusions with the peptide thioredoxin, were isolated using a three-step purification strategy involving nickel-Sepharose chromatography, enterokinase cleavage, and cationic exchange chromatography. The identity of the peptides was confirmed by SDS-PAGE. Antimicrobial activity assays demonstrated that all the recombinant peptides had strong bactericidal properties and that the HD5 and Mytilin-1 fusion protein displayed higher activity against E. coli and Staphylococcus aureus. The results of this study provide a platform for the development of novel cationic peptides for biological studies.

Miran Jeong, Do-Won Jeong, Jong-Hoon Lee. Safety and biotechnological properties ofEnterococcus faecalis and Enterococcus faecium isolates from Meju. J. Korean Soc. Appl. Biol. Chem. 58(6):813-820We assessed the safety and biotechnological properties of 17 Enterococcus faecalis and 18Enterococcus faecium strains isolated from Meju to select starter candidates for

quality Mejuproduction. Minimum inhibitory concentration assays showed that all strains were susceptible to ampicillin, chloramphenicol, ciprofloxacin, erythromycin, gentamicin, penicillin G, and tetracycline, as judged by the breakpoint values defined at the species level by the European Committee on Antimicrobial Susceptibility Testing, whereas two E. faecalis strains were resistant to vancomycin, which could be ascribed to acquired resistance. None of the strains exhibited α- or β-hemolytic activities or biofilm formation. Both species showed similar levels of biogenic amine production, and noticeable amounts of tyramine (3397.4 ± 172.4 ppm) were produced by all strains in the presence of precursors (histidine, lysine, ornithine, and tyrosine). All strains exhibited protease, lipase, and acid production, with decreased activity observed with increasing NaCl concentrations. This study confirmed the necessity of antibiotic resistance screening of Enterococcus species for food production, with the identification of vancomycin-resistant strains from Meju.

Chan Hyeok Kwon, Young Deuk Lee. Distribution and dissipation of deltamethrin, dimethomorph, imidacloprid, mepanipyrim, and metalaxyl applied to chili peppers. J. Korean Soc. Appl. Biol. Chem. 58(6):821-829Field trials were carried out to investigate the dissipation rates of pesticides as well as to measure the terminal residues in chili peppers. Deltamethrin, dimethomorph, imidacloprid, mepanipyrim, and metalaxyl were applied to chili peppers 15 days prior to harvest. Chili pepper samples were collected at specified intervals and were subjected to residue analysis and dislodgeable residue study. In most cases, initial residue levels were rapidly dissipated with time. The dissipation pattern of pesticide residues fits well to first-order kinetics. Biological half-lives, as calculated from regression curves, of most pesticides were estimated to be <4 days. In the fast growing chili pepper, a dilution effect was considerably involved in the rate of dissipation. As a result, it was possible to predict the terminal residue at harvested crops from the initial residue level and declining curve of pesticides.

Ok Kyung Koo, Seung Min Kim, Sun-Hee Kang. Antimicrobial potential of Leuconostocspecies against E. coli O157:H7 in ground meat. J. Korean Soc. Appl. Biol. Chem. 58(6):831-838Ground beef is risky by foodborne pathogens such as E. coli O157:H7 due to the cross-contamination during grinding. The objective of this study was to evaluate the antagonistic activities of Leuconostoc species isolated from ground beef product in order to limit the growth of E. coliO157:H7. While Leuconostoc has been known as spoilage bacteria, the Leuconostoc isolates showed antimicrobial activity on foodborne pathogens such as E. coli O157:H7, Salmonella,Staphylococcus aureus, Listeria monocytogenes, and meat-spoilage bacteria Brochothrix thermosphacta. Antimicrobial activity of cell-free supernatant (CFS) was evaluated by heat, enzyme, and pH adjustment and antagonistic activity by cell competitive growth. In addition, CFS concentration was screened for the presence of organic acids for the antimicrobial properties. Regulation of virulence genes was investigated by inhibition of locus of enterocyte effacement expression by CFS. The result showed that Leuconostoc isolates inhibited the growth of E. coliO157:H7 by active antimicrobial agent including organic acids and the growth of background microorganisms, and E. coli O157:H7 in ground beef was inhibited with the supplement of the CFS.

Angeline Atieno Ochung’, Lawrence Arot Oyango Manguro, Phillip Okinda Owuor, Isaac Ogoche Jondiko, Regina Achieng’ Nyunja, Hosea Akala, Pauline Mwinzi, Sylvia Awino Opiyo. Bioactive carbazole alkaloids fromAlysicarpus ovalifolius (Schumach). J. Korean Soc. Appl. Biol. Chem. 58(6):839-846Phytochemical and biological evaluation of the stem bark of Alysicarpus ovalifolius led to the isolation of three carbazole alkaloids identified as mohanimbine (1), koenimbine (2) and koenidine (3) along with quercetin 3-O-glucoside (4), kaempferol 7-O-glucoside (5), orientin (6), apigenin (7), quercetin (8), plumbagin (9) and stigmasterol (10). The structures of these compounds were elucidated using physical and spectroscopic methods as well as comparison with the literature data. Compound 3 showed strong

activity against chloroquine-sensitive strain I (D6) and the multi-drug resistant Indochicha I (W2) of Plasmodium falciparum with IC50 values of 63.07 ± 0.01 and 54.19 ± 0.04 ng/mL, respectively. Compound 1 on the other hand exhibited moderate larvicidal against Anopheles gambiae larvae as well as antimicrobial activities against Candida albicans and gram-positive Staphylococcus aureus, respectively.

Khalid Abdallah Hussein, Jin Ho Joo. Isolation and characterization of rhizomicrobial isolates for phosphate solubilization and indole acetic acid production. J. Korean Soc. Appl. Biol. Chem. 58(6):847-855Recently, public concerns regarding the use of agrochemicals have increased due to the environmental impacts and potential risks to human health. The application of beneficial microorganisms is a novel technology to improve plant health and productivity and has therefore been extensively studied as an alternative strategy for biocontrol. In our study, 122 microbial isolates were obtained from the rhizosphere of Panax ginseng and subsequently tested in vitro for phosphate solubilization and indole acetic acid (IAA) production. Pikovskaya’s medium was used to estimate rhizomicrobial isolates to solubilize tricalcium phosphate [Ca3 (PO4)2]. Among all the investigated strains, 82 % of rhizospheric fungi showed phosphate solubilization activity; however, only 57.1 % of the rhizobacteria isolates showed phosphate solubilization ability. For IAA production, 64.7 % of the tested rhizofungi isolates were able to produce the phytohormone; however, only 47.62 % of the rhizobacteria isolates exhibited IAA production. Among all investigated species, Pseudomonas fluorescence and Azotobacter chroococcum showed the highest phosphate solubility demonstrating 885.4 and 863.4 μg mL−1, respectively. Mucor sp. produced 42.3 μg mL−1 of IAA in Czapek’s tryptophan medium, and the highest fungal species to solubilize the inorganic phosphate (237.5 μg mL−1) was estimated by Penicillium sp. Rhizobacteria were more effective than rhizofungi in phosphate solubilization and IAA production. This study introduces some potent species in terms of phosphate solubilization and IAA production which could be likely to improve soils’ quality and promote plant growth.

Seockmo Ku, Hyun Ju You, Myeong Soo Park, Geun Eog Ji. Effects of ascorbic acid on α-l-arabinofuranosidase and α-l-arabinopyranosidase activities fromBifidobacterium longum RD47 and its application to whole cell bioconversion of ginsenoside. J. Korean Soc. Appl. Biol. Chem. 58(6):857-865Bifidobacterium longum RD47 was cultured in 24 kinds of modified MRS broths containing various ingredients to select the most promising source that induces microbial enzymes. Among the various ingredients, ascorbic acid significantly enhanced α-l-arabinofuranosidase and α-l-arabinopyranosidase activities in Bifidobacterium longum RD47. Addition of 2 % ascorbic acid (w/v) to MRS showed the maximum enzyme activities. Both whole cell and disrupted cell homogenates showed efficient ρ-nitrophenyl-β-d-glucopyranoside and ρ-nitrophenyl-β-d-glucofuranoside hydrolysis activities. The initially enhanced α-l-arabinopyranosidase and α-l-arabinofuranosidase activities by ascorbic acid were maintained over the cell disruption process. The optimal pH of α-l-arabinofuranosidase and α-l-arabinopyranosidase was 5.0 and 7.0, respectively. Both enzymes showed the maximum activities at 40.0 °C. Under the controlled condition using Bifidobacterium longum RD47, ginsenoside Rb2, and Rc were converted to ginsenoside Rd.

Min Young Lee, Hoon Kim, Kwang Soon Shin. In vitro and in vivo effects of polysaccharides isolated from Korean persimmon vinegar on intestinal immunity. J. Korean Soc. Appl. Biol. Chem. 58(6):867-876The in vitro and in vivo effects of crude polysaccharide isolated from Korean persimmon vinegar (KPV-0) on the intestinal immunostimulation were examined. KPV-0 was not cytotoxic to intestinal epithelial Caco-2 cells and could be transported through a Caco-2 cell monolayer in an in vitro coculture system. For and in vitro experiment, KPV-0 treatment significantly increased IgA production by Peyer’s patch (PP) cells and triggered an increase in transforming growth factor (TGF)-β1 and interleukin (IL)-6 levels. To investigate the in vivo effects of KPV-0 treatment, KPV-0 was administered to mice orally

at different doses for 20 days. Oral administration of KPV-0-induced IgA and cytokines (IL-6, granulocyte macrophage-colony-stimulating factor, and TGF-β) production by PP cells and significantly increased the IgA levels in intestinal fluids and feces. These results suggested that the polysaccharides isolated from persimmon vinegar appear to modulate the intestinal immune system and could be beneficial to human health.

Minae Joo, Hye-Gyeong Yoo, Hyun-Ju Kim, Hyung-Jin Kwon. ToxB encodes a canonical GTP cyclohydrolase II in toxoflavin biosynthesis and ribA expression restored toxoflavin production in a ΔtoxB mutant. J. Korean Soc. Appl. Biol. Chem. 58(6):877-885Burkholderia glumae synthesizes toxoflavin, a phytotoxin that contributes the virulence of this phytopathogen. The toxoflavin biosynthetic gene cluster contains a tox operon composed of five genes, toxABCDE, and toxB is predicted to encode a GTP cyclohydrolase II, which is the first enzyme (RibA) in riboflavin biosynthesis. ToxE is also homologous to RibD, the bifunctional deaminase/reductase acting on the RibA product. This suggests that toxoflavin and riboflavin biosynthesis share the first two steps in their respective biosyntheses. In this study, we demonstrated that ToxB and B. glumae RibA (Bglu-RibA) both displayed GTP cyclohydrolase II activity with comparable kinetic parameters. When toxB was inactivated, toxoflavin production was abolished, and introduction of a plasmid copy of Bglu-ribA restored toxoflavin production in a ΔtoxBmutant. ToxB and Bglu-RibA can thus be defined as GTP cyclohydrolase II isozymes, even thoughBglu-ribA is not a genetic equivalent of toxB because the chromosomal copy of Bglu-ribA is unable to support toxoflavin production in the absence of toxB. In LB agar culture, toxAB is incapable of complementing ΔtoxB though toxABC induced toxoflavin accumulation in the ΔtoxB mutant up to 80 % of the WT level. This indicates that toxBC co-expression is a critical factor for toxoflavin biosynthesis in this condition, suggesting that the WD repeat protein ToxC acts as a scaffolding protein in a toxoflavin biosynthetic metabolon. In LB liquid culture, toxAB successfully restored toxoflavin production, suggesting that a role of toxBC co-expression is dependent upon growth condition.

Jingmei Li, Yeun Hong, Jae-Hwan Kim, Pei Qin, Mi-Ju Kim, Hae-Yeong Kim. Multiplex PCR for simultaneous identification of turkey, ostrich, chicken, and duck. J. Korean Soc. Appl. Biol. Chem. 58(6):887-893To discriminate economically motivated adulteration in poultry meats and products, species-specific oligonucleotide primer sets for multiplex polymerase chain reaction (PCR) of four poultry meats were newly designed. Sequences for species-specific primers were generated based on the nucleotide variation in mitochondria cytochrome b gene through sequence comparisons. The sizes of species-specific PCR products for turkey, ostrich, chicken, and duck were 217, 330, 516, and 820 bp, respectively. By adjusting PCR conditions and the concentrations of primers, a simultaneous discrimination of four poultry meats was successfully performed. The limits of detection of both single and multiplex PCR for turkey were 0.05 and 0.005 ng for other species, respectively. The developed multiplex PCR method was successfully applied to identify the target species in 34 of poultry meat products and thus was considered to be a useful tool for monitoring poultry meat products.

Yoon-Su Baek, Jae-Woo Jung, Sung-Hee Lee, Nam-In Baek, Ji-Hae Park. A new pregnane hexaglycoside fromAdonis multiflora. J. Korean Soc. Appl. Biol. Chem. 58(6):895-899A new pregnane hexaglycoside, named amurensioside L (1), was isolated from the whole plant ofAdonis multiflora. Amurensioside L (1) emerged to be new pregnane hexaglycoside which we fully spectroscopically characterized. The chemical structure of compound 1 was established as 14β,20R-epoxy-3β-hydroxypregna-4,6-dien-18-oic acid-20-lactone 3-O-β-d-glucopyranosyl-(1 → 4)-O-β-d-diginopyranosyl-(1 → 4)-O-β-d-cymaropyranosyl-(1 → 4)-O-β-d-cymaropyranosyl-(1 → 4)-O-β-d-oleandropyranosyl-(1 → 4)-O-β-d-oleandropyranoside on the basis of spectroscopic analyses including nuclear magnetic resonance spectrometry, high-resolution fast atom bombardment mass

spectrometry, and infrared spectroscopy.

Apinun Kanpiengjai, Saisamorn Lumyong, Pairote Wongputtisin, Dietmar Haltrich, Thu-Ha Nguyen, Chartchai Khanongnuch. Efficient secretory expression of gene encoding a broad pH-stable maltose-forming amylase from Lactobacillus plantarum S21 in food-grade lactobacilli host. J. Korean Soc. Appl. Biol. Chem. 58(6):901-908The native and the N-terminal signal peptide sequence deleted gene encoding for α-amylase fromLactobacillus plantarum S21 were cloned into the inducible lactobacilli expression vectors pSIP409 and pSIP609 and expressed in L. plantarum WCFS1 and food-grade L. plantarum TGL02, respectively. Only the native amylase gene was expressed and secreted extracellular amylase at a level of approximately 2000 U/L with 90 % secretion efficiency from both hosts. The purified extracellular amylase from the L. plantarum TGL02 retained unique properties of the wild-type enzyme, particularly the broad pH stability (4.0–8.0) and maltose-forming activity. The results indicate high compatibility of L. plantarum S21 signal peptide sequence to both recombinant lactobacilli hosts. The recombinant lactobacilli exhibited high efficiency for direct lactic acid production from starch as found with L. plantarum S21. The efficient compatible signal peptide is also expected to be applied in secretory expression for production of valuable proteins in food-grade lactobacilli host.

Tae Jin Kim, Kyoung Bok Lee, Seung-A Baek, Jaehyuk Choi, Sun-Hwa Ha, Sun-Hyung Lim, Soo-Yun Park, Yunsoo Yeo, Sang Un Park, Jae Kwang Kim. Determination of lipophilic metabolites for species discrimination and quality assessment of nine leafy vegetables. J. Korean Soc. Appl. Biol. Chem. 58(6):909-918The lipophilic compounds in nine vegetables consumed in Korea were characterized for diversity in phytochemical content. We also analyzed the relationships among these compounds in terms of their contents. The profiles of 18 lipophilic compounds in the leaves were subjected to data-mining processes, including principal component analysis (PCA), Pearson’s correlation analysis, hierarchical clustering analysis (HCA), and partial least squares discriminant analysis (PLS-DA). These species could be distinguished by means of the PCA results. HCA of these phytochemicals resulted in clusters derived from closely related biochemical pathways. PLS-DA showed significant separation among extracts from the following four families: Amaranthaceae, Asteraceae, Brassicaceae, and Malvaceae. The major metabolites that facilitated differentiation in the PLS-DA model were campesterol, β-sitosterol, β-amyrin, stigmasterol, cholesterol, octacosanol (c28), α-amyrin, and hexacosanol (c26). Chard contained high levels of triacontanol (c30), which was positively correlated with the c28 content (r = 0.746, p < 0.0001). Stigmasterol, α-amyrin, and β-amyrin contents were higher in Asteraceae species including chicon, lettuce, and oak than other plant families. These results demonstrate the utility of metabolic profiling, combined with multivariate analysis, for discrimination of vegetable species as well as evaluation of food quality.

Jiayu Li, Qi Zhang, Wenwen Hu, Xiaoyan Yang, Haibin He. Stability of phenolic acids and the effect on weed control activity. J. Korean Soc. Appl. Biol. Chem. 58(6):919-926Phenolic acid is a very important class of allelochemicals with allelopathic weed control activity. In this study, three benzoic acid derivatives (syringic, 4-hydroxybenzoic, and vanillic acids), three cinnamic acid derivatives (cinnamic, 4-hydroxycinnamic, and ferulic acids) were tested, and high-performance liquid chromatography was used to conduct a dynamic analysis on the changes in the concentration of phenolic acids in a bioassay based on the initial concentration and test time. The results showed that the concentration of individual phenolic acids and a solution of mixed phenolic acids decreased to a certain extent irrespective of environment, i.e., bioassay (4–7 days) or a rice-growing environment, and a significant decrease in concentration was measured after 48 h. Based on the above results, the laboratory bioassay was conducted using a fresh solution of phenolic acids every 48 h. The results showed that the instability of

phenolic acid could affect its weed control activity, and this effect was more significant for high concentrations of phenolic acids. On the other hand, changing the solution did not have a significant impact on the weed control activity of phenolic acids in the natural environment (pH 6.50), in which allelopathic rice release phenolic acids. These results reveal the instability of phenolic acids could significantly reduce the inhibition rate on the growth index for receptor plants in an indoor bioassay.

Liang Pang , Kai Zhu , Xia Feng, Weiwei Liu, Deguang Peng, Lihua Qiu, Xiang Gao, Jiang Deng, Ying Li , Xin Zhao. Protective effect of liensinine on periodontitis through its antioxidant effect in mice. J. Korean Soc. Appl. Biol. Chem. 58(6):927-936Liensinine is the most important functional compound in lotus seeds; its protective effect on periodontitis was determined and the mechanism of its antioxidant effect was also explained. During the experiment, we could observe that liensinine could reduce the gingival index in periodontitis-induced mice, and these effects were similar to that during the treatment of periodontitis with the metronidazole drug. After the treatment with liensinine, the serum Immunoglobulin G1 (IgG1) level was found to increase and IgG2a level was found to decrease compared to the periodontitis control mice. Liensinine and metronidazole could increase the superoxide dismutase (SOD) serum levels in the periodontitis-induced mice, and both the drugs at the same concentration (200 mg/kg) showed similar SOD activities, which was found to increase. Liensinine also increased the oxidation-related factor of catalase (CAT), glutathione peroxidase, and decreased nitric oxide, malondialdehyde, and endothelin in the serum of mice, compared to the periodontitis control mice. The serum cytokine levels of Interleukin-6, tumor necrosis factor-α, and Interferon-γ in liensinine-treated mice were lower than the control mice, but higher than the normal mice. The serum IL-4 cytokine level of liensinine-treated mice was reversed, i.e., higher than the control mice. In the periodontal tissue, the mRNA and protein expressions of inhibitor of kappa B-α, transforming growth factor beta 1, Interleukin-10, Mn-SOD, Gu/Zn-SOD, and CAT were increased by the treatment with liensinine compared to the control group mice, and nuclear factor kappa B expressions showed the opposite trend. Liensinine is a good functional compound in the treatment of periodontitis, similar to metronidazole drug. Higher concentration of liensinine had better effects.

Eun-Jung Lee, Joo-Sung Kim, Yun-Ji Kim. Addition of thiamine dilaurylsulfate to reduce the intensity of hydrostatic pressure treatment for microbial safety of Korean Jogaejeot- muchim (salted-fermented-seasoned short-neck clam). J. Korean Soc. Appl. Biol. Chem. 58(6):937-944This study investigated hydrostatic pressure (HP) treatment with the addition of thiamine dilaurylsulfate (TDS) for the microbial safety of the final product (salted-fermented-seasoned short-neck clam) during Jogaejeot-muchim manufacturing process. HP treatment (100–600 MPa) was performed at each step (raw, salted-fermented, and salted-fermented-seasoned short-neck clam) of the Jogaejeot-muchim manufacturing process. The reduction effect on Staphylococcus aureus in the salted-fermented-seasoned short-neck clam step was 1.2 log after 400 MPa treatment, and it was lower than those of raw and salted-fermented short-neck clam steps (4.3 and 4.9 log, respectively) after 400 MPa treatment. To improve the microbial safety of the salted-fermented-seasoned short-neck clam step, TDS (1 g/kg) was added, and 1 log reduction was obtained. In the salted-fermented-seasoned short-neck clam step, HP treatment with TDS addition showed a synergistic effect, and the HP intensity for a 2 log reduction was reduced from 500 to 100 MPa treatment with TDS addition, and a 4 log reduction was achieved by the 300 MPa treatment with addition of TDS. This study concluded that the TDS addition contributed to reducing the intensity of HP treatment required for 99 % inactivation of S. a ureus, and that combined treatment (300 MPa treatment with TDS) can be used to provide microbial safety assurance in Korean Jogaejeot-muchim.

Sook-Young Lee, Dae-Yoon Cho, Min-Kyoung Lee. Quality characteristics of fermented cheese analogs produced using roasted soy flour and different

fermentation times with rice straw. J. Korean Soc. Appl. Biol. Chem. 58(6):945-954To improve the nutritional and physiological functionality of cheese analogs, fermented cheese analogs treated with Flavourzyme® were developed and the effects of roasting soy flour and fermentation time (1, 2, and 3 days) with rice straw on their quality characteristics, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), isoflavone content, and antioxidant activity were examined. Heat coagulation was the highest for the unroasted and roasted soy flour groups fermented for 1 day, with the unroasted group having higher values than the roasted group, regardless of fermentation time. The daidzein and genistein contents were the highest in the soy flour and roasted soy flour groups fermented for 2 days and 1 day, respectively. The antioxidant activity and heat extension were the highest for the soy flour group and roasted soy flour group fermented for 1 day, respectively. In the SDS-PAGE patterns, protein subunits of P26 in roasted soy flour groups were consistently decomposed by increasing the fermentation time. The yield and pH in both groups decreased as fermentation time increased. The adhesiveness of the soy flour group was highly increased by fermentation, and the cohesiveness, springiness, and appearance of the roasted soy flour group were improved by fermentation. The optimum fermentation time for the fermented cheese analogs was 1 day, as higher yields, heat coagulation, heat extension, lightness, dE, cohesiveness, and springiness were obtained. Between the two groups fermented for 1 day, the soy flour group showed better heat coagulation, genistein content, antioxidant activity, cohesiveness, springiness, and adhesiveness; however, the roasted soy flour group showed better yield and heat extension.

Min-Gi Kim, Hoi-Seon Lee. Insecticidal toxicities of naphthoquinone and its structural derivatives. Appl. Biol. Chem. 59(1):3-8Insecticidal toxicities of bioactive constituent from Diospyros kaki roots and its structural derivatives were evaluated against the larvae of Aedes aegypti, Culex pipiens pallens, and Ochlerotatus togoi. Bioactive constituent of D. kaki roots was isolated by some chromatographic methods and identified as 5-hydroxy-2-methyl-1,4-naphthoquinone. Based on the LC50 values of 5-hydroxy-2-methyl-1,4-naphthoquinone structural derivatives, 2,3-dichloro-5,8-dihydroxy-1,4-naphthoquinone (0.89, 0.80, and 1.04 μg/mL) had the most potent insecticidal activity, followed by 2,3-dibromo-1,4-naphthoquinone (1.32, 1.28, and 1.94 μg/mL), 5,8-dihydroxy-1,4-naphthoquinone (3.57, 3.34, and 5.04 μg/mL), 2,3-dichloro-1,4-naphthoquinone (4.76, 3.89, and 5.33 μg/mL), 5-hydroxy-2-methyl-1,4-naphthoquinone (5.66, 5.43, and 6.09 μg/mL), and 2,2′-bi(3-hydroxy-1,4-naphthoquinone) (33.93, 32.82, and 36.17 μg/mL) against Ae. aegypti, Cx. pipiens

pallens, and Oc. togoi, respectively. In this regard, 5-hydroxy-2-methyl-1,4-naphthoquinone and its structural derivatives could be suitable as insecticidal agents to control the larvae of the three mosquito species.

Hyeri Lee, Eenhye Kim, Yongho Shin, Jong-Hwa Lee, Hor-Gil Hur, Jeong-Han Kim. Identification and formation pattern of metabolites of cyazofamid by soil fungus Cunninghamella elegans. Appl. Biol. Chem. 59(1):9-14This study was performed to investigate the formation of microbial metabolites from cyazofamid by the soil fungus Cunninghamella elegans. The incubation of cyazofamid with C. elegans was conducted for 10 days. Cyazofamid disappeared after 7 days of incubation, producing three metabolites. Metabolites identified by liquid chromatography–tandem mass spectrometry were 4-chloro-5-(4-(hydroxymethyl)phenyl)-imidazole-2-carbonitrile (CHCN), 4-(4-chloro-2-cyanoimidazole-5-yl)benzoic acid (CCBA) and 4-chloro-2-cyano-5-(4-(hydroxymethyl)phenyl)N,N-dimethyl-1H-imidazole-1-sulfonamide (CCHS). A new metabolite, CCHS, was further confirmed by 1H-13C HSQC (heteronuclear single-quantum correlation) using nuclear magnetic resonance. As a possible metabolic pathway, cyazofamid could be oxidized to CCHS, degraded to CHCN and further oxidized to CCBA. The metabolic system of C. elegans would be a powerful tool for predicting and identifying phase I metabolites that could be formed in mammalian systems.

Xiao-Ding Li, Jie Li, Meng Wang, Hong Jiang. Copigmentation effects and thermal degradation kinetics of purple sweet potato anthocyanins with metal ions and sugars. Appl. Biol. Chem. 59(1):15-24The copigmentation effects of purple sweet potato anthocyanins (PSPAs) with metal ions and sugars were investigated in model solutions at pH 4. The thermal stability of PSPAs was also explored in the presence of sugars and 5-hydroxymethylfurfural (5-HMF). Copigmentation are characterized by hyperchromic effect and bathochromic shift. The hyperchromic effect of Fe3+ reached 25.15 % even at a very low concentration 0.005 mol L−1, while the values of the other metal ions at 0.05 mol L−1 were ranked in the following ascending order: K+ < Ca2+ < Mg2+ < Zn2+ < Cu2+ < Fe2+ < Al3+. The bathochromic shift was not observed in all sugar reaction solutions; glucose showed the highest values of hyperchromic effect at the concentration range 150–300 g L−1, followed by fructose and sucrose; konjac glucomannan showed the highest effect even at a much lower concentration among the macromolecular sugars. Especially, the konjac glucomannan exhibited a much better color enhancement than glucose. Small molecular sugars accelerated the thermal degradation of PSPAs, whereas macromolecular sugars showed a protective effect particularly at high temperatures. Moreover, 5-HMF resulted in the deterioration of the thermal stability of PSPAs and was suggested to be an important labile factor for copigmented PSPA solutions.

Hyun-Jin Kim, Seong-Gook Kang, Lily Jaiswal, Jinglei Li, Ju-Hee Choi, Sang-Mi Moon, Jeong-Yong Cho, Kyung-Sik Ham. Identification of four new angiotensin I-converting enzyme inhibitory peptides from fermented anchovy sauce. Appl. Biol. Chem. 59(1):25-31The inhibitory activity of an angiotensin I-converting enzyme (ACE), a key regulatory enzyme of blood pressure from the fermented anchovy sauce, was evaluated, and ACE inhibitory peptides were purified. The ACE activity significantly increased with an increase in salt concentration. In addition, the ACE inhibitory activity of the fermented anchovy sauce containing high salt content (25 %) significantly increased with an increase in fermentation time. The maximum activity (96 %) was reached after 15 months of fermentation. Four ACE inhibitors [Pro-Lys (PK), Gly-Cys-Lys (GCK), Asn- His-Pro (NHP), and Asp-Gly-Gly-Pro (DGGP)] in the fermented anchovy sauce were purified through various chromatographic techniques and identified by electrospray ionization tandem mass spectrometer analysis. Four newly identified peptides were synthesized and analyzed for ACE inhibitory activity in order to confirm that the purified peptides were actually ACE inhibitors. The IC50 values for ACE inhibitory activities of synthesized peptides DGGP, GCK, NHP, and PK were 164, 178, 1172, and 4092 μM, respectively.

Sunyoung Park, Han Sub Kwak, Myeonggeun Oh, Youngseung Lee, Yoonhwa Jeong, Misook Kim. Physicochemical, microbiological, and sensory characteristics of soy sauce fermented in different regional ceramics. Appl. Biol. Chem. 59(1):33-41Korean traditional soy sauce was fermented for 120 days in ceramics obtained from five different regions in Korea. The average internal and external temperatures of soy sauce in ceramics were 20.0 and 18.3 °C, respectively. Physicochemical, microbiological, and sensory characteristics were analyzed. Ceramics from different regions generated differences in physicochemical characteristics. Soy sauce fermented in the southwest regional ceramic, which has a low height and wide circumference, had significantly lower salt content, higher reducing sugar content, and protease activity rather than others. In descriptive analysis, soy sauce fermented in the ceramic from Seoul capital region had a lower intensity of saltiness and biting taste, whereas the soy sauce in the southwest ceramic had comparable saltiness and biting taste attributes as those in the southeast and central regions, and Jeju Island ceramics. Five regional ceramics were categorized into three groups by principal component analysis. Based on the physicochemical characteristics, soy sauces from southwest and Seoul capital regions had distinctive characteristics in comparison with the soy sauces from Jeju Island, southeast, and central regions.

Md. Humayun Kabir, A. M. Abd El-Aty, So Jeong Im, Md. Musfiqur Rahman, Sung-Woo Kim, Waziha Farha, Jeong-Heui Choi, Da-I Jung, Young-Jun Lee, Troung Lieu, Ho-Chul Shin, Geon-Jae Im, Su-Myeong Hong, Jae-Han Shim. Determination of residual levels of metrafenone in lettuce grown under greenhouse conditions using gas chromatography with a micro-electron capture detector. Appl. Biol. Chem. 59(1):43-49The current study was carried out to detect metrafenone residues and estimate the half-lives in lettuce grown under greenhouse conditions using gas chromatography with a micro-electron capture detector. Metrafenone was applied in two and three doses (with a 7-day interval) to two different sites in Naju (distance between sites 1 and 2 = 6.5 km) in different seasons (spring and autumn). Samples were extracted with acetone and matrix components were purified using a silica solid-phase extraction (SPE) cartridge. The calibration curve over the concentration range of 0.02–5.0 mg/L was linear with a determination coefficient (R2) = 0.9993. The limits of detection and quantification were 0.003 and 0.01 mg/L, respectively. Recoveries at two fortification levels (0.1 and 0.5 mg/kg) ranged from 78.5 to 113.3 % with a relative standard deviation <8 %. The calculated half-lives were 2.3 and 2.2 days and 4.9 and 5.0 days for the double and triple doses at sites 1 and 2, respectively. These results suggest that the method is effective and sensitive for the determination of metrafenone residues in field-incurred lettuce samples, and the difference in the reported half-lives is due to seasonal variation.

Dara Doeun, Hye-Sun Shin, Myung-Sub Chung. Effects of storage temperatures, vacuum packaging, and high hydrostatic pressure treatment on the formation of biogenic amines in Gwamegi. Appl. Biol. Chem. 59(1):51-58This study investigated the effects of storage temperatures, vacuum packaging, and high hydrostatic pressure (HHP) on the formation of biogenic amines (BAs) in Gwamegi. The Gwamegi samples were subjected to 400 MPa pressure for 10 min at 20 °C or vacuum packaging, and then stored at −16, 5, or 25 °C for 28 days. The amounts of six BAs (histamine, tyramine, cadaverine, putrescine, spermine, and spermidine) and the total plate counts were checked weekly. Compared with the control at 5 °C, the total plate counts were 2–3 log CFU lower for the HHP-treated samples and 1–3 log lower for the vacuum-packaged samples. Tyramine, spermine, and spermidine were found in Gwamegi stored at −16 °C and tyramine, cadaverine, spermine, and spermidine at 5 °C, whereas all six BAs were found at 25 °C. Histamine was relatively low in amount in all the treated samples. The BAs reached a high level during storage at 25 °C, with the total concentration reaching 1099 ppm in 28 days. After HHP treatment or vacuum packaging, the concentration of cadaverine and spermidine decreased, whereas that of tyramine

and spermine increased. Tyramine was the dominant BA in all the samples and during storage at 5 °C in the control (0–56.67 ppm), HHP-treated (0–105 ppm), and vacuum-packaged (0–90 ppm) samples. Both HHP treatment and vacuum packaging effectively reduced the formation of some types of BAs and improved the microbial quality in Gwamegi. However, simultaneously, they also promoted the formation of others types of BAs such as tyramine and spermine.

Yunhee Jo, Hyun-Kyu Kyung, Hae-Jun Park, Joong-Ho Kwon. Irradiated fruits can be identified by detecting radiation-induced markers with luminescence and ESR analyses for different trading fruits. Appl. Biol. Chem. 59(1):59-65Photostimulated luminescence (PSL) or thermo-luminescence (TL) analysis and electron spin resonance (ESR) spectroscopy were performed to detect radiation-induced markers in various trading fruits, such as oranges, grapefruits, mandarins, lemons, limes, bananas, and pineapples. All the unknown samples were identified as non-irradiated fruits, and gamma irradiation at 1 kGy permitted dose by Codex and US FDA was applied to investigate radiation-induced markers. The photon counts for all calibrated PSL samples revealed higher than 5000 (positive) except banana. The ESR triplet signals were detected as a radiation-induced marker resulting from cellulose existing in irradiated fruits excluding banana. The unambiguous identification of irradiated banana was impossible by both techniques. However, isolated minerals from all fruit samples showed radiation-induced typical TL glow curves through the normalization step, confirming the feasible application of TL analysis for identifying irradiation status of all the subjected trading fruits.

Changhoon Chai, Haejin Jang, Se-Wook Oh. Real-time PCR-based quantification of Shigella sonnei in beef and a modified Gompertz equation-based predictive modeling of its growth. Appl. Biol. Chem. 59(1):67-70Shigella sonnei shares many physiological aspects with Escherichia coli; thus, so far no culture-based method has been developed to detect and quantify S. sonnei separately from E. coli. Therefore, little information is available for the growth characteristics of S. sonnei in food. This study aimed to address a systematic scheme to quantify S. sonnei in beef separately from E. coli using quantitative real-time polymerase chain reaction (qRT-PCR) and subsequently predict its growth characteristics. The use of S. sonnei-specific primers in qRT-PCR allowed to obtain growth curves of S. sonnei in beef at different temperatures, and the fitting of curves into a modified Gompertz model let us analyze the growth characteristics such as the lag time, maximum growth rate, and maximum quantity of S. sonnei in beef at different temperatures. A systematic scheme for RT-PCR-based quantification and a predictive modeling described in this study may be a useful means to analyze S. sonnei growth in food.

Gyeonghui Yu, Heesun Jung, Hyejung Mok. Indocyanine green-incorporated exosomes for improved in vivo imaging of sentinel lymph node. Appl. Biol. Chem. 59(1):71-76For improved physical stability and fast accumulation in sentinel lymph nodes in vivo, indocyanine green (ICG) was incorporated into exosomes (EXOs) to form a novel ICG-based near-infrared fluorescence probe. Compared with free ICG, prepared ICG-TBAI-encapsulated EXOs (ICG/EXOs) smaller than 50 nm had greater physicochemical stability in aqueous solution when exposed to light at 37 °C. In addition, sentinel lymph nodes were visualized within minutes with ICG/EXOs at a low ICG dose (0.05 mg/kg) owing to its well-controlled size distribution. Taken together, the results show that ICG/EXOs are effective biocompatible and sensitive near-infrared probes for real-time sentinel lymph node imaging in vivo.

Sang Joon Mo, Joo-Won Suh. Elucidation of first step of the allylmalonyl-CoA biosynthetic pathway by expression of heterologous KSIII gene and enhancement of 36-methyl-FK506 production by genetic and chemical engineering. Appl. Biol. Chem. 59(1):77-88We identified another tcsABCD genes encoding biosynthesis of the allylmalonyl-CoA in

new FK506 producing strain, Streptomyces sp. KCCM 11116P. The functionality of tcsABCD is confirmed by gene deletion. To validate the functionality of these genes, tcsABCD genes were heterologously expressed in Streptomyceshygroscopicus ATCC 14891 using pSE34. The resulting strain produced the FK506 which showed a molecular ion [M+NH4]+ at m/z 821 as verified by LC/ESI–MS. To address the function of TcsB in the allylmalonyl-CoA pathway, we pursued the construction of two different combinations: the heterologous FabH-like β-ketoacyl-acyl carrier protein synthase III gene expression and chemical complementation. Plasmid-based expression of the fabH homolog dpsC in ΔTcsB using pSE34 and pSET152(ermE*) led to restoration of approximately 32.2 and 28.7 % of the levels of FK506 relative to Streptomyces sp. KCCM 11116P, respectively. Furthermore, the addition of propionic acid to the culture of ΔTcsB restored FK506 production up to 35.6 % of overall FK506 levels in Streptomyces sp. KCCM 11116P. We have engineered production of 36-methyl-FK506 in ΔTcsB. The heterologous expression of Syteptomycesglaucescens FabH in ΔTcsB gave rise to 5.9-fold increase in the 36-methyl-FK506 level as compared with mutasynthetic production by 4-methylpentanoic acid. Supplementing the culture broth of ΔTcsB expressing S. glaucescens FabH with valine resulted in an additional 2.5-fold increase in the 36-methyl-FK506 titer. Overall, this study shows that TcsB was found to maintain a very high-specific activity for propionyl-CoA in allylmalonyl-CoA biosynthetic work and S. glaucescens FabH which use isobutyryl-CoA as substrate can initiate methyl side chain biosynthesis in 36-methyl-FK506 in ΔTcsB.

Jaemin Lee, Myung Hee Lee, Eun Ju Cho, Sanghyun Lee. High-yield methods for purification of α-linolenic acid from Perilla frutescens var. japonica oil. Appl. Biol. Chem. 59(1):89-94This study was conducted to investigate the purification methods for high-yield of α-linolenic acid (ALA) from Perilla frutescens var. japonica oil (PO). PO was treated with 2 g of urea, cooled for 24 h, and 81.75 % of ALA was obtained after gas chromatography–flame ionization detector analysis. Urea complexation at low temperature (refrigeration) is a prospective method for the extraction of unsaturated fatty acids in high yields. Our results suggest that urea treatment with cooling at 10 °C could be used for purification of ALA from PO. It will be enhanced for the development of nutritional, medical, and cosmetic value.

Eun-Ae Kim, Sun-Young Lee, Sook-Young Lee. Quality characteristics of steamed rice bread prepared with different contents of proteolytic enzyme. Appl. Biol. Chem. 59(1):95-102The aim of this study was to investigate the quality characteristics of steamed rice bread containing the various contents of protease (0, 0.5, 1.0, and 1.5 % Flavourzyme®), pH, and expansion of the dough during different fermentation times (0, 20, and 30 min), and hardness after different storage times (0, 1, 2, and 3 days). When the Flavourzyme® contents increased, pH of the dough gradually decreased, but expansion increased up to 30 min of the fermentation. Specific loaf volume was the greatest for steamed rice bread prepared with 1.5 % Flavourzyme®. In SDS electrophoretic patterns, the band intensity of wheat gluten (P37) decreased by 1.0 and 1.5 % Flavourzyme® treatments. The α-glutelin of rice protein clearly degraded in the samples prepared with 1.0 and 1.5 % Flavourzyme®, whereas the band intensity of β-glutelin decreased only in the sample with 1.5 % Flavourzyme®. The band intensity of soy lipoxygenase and the 7S globulin fraction (P54) of soy protein decreased in the sample with 0.5 % Flavourzyme®. The latter showed the highest water-holding capacity and the highest hardness but lowest rate of retrogradation during 3 days of storage. In microstructure of the control, pore size was the smallest and the fewest pores were observed. The greater the contents of Flavourzyme®, the greater the size and number of pores were observed. Steamed rice bread prepared with 0.5 % Flavourzyme® was thought to be optimal due to retarded retrogradation because of the highest water-holding capacity, highest scores of mouthfeel, and overall quality due to greater springiness and cohesiveness.

Sung-Hee Kim, Yeo Cho Yoon, Jin-Taek Hwang, Mi Jeong Sung, Myung-Sunny Kim, Haeng Jeon Hur, Mee-Ra Rhyu, Jae-Ho Park. Tangeretin synergistically

enhances odorant-induced increases in cAMP and calcium levels and CREB phosphorylation in non-neuronal 3T3-L1 cells. Appl. Biol. Chem. 59(1):103-108Tangeretin is a natural flavone found mainly in citrus peels. Although emerging evidence suggests the beneficial health effects of tangeretin, little is known about the biological role of tangeretin in the odorant-induced signal transduction pathway. In this study, the effects of tangeretin on odorant-stimulated non-neuronal 3T3-L1 cells were evaluated. Here, we present the first evidence that an olfactory receptor for lyral, an odorant, is expressed in 3T3-L1 cells and that it responds to its ligand. Stimulation with lyral increased the cyclic adenosine monophosphate (cAMP) and Ca2+ levels as well as the phosphorylation of cAMP response element binding protein (CREB). Pretreatment with tangeretin synergistically enhanced the effects of stimulation with lyral: up-regulation of cAMP and Ca2+ levels and CREB phosphorylation. These data collectively suggest that tangeretin synergistically enhances the odorant-induced signaling pathway through modulation of cAMP and CREB signal transduction in non-chemosensory cells.

Eun-Jin Kim, Heyn Lee, Ju-Hoon Lee, Sangryol Ryu, Jong-Hyun Park. Morphological features and lipopolysaccharide attachment of coliphages specific to Escherichia coli O157:H7 and to a broad range of E. coli hosts. Appl. Biol. Chem. 59(1):109-116The objective of the present study was to analyze host-phage adsorption of bacteriophages infecting Escherichia coli O157:H7 and the other E. coli strains. Out of 55 coliphage strains, we selected seven coliphages infectious only to 23 E. coli O157 and seven other coliphages of broad specificity to E. coli O157:H7 and other 61 E. coli. Escherichia coli O157-specific phages and the broadly specific phages all belonged to the Siphoviridae and Myoviridae family, respectively. Escherichia coli O157-specific phages infected E. coli O157:H7, but not E. coli O157:H7△rfaL, deletion mutant of O-antigen ligase gene for lipopolysaccharide. Five coliphages among the broadly specific phages infected E. coli O103, but not E. coli O103△rfaG, deletion mutant of the glycosyltransferase gene. E. coli O157:H7-specific phages among Siphoviridae recognized O-antigen of E. coli O157, but the broadly specific coliphages of Myoviridae may recognize O-antigen and/or a part of the lipopolysaccharide core as an adsorption site in various E. coli. The receptor of the two coliphage groups interacts with some part of lipopolysaccharide, and the tail morphology of the coliphages may be related to their adsorption to and recognition of a different part of lipopolysaccharide. In particular, specificity of E. coli O157:H7-specific phages carrying the long tail of Siphoviridae for O-antigen as a receptor seems to be high.

Azhar U. Khan, Mahboob Alam, Dong-Ung Lee. A bench-top catalyst: BF3·SiO2-assisted synthesis, biological assay, and computational simulations of azacholestanes. Appl. Biol. Chem. 59(1):117-127A series of steroidal N-2′-hydroxy ethenyl-7a-aza-B-homo-5α-cholestane-7-one and its analogs were prepared by the reaction of steroidal α,β-unsaturated ketone with 2-azidoethanol in the presence of BF3.SiO2 as a heterogeneous catalyst in good yields. The structure of the compounds have been established by elemental and spectral data analyses. In vitro evaluation of antimicrobial and antioxidant activity of the synthesized compounds was reported. The activity of compound 6 was further evaluated based on structural and molecular modeling and docking studies. Moreover, the structure–biological activity relationship was investigated using physicochemical and molecular descriptors with the WebMO job manager employing semi-empirical PM3, basic level method, and molinspiration tools, respectively.

Bo Ram Shin, Hye-Won Song, Joon-Goo Lee, Hae-Jung Yoon, Myung-Sub Chung, Young-Suk Kim. Comparison of the contents of benzo(a)pyrene, sesamol and sesamolin, and volatiles in sesame oils according to origins of sesame seeds. Appl. Biol. Chem. 59(1):129-141The objective of the present study was to compare the contents of antioxidants (sesamol and sesamolin) and benzo(a)pyrene (BaP), as well as the volatile profiles in sesame oil obtained from sesame seeds cultivated in four different areas in Korea, and those

cultivated in other countries, including Ethiopia, India, and China. The sesamol content was lower in Korean sesame oils than in Ethiopian, Indian, and Chinese sesame oils, whereas sesamolin content was higher in sesame oils derived from Korea than those in sesame oils derived from other countries. There was also a difference in the contents of BaP in the sesame oils obtained from seeds in different cultivated areas. The volatiles that allowed Korean samples to be discriminated from those of other countries were mainly pyrroles and sulfur-containing compounds such as 1-ethyl-1H-pyrrole, 2-ethyl-4-methyl-1H-pyrrole, 2,4-dimethylthiazole, and 5-ethyl-4-methylthiazole.

Xiao Meng, Yu Qian, Li-Shi Jiang, Jin-Mei Kang, Yan Chen, Juan Wang, Shu-Kun Liu, Zhen-Ming Che, Xin Zhao. Effects of Lactobacillus plantarum SCS2 on blood glucose level in hyperglycemia mice model. Appl. Biol. Chem. 59(1):143-150In this study, the hyperglycemia mice model was established with 1-week high sugar and fat diet plus with 70 mg/kg body weight of streptozotocin injection for 3 days. Sixty male Kunming mice of 3 weeks old in a specific-pathogen-free grade were divided into six groups randomly, which includes normal group (NG), prevention group (PG), treatment group for low dose (TGL), middle dose (TGM), high dose (TGH), and model group (MG). NG and MG mice were fed with sterile physiological saline (10 mL/kg body weight). PG mice were fed with the concentration of 6.0 × 109 CFU/mL L. plantarum SCS2 suspensions from the second to third week. TGL, TGM, and TGH mice were fed with the concentration of 2.0 × 109, 4.0 × 109, and 6.0 × 109 CFU/mL L. plantarum SCS2 suspensions (10 mL/kg body weight), respectively from fourth to tenth week. The results showed that the fasting and postprandial 2 h blood glucose levels of TGH mice were reduced by L. plantarum SCS2 significantly (p < 0.05) as compared with MG. The body weight of TGH mice came to normal level at tenth week. Content of K+ in plasma of TGH mice was increased and contents of Na+ and Cl− in the plasma of TGH mice were decreased as compared with MG. Meanwhile, content of glycogen in TGH mice was reduced. However, the effect of L. plantarum SCS2 on the prevention of hyperglycemia in PG mice was not significant as compared with NG mice during the experiment. These results suggested that L. plantarum SCS2 showed a hypoglycemic effect on hyperglycemic mice model.

Nam Tuan Vu, Juan Manuel Pardo, Elizabeth Alvarez, Ham Huy Le, Kris Wyckhuys, Kim-Lien Nguyen, Dung Tien Le. Establishment of a loop-mediated isothermal amplification (LAMP) assay for the detection of phytoplasma-associated cassava witches’ broom disease. Appl. Biol. Chem. 59(2):151-156Cassava (Manihot esculenta Crantz) is one of the most important food crops in the tropics; however, bacterial phytopathogens pose a serious threat to its farming. Cassava Witches’ Broom Disease (CWB) is caused by the infection of phytoplasma and is manifested as reduction in tuber yield and starch content at harvest of 10 and 30 %, respectively. Although polymerase-chain reaction provides the gold standard in diagnostics, this method requires significant investments in infrastructure and training. Here, we developed a loop-mediated isothermal amplification (LAMP) assay that allows specific detection of phytoplasma from field-collected samples. Three primer sets were designed, of which two detected phytoplasma DNA sequence encoding 16S rRNA (16S rDNA), the other detected cassava actin. Following a 1 h LAMP reaction at 63 °C, a positive reaction can be visualized by agarose gel electrophoresis, hydroxynaphthol blue color change, or the presence of a precipitate. In a pilot field study, the assay was able to rapidly distinguish between healthy and CWB-infected cassava. With further development, a LAMP for routine on-site screening of cassava crops can be envisioned.

Ji Young Kim, BoRa Yi, Chankyu Lee, Seo-Yeong Gim, Mi-Ja Kim, JaeHwan Lee. Effects of pH on the rates of lipid oxidation in oil–water system. Appl. Biol. Chem. 59(2):157-161The effect of pH on the oxidative stability in oil–water systems was determined at 40 °C for 18 days by monitoring headspace oxygen depletion and concentration of conjugated dienoic acids (CDAs). Two types of pH solutions were prepared: a buffer solution and a mixture of NaOH and HCl. When the pH of the mixture of NaOH and HCl increased from 1

to 13, the oxidative stability in oil decreased significantly in the order of pH 1 > 10 > 7 > 4 > 13. In case of the buffer solution, the oxidative stabilities of oils were in the order of pH 4 = 7 > 10. Depending on the pH and buffering capacities, oxidative stability of oils were greatly influenced.

Hee Rang Yun, Yeong Gil Rim, Jae Bok Heo. Rice small GTPase Rab11 is required for intracellular trafficking from the trans-Golgi-network to the plasma membrane and/or prevacuolar compartments. Appl. Biol. Chem. 59(2):163-171Members of the Rab family of small GTPases play important roles in membrane trafficking along the exocytic and endocytic pathways, their function being dependent on their localization. Here, we show the vital roles of the Oryza sativaRab11 (OsRab11) in intracellular trafficking using a dominant-negative mutant approach based on a protoplast-trafficking assay. GTP-binding and GTP hydrolysis assays revealed that OsRab11 is a small GTP-binding protein complementing △ypt32/ypt31. Green fluorescent protein (GFP)-OsRab11 has been localized to both the trans-Golgi-network (TGN) and the endosomes/prevacuolar compartments (PVC) in Arabidopsis protoplasts. The protoplast transformation with the dominant-negative mutant OsRab11(S28N) revealed that the trafficking of plasma membrane marker proteins [H+-ATPase-GFP and Ca2+-ATPase8-GFP (ACA8-GFP)] and central vacuole marker proteins [Arabidopsis aleurain-like protein (AALP-GFP) and sporamin (Spo-GFP)] was inhibited. Moreover, overexpression of Arabidopsis AtRha1 did not recover the trafficking inhibition of marker proteins from the central vacuole. These results strongly indicate that OsRab11 localizes to either the TGN or the PVC, and plays a significant role in the intracellular trafficking from the TGN to the PM and/or to the PVC in planta.

Dae Gyuun An, Mi Na Cha, Saravanan Prabhu Nadarajan, Bong Gyu Kim, Joong-Hoon Ahn. Bacterial synthesis of four hydroxycinnamic acids. Appl. Biol. Chem. 59(2):173-179Hydroxycinnamates are a class of phenolic compounds that have a C6-C3 carbon backbone. Hydroxycinnamic acids are derived from cinnamic acid via hydroxylation or methylation and are found in foods such as pears, coffee beans, and dandelions. They are involved in protection against chemotherapy side effects and the prevention of cardiovascular disease and cancer. We synthesized four types of hydroxycinnamates (p-coumaric acid, cinnamic acid, caffeic acid, and ferulic acid) from glucose in Escherichia coli by introducing different combination of four genes: tyrosine ammonia lyase, phenylalanine ammonia lyase, S.espanaensis monooxygenase, and Oryza sativaO-methyltransferase. The final yields of hydroxycinnamic acids were increased by engineering the metabolic pathway of E. coli. Using these strategies, 100.1 mg/L p-coumaric acid, 138.2 mg/L caffeic acid, 64 mg/L ferulic acid, and 1072.3 mg/L cinnamic acid were synthesized.

Mohammad Sayed Alam, Junaid Uddin Ahmed, Dong-Ung Lee. Biological features, drug-likeness, pharmacokinetic properties, and docking of 2-arylidenehydrazinyl-4-arylthiazole analogues. Appl. Biol. Chem. 59(2):181-192Thiazoles are an important class of heterocyclic compounds that possess a sulfur and nitrogen containing five-membered ring, which acts as a pharmacophore, and show a wide range of complex biological activities. A series of sixteen 2-arylidenehydrazinyl-4-arylthiazole analogues (3a–p) were evaluated for cytotoxic activity against brine shrimp (Artemia salina) nauplii and their minimum inhibitory concentrations were determined against two Gram-positive (Listeriamonocytogenes and Enterococcus faecalis) and two Gram-negative bacterial strains (C.sakazakii and E. coli). Of the tested compounds, 3g demonstrated highest cytotoxicity with a LC50 value of 54 ppm followed by compound 3h(LC50 = 85 ppm), in a short-term bioassay using A. salina, whereas compound 3i exhibited the most potent antibacterial activities against L.monocytogenes, E. faecalis, and C.sakazakii with MIC values ranging from 50 to 100 μg mL−1. Compound 3g showed highest antibacterial activity against E. coli (MIC = 50 μg mL−1). In silico drug-likeness, pharmacokinetic (ADME) properties, toxicity effects, and drug scores were also evaluated, and none of the sixteen compounds

were found to violate Lipiniski’s rule of five or Veber’s rule, indicating potential for development as oral drug candidates. In addition, a docking study of compound 3i into the active site of E. coli FabH receptor, an attractive target for the development of new antibacterial agents, showed it has good binding properties.

Kyungho Lee, Da Hyun Lee, You Jung Jung, Soon Young Shin, Young Han Lee. The natural flavone eupatorin induces cell cycle arrest at the G2/M phase and apoptosis in HeLa cells. Appl. Biol. Chem. 59(2):193-199Eupatorin (5,3′-dihydroxy-6,7,4′-trimethoxyflavone) is a natural flavone exhibiting antiproliferative activity in diverse cancer cell lines. In the present study, we characterized the cytotoxic effect of eupatorin in HeLa cervical carcinoma cells. Treatment with eupatorin induced cell cycle arrest at the G2/M phase within 12 h, followed by the induction of apoptosis. Eupatorin down-regulated cyclin D1 within 3 h, whereas up-regulated cyclin B1 levels after 12 h. Eupatorin also accumulated p53, p21, and Bax levels and stimulated the cleavage of caspase-3, caspase-7, and poly(ADP-ribose) polymerase after 12 h of treatment. In addition, we found that eupatorin-induced p21 expression was p53-dependent, while Bax expression was Egr-1-dependent pathway. Our results suggest that eupatorin induces G2/M cell cycle arrest through dysregulation of cell cycle regulatory proteins and triggers apoptosis through activation of the p53-dependent and p53-independent pathways.

Jun-Cheol Moon, Ju-Hee Kim, Cheol Seong Jang. Development of multiplex PCR for species-specific identification of the Poaceae family based on chloroplast gene, rpoC2. Appl. Biol. Chem. 59(2):201-207In this study, we report the development of a multiplex polymerase chain reaction (PCR) method using species-specific primers for the simultaneous detection of Poaceae members, including adlay, barley, maize, rice, and wheat, based on sequence polymorphism in DNA-directed RNA polymerase beta-prime chain genes (rpoC2). Species-specific primer pairs were constructed with a common forward primer and reverse primers differing by sequence polymorphisms and gene location. Each primer pair was designed to PCR-amplify products of five chloroplast genes of 443, 346, 278, 221, and 96 bp for rice, barley, adlay, wheat, and maize, respectively. Multiplex PCR with a series of template DNA concentrations (0.01–10 ng/μL) was used to optimize amplification of fragments from pooled Poaceae. In addition, species-specific primers were used to detect components of seven commercial flour-mixed products. This combination of the sensitivity of multiplex PCR with the specificity of primers designed to detect unique species has broad applications in the processed food industry.

Kyu-Won Hwang, Daniel Son, Hyeong-Wook Jo, Chun Hwan Kim, Ki Cheol Seong, Joon-Kwan Moon. Levels of curcuminoid and essential oil compositions in turmerics (Curcuma longa L.) grown in Korea. Appl. Biol. Chem. 59(2):209-215Levels of curcuminoids and essential oil compositions from eighteen turmerics (Curcuma longaL.) grown in Jeju were analyzed. 0.5 g of turmeric samples was extracted with 20 mL of methanol for 7 h by shaking and was ultra-sonicated for 30 min. The extracts were analyzed by high-pressure liquid chromatography-diode array detector using Brownlee SPP C18 column (4.6 × 100 mm, 2.7 µm) after filtration. Curcumin (CUR), demethoxycurcumin (DEM), and bisdemethoxycurcumin (BIS) were monitored at the wavelength of 420 nm. Total curcuminoids in turmerics ranged from 53.9 ± 1.0 to 5396.0 ± 101.8 µg/g dry turmeric weight. In decreasing order, curcuminoids found were CUR (4669.7 µg/g) > DEM (565.3 µg/g) > BIS (160.9 µg/g). K145023 variety contains the highest amount of curcuminoids and variety number 10 contains the least amount. Essential oils were extracted with steam distillation method and analyzed with GC–MS. The major components of turmeric oil were α-zingiberene (27.70–36.75 %), aromatic-turmerone (19.54–32.24 %), β-sesquiphellandrene (13.14–18.23 %), α-turmerone (3.72–6.50 %), β-turmerone (2.86–5.60 %), and β-bisabolene (2.50–3.46 %).

Hyeryoung Jung, Seunghyun Ahn, Beum Soo Kim, Soon Young Shin, Young Han

Lee, Yoongho Lim. Isoflavones as modulators of adenosine monophosphate-activated protein kinase. Appl. Biol. Chem. 59(2):217-225Adenosine monophosphate-activated protein kinase (AMPK) is expressed in all eukaryotic cells and can therefore be found in vertebrates, invertebrates, and plants. Since AMPK participates in the regulation of homeostasis on various levels, small compounds that can modulate AMPK activity could be valuable research tools. Several flavonoids can modulate AMPK. Here we investigated the modulatory effect of 37 isoflavones on AMPK activity using an in vitro kinase assay. Because the relationship between the structural properties of flavonoids and their modulatory activities has not been elucidated yet, we used comparative molecular field analysis to derive the structural conditions for modulation of AMPK activity. The molecular binding mode of isoflavones to AMPK was elucidated using in silico docking studies. The findings presented here can aid in the design of new modulators with better specificity for AMPK.

Behrooz Yahyaei, Niloofar Peyvandi, Hamed Akbari, Sepideh Arabzadeh, Sima Afsharnezhad, Hatef Ajoudanifar, Parastoo Pourali. Production, assessment, and impregnation of hyaluronic acid with silver nanoparticles that were produced by Streptococcus pyogenes for tissue engineering applications. Appl. Biol. Chem. 59(2):227-237Hyaluronic acid (HA) is a mucopolysaccharide with different applications in the medicinal field. Since HA does not have antibacterial property, in this research a bacterium with both abilities of HA and silver nanoparticles (SNPs) production was used; HA containing SNPs (HA/SNPs) was obtained and the toxic effect of HA/SNPs was achieved using Limulus Amebocyte Lysate (LAL) and 3-(4,5 dimethyl thiazol-2yl)-2,5 diphenyl tetrazolium bromide (MTT) assays. HA and SNPs were produced separately by Streptococcus pyogenes. Then HA was analyzed using fourier transform infrared (FTIR) spectroscopy and transmission electron microscope (TEM), and SNPs were analyzed using TEM and X-ray diffraction analysis (XRD). In order to produce HA/SNPs complex, SNPs were added to the supernatant containing HA and the complex was precipitated using isopropyl alcohol. HA, SNPs, and HA/SNPs were used for the antibacterial assay. Results proved the formation of HA and SNPs in the bacterial culture medium. SNPs sizes were around 5–30 nm and entered in the structure of HA. Antibacterial test showed that unlike the pure HA, SNPs and HA/SNPs had antibacterial activity. LAL and MTT assays results showed that HA/SNPs had low and dose-dependent toxic effects. In future, the use of HA/SNPs scaffold in tissue engineering field is expected.

Furkan Özen, Suat Tekin, Kenan Koran, Süleyman Sandal, Ahmet Orhan Görgülü. Synthesis, structural characterization, and in vitro anti-cancer activities of new phenylacrylonitrile derivatives. Appl. Biol. Chem. 59(2):239-248The present study was designed to both synthesize phenylacrylonitrile compounds (1a–k) and their anti-tumor activities on human breast cancer cell line (MCF-7) were determined. The structures of all the compounds were defined by melting point, elemental analysis, FT-IR, 1H, 13C, 13C-APT, and HETCOR-NMR spectroscopy. Anti-tumor activities of these compounds on cell viability were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay against MCF-7. The MCF-7 cell lines were treated with 1, 5, 25, 50, and 100 μM concentrations of phenylacrylonitrile compounds for 24 h. At the end of the experiments, 1a, 1b, 1c, 1g, and 1h compounds reduced cell viability (p < 0.01). Additionally, the anti-cancer activities of these compounds on MCF-7 were investigated by comparing IC50 values. In conclusion, while some of the synthesized phenylacrylonitrile compounds (1a, 1b, 1c, 1g, and 1h) have anti-tumor activity, other phenylacrylonitrile compounds (1d, 1e, 1f, 1k, and 1h) have no effect on human breast cell lines.

Kyungho Lee, Da Hyun Lee, You Jung Jung, Soon Young Shin, Dongsoo Koh, Young Han Lee. A methoxyflavanone derivative, 2′,3′,4′-trimethoxy-5,6-naphthoflavanone, inhibits proliferation of HCT116 human colon cancer cells by inducing G2/M cell cycle arrest and apoptosis. Appl. Biol. Chem. 59(2):249-253

A methoxylated flavanone derivative, 2′,3′,4′-trimethoxy-5,6-naphthoflavanone (TMNF), was synthesized, and its anti-proliferative activity was evaluated. In HCT116 colon cancer cells, TMNF inhibited cellular proliferation and triggered cytotoxicity. A flow cytometry assay showed that TMNF caused cell cycle arrest at the G2/M phase with a concomitant decrease in the population of G1 phase cells. Furthermore, TMNF caused accumulation of p53 and its downstream target, Bax, and stimulated cleavage of caspase-7 and poly(ADP-ribose) polymerase. These data suggest that methoxylated naphthoflavanone can be an attractive pharmacophore for further development of therapeutic adjuvants.

Jeong Min Lee, Jaemin Lee, Seon Haeng Cho, Sang-Won Lee, Young-Ock Kim, Ik-Hyun Cho, Hak-Jae Kim, Sanghyun Lee. Chiisanenside, a new triterpene glycoside from the fruits of Acanthopanax chiisanensis. Appl. Biol. Chem. 59(2):255-258A new triterpene glycoside was isolated from the fruits of Acanthopanax chiisanensis and identified as 3-O-β-d-[(6-carboxymethyl)-glucopyranosyl-(1 → 3)-β-d-glucuronopyranosyl]-olean-12-en-28-oic acid by spectral analyses. This compound was isolated for the first time from nature, and named chiisanenside.

Kwang-Su Park, Youhoon Chong, Mi Kyoung Kim. Myricetin: biological activity related to human health. Appl. Biol. Chem. 59(2):259-269Myricetin (3,5,7,3′,4′,5′-hexahydroxyflavone) is one of the natural flavonols from fruit, vegetable, tea, and medicinal plants. A great deal of attention has been paid to this compound owing to its various health-promoting effects. In this review, we provide a comprehensive discussion of the therapeutic potential of myricetin by focusing on its biological activity and pharmacokinetic properties.

Jinrui Wei, Kun Liu, Faquan Lin, Chunping He, Xuan Luo, Jiajia Zou, Wei He, Wenqian Nong, Cuiwu Lin. Synthesis, and antioxidant, thrombin-inhibitory, and anticancer activities of hydroxybenzamide derivatives. Appl. Biol. Chem. 59(2):271-281Four hydroxybenzamide derivatives were synthesized from protocatechuic acid. The antioxidant activities of these derivatives were evaluated in vitro using DPPH radical scavenging and reducing power assays. In addition, these compounds were subjected to a chromogenic thrombin-inhibitory assay using Chromozym-TH as the substrate. The anticancer effects of these derivatives were also investigated using the MTT assay in three human cancer cell lines. Four derivatives exhibited excellent DPPH radical scavenging activity and redox potential. Their highly inhibitory effect against thrombin may allow them to be utilized as novel micromolecule thrombin inhibitors. Furthermore, four derivatives demonstrated different inhibitory activities in three human cancer cell lines but showed minimal harmful effects on normal HUVEC cells. Therefore, hydroxybenzamide derivatives indicate the superior antioxidant, thrombin-inhibitory, and anticancer activities along with the excellent biocompatibility, which have the potential application in therapeutic interventions and disease prevention of cancer, thrombotic diseases, and aging.

Jeong Hwa Kim, Sang Min Kim, Kwang Hyun Cha, Il-Kyoon Mok, Song Yi Koo, Cheol-Ho Pan, Jae Kwon Lee. Evaluation of the anti-obesity effect of the microalga Phaeodactylum tricornutum. Appl. Biol. Chem. 59(2):283-290Phaeodactylum tricornutum (PT) is a classical model diatom widely used in ecology, physiology, biochemistry, and molecular biology studies. We evaluated the anti-obesity effects of PT powder based on a number of metabolic parameters in a model of diet-induced obesity. We fed C57BL/6 mice a high-fat diet supplemented with PT powder (15 or 30 % w/v) for 12 weeks, and determined energy intake, weight loss, and lipid profiles each week. PT powder significantly reduced body weight gain, and epididymal and perirenal adipose tissue weight via activation of AMPK and HMGCR pathways. Moreover, we found that fucoxanthin, the putative anti-obesity compound in PT, was effectively micellized and transferred to the soluble fraction at the ileum in an in vitro simulated digestion system. Our results indicate that PT powder has an anti-obesity effect and

suggest that it is a candidate substance for the development of anti-obesity foods, supplements, and even drugs.

Yoon Young Kang, Inseong Choi, Youhoon Chong, Woon-Seok Yeo, Hyejung Mok. Complementary analysis of curcumin biodistribution using optical fluorescence imaging and mass spectrometry. Appl. Biol. Chem. 59(2):291-295In this study, a complementary analysis was performed to improve the precision of the determination of in vivo curcumin biodistribution after intravenous administration. Overall, similar curcumin biodistribution profiles were obtained using optical fluorescence imaging and mass spectrometry. Poor curcumin accumulation was observed in the heart, spleen, and kidney. However, noticeable accumulation of curcumin in the brain was only observed using fluorescence imaging, probably owing to the insufficient extraction of curcumin from the brain for mass spectrometry. In addition, an exact and reliable measurement of curcumin accumulation in tissues such as the liver, gallbladder, and pancreas was performed using mass spectrometry because of high autofluorescence. Taken together, complementary analysis using optical fluorescence imaging and mass spectrometry allowed the precise determination of curcumin in each tissue. Furthermore, this complementary analytical strategy could be used to elucidate the in vivo distribution of a wide range of fluorescent polyphenols.

Jin-A Park, Daun Jeong, Dan Zhang, Seong-Kwan Kim, Sang-Hyun Cho, Soo-Min Cho, Hee Yi, Jae-Han Shim, Jin-Suk Kim, A. M. Abd El-Aty, Ho-Chul Shin. Simple extraction method requiring no cleanup procedure for the detection of minocycline residues in porcine muscle and milk using triple quadrupole liquid chromatography-tandem mass spectrometry. Appl. Biol. Chem. 59(2):297-303A versatile analytical method was developed for simple detection of minocycline residues in porcine muscle and milk using liquid chromatography-triple quadrupole tandem mass spectrometry (LC/MS/MS) with electrospray ionization. Samples were extracted with a mixture of acetonitrile and ethyl acetate (2:1, v/v), and then defatted using n-hexane. No cleanup procedure was deemed necessary. Minocycline was separated on a reversed-phase analytical column using a combination of 0.1 % formic acid in water (A) and 0.1 % formic acid in acetonitrile (B) as the mobile phase. Matrix-matched calibration showed good linearity over a concentration range of 10–60 μg/kg with a determination coefficient (R2) of 0.9727. Fortified porcine and milk samples having concentrations equivalent to and double the limit of quantification (LOQ = 10 ng/g), respectively, yielded recovery ranges between 83.02 and 8.03 % and relative standard deviations <18 %. Samples collected from a large market located in Seoul, Korea, tested negative for minocycline residue. These results show that a combination of acetonitrile and ethyl acetate can effectively extract minocycline from porcine muscle and milk without solid-phase extraction, a step usually required for cleanup before analysis. The developed method is simple, sensitive, and can be extrapolated to other food animal products that are rich in protein and fats.

Ji Yeon Seo, Jin Ho Jang, Jae-Sik Kim, Eun-Jeong Kim, Jong-Sang Kim. Development of low-sugar antioxidant jam by a combination of anthocyanin-rich berries. Appl. Biol. Chem. 59(2):305-312A major challenge for the food industry is to satisfy the consumer demand for health benefits as well as taste. In this study, we investigated the antioxidant activities of extracts from four anthocyanin-rich jams, as well as their low-sugar versions, which used white grape juice concentrate as a white refined sugar substitute. It was determined that blueberry + Korean black raspberry jam dramatically scavenged 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid), peroxyl radicals, and reduced ferric ions in the tested jams and contained high amounts of phenols, flavonoids, and anthocyanins. In addition, the antioxidant capacity and content of phenolics, flavonoids, and anthocyanins were retained during the processing of berry jams. Berry jams and their low-sugar jams exhibited inhibitory effects on reactive oxygen species production in HepG2 cells and lipid peroxidation in the liver and brain homogenates. In conclusion, blueberry + Korean black raspberry and

raspberry + strawberry jams have strong antioxidant capacities, which are retained in jams containing sugar substitutes. These results suggest that anthocyanin-rich berry jams with natural sugar replacements could be developed as high functional foods and also decrease the risk for metabolic diseases.

Min-Soo Kim, Mi-Hee Woo, Young-Hyo Chang, Namhyun Chung, Joong-Su Kim. Biochemical characterization of a noble xylanase from Paenibacillus sp. EC116. Appl. Biol. Chem. 59(2):313-320In our present study, a new Endo-1,4-β-xylanase (EC 3.2.1.8) was isolated, which belongs to glycosyl hydrolase family 10 member. The new enzyme designated as xynA was expressed for biochemical characterization. The xynA xylanase was identified in Paenibacillus sp. EC116. The novel gene had the size of 2673 base pairs, which correspond to 891 amino acid residues. The expressed enzyme has a size of ~100 kDa. The amino acid sequence was similar to that of endo-1,4-beta-xylanase from Paenibacillus sp. FSL R5-192 (ETT36211.1) (96 % identical). The xylanase has an optimal temperature of 40 °C and the pH optimum of 6.0. The specific activity of the xylanase toward birchwood xylan was about 0.69 μmol min−1 mg−1, the Vmax value of 1.639 μmol mg−1 min−1, and the Km value of 35.1 mg ml−1. The EC116 xylanase was relatively stable up to 60 °C. Xylanase enzymes have numerous industrial applications, which need to meet the specific requirements for each application. We hope that our EC116 xylanase is one of the candidates for commercial applications.

Cheng-Yi Zhang, Ki-Hoon Lee, Shin Young Park, Minhui Kim, Myung-Sub Chung, Sook Young Lee, Young-Gil Jeong, Sang-Do Ha. Evaluation of the quality change and estimation of the shelf life of instant Bibimbap (Korean Mixed Rice) during storage. Appl. Biol. Chem. 59(2):321-326This study estimates the shelf life of four different instant bibimbaps. Each product was stored at 25, 35, or 45 °C for 180 days, and the changes in the physiochemical, microbiological, and sensory parameters were analyzed during storage. The total bacteria and coliforms for the bibimbaps were up to 4 and 1 log CFU/g, respectively. The highest population of yeast and mold was 1.98 log CFU/g, and Bacillus cereus was not detected in all bibimbaps. The moisture content and acid value were used to estimate the shelf life of bibimbaps. The shelf life of the four types of bibimbaps was over 4 years.

MyoungLae Cho, Seok-Beom Ko, Jae-Min Kim, Ok-Hwan Lee, Dae-Won Lee, Jong-Yea Kim. Influence of extraction conditions on antioxidant activities and catechin content from bark of Ulmus pumila L.. Appl. Biol. Chem. 59(3):329-336This study investigated the influence of extraction conditions on antioxidant activities and catechin content from bark of Ulmus pumila. The U. pumila was extracted using various concentrations of ethanol (30, 50, 80, and 99 %) and different extraction times (1, 2, and 3 h) at 60 °C. The ethanol extracts were evaluated for their antioxidant activities [1,1-diphenyl-1-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activities and reducing power], total phenolic (TP), and proanthocyanidin (TA) contents. In addition, the antioxidant activities were correlated with TP and TA contents, and the catechin content was determined using high-performance liquid chromatography. The 99 % EtOH for 3-h extracts exhibited the highest antioxidant abilities with the highest phenolic and proanthocyanidin contents. TP content showed little correlation (R2 = 0.31–0.61) with their antioxidant activities; however, a significant correlation (R2 = 0.65–0.91) was observed with proanthocyanidin content. The catechin content of the 99 % EtOH for 3-h extracts was greater than that of other EtOH extracts. Therefore, these results indicate that the antioxidant capacity of EtOH extracts from bark of U. pumila may be due to catechin, and the optimal extraction condition was 99 % EtOH for 3 h.

Rupak Chakraborty, Ji Hyeong Baek, Eun Young Bae, Woe-Yeon Kim, Sang Yeol Lee, Min Gab Kim. Comparison and contrast of plant, yeast, and mammalian ER stress and UPR. Appl. Biol. Chem. 59(3):337-347The endoplasmic reticulum (ER) is a well-characterized protein folding mechanism in

eukaryotic organisms. Many secretory and membrane proteins are folded in the ER before they are translocated to their functional destination. Various conditions, such as biotic, abiotic, or physiological stresses, lead to the accumulation of unfolded and misfolded proteins in the ER, resulting in ER stress. In response to ER stress, cells initiate a protective response called the unfolded protein response (UPR) to maintain cellular homeostasis. Previous studies suggest that inositol-requiring kinase 1 (IRE1) is a universal ER stress sensor in yeast, mammals, and plants. IRE1-mediated splicing of UPR transducers, such as HAC1, XBP1, and bZIP60, triggers the UPR in yeast, mammals, and plants, respectively. In mammals, activated transcription factor 6 and double stranded RNA-activated protein kinase-like ER kinases are involved in the UPR. In plants, the additional UPR transducers bZIP28 and bZIP17 are activated by Golgi-localized S1P and S2P proteases. Subsequently, these UPR transducers are exported to the nucleus and upregulate the expression of UPR-responsive genes encoding BiP, calreticulin, calnexin, protein disulfide isomerase, and glucose-regulated protein 94 to decrease the amount of misfolded proteins and induce endoplasmic reticulum-associated degradation. In plants, the UPR signaling pathway plays an important role in ER homeostasis and normal biological processes; however, the molecular mechanisms of the UPR in plants remain poorly understood. This paper provides an overview of the regulatory and signaling mechanisms of the UPR across kingdoms. In addition, the emerging role of the UPR in plant physiology and defense response will be discussed.

Heeok Hong, Kai-Min Niu, Jun-Hyeong Lee, Sangbuem Cho, Sung-Gu Han, Soo-Ki Kim. Characteristics of Chinese chives (Allium tuberosum) fermented by Leuconostoc mesenteroides. Appl. Biol. Chem. 59(3):349-357This study was conducted to investigate the characteristics of bioactive compounds in Chinese chives juice (FC) fermented with Leuconostoc mesenteroides SK1962, a lactic acid bacteria isolated from Kim-chi. L. mesenteroides SK1962 only (LO) was used as comparison. The pH of FC gradually decreased from 6.21 to 4.23 during the 48-h incubation period, which was similar to that of LO. The growth of L. mesenteroides SK1962 in FC was higher compared with that in LO for various incubation times, with the exception of the 8-h incubation. Total polyphenol and flavonoid contents in FC were higher as compared with those in LO during incubation, leading to increased antioxidant activity in FC at different incubation times. Moreover, FC was more effective than LO in reduction of superoxide free radical production in primary bovine mammary alveolar cells. In addition, FC demonstrated antibacterial properties against pathogenic bacteria such as Listeria monocytogenes, Pantoea agglomerans, Haemophilus parasuis, Salmonella gallinarum, Escherichia coli O157, and Burkholderia. sp. Although LO also showed antibacterial effects against the above-mentioned pathogenic bacteria, its antibacterial activities were generally lower compared with those of FC. The results show that the antioxidant and antibacterial activities in Chinese chives was induced by fermentation with L. mesenteroides SK 1962. In conclusion, fermentation may lead to an increase in bioactive compounds including total polyphenol and flavonoid.

Hyun-Joo Chang, Jeil Hong, Nari Lee, Hyang Sook Chun, Hwa Yeon Kim, Sung-Wook Choi, Gyeongsik Ok. Growth inhibitory effect of bacteriophages isolated from western and southern coastal areas of Korea against Vibrio parahaemolyticus in Manila clams. Appl. Biol. Chem. 59(3):359-365Vibrio parahaemolyticus is one of the leading causes of seafood-related illnesses in Korea and the whole world. The growth inhibitory effect of V. parahaemolyticus-specific phages (Vpp) in clam samples was investigated. Six bacteriophages specific to V. parahaemolyticus were isolated by the agar overlay method using sediment and seafood samples from the western and southern coastal areas of Korea. Host range, restriction digestion pattern, and transmission electron microscope images of phage isolates were examined. The most effective phage, Vpp2, at a multiplicity of infection (MOI) of 10 showed 4.2 log CFU/mL reduction in V. parahaemolyticus growth at 30 °C and 6 log CFU/mL reduction after the incubation at 37 °C for 6 h. In a food application study, Vpp2 at an MOI of 100 demonstrated a 2.1 log CFU/mL reduction at 25 °C after a 24-h

incubation in shell-less Manila clams spiked with V. parahaemolyticus. Additional studies are needed to characterize the newly discovered Vpp and their potential applications as biocontrol agents in food.

Sora Han, Young Yang. Phenotypic and functional dissection of myeloid-derived suppressor cells. Appl. Biol. Chem. 59(3):367-371Myeloid-derived suppressor cells (MDSCs) are originated and differentiated population from common hematopoietic progenitor cells. Generally, in the late stage of inflammation, MDSCs differentiation and expansion are promoted to suppress the over-activated immune system so that the immune system can maintain the homeostasis. Recently, it has been revealed that MDSCs accumulate in cancer patients and tumor-bearing experimental animals, and these tumor-derived MDSCs suppress anti-tumor immunity by secreting immunosuppressive cytokines including reactive oxygen species and inducible nitric oxide synthase. This fact prompts scientists to shed light on MDSCs as significant targets for anti-cancer immunotherapy. However, due to morphological, phenotypic, and functional heterogeneities of MDSCs, it is not easy to develop therapeutic strategies targeting MDSCs. In this review, we will summarize recent progress on defined subsets of MDSCs and their strategies to suppress T cell-mediated anti-tumor immunity.

Pitchaporn Wanyo, Niwat Kaewseejan, Naret Meeso, Sirithon Siriamornpun. Bioactive compounds and antioxidant properties of different solvent extracts derived from Thai rice by-products. Appl. Biol. Chem. 59(3):373-384We investigated the contents and compositions of bioactive compounds in by-products of rice as affected by extraction with different solvents. Free and bound phenolic compounds and their antioxidant activities of rice bran and husk extracted by acetone, ethanol, and water at different temperatures (50, 60, 70, and 80 °C) were evaluated. Overall, the heated water extract provided the highest 2,2-diphenyl-1-picrylhydrazyl radical scavenging activities, ferric-reducing antioxidant power, total phenolic content, and total flavonoid content compared to the other solvents of all the samples studied. The antioxidant activities increased when the water temperature increased from 50 to 70 °C but decreased at 80 °C. The contents of bound phenolics were greater than those of free phenolics, including phenolic acids and flavonoids, in all the samples studied. Acetone gave the highest amounts of γ-oryzanol and tocopherols in all samples. With a reduction in particle size of the rice husk, there was a significant increase in extracted phenolic acids, flavonoids, and antioxidant properties.

Saksri Sanyacharernkul, Sirinapa Nantapap, Kallaya Sangrueng, Narong Nuntasaen, Wilart Pompimon, Puttinan Meepowpan. Antifungal of modified neolignans from Mitrephora wangii Hu. Appl. Biol. Chem. 59(3):385-389Chromatographic separation of leaves and twigs from M. wangii had led to the isolation of conocarpan (1) and 3′-methoxyconocarpan (2). These structures were assigned on the basis of spectroscopic methods. Compound 1 was modified by organic reactions and the modified compound 5 exhibited potent antifungal, Pyricularia oryzae. The method used poisoned food technique with mycelial growth inhibition for 14 days, 57 % (100 ppm). Compound 5 was stronger than mancozeb which the fungi toxicity drug used as a positive control.

Kyungho Lee, Da Hyun Lee, Ji-Ho Kim, You Jung Jung, Soon Young Shin, Dongsoo Koh, Young Han Lee. The chalcone derivative HymnPro generates reactive oxygen species through depletion of intracellular glutathione. Appl. Biol. Chem. 59(3):391-3962-Hydroxy-4-methoxy-2′,3′-benzochalcone (HymnPro), a chalcone derivative, induces cell cycle arrest at the G2/M phase followed by caspase-dependent apoptosis. Structurally, HymnPro contains a α-β olefin that can act as a Michael acceptor, and that is involved in the depletion of cellular glutathione. However, the effect of HymnPro on the generation of reactive oxygen species (ROS) is still unknown. In the present study, we observed that treatment of Capan-1 pancreatic cancer cells with HymnPro resulted in a dose-dependent

accumulation of ROS, owing to the depletion of intracellular glutathione. Treatment with ROS scavenger N-acetylcycteine prevented HymnPro-induced caspase activation and cell death, but had little effect on G2/M cell cycle arrest and microtubule assembly. Our results suggest that HymnPro causes tubulin binding-mediated cell cycle arrest at the G2/M phase as well as ROS-mediated caspase activation.

Dongfang Chen, Junling Shi, Xinzhong Hu. Enhancement of polyphenol content and antioxidant capacity of oat (Avena nuda L.) bran by cellulase treatment. Appl. Biol. Chem. 59(3):397-403In this work, cellulase was used to treat with oat bran, and its effect on the total polyphenol content and phenolic profiles were investigated, with heating-only treatment as the control. Antioxidant capacity of the phenolic extracts from oat bran was assessed by scavenging of 2,2′-azinobis (3-ethylbenzothiazoline-6- sulphonic acid), 2, 2-diphenyl-1-picrylhydrazyl radical cation, the ferric reducing antioxidant power, and protein oxidative damage protection assays. As a result, cellulase treatment significantly increased the total phenolic content, total antioxidant capacity, and the ability on the protection of protein from oxidative damage of the oat bran compared with heating-only treatment. Furthermore, cellulase treatment significantly increased availability of most phenolic compounds, except of gallic acid, such as caffeic acid by 97 % (5.33 vs. 10.51 μg g−1), vanillin by 28 % (3.32 vs. 4.24 μg g−1), p-coumaric acid by 105 % (5.55 vs. 11.35 μg g−1), and ferulic acid by 914 % (12.33 vs. 124.03 μg g−1). However, heating-only treatment had no significant influence on the five detected phenolic compounds. Therefore, the study revealed that cellulase treatment would produce polyphenol-rich oat products with increased antioxidant activity.

Kamel Chaieb, Mohamed Hagar, Nagi R. E. Radwan. Biodegradation and decolorization of azo dyes by adherent Staphylococcus lentus strain. Appl. Biol. Chem. 59(3):405-413A Staphylococcus lentus strain, isolated from Red sea water, was tested for decolorization capacity of Congo red, Evans blue, and Eriochrome Black T azo dyes. Biodegradation (100 mgl−1) of these dyes was studied within 24 h in Mineral Salt Medium solution containing 0.10 % (w/v) yeast extract and 7 mM of glucose at a pH of 7.2 and a temperature of 37 °C. Using a 2.2 × 106 CFU/mL inoculum size, S. lentus was able to decolorize these azo dyes with different degree. The staphylococcal biomass achieves approximately 100 % decolorization of Congo red and Eriochrome Black T by the end of treatment. FTIR and UV–Vis analyses confirm biodegradation potential of the tested strain. Moreover, the phytotoxicity of the dye solutions resulting from this treatment shows lower toxic nature compared to untreated solution of the respective dyes.

Chong-Kyu Lee, Min-Yeong Jang, Hwa Rang Park, Gab-Chul Choo, Hyun Seo Cho, Sam-Bong Park, Ki-Cheol Oh, Jong-Bin An, Bong-Gyu Kim. Cloning and characterization of xylanase in cellulolytic Bacillus sp. strain JMY1 isolated from forest soil. Appl. Biol. Chem. 59(3):415-423Microbes play an important role in carbon turnover in forest ecosystems by producing polysaccharide-degrading enzymes such as cellulase, xylanase, and β-glucosidase. In the present study, we isolated a bacterial strain producing cellulase and xylanase from the Forest Park at Gyeongnam National University of Science and Technology using LB agar plates containing 0.5 % carboxymethyl cellulose and 0.01 % trypan blue. Based on 16S rRNA sequencing and API analysis, the isolated strain was identified as a Bacillus species and named Bacillus sp. JYM1. The optimal growth temperature of Bacillus sp. JYM1 was 37 °C. The maximal activities of carboxymethyl cellulase (CMCase) and xylanase were obtained after a 24-h cultivation. The optimal pH and temperature were 6.0 and 50 °C for CMCase and 5.0 and 50 °C for xylanase, respectively. The gene responsible for the xylanase activity in Bacillus sp. JYM1 was cloned and expressed in Escherichia coli. The expressed recombinant protein showed similar biochemical properties to the xylanase of Bacillus sp. JYM1. Therefore, our results confirmed that the gene cloned from Bacillus sp. JYM1, herein named Bxyn, encodes xylanase.

T. Y. Gorgulu, O. D. Ozdemir, A. S. Kipcak, M. B. Piskin, E. M. Derun. The effect of lemon on the essential element concentrations of herbal and fruit teas. Appl. Biol. Chem. 59(3):425-431In this study, the effect of lemon addition to the element content of several herbal and fruit teas is investigated. Lemon addition to tea is a traditional cultural practice in many countries. For this purpose, fennel, mint, and sage are selected as herbal teas, and apple, lemon, and rosehip are selected as fruit teas. The essential elements of calcium, cobalt, copper, iron, potassium, magnesium, sodium, phosphorus, selenium, and zinc are determined in the aforementioned teas with and without lemon addition by using an inductively coupled plasma optical emission spectrometer. From the results of the experiments, the lemon addition caused different changes with respect to the tea type. Potassium (1178 ppm) in apple tea with lemon, sodium (215.1 ppm) in fennel tea with lemon, and calcium (81.88 ppm) and magnesium (53.83 ppm) in mint tea with lemon are found to be the four major essential elements in the teas. In general, the elemental contents are increased with the lemon addition for all of the tea types, except for Na in the sage tea.

Mehdi Khoshneviszadeh, Amirhossein Sakhteman. Exploring quantitative structure–activity relationship (QSAR) models for some biologically active catechol structures using PC-LS-SVM and PC-ANFIS. Appl. Biol. Chem. 59(3):433-441Exploring predictive QSAR models for dopamine catechol structures could be used in designing more potent ligands. In this study, efforts were taken to find out the most important molecular features responsible for the biological activity of catechol structures. All 2D descriptors of Dragon including constitutional, topological, molecular walk counts, BCUT descriptors, Galvez topological, 2D autocorrelations, functional groups, atom-centred fragments, empirical descriptors and properties were calculated for the structures. Two non-linear modelling methods (PC-LS-SVM and PC-ANFIS) were used and compared in this QSAR study. The results revealed the more predictive ability of PC-LS-SVM in the QSAR analysis of the compounds with catechol substructure. The roles of topological properties and number of hydrogen bond donors group as molecular features responsible for the activity of the compounds were discussed. The obtained QSAR models can be used in future studies of drug development for human dopamine D2 receptor.

Mihyang Kim, Nayoung Kim, Jaehong Han. Deglycosylation of flavonoid O-glucosides by human intestinal bacteria Enterococcus sp. MRG-2 and Lactococcus sp. MRG-IF-4. Appl. Biol. Chem. 59(3):443-449Daidzin, daidzein 7-O-glucoside, is a major isoflavone in soybean and acts as a phytoestrogen. By intestinal bacteria dietary, daidzin is hydrolyzed to the aglycone daidzein and further converted to the more reduced metabolites, such as dihydrodaidzein, tetrahydrodaidzein, and equol. Human intestinal bacteria Enterococcus sp. MRG-2 and Lactococcus sp. MRG-IF-4, which convert daidzin to daidzein, were isolated under anaerobic condition, and identified by 16S rRNA gene sequence analysis. Changes of OD600 and pH were measured during the anaerobic growth, and deglycosylation kinetics of daidzin and genistin were measured by HPLC. Both bacteria also converted other isoflavone 7-O-glucosides, glycitin, ononin, and sissotrin, to their aglycones, glycitein, formononetin, and biochanin A, respectively. Apigetrin was deglycosylated to apigenin by these bacteria too. However, rutin, hesperidin, and naringin were not converted to the aglycones. Phylogeny analysis of the isolated strains also found that bacterial species identified by 16S rRNA gene sequence was not correlated with its metabolic ability of flavonoid biotransformation.

Bae Jin Kim, Hee Kyoung Jung, Yoo Seok Jeong, Su Jin Yang, Joo-Heon Hong. Effect of microencapsulated Bacillus subtilis strain CBD2-fermented grain on loperamide-induced constipation in mice. Appl. Biol. Chem. 59(3):451-462Constipation is a common cause of discomfort that affects the quality of life. The aim of the present study was to evaluate the ameliorative effects of microencapsulated food products using Bacillus subtilis CBD2 strain-fermented grain in mice with loperamide-

induced constipation. Microcapsules prepared by spray-drying with 2 % pectin and 2 % alginic acid (SD-P2A2) showed higher adhesion and dissolution of amylase than those prepared by freeze-drying. Experimental mice were fed a diet containing SD-P2A2 for 7 days prior to loperamide treatment. The ameliorative effects of an experimental diet containing SD-P2A2 on constipation were compared with those observed in mice fed a control diet containing non-fermented grain. Following the induction of constipation in mice, a marked alteration was observed in fecal parameters (frequency of defecation, fecal weights, fecal water content, and fecal pellet numbers in the distal colon), gastrointestinal transit ratios, harmful enzyme activities, and gene expression differences related to intestinal muscle contraction. These symptoms were significantly ameliorated in mice fed the experimental diet with SD-P2A2 versus constipated control mice. Thus, the intake of microencapsulated food products using B. subtilis CBD2 strain-fermented grain exhibited probiotic effects; these products may be used as a functional material for reducing constipation.

Mohammad Saber Araghi, Mohammad Ebrahim Olya, Reza Marandi, Seyed Davar Siadat. Investigation of enhanced biological dye removal of colored wastewater in a lab-scale biological activated carbon process. Appl. Biol. Chem. 59(3):463-470In this study, the influence of bacteria and activated carbon adjacent was investigated on treatment and decolorization of synthetic wastewater in a Biological Activated Carbon process. Pseudomonas putida was selected as a bacterial strain capable of degrading aromatic compounds and Acid Blue 92 was used as model azo dye. The optimum conditions for environmental factors affecting decolorization of azo dye was achieved from series of batch experiments. At 25 °C and pH7 the highest efficiency was obtained. At concentration of 20 mg L−1 in batch process, more than 90 % of Acid Blue 92 was reduced in color within just 3 h under optimum conditions. Dual-beam UV–Visible spectrometry and plate assay confirmed biodegradation of the dye by P.putida. The optimum values were then used for continues process. It was found that flow rate and thereby retention time has a critical role in biofilter efficiency. A lab-scale submerged downflow GAC column was sustained by bacterial strain to form the biofilter. About 70 % decolorization was obtained in continuous biological activated carbon process within 2 h with 60 mL h−1 flow rate while adsorption of dye reached the equilibrium and no further dye removal observed. Our results suggest the potential use of P. putida in azo dye decolorization. The combination of activated carbon adsorption and biodegradation by bacterial strains capable of degrading xenobiotic proved to be a promising technique for treatment of dye-contaminated wastewater.

Ju Won Park, So Hee Kim, Seung Jun Choi. Effect of cosolvent and surfactant on the solubility and stability of citral in a beverage model. Appl. Biol. Chem. 59(3):471-474To evaluate how triacetin and surfactant affect the solubilization and stability of citral, solutions containing citral were prepared with polyoxyethylene stearyl ethers and triacetin. Triacetin had a significant impact on the increment of saturation concentration of citral, and the citral solubility increased as surfactants were added. However, an interaction effect between triacetin and surfactant on citral solubility was not observed. At pH 3, the degradation rate constant of citral gradually decreased with a proportion of the triacetin concentration. Additionally, the chemical degradation of citral was considerably retarded in micelles built with surfactant. The surfactant with the smaller hydrophilic head exhibited higher citral stabilization ability than the one with the larger hydrophilic head; however, a negative interaction effect between triacetin and surfactant on citral stability in an acidic environment was observed.

Ngoc-Hoa Truong, Suyong Lee, Soon-Mi Shim. Screening bioactive components affecting the capacity of bile acid binding and pancreatic lipase inhibitory activityl. Appl. Biol. Chem. 59(3):475-479The hypothesis of the current study was that bioactive components including vitamins, dietary fibers, and polyphenols may have both the ability to bind to bile acid and inhibit

pancreatic lipase activity. ß-carotene, vitamin C, and vitamin U as vitamin, cellulose, ß-glucan, and pectin as dietary fibers, and apigenin, curcumin, hesperidin, kaempferol, and quercetin as polyphenols were chosen. The bile acid binding capacity was measured via the furfural–H2SO4 test at the concentration of 2.5 mg/mL with cholestyramine resin used as a positive control (binding value of 87.17 %). The pancreatic lipase inhibitory ability of bioactive components was measured by following the hydrolysis of p-nitrophenyl butyrate. Among the compounds tested, pectin, vitamin C, and ß-glucan had the highest binding capacity to bile acid, at 82.7, 67.8, and 33 %, respectively. The results suggest a positive correlation between water solubility and the bile acid binding capacity of the tested bioactive components. At 20 mg/mL concentration, the inhibiting values of all components were relatively lower than the positive control (orlistat, 54.4 %), ranging from 1.3 to 7.6 % of the negative control. Among them, curcumin and cellulose are the highest pancreatic lipase inhibitors with inhibiting values of 7.6 and 7.5 %, respectively. These results imply ß-glucan, pectin, vitamin C, cellulose, and curcumin as potential components in reducing the uptake of hydrophobic toxic components through inhibiting micelle formation under human digestion fluid.

Da Ye Han, Hye Rin Lee, Bong Gyu Kim, Joong-Hoon Ahn. Biosynthesis of ferulic acid 4-O-glucoside and feruloyl glucoside using Escherichia coli harboring regioselective glucosyltransferases. Appl. Biol. Chem. 59(3):481-484Twelve candidate uridine diphosphate-dependent glucosyltransferases (UGTs) from Arabidopsis thaliana were screened to identify UGTs that synthesized ferulic acid 4-O-glucoside or feruloyl glucoside, respectively. We used biotransformation using E. coli harboring each UGT. E. coli harboring AtUGT71C1 could synthesize feruloyl glucoside with a conversion rate of 1.8 μM/h. E. coli harboring AtUGT72E2 showed the best conversion rate for converting ferulic acid into ferulic acid glucoside, at 15.8 μM/h. Molecular docking analysis of ferulic acid into the modeled structures of AtUGT71C1 and AtUGT72E2 was used to elucidate the different conversion rates of ferulic acid into corresponding glucoside.

Cho Hi Park, Mi Ja Ahn, Gyu Seon Hwang, Sang Eun An, Wan Kyunn Whang. Cosmeceutical bioactivities of isolated compounds from Ligularia fischeri Turcz leaves. Appl. Biol. Chem. 59(3):485-494Isolated compounds from the leaves of Ligularia fischeri Turcz were evaluated for their cosmeceutical bioactivities. Four novel compounds, quercetin-3-O-β-d-glucoside (LF-4), quercetin-3-O-rutinoside (LF-5), ethyl-β-d-galactopyranoside (LF-7), and pterodontriol-6-O-β-d-glucopyranoside (LF-8), were isolated for the first time from this plant. Additionally, the four known compounds, 3-O-caffeoylquinic acid (LF-1), 5-O-caffeoylquinic acid (LF-2), 3, 5-di-O-caffeoylquinic acid (LF-3), and quercetin-3-O-galactoside (LF-6), were also isolated. To demonstrate the value of this plant as a cosmeceutical resource, the antioxidant activity and the inhibitory activity on tyrosinase and elastase were evaluated. The present study is the first time that the elastase inhibition of isolated compounds from L. fischeri Turcz has been investigated using the elastase inhibitory assay for anti-wrinkle effect. The ethyl acetate fraction had the highest activities in all the evaluated assays compared with the other fractions. At the compound level, LF-3 had the greatest antioxidant and elastase inhibition effect, and LF-2 showed a high level of tyrosinase inhibition. In addition, quantitative analysis of the fractions showed that the ethyl acetate fraction had the highest amount of isolated cosmeceutical bioactive compounds: LF-2, 0.9 %; LF-3, 5.3 %; LF-5, 2.8 %; and LF-6, 12.8 %.

Jung-Hyun Yun, Young-Jun Kim, Kyung-Hee Koh. Investigation into factors influencing antioxidant capacity of vinegars. Appl. Biol. Chem. 59(4):495-509Thirty commercial vinegars were investigated in order to study the factors influencing antioxidant capacity. Total phenols and flavonoids, 1,1-diphenyl-2-picrylhydrazyl, and 2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical scavenging capacity, nitrite scavenging capacity, reducing capacity, and lipid oxidation inhibition method were used. Factor and k-means cluster analysis partitioned the vinegars into three clusters according to antioxidant capacity. Traditional balsamic vinegar (TBV), balsamic vinegar, and tomato

vinegar were grouped as cluster 2, which contained the highest antioxidant capacity. Especially, TBV exhibited the highest antioxidant capacity. In high-performance liquid chromatography analysis, cluster 2 also had a high content of polyphenols and 5-hydroxymethylfurfural. These compounds presented positive correlation with the antioxidant capacity. The results suggested that polyphenols and 5-hydroxymethylfurfural were important factors influencing the antioxidant capacity of vinegars.

Eunah Kang, Da Hyun Lee, You Jung Jung, Soon Young Shin, Dongsoo Koh, Young Han Lee. α-Pinene inhibits tumor invasion through downregulation of nuclear factor (NF)-κB-regulated matrix metalloproteinase-9 gene expression in MDA-MB-231 human breast cancer cells. Appl. Biol. Chem. 59(4): 511–5162,6,6-Trimethylbicyclo[3.1.1]hept-2-ene (α-Pinene) is an organic compound of the terpene class found in the essential oil of many plants. In this study, the inhibitory effect of α-pinene on tumor invasion in highly metastatic MDA-MB-231 human breast cancer cells was evaluated. α-Pinene inhibited tumor necrosis factor (TNF)-α-induced invasiveness of MDA-MB-231 cells as revealed by three-dimensional spheroid invasion assay. Further analysis showed that α-pinene reduced TNFα-induced matrix metalloproteinase-9 gene promoter activation and mRNA expression in a dose-dependent manner. In addition, α-pinene treatment attenuated TNFα-induced nuclear factor κB (NF-κB) activation and NF-κB-dependent transcriptional activity. These results suggest that α-pinene has a significant effect on the inhibition of tumor invasion and may potentially be developed into an anti-metastatic drug.

Ju Hwan Hyun, Hyun-Wook Choi, Dong-Ho Seo, Jong-Dae Park, Jun-Suk Kum, Hyungjae Lee, Byung-Yong Kim, Moo-Yeol Baik. Optimization of thermal processing conditions for brown rice noodles. Appl. Biol. Chem. 59(4): 517–524Based on the current thermal processing conditions for rice noodles (80–85 °C for 20–30 min), we used response surface methodology to find brown rice (BR) noodle processing conditions that maximize the noodles antioxidant activity, digestibility, and gelatinization. The experiments were designed according to the central composite design, including two independent variables (temperature and time) and six dependent variables [total phenolic contents (TPC), total flavonoid contents (TFC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activities, digestibility, and gelatinization]. Antioxidant activities decreased but digestibility and gelatinization increased with increasing temperature and time. All dependent variables suggested a quadratic model except TFC, but the probability of TPC was not applicable (0.1505), and DPPH radical scavenging activity showed a relatively low R2 (0.6750). Therefore, we applied the other three dependent variables (ABTS radical scavenging activity, digestibility, and gelatinization) to the 3D response surface and proposed three optimum conditions: (1) for antioxidant activity (70 °C, 22.95 min), (2) for digestibility and gelatinization (88.18 °C, 34.89 min), and (3) for all three variables (88.5 °C, 40 min). In a validation test, all dependent variables showed values within a 5 % error range except TFC. These results show the optimum processing conditions for BR noodles to maximize antioxidant activity and provide sustainability in BR noodle processing.

Ji Yeon Kim, Doram Kim, Oran Kwon. Effective screening for the anti-hypertensive of selected herbs used in the traditional Korean medicines. Appl. Biol. Chem. 59(4): 525–532In this study, Korean medicinal herb extracts were investigated for their in vitro effect on anti-hypertensive activities, including nitric oxide (NO) production in the human umbilical vein endothelial cell line EA.hy529 and angiotensin-converting enzyme (ACE) activity. Traditional remedies related to the treatment of hypertension were identified using DongUiBoGam, the most comprehensive source for Korean traditional pharmacopoeia. Herbal remedies were selected by translating the symptom terminology into westernized medicinal terms using the Korean classification of disease in Oriental medicine and Korea Traditional Knowledge Portal. Seven types of herb extracts were selected and screened for in vitro anti-hypertensive activities. Among the extracts, those

of Evodiae fructus, Bambusae caulis in Taeniam, Sophorae fructus, Melonis calyx, and Schizonepeta rhizome produced significant increases in NO production, although their ability to inhibit ACE were weaker. Using Fourier transform ion cyclotron resonance mass spectrometry, the major flavonoids exhibiting anti-hypertensive activities were identified: quercetin, apigenin, and luteolin. Our results indicate that these medicinal herbs are potent natural anti-hypertensive agents that can be developed for clinical therapies. In addition, our systematic approach to identifying candidate herbal remedies in the traditional Korean pharmacopoeia can be used to search for additional traditional remedies, such as for hypoglycemia and obesity.

Lobna Mejri, Mnasser Hassouna. Characterization and selection of Lactobacillus plantarum species isolated from dry fermented sausage reformulated with camel meat and hump fat. Appl. Biol. Chem. 59(4): 533–542A total of 29 strains of Lactobacillus plantarum, isolated from traditional dry fermented camel sausages and identified by biochemical and molecular methods, were characterized by their technological and safety properties including acidifying capacity, proteolytic activity, and antimicrobial and antibiotic resistance in order to select the most suitable for use as starter cultures in combination with coagulase-negative staphylococci to improve and standardize the safety and quality of sausages. All the Lb. plantarum were able to hydrolyze casein and some strains showed very weak lipolytic activity. Moreover, all isolates showed acidifying activity by reducing pH to less than 4.3 at 15 and 25 °C after 72 h. Related to safety aspects, the totality of Lb. plantarum was resistant to tetracycline, gentamicin, and kanamycin. A large majority of strains exhibited antimicrobial activities against Listeria monocytogenes, Staphylococcus aureus, and Aeromonas hydrophila.

Miyoung Nam, Cheol-Hee Kim, Dong-Uk Kim, Sook-Jeong Lee, Kwang-Lae Hoe. h-Prune as a novel binding protein of NS5A that regulates ERK1/2 activation. Appl. Biol. Chem. 59(4): 543–551Hepatitis C virus (HCV) non-structural 5A (NS5A) protein is associated with a wide variety of host signaling pathways by binding to C-terminal polyproline (PxxP) motifs of various proteins. In this study, we used yeast two-hybrid analysis and a GST pull-down assay to screen a novel NS5A interacting protein and elucidate the binding site and cellular signaling by focusing on recombinant human epidermal growth factor (rhEGF)-mediated ERK1/2 activation. Screening a liver cDNA library revealed that h-prune, a member of the DHH (Asp-His-His) protein superfamily, directly interacted with HCV NS5A C-terminus. In particular, a mutation of five proline amino acids to alanine in this region revealed that these two proteins produced strong interaction through this domain. It is known that h-prune possesses a highly conserved DHH motif, which has exopolyphosphatase activity that accelerates hydrolysis of inorganic polyphosphate. A time-chasing analysis after rhEGF treatment demonstrated that h-prune overexpression almost restored NS5A-mediated attenuation of ERK1/2 phosphorylation, but h-prune itself did not alter this signaling. Although the detailed mechanisms need to be clarified, this study demonstrates that h-prune interacts directly with the PxxP motif of the HCV NS5A C-terminus and that this binding alters the rhEGF-mediated ERK1/2 signaling cascade in liver cells.

Yoon Kyung Kwon, Soo Jung Choi, Cho Rong Kim, Jae Kyeom Kim, Youn-Jung Kim, Jong Hun Choi, Sang-Wook Song, Chang-Ju Kim, Gwi Gun Park, Cheung-Seog Park, Dong-Hoon Shin. Antioxidant and cognitive-enhancing activities of Arctium lappa L. roots in Aβ1-42-induced mouse model. Appl. Biol. Chem. 59(4): 553–565Many studies have shown that oxidative stress induced by hydrogen peroxide can lead to neuronal cell death. In this study, protective effects of the ethanol extract of A. lappa L. roots against oxidative stress in PC12 cells were measured by 2′, 7′-dichlorofluorescein diacetate and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Oxidative stress contributes to the memory deficits induced by Beta-amyloid1-42 in mice. Treatment with the ethanol extract of A. lappa L. roots improved working and reference

memory in mice in the Y-maze and passive avoidance tests. To detect the lipid peroxidation levels, malondialdehyde was measured. To purify the bioactive compound, active fractions and components of A. lappa L. roots were isolated by partitioning, open column chromatography, TLC, high-performance liquid chromatography, electron ionization mass spectrometry, and 1H/13C-nuclear magnetic resonance. We identified the bioactive compound as quinic acid, which is a powerful antioxidant agent. Ultimately, the roots of A. lappa L. may become available as an antioxidant food for Alzheimer’s disease patients and those with other oxidative stress-induced disorders.

Eun-Ji Oh, Jung-Hwa Kwon, Su-Yeon Kim, Seo-Ji In, Dong-Geol Lee, Mi-Yeon Cha, Hee-Cheol Kang, Jeon Hwang-Bo, Youn-Hyung Lee, In-Sik Chung, Nam-In Baek. Red pigment produced by Zooshikella ganghwensis inhibited the growth of human cancer cell lines and MMP-1 gene expression. Appl. Biol. Chem. 59(4): 567–571A red pigment produced by Zooshikella ganghwensis, was purified by solvent extraction and fractionation. The pigment was identified to be prodigiosin on the basis of spectroscopic data such as 1H-NMR, 13C-NMR, DEPT, 2D-NMR, IR, and FAB-MS. Prodigiosin inhibited matrix metalloproteinase-1 gene expression and the growth of human cancer cell lines, SK-BR-3, HCT-116, SK-OV-3, HeLa, HepG2, and SK-MEL-5.

Hyun Jung Kim, Minseon Koo, Daekeun Hwang, Jeong Hee Choi, Seung Min Kim, Se-Wook Oh. Contamination patterns and molecular typing of Bacillus cereus in fresh-cut vegetable salad processing. Appl. Biol. Chem. 59(4): 573–577Microbiological contamination of fresh vegetables is a recent food safety concern. Bacillus cereus has been recognized as the most frequently detected foodborne pathogen in fresh-cut salads in Korea. Prevalence and level of contamination of B. cereus was determined in samples from processing lines of packaged fresh-cut vegetable salad manufacturing companies (A, B, C, and D). B. cereus was detected in the 27.3–30.8 % of food samples (raw material, salads in washing steps, and final products) and the contamination levels were up to 9.5 × 102 CFU/g, while detection rates in environmental samples were very low. Molecular subtyping of B. cereusstrains detected in the processing environments (company A) and retail products (companies A and E) was conducted to reveal the contamination sources during processing. High genetic similarity was found in the bacterial strains obtained from the processing lines and retail products of company A. This result suggested that B. cereus strains of same clone may have circulated in the products prepared in the same company. Genetic similarities were also observed among part of the B. cereus isolates obtained from the processing line of company A and from the retail products of company E, suggesting that some of the B. cereus clones may have originated from the raw materials. The identified information can be used to develop the intervention technology for fresh-cut vegetable processing.

Jungchan Nam, Woon-Seok Yeo. Controlled drug release using ascorbate-responsive quercetin-conjugated alginate hydrogels. Appl. Biol. Chem. 59(4): 579–584In this paper, we report a new strategy for controlled cargo release using quercetin-conjugated alginate (Alg) hydrogels that has enhanced hydrogel stability and can release cargo molecules through on-demand metal cation reduction by a biological reducing agent, ascorbate. By conjugating hydrophobic quercetin to the Alg backbone, hydrogel stability was increased. The encapsulated cargos were released on demand through reduction of cross-linking Fe(III) ions by ascorbate, harnessing the differences in cross-linking capability between Fe(II) and Fe(III). We found that encapsulated fluorescein in quercetin-conjugated Alg hydrogels was released much slower than that in hydrogels without quercetin conjugation. In addition, the stability of the hydrogels and the release of encapsulated fluorescein were controlled by modulating oxidation state of Fe using ascorbate. We validated our strategy by demonstrating the enhanced cancer cell killing of the doxorubicin-encapsulated hydrogels upon treatment with ascorbate. We believe that the strategy would be used as an effective tool for the anticancer drug delivery that can be controlled by ascorbate as overcoming the drawback of Alg hydrogels.

Dingyi Yang, Xiaochuan Zou, Ruokun Yi, Weiwei Liu, Deguang Peng, Xin Zhao. Neferine increase in vitro anticancer effect of dehydroepiandrosterone on MCF-7 human breast cancer cells. Appl. Biol. Chem. 59(4): 585–596This study was conducted to investigate the in vitro anticancer reinforcing effects of neferine (Nef) on dehydroepiandrosterone (DHEA) and the mechanism was also determined during the investigation. By the growth effects of Nef and DHEA on MCF-7 human breast cancer cells, 8 mg/mL Nef was a non-virulent concentration in MCF-7 cells, and this concentration was used for further experiment. In 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide assay, 30 mg/mL DHEA showed 49.4 % growth inhibitory effect in MCF-7 cells, whereas Nef (8 mg/mL) + DHEA (30 mg/mL) treatment had the higher effect at 67.8 %. The flow cytometry analysis results showed that 15 and 30 mg/mL DHEA-treated MCF-7 cells had 12.2 and 21.6 % apoptotic cells, respectively, Nef + DHEA could raise the apoptotic cells to 36.7 %. Reverse transcription-polymerase chain reaction assay shows remarkable results according to which DHEA could significantly increase caspase-3, caspase-8, caspase-9, Bax, p53, p21, E2F1, Fas, FasL mRNA expressions and decrease Bcl-2, Bcl-xL, HIAP-1, HIAP-2, survivin expressions as compared to the untreated control cancer cells. Moreover, these effects depend on the concentration of DHEA, and Nef which could further strengthen these effects. From these results, low concentration of Nef could not influence the growth of MCF-7 cells, but using its sensitization effect, Nef raised the in vitro effects of DHEA. Nef could be got easily. With these results we can accomplish that Nef + DHEA might be used as the new anticancer materials combination.

Yeon Bok Kim, Soo-Yun Park, Chang Ha Park, Woo Tae Park, Sun-Ju Kim, Sun-Hwa Ha, Mariadhas Valan Arasu, Naif Abdullah Al-Dhabi, Jae Kwang Kim, Sang Un Park. Metabolomics of differently colored Gladiolus cultivars. Appl. Biol. Chem. 59(4): 597–607Gladiolus (Gladiolus grandiflora Hort.) plants are a perennial favorite, known for their beautiful flowers of many different colors. In this study, we determined metabolic differences between seven Gladiolus cultivars with differently colored flowers by monitoring anthocyanin, carotenoid, and hydrophilic metabolites. We detected nine anthocyanins in the seven cultivars. Only the ‘Violetta’ cultivar contained all nine anthocyanins and it exhibited the highest anthocyanin content, whereas no anthocyanins were detected in the ‘New Wave’ (white color) or ‘Limoncello’ (yellow color) cultivars. In addition, we detected seven carotenoids, the contents of which varied significantly among the cultivars depending on the flower color. ‘Limoncello’ exhibited the highest levels of carotenoids. Of the seven carotenoids, β-carotene and lutein accumulated in the most cultivars. In addition, we identified 43 metabolites using gas chromatography–time-of-flight mass spectrometry. The levels of organic acids and sugars in the ‘New Wave’ cultivar differed significantly from those in the ‘Violetta’ and ‘Limoncello’ cultivars with a P value < 0.01. Thus, our results may help in understanding the metabolic differences between differently colored Gladiolus cultivars and should be useful in future databases.

Hwa-Won Lee, Sang-Guei Lee, Hoi-Seon Lee. Active component isolated from Eugenia caryophyllata leaves and its structural analogues show insecticidal properties against Pochazia shantungensis. Appl. Biol. Chem. 59(4): 609–614The purpose of this study was to isolate an active constituent from the essential oil of Eugenia caryophyllata leaves and to evaluate its insecticidal activity against the nymph and adults of Pochazia shantungensis. According to some chromatographic methods and spectroscopic analyses, the active constituent of E. caryophyllata leaves was identified as eugenol. Based on the LC50 values of eugenol and its structural analogues against the nymph and adults of P. shantungensis, isoeugenol (LC50, 83.29 and 91.03 mg/L) exhibited the highest insecticidal activity, followed by methyl isoeugenol (105.61 and 114.48 mg/L), eugenol (124.44 and 143.24 mg/L), methyl eugenol (126.31 and 143.84 mg/L), and acetyl eugenol (165.11 and 170.06 mg/L). Insecticidal activity against P. shantungensis was dependent on the presence of a functional group in 4-ally-

2-methoxyphenol. In conclusion, E. caryophyllata oil and eugenol analogues might be suitable alternative synthetic insecticides.

Hyun Jun Kim, Moon Sea Song, Hyun Soo Kim, Se Ik Park, Sang Sub Han, Sang Hyun Lee. Carbon dioxide absorption for Liriodendron tulipifera using fertilization. Appl. Biol. Chem. 59(4): 615–621We conducted this study to obtain the basic information for forest biomass estimation in a national unit by predicting the carbon dioxide absorption of the early growth of Liriodendron tulipifera based on several fertilization methods. The study site, located in Gimje-si, Jeollabuk-do, consisted of 11 fertilizer treatment plots, repeated three times by species. Five sample trees were planted in each plot, and a total of 165 sample trees of each species were planted. Of these sample trees, every two samples of the L. tulipifera were cut down in order to estimate the biomass. The results were similar to the analysis results for the mean volume. In comparison to the control group with a biomass of 310.35 g, the biomass of the LLF and HLF groups was relatively high, whereas the biomass of the CF and CP groups was relatively low. As a result, the LLF-150 group had the highest biomass at 96.36 g, and all of the treatment groups had higher biomasses than the control group (16.14 g) in the case of branches. In the case of leaves, the HLF-150 group had the highest biomass at 239.48 g and the CP group had the lowest biomass at 115.60 g. In the case of roots, the LLF-150 group had the highest biomass at 495.45 g, and only the CP group (186.82 g) had a lower biomass than the control group (194.90 g).

Eun Ji Kwon, Min Kyoung Lee, Hyesung Jeon, Jae Min Hwang, Ju-Kon Kim, Minkyun Kim. Analysis of OsmiR399 expression and down-regulation of LTN1 by the multiple members of OsmiR399 family in rice. Appl. Biol. Chem. 59(4): 623–630Among the many miRNAs involved in plant stress responses, miR399 is well known to be involved in controlling phosphate homeostasis by down-regulating the expression of PHO2,which encodes the ubiquitin-conjugating E2 enzyme, UBC24, in Arabidopsis. In this study, to understand the expression of the rice OsmiR399 genes under abiotic stress conditions, the expression of the eleven OsmiR399 (a-k) genes was studied by analyzing the levels of their precursor transcripts (pre-miRNAs) in the roots and shoots of rice seedlings subjected to the stress-responsive phytohormone abscisic acid (ABA). We found that the OsmiR399 genes showed different patterns in pre-miRNA accumulation. In particular, OsmiR399b, OsmiR399d, OsmiR399e, and OsmiR399f showed high and steady accumulation in both the roots and shoots regardless of ABA treatments. However, OsmiR399c and OsmiR399k showed ABA-induced expression in the whole plant body or aerial part of the rice seedlings. In addition, to test the possibility that the putative rice PHO2 ortholog of Arabidopsis, also known as LEAF TIP NECROSIS 1 (LTN1), might be down-regulated by the multiple OsmiR399s with certain sequence divergences, four different lines of transgenic rice plants that overexpress either the constitutively expressed OsmiR399s (OsmiR399d and OsmiR399f) or the ABA-inducible OsmiR399s (OsmiR399c and OsmiR399k) were produced and the levels of OsmiR399 pre-miRNAs and LTN1 transcripts were analyzed. A significant decrease in the accumulation of LTN1 transcripts and an increase in the OsmiR399 pre-miRNAs levels were found in all of the transgenic plants. Based on these results, we concluded that LTN1 is down-regulated by multiple OsmiR399 genes in rice.

Kyung Hee Roh, Byung Wook Kang, Ho Bang Kim, Sang Ho Lee, Yoonkang Hur, Hankuil Yi. Physiological importance and possible applications of β-substituted alanine synthase in plants. Appl. Biol. Chem. 59(4): 631–647Cysteine, as well as its precursors and derivatives, plays important roles in plant development and stress responses. In plants, a diverse range of reactions affecting cysteine content are catalyzed by the β-substituted alanine synthase (BSAS) enzyme family. Individual BSAS family members use similar reaction mechanisms involving pyridoxal phosphate cofactors and show catalytic preferences for biosynthesis, degradation, or modification of the cysteine amino acid. In Arabidopsis thaliana (Arabidopsis) of the Brassicaceae family, four distinct biochemical activities are

characterized at the gene level, namely, O-acetylserine sulfhydrylase, β-cyanoalanine synthase, l-cysteine desulfhydrase, and S-sulfocysteine synthase activities. Reverse genetic approaches in Arabidopsis were used to elucidate the physiological roles of metabolites of cysteine metabolism (O-acetylserine, sulfide, cysteine, cyanide, and S-sulfocysteine) during the processes of root hair development, pollen tube germination, heavy metal tolerance, defense responses, stomatal closure, and autophagy. Key catalytic residues determining reaction specificities in different BSAS enzymes are being identified, along with the roles of macromolecular complexes involving BSAS. The biochemical properties of BSAS active sites are being investigated in various organisms, including plants, for possible application to the development of new biological materials and drugs. Systematic and comparative genomic studies of BSAS enzymes in Brassica plants, close relatives of Arabidopsis, requiring high cysteine production for optimum growth and disease resistance, will be useful for the future study of the diversification of BSAS and the biotechnological improvement of these important crop plants.

H. Lee, J.-H. KimE. Kim, Y. Shin, J.-H. Leem H. Jung, Y. Lim, H. S. Lee, J.-H. Kim. Biotransformation and molecular docking of cyazofamid by human liver microsomes and cDNA-expressed human recombinant P450s. Appl. Biol. Chem. 59(4): 649–653The purpose of this study was to understand the formation of metabolites from the metabolic reaction of cyazofamid with human liver microsomes. Human liver microsomal incubation of cyazofamid in the presence of NADPH produced one metabolite, 4-chloro-2-cyano-5-(4-(hydroxymethyl)phenyl)N,N-dimethyl-1H-imidazole-1-sulfonamide (CCHS). An incubation study using cDNA-expressed human recombinant P450s (rCYPs) demonstrated that cyazofamid-derived CCHS is mediated by CYP2B6, 2C9, and 2C19 at different reaction rates. The crystal structure of cyazofamid was obtained using single-crystal X-ray diffraction. According to a molecular modeling study of the crystal structure of cyazofamid with the rCYPs 2B6, 2C9, 2C19, and 3A4, the metabolic reactivities (2B6 > 2C19 > 2C9) were well-correlated to the distances between heme irons of CYPs and 4-methylphenyl group of cyazofamid.

Leesun Kim, Jin-Woo Jeon, Yun-Se Lee, Hwang-Ju Jeon, Byung-Jun Park, Hoi-Seon Lee, Sung-Deuk Choi, Sung-Eun Lee. Monitoring and risk assessment of polychlorinated biphenyls (PCBs) in agricultural soil collected in the vicinity of an industrialized area. Appl. Biol. Chem. 59(4): 655–659Samples of agricultural soils possibly contaminated by polychlorinated biphenyls (PCBs) were collected from Anseong, Korea. The agricultural areas chosen have possibly been contaminated by nearby factories, either directly or via atmospheric deposition, and require monitoring. The concentrations of 29 out of 209 PCB congeners were determined at five sites using high-resolution gas chromatography/high-resolution mass spectrometry. Total concentrations of the 29 PCB congeners in soil samples obtained from each study site were 106.65, 149.15, 222.67, 166.15, and 118.28 pg g−1 dry weight, respectively, with pentaCBs and hexaCBs giving the highest concentrations of the congeners studied. The total toxic equivalent (TEQ) for 12 PCBs from each site was 0.05, 0.11, 0.08, 0.05, and 0.04 ng kg−1 dry weight, respectively. These values were mainly due to high TEQ values for PCB 126 (3,3′,4,4′,5-pentachlorobiphenyl) from each site (at 0.05, 0.10, 0.07, 0.04, and 0.03 ng kg−1 dry weight, respectively). The TEQ values are much lower than the action level of 20 ng WHO–TEQ kg−1 recommended by the World Health Organization, but PCB concentrations in this area should be monitored nonetheless, since these manmade compounds bioaccumulate, thus threatening the ecosystem and human health.

Sunghoon Kim, Youngdae Yoon. Assessing bioavailability and genotoxicity of heavy metals and metallic nanoparticles simultaneously using dual-sensing Escherichia coli whole-cell bioreporters. Appl. Biol. Chem. 59(4): 661–668Genetically engineered bacterial strains called whole-cell bioreporters (WCBs) generated by fusing the promoter region of stress-responsive genes and reporter genes have been

widely used as biosensors to detect toxic materials in the environment. In this study, we report a dual-sensing WCB harboring recAp::egfp and zntAp::mcherry to measure the genotoxicity and bioavailability of heavy metals and metallic nanoparticles (NPs) simultaneously. Since the dual-sensing WCB harbored recAp::egfp and zntAp::mcherry, the genotoxicity and bioavailability of heavy metals and metallic NPs that activate ZntR would be assessed by measuring the fluorescence signal of enhanced green fluorescent protein (eGFP) and mCherry, respectively. Among the tested heavy metal(loid)s, only Cr induced both eGFP and mCherry expression, and some of them only induced mCherry, thereby suggesting that Cr is genotoxic. In case of the tested metallic NPs, Ti2O NPs, ZnO NPs, and Au NPs showed weak inhibitory effects on growth, but the eGFP was not induced. It was inferred that the tested NPs were not genotoxic and the inhibitory effects would not be related to direct DNA damage pathways. In addition, it was observed that ZnO NPs induced mCherry expression, indicating that the Zn ion was dissolved from the NPs. Although the dual-sensing WCB described here was limited to ZnO NPs, WCBs would be an alternative tool to investigate the dissolution of metallic NPs when the corresponding metal ion sensing systems were available.

Kyeong-Ryeol Lee, Eun-Ha Kim, Kyung Hee Roh, Jong-Bum Kim, Han-Chul Kang, Young Sam Go, Mi Chung Suh, Hyun Uk Kim. High-oleic oilseed rapes developed with seed-specific suppression of FAD2 gene expression. Appl. Biol. Chem. 59(4): 669–676Vegetable oil is not only important for its edibility but for industrial purposes. High-oleic vegetable oil is especially useful for making biodiesel because it is highly stable against oxidation. Transgenic oilseed crops with modified fatty acid compositions have been developed with several biotechnological gene-silencing methods. The seed oils with the most successfully altered fatty acid compositions are produced from high-oleic oilseed crops in which FAD2 gene expression is suppressed. Vegetable oil from oilseed rape (Brassica napus) is one of the most commonly used vegetable oils throughout the world and its oleic acid content is moderately high (about 65 %). Therefore, oilseed rape can be modified to produce high-oleic oilseed crops. Oilseed rape has four FAD2 genes, originating from B. rapa and B. oleracea, with nucleotide identities of 88–97 %. To produce transgenic high-oleic oilseed rape, plant transformation vectors were constructed using antisense RNA and RNA interference (RNAi) to modify the BrFAD2-1 gene, and canola-type cultivar Youngsan was transformed with Agrobacteriumcarrying the vectors. The transgenic lines generated, AS9A, HP15, and HPAS29, showed high-oleic phenotypes, which were stably inherited. Their oleic acid contents increased from 67 (Youngsan) to 78, 85, and 86 %, respectively, and their polyunsaturated fatty acid (PUFA) contents decreased from 24 (Youngsan) to 13, 8, and 6 %, respectively. HPAS29, developed with a combined antisense RNA–RNAi method, produced seed oil with the highest oleic acid and lowest PUFA contents. These transgenic high-oleic oilseed rapes could be useful in the manufacture of high-temperature frying oils and high-quality biodiesel fuel.

Ji-Hwan Yook, Choong-Min Kang, Woo-Yeon Kim. Phosphate-buffered native gel electrophoresis of E. Coli H-NS. Appl. Biol. Chem. 59(5): 677–679Histone-like nucleoid structuring protein (H-NS) is a global regulator in enteric bacteria and other bacteria. It has been reported that H-NS has no modifications for its activity. Even though H-NS is called histone-like protein, it is acidic, contrary to the basic eukaryotic histones. Here, we report that there were mobility differences for various sources of H-NS seen in sodium phosphate-buffered native gel electrophoresis. We examined various native gel electrophoresis conditions using the wild-type E. coli (MG1655), three H-NS mutants (two were H-NS knockout mutants), and the E. coli BL21 overexpressing H-NS (BL21 (DE3)/pPD3).

Ha Duc Chu, Kim-Lien Nguyen, Yasuko Watanabe, Dung Tien Le, Lam-Son Phan Tran. Expression analyses of soybean genes encoding methionine-R-sulfoxide reductase under various conditions suggest a possible role in the adaptation to stress. Appl. Biol. Chem. 59(5): 681–687

Under stress, plant cellular proteins can be oxidized at multiple amino acid residues causing protein dysfunction that may lead to reduced viability of plants. One of the oxidized amino acids, methionine sulfoxide (MetO), was frequently found in stressed plants. In soybeans, there are five methionine-R-sulfoxide reductases (GmMSRBs) responsible for the reduction of Met-R-O, one of the two MetO isomers. To identify GmMSRBs that may be involved in repairing Met-R-O under different stress conditions, we determined transcript levels of GmMSRBs in various tissues subjected to dehydration/drought, high salinity, and abscisic acid (ABA) during different developmental stages. Under normal conditions, expression levels were the highest in leaves, followed by roots, and lowest in seeds and seed pods. Among the GmMSRBs, transcripts of GmMSRB1 in the leaves were the highest; at the same time, GmMSRB5 was shown to be expressed at the lowest levels. Expression of GmMSRBs were then determined under stress-inducing conditions. In seedling shoots, GmMSRB2 and GmMSRB5 were expressed in response to drought conditions. In vegetative V6 trifolia, only GmMSRB3 was induced under drought. In reproductive R2 trifolia, the expression of GmMSRB2 and GmMSRB5 were induced by drought. However, expressions of all five GmMSRBs in the roots were not affected by the any stress-inducing conditions. Under salt stress, GmMSRB1 was down-regulated in seedling shoots and GmMSRB5 was up-regulated in seedling roots. Treatment with ABA did not affect the transcript levels of any GmMSRBs in seedling shoots. However, this treatment up-regulates GmMSRB2 in seedling roots. Our data suggested that with the exception of GmMSRB4, all the remaining four GmMSRBs play a role in soybean responses to multiple environmental stresses and that genes encoding cytosolic and plastidic GmMSRBs respond differently under stress.

A Ra Cho, Dae Gyun An, Youngshim Lee, Joong-Hoon Ahn. Biotransformation of quercetin to quercetin 3-O-gentiobioside using engineered Escherichia coli. Appl. Biol. Chem. 59(5): 689–693Various flavonoid O-diglycosides are found in nature but the biological activities of only a few compounds have been explored due to difficulty in obtaining samples. In order to circumvent the need for extraction and purification of the natural compounds from plants, we used engineered Escherichia coli strain that harbors two uridine diphosphate-dependent glycosyltransferases (UGTs; BcGT1 and CaUGT) and two nucleotide sugar biosynthetic genes (pgm and galU). Using this strain, we synthesized quercetin 3-O-gentiobioside. After optimization of induction temperature, cell density, and reaction temperature, approximately 46.2 mg/L quercetin 3-O-gentiobioside was synthesized from quercetin with 49 % conversion efficiency.

Dong-Jin Kim, Hye-Jin Shin, Byung-Koo Ahn, Jin-Ho Lee. Competitive adsorption of thallium in different soils as influenced by selected counter heavy metals. Appl. Biol. Chem. 59(5): 695–701This study was conducted to investigate thallium (Tl) adsorption in different soils, including acidic Jeonju (JJ), neutral Iksan (IS), and alkaline Danyang (DY) soils treated with various single counter metal ions and to examine the competitive adsorption of Tl with other metals, including cadmium (Cd), copper (Cu), nickel (Ni), lead (Pb), and zinc (Zn), in multicomponent systems. Thallium buffering capacity of the soils was ordered as IS > DY > JJ soil in the single-component systems, whereas it was DY > IS > JJ soil in the multicomponent systems. In the competitive systems of Tl with different single counter metals, the Tl adsorption capacity of the soils ranged from 20.33 to 88.38 and Tl bonding energy values ranged from 0.369 to 0.731. Thallium adsorption capacity and bonding energy were negatively correlated. Selectivity sequences of metals in multicomponent systems were Pb > Tl > Cu > Ni > Zn > Cd in the JJ soil, Pb > Cu > Tl > Zn > Cd > Ni in the IS soil, and Pb > Cu > Tl ≥ Zn > Cd > Ni in the DY soil. The metal buffering capacity in the JJ, IS, and DY soils varied from 293.2 to 1125.2. The values were ordered as Tl ≫ Pb ≥ Zn ≥ Ni > Cu ≥ Cd in the JJ soil, Pb > Cu > Tl ≫ Ni > Zn ≥ Cd in the IS soil, and Pb > Cu ≫ Tl > Ni > Cd ≥ Zn in the DY soil.

Ha Danh Duc. Biodegradation of 3-chloroaniline by suspended cells and biofilm

of Acinetobacter baumannii GFJ1. Appl. Biol. Chem. 59(5): 703–709Acinetobacter baumannii strain GFJ1 was isolated from soil using 3-chloroaniline (3CA) as sole of carbon, nitrogen, and energy source under both aerobic and anaerobic conditions. The investigation of aerobic utilization profile showed that the utilization kinetics of 3CA followed the Edward model with a maximum specific degradation as 3.45 ± 0.33 µM.h−1.mg cell protein−1, and apparent half-saturation coefficient value was 0.062 ± 0.01 mM. The aerobic utilization toward 3CA was stimulated with the addition of sodium nitrate and citrate. Under anaerobic conditions, A. baumannii GFJ1 was able to utilize 3CA linked with nitrate reduction. The investigation of biofilm formation showed that biofilm formation was affected by cosubstrates and 3CA concentrations. Biofilm formation enhanced with the presence of cosubstrates, especially nitrogen sources. The biofilm formation and chemical degradation by biofilm increased in the following intervals of incubation with the supply of fresh medium. The results indicate that A. baumannii GFJ1 has a potential for the application to clean up 3CA.

Chang-Hee Lee, Sang-Hyun Seo, O-Jun Kwon, Man Park, Won-Chan Kim, Sang-Jae Kang. Functional characterization of a chemical defoliant that activates fruit cluster Leaf defoliation in ‘Fuji’ apple trees. Appl. Biol. Chem. 59(5): 711–720The removal of fruit cluster leaves was shown to be a valuable method for controlling fruit quality and producing high-grade apples. A chemical defoliant that functions as an activation switch was shown to turn on the genes necessary for fruit cluster leaf defoliation. Elucidating the mechanism involved in leaf defoliation is crucial to our understanding of the use of chemical defoliants in fruit trees. To gain insight into chemical defoliant-mediated leaf defoliation, we first confirmed the occurrence of ethylene production by applying a chemical defoliant on fruit cluster leaves. Then, we used RNA-seq analysis to obtain a series of transcriptome profiles for genes and proteins involved in leaf senescence induction. Within 2 days of applying the chemical defoliant to apple trees, 1-aminocyclopropane carboxylic acid (ACC) oxidase, ACC synthase, a chlorophyll binding protein, and polygalacturonase-related genes were up-regulated at least sixfold. An in vitro enzyme assay showed that lanolin oil activates ACC synthase activity, a key regulatory enzyme in the ethylene pathway. We also showed that chemical defoliant decreased the light saturation point and total chlorophyll content. Then, we used a polygalacturonase activity assay to confirm the effects of chemical defoliant on leaf senescence in vivo. Furthermore, treatment with chemical defoliant resulted in a significant increase in the chromaticity value of a*, whereas L* and b* decreased in the apple fruit. Taken together, we conclude that chemical defoliant could selectively affect fruit cluster leaves, which suggested that it can be used as a selective defoliant.

Ji Hyun Kim, Jaemin Lee, Sanghyun Lee, Eun Ju Cho. Quercetin and quercetin-3-β-d-glucoside improve cognitive and memory function in Alzheimer’s disease mouse. Appl. Biol. Chem. 59(5): 721–728We investigated the protective activities of quercetin (Q) and quercetin-3-β-d-glucoside (Q3G) under Aβ25–35-injected Alzheimer’s disease (AD) model mice. To induce an AD-like disease in an in vivo model, mice were injected with Aβ25–35 via the intracerebroventricular route. Q and Q3G were administered orally for 14 days at 30 mg/kg/day. Learning and memory functions were evaluated using behavioral experiments that comprise tests like T-maze, Morris water maze, and novel object recognition. The administration of Q and Q3G improved memory and cognitive function, compared with Aβ25–35-injected control mice in the T-maze and object recognition test. Q and Q3G administration decreased time in order to get to the platform during the Morris water maze test. The group administered Q experienced higher protective effects from long-term spatial memory and learning ability impairments than the group administered Q3G. Furthermore, lipid peroxidation and NO formation in the brain were significantly elevated in Aβ25–35-injected control mice, and this was inhibited by Q and Q3G. These results demonstrated that Q and Q3G improved Aβ25–35-induced memory deficits and cognitive impairment and promoted protection in the brain against oxidative stress.

M. Taufique Arefin, M. Mokhlesur Rahman, M. Wahid-U-Zzaman, Jang-Eok Kim. Appraisal of heavy metal status in water for irrigation usage of the Bangshi River, Bangladesh. Appl. Biol. Chem. 59(5): 729–737An attempt was made to assess the status of heavy metals in the Bangshi River water for irrigation usage. River water was sampled from 20 sampling points for chemical analysis of physicochemical parameters and metal ions. Our analyses revealed that water samples were slightly alkaline to alkaline. All samples had reasonable electrical conductivity, and these samples were from low to medium salinity classes as being fit for irrigation on soils having moderate level of permeability. Water samples were categorized freshwater as per total dissolved solids (TDS). All water samples were excellent indicating low alkalinity hazard, based on sodium adsorption ratio (SAR), but most of the samples were doubtful to unsuitable for soluble sodium percentage (SSP). All samples were free from residual sodium carbonate (RSC) suggesting suitability for irrigation purpose. For water hardness, samples were rated as moderately hard, hard, and very hard. The status of Fe, Zn, Pb, Cd, and Ni in water samples were within permissible levels but Cr, Cu, and Mn content in water samples were in excess of recommended limits and these ions would be considered toxic in long-term irrigation system. Significant relationships existed between EC versus TDS, SAR versus SSP, SSP versus RSC, and SAR versus RSC. There were significant correlations between metal ions: Fe–Cu, Fe–Pb, Cu–Pb, Fe–Mn, Cu–Mn, and Mn–Pb. This finding showed that Cr, Cu, and Mn ions were considered contaminants in river water used for irrigation because the present status of these metals may pose a harmful impact on human health.

Byoung Kwon Park, Su In Lee, Younghee Lee, Sooyoung Cho, Young Seek Lee, Hyung-Joo Kwon. Effect of a monoclonal antibody against human relaxin-2 on cancer cell growth inhibition. Appl. Biol. Chem. 59(5): 739–746Relaxin-2 has gained much attention because of its importance in cancer cell proliferation, invasion, and angiogenesis. However, the potential of relaxin-2 as a target for immunotherapy has never been directly validated before. In the present study, we produced a human relaxin-2-specific antibody by immunizing mice using a specific liposome complex containing relaxin-2 epitope peptide and CpG-DNA. We isolated a hybridoma clone, 1B11F12, producing the anti-relaxin-2 monoclonal antibody. We confirmed with immunoprecipitation analysis that the antibody specifically recognizes native human relaxin-2. Treatment with the antibody suppressed the growth of some cancer cell lines. Therefore, we suggest that relaxin-2 could be a reasonable target of cancer therapeutics, and the monoclonal antibody could be used to treat cancer in humans.

Ah Young Lee, Sullim Lee, Hyun Young Kim, Sanghyun Lee, Eun Ju Cho. Anti-inflammatory effects of luteolin and luteoloside from Taraxacum coreanum in RAW264.7 macrophage cells. Appl. Biol. Chem. 59(5): 747–754The effects of luteolin (LT) and luteoloside (LS) from Taraxacum coreanum, using lipopolysaccharide (LPS)/interferon-gamma (IFN-γ)-induced RAW264.7 macrophage cells, on anti-inflammation were investigated. Our study was focused on the ethyl acetate fraction from T. coreanum (ETC) and its active compounds and their protective role against inflammation. The ETC and its active compounds, LT and LS, showed dose-dependent inhibitory activity against the production of nitric oxide (NO) and reactive oxygen species (ROS) in LPS/IFN-γ-stimulated RAW264.7 cells. In addition, ETC and its active compounds inactivated nuclear factor-kappa B and down-regulated inflammatory mediators. The results also showed that treatment with ETC, LT, and LS decrease pro-inflammatory cytokines, tumor necrosis factor-alpha, and interleukin-6. In conclusion, our studies indicated that ETC has anti-inflammatory activity owing to inhibition of NO/ROS generation and down-regulation of inflammatory mediators and cytokines. Moreover, LT and LS are bioactive compounds of ETC with protective effects against inflammation.

Kwang-Su Park, Woong Jung, Youhoon Chong, Mi Kyoung Kim. Quercetin 7-O-glutamate sensitizes Escherichia colito vancomycin. Appl. Biol. Chem. 59(5): 755–758

We have reported previously that conjugation of glutamic acid at the 7-O position of quercetin could significantly enhance its biological activity. In this study, as a part of our ongoing efforts to investigate the therapeutic potential of this novel quercetin–glutamate conjugate (1), we evaluated the effect of its combination with vancomycin against Gram-negative Escherichia coli, which are not normally affected by vancomycin alone. Quercetin conjugate 1 was shown to potentiate the antibacterial activity of vancomycin against E. coli, which could be attributed to its membrane perturbation activity, but not to facilitation of the efflux pump-mediated penetration of vancomycin.

Heesun Jung, Gyeonghui Yu, Hyejung Mok. CpG oligonucleotide and α-d-mannose conjugate for efficient delivery into macrophages. Appl. Biol. Chem. 59(5): 759–763For improved intracellular uptake of CpG ODN into macrophages, we developed CpG ODN and mannose conjugate (CpG–Man conjugate) via simple conjugation of α-d-mannose to CpG ODN. CpG–Man conjugate showed greatly enhanced intracellular uptake by 2.1-fold into macrophages, compared to CpG ODN in a TLR-9 receptor-specific manner. In addition, internalized CpG–Man conjugate successfully triggered TNF-α release in macrophages. Taken together, the CpG–Man conjugate can serve as the promising delivery systems of CpG ODN into immune cells as a candidate adjuvant.

Joungdu Shin, Seung Gil Hong, Sung-Chul Kim, Jae Eui Yang, SangRyong Lee, Fan-Zhu Li. Estimation of potential methane production through the mass balance equations from agricultural biomass in Korea. Appl. Biol. Chem. 59(5): 765–773Biomass is a renewable energy resource derived from all organic materials produced by both human and natural activities. Total biomass is amounted to be 58,010 Gg yr−1 from agricultural sector during 2013 in Korea: livestock manure, crop residues, and agro-industrial wastes. Potential methane production from agricultural biomass was calculated based on IPCC guidelines using manipulated equations. The main parameters were emission factor, total waste amount, and physico-chemical properties of each waste to estimate methane production. Calculated total potential methane production from the different categories for livestock, crop residues, and agro-industrial wastes was 502 Gg yr−1 in Korea. Poultry waste generated the highest methane potential with 227 Gg yr−1 followed by 80 Gg yr−1 from cattle waste. For crop residues and agro-industrial wastes, estimated methane production was 1 and 126 Gg yr−1, respectively. Results of this study show that livestock manure gave the highest methane emission in the agricultural sector. With this, more effective management of livestock wastes is necessary to develop and maximize technology on harnessing methane as alternative energy.

Jang-Eun Lee, So Rim Park, Seong Il Lim. Mass spectrometry-based metabolomic study of traditional Doenjang effects against hepatic fibrosis. Appl. Biol. Chem. 59(5): 775–780Korean traditional Doenjang are currently certified as traditional food. Certified Doenjang are valuable as a traditional food, however, only a few studies have evaluated and researched their quality characteristics and functionality. In the present study, we investigated the metabolite profile of Doenjang according to its α-SMA expression suppressing effect through mass spectrometry-based metabolomics. Also, the relationship between Doenjang metabolites and the suppression of α-SMA expression was identified. In the efforts to find traditional Doenjangmetabolites related to liver functions, branched-chain amino acids, p-aminobenzoic acid, 7,4′-dihydroxyflavone, soyasaponin, and isoflavone aglycone were found to be higher in the Doenjang group and showed higher α-SMA expression suppression activity. The present study highlights the fact that comprehensive metabolite analysis of traditional Doenjang provides information on its ability to improve liver function as well as other useful information for better understanding of the factors related to the effects of Doenjang metabolites against hepatic fibrosis.

Jin Yoo, Sun-Hee Woo, Ki-Do Park, Keun-Yook Chung. Effect of no-tillage and conventional tillage practices on the nitrous oxide (N2O) emissions in an upland soil: soil N2O emission as affected by the fertilizer applications. Appl. Biol. Chem. 59(6): 787–797The effect of no-tillage and conventional tillage practices on the nitrous oxide (N2O) emissions from the upland soil was evaluated in the cultivation of soybean in the temperate climate from June 2014 to September 2015 in Korea. In addition, we investigated the links between N2O emitted from field soil and different kinds of fertilizers. An experimental plot was composed of two main sectors that were no-tillage and conventional tillage, and then they were subdivided into four plots according to types of fertilizers: CF, chemical fertilizer, LP, liquid pig manure, HV, hairy vetch, and NF, non-fertilizer. The monthly averages of N2O emissions were significantly different from each other during the growing seasons of soybean; in July, N2O emission was significantly the highest, whereas, in September, its emission was the lowest (LSD, p = 0.05). In 2015, compared to those treatments in conventionally-tilled soils, the cumulative N2O emissions in NK, CF, HV, and LP of no-tilled soils were reduced by 20, 28.7, 35.7, and 28.1 %, respectively (LSD, p = 0.05). It was shown that N2O emission was significantly reduced in the different fertilizer treatments of no-tilled soils, compared to those of conventionally-tilled soils, respectively. Furthermore, the cumulative N2O emission in no-tilled soils was reduced by 0.03–0.09 kg N2O compared to that in tillage soils. It was found that soil N2O emission was about 11 % less in LP than in CF. Results obtained from our study indicate that the use of no-tillage practice and liquid pig manure, rather than tillage practice and chemical fertilizer, can decrease the N2O emission.

Jo-Won Lee, Bo-Ra Kim, Yena Heo, Gui-Seck Bae, Moon Baek Chang, BoKyung Moon. Feasibility of using kimchi by-products as a source of functional ingredients. Appl. Biol. Chem. 59(6): 799–806Baechu and radish are the major by-products generated from kimchi factories. Because vegetable and fruit by-products have abundant functional ingredients, the availability of them as a source of bioactive compounds is gaining attention. The objective of this study was to determine the feasibility of using kimchi by-products as sources of functional ingredients. For this purpose, 13 by-product samples were collected from different kimchi factories and analyzed for their sugar contents, total dietary fiber (TDF), antioxidant capacity, water holding capacity (WHC), viscosity, and specific gravity. The dietary fiber contents in kimchi by-product cabbage powder and radish powder were 27.37–49.54 and 30.19–62.81 %, respectively. TDF, WHC, and antioxidant capacities of kimchi by-products were equally as high as those of other by-products that are reused in industry. These results suggested that kimchi by-products have the potential as a new agro-industrial by-product that could be recycled as valuable sources of dietary fibers.

Md Nazmul Huda, Da Hye Kim, Erdenedolgor Erdene-Ochir, Yoon Suk Kim, Cheol-Ho Pan. Expression, phosphorylation, localization, and microtubule binding of tau in colorectal cell lines. Appl. Biol. Chem. 59(6): 807–812Tau is a microtubule-associated protein that causes proper role of neuron by assembling and stabilizing microtubules. The amount and post translational modification of tau can change its function as a stabilizer of microtubules. The aim of the study was to look for the expression of tau in colorectal cancer and normal cells, along with phosphorylation and microtubule binding properties of tau expressed in colorectal cancer cell. Two colorectal cancer cells (SW480 and HCT 116) expressed tau that was also phosphorylated, whereas the others (Caco-2, HT-29 and DLD1) did not. Colorectal normal cell (CCD-18Co) expressed very tiny amount of tau that was not phosphorylated. A big fraction of tau in HCT 116 did not bind to microtubule. The results suggest that some colorectal cancer cells express hyperphosphorylated tau as found in Alzheimer’s disease. So, tau in colorectal cancer cells does not look like same as tau in normal adult brain; rather it works nearly same as tau in neurodegenerative disease.

Dursun Kısa, Mahfuz Elmastaş, Lokman Öztürk, Ömer Kayır. Responses of the phenolic compounds of Zea maysunder heavy metal stress. Appl. Biol. Chem.

59(6): 813–820Heavy metal toxicity is one of the major abiotic stresses caused by physiological and biochemical changes. Plants have evolved various phytochemical defense mechanisms to cope with this abiotic stress conditions. Phenolic compounds are one of the stress responses and have multiple roles in respect to adaptation of plants to the environment. In the present study, we aimed to evaluate the differential accumulation of various phenolics with HPLC in the leaves of corn exposed to increasing heavy metal doses in the plant growth medium. The application of Cd, Cu, and Pb increased the total phenolics in all treatments compared to control groups. Chlorogenic acid and rutin were the main phenolic compounds in respect to quantifying. However, the contents of caffeic acid, ferulic acid, and vanillic acid were comparatively lower than chlorogenic acid and rutin in all samples. The content of chlorogenic acid significantly increased and rutin slightly increased in the treatment of the heavy metals. The levels of caffeic acid and ferulic acid significantly decreased in all exposures of heavy metals compared to control groups. The content of vanillic acid changed according to heavy metal types and doses in the leaves of corn, and the low doses of Pb and Cd increased the level of vanillic acid. We show that there is a positive correlation with the total phenolic content and chlorogenic acid when the corn is exposed to Pb. Moreover, there are negative correlations between total phenolic compound and caffeic acid, ferulic acid in the application of Cu and Cd.

Hee-Kwon Lee, Hoi-Seon Lee. Toxicities of active constituent isolated from Thymus vulgaris flowers and its structural derivatives against Tribolium castaneum (Herbst). Appl. Biol. Chem. 59(6): 821–826Insecticidal activity of 2-isopropyl-5-methylphenol isolated from Thymus vulgaris oil and its derivatives against Tribolium castaneum adults and larvae were evaluated using contact and fumigant bioassays. To elucidate the structure–activity relationship, 2-isopropyl-5-methylphenol and its derivatives were determined. Based on the LD50 values, 2-isopropylphenol was most toxic to T. castaneum adults and larvae, followed by 4-isopropylphenol, 2-methylphenol, 3-methylphenol, and phenol in contact and fumigant bioassays. These results indicate that the introduction of a functional group such as methyl and isopropyl group into the phenol skeleton has an important influence on contact and fumigant toxicities. Furthermore, 2-isopropyl-5-methylphenol and its derivatives could be used as an alternative to control stored-grain insects.

Yong Hoon Joo, Do Hun Lee, Nam Taek Lee, Namhyun Chung. Model development for prediction of the allergic response to the wheat proteins ω-5 gliadin and HMW-glutenin. Appl. Biol. Chem. 59(6): 827–831As one of the eight foods that account for 90 % of food allergies, wheat must be excluded from the diet in patients suffering from wheat allergies. From studies of wheat-dependent exercise-induced anaphylaxis (WDEIA), which has been used as a model to develop hypoallergenic wheat, we now know that the gluten fraction of wheat protein, particularly ω-5 gliadin and high-molecular-weight (HMW)-glutenin, is responsible for the allergic response. However, studies of allergic responses with WDEIA have been performed with a single wheat cultivar. Thus, in an effort to provide more information for the development of hypoallergenic wheat, we compared various cultivars with different countries of origin and characteristics. For the first step, we compared the allergen contents (ω-5 gliadin and HMW-glutenin) in each cultivar and the allergic response caused by each cultivar. Domestic wheat cultivars had lower contents of ω-5 gliadin and HMW-glutenin than those of imported wheat cultivars. Additionally, some cultivars caused varying allergic responses due to their allergen components. From regression analysis of allergen contents and allergic responses in vivo, we suggest a prediction model to estimate the extent of allergic response based on the ω-5 gliadin and HMW-glutenin contents. Further studies are needed to analyze the biological interactions between allergens from various cultivars and allergic response factors.

Ryeo Jin Kim, Mi Chung Suh. The GxSxG motif of Arabidopsis monoacylglycerol lipase (MAGL6 and MAGL8) is essential for their enzyme activities. Appl. Biol. Chem. 59(6): 833–840

Monoacylglycerol lipase (MAGL) catalyzes the hydrolysis of monoacylglycerols (MAG) to free fatty acids and glycerol, which is the last step of triacylglycerol breakdown. Among sixteen members, Arabidopsis thaliana MAGL6 (AtMAGL6) and AtMAGL8 showed strong lipase activities, but several AtMAGLs including AtMAGL16 displayed very weak activities (Kim et al. in Plant. J 85:758–771, 2016). To understand the internal factors that influence Arabidopsis MAGL activities, this study investigated the significance of ‘GxSxS motif,’ which is conserved in MAGLs. First, we observed that the presence of a serine protease inhibitor, phenylmethylsulfonyl fluoride, decreased the enzyme activity of AtMAGL6 and AtMAGL8 by IC50 values of 2.30 and 2.35, respectively. Computational modeling showed that amino acid changes of the GxSxG motif in AtMAGL6 and AtMAGL8 altered the nucleophilic elbow structure, which is the active site of MAGLs. Mutating the GxSxG motif in the recombinant maltose binding protein (MBP):AtMAGL6 and MBP:AtMAGL8 proteins to SxSxG, GxAxG, and GxSxS motifs completely demolished the activities of the mutant MAGLs. In contrast, no significant differences were observed between the activities of AtMAGL16 wild type form harboring the SxSxG motif, and mutant AtMAGL16 containing the GxSxG motif. These results revealed that the glycine and serine residues of the GxSxG motif are essential for AtMAGL6 and AtMAGL8 enzyme activities, and that AtMAGL16 may not be involved in the hydrolysis of lipid substrates.

Cheol Woo Min, Yu Ji Kim, Ravi Gupta, So Wun Kim, Won Young Han, Jong Min Ko, Hang Won Kang, Won Byong Yoon, Myoung Gun Choung, Yong Chul Kim, Sun Tae Kim. High-throughput proteome analysis reveals changes of primary metabolism and energy production under artificial aging treatment in Glycine max seeds. Appl. Biol. Chem. 59(6): 841–853This study was conducted to obtain basic information on protein profile changes by artificial aging in soybean seeds. Seed proteins were extracted using the protamine sulfate precipitation method, which improves the detection of low-abundance proteins (LAPs) by depleting the major seed storage proteins . Isolated proteins were separated by high-resolution two-dimensional gel electrophoresis (2-DE), and differentially modulated protein spots were identified by MALDI-TOF/TOF. A total of 33 differential proteins were identified of which 31 and 2 showed decreased and increased abundances, respectively. Functional annotation of the identified proteins revealed that proteins were mainly associated with primary metabolism (55%) and response to stimulus (20.9%). Proteins with increased abundance were associated with nutrient reservoir activity (spots 5, 10), while the decreased abundance proteins were mainly involved in the primary metabolism such as carbohydrate metabolic process (spots 1–3, 11), protein folding (spots 6–9, 33), glucose metabolic process (spot 25) oxidoreductase activity (spots 19–24), UDP-glucose pyrophosphorylase activity (spots 12, 13). These results provide information about proteome changes, especially, LAPs during artificial seed aging treatment.

Nguyen Hoai Nguyen, Chan Young Jeong, Won Je Lee, Hojoung Lee. Identification of a novel Arabidopsis mutant showing sensitivity to histone deacetylase inhibitors. Appl. Biol. Chem. 59(6): 855–860In the eukaryote nucleus, DNA wraps around histone cores and each core complex contains eight proteins including four pairs of H2A, H2B, H3, and H4 protein. The histone tail can be modified to alter the interaction strength between the histone core and a given DNA region, resulting in a change in gene expression. Various types of histone modifications are known to be involved in a number of cellular responses. In this study, we identified and characterized a novel mutant based on a growth assay using histone deacetylase inhibitors in combination with auxin, and we have named the mutant sensitive to histone deacetylase inhibitors 1. Our results imply that histone acetylation plays an important role in shoot morphology and leaf production in Arabidopsis.

Erdenedolgor Erdene-Ochir, Bok-Kyu Shin, Md Nazmul Huda, Da Hye Kim, Eun Ha Lee, Dae-Geun Song, Yoon-Mi Kim, Sang Min Kim, Cheol-Ho Pan. Cloning of a novel endogenous promoter for foreign gene expression in Phaeodactylum tricornutum. Appl. Biol. Chem. 59(6): 861–867

haeodactylum tricornutum is a model diatom, and its genomic sequence data and expressed sequence tag databases are available. This study was to discover a new endogenous promoter that drives strong constitutive expression of a protein of interest in P. tricornutum. To find promoter candidates, the intracellular proteins of P. tricornutum grown to stationary phase were extracted and identified by LC–MS/MS. Glutamine synthetase (GLNA) was one of the most abundantly expressed proteins during the stationary phase. Promoter is usually located on 5′ upstream region of open reading frame of the gene. Thus, two fragments of 5′ upstream region of the GLNA gene as putative promoters, 501 and 997 bp long, were amplified and cloned into enhanced Green Fluorescent Protein (eGFP) reporter systems. The constructed reporter systems were transformed into P. tricornutum and the eGFP expression levels were compared to those of reporter systems using the promoters of fcpA (fucoxanthin chlorophyll a/c binding protein A) and CIP1 (putative replication-associated proteins of a Chaetoceros lorenzianus-infecting DNA virus) as controls. The expression of eGFP driven by either GLNA promoter (501 and 997 bp) was linearly related to cell density, and eGFP was expressed constitutively regardless of the cultivation phase. The eGFP expression level driven by the GLNA promoters was at least 4 times higher than the fcpA-driven eGFP expression level at the stationary phase. The 501 and 997 bp regions of the GLNA promoter had similar activity patterns for transcribing the downstream gene. These results indicate that at least 501-bp region of the GLNA promoter can be used as a strong constitutive promoter in genetic engineering of P. tricornutum.

Young Mee Kim, You Jin Jeon, Jung Sik Huh, Sung Dae Kim, Kyung Kgi Park, Moonjae Cho. Effects of enzymatic hydrolysate from seahorse Hippocampus abdominalis on testosterone secretion from TM3 Leydig cells and in male mice. Appl. Biol. Chem. 59(6): 869–879Protein hydrolysates, the so-called bioactive peptides, are specific protein fragments that have positive effects on several body functions and may improve human health. Marine organism-derived protein hydrolysates and bioactive peptides have shown to possess many physiological functions. Seahorses, used in traditional medicine, are generally ground to powder form either for direct application or for application following dissolution in warm water. In this present study, we purified, hydrolyzed, and characterized two bioactive peptides (ALC and PEP) obtained from seahorse (Hippocampus abdominalis). The hydrolysates derived from seahorse significantly upregulated the expression of cyclin D and cyclin E and increased cell proliferation and testosterone level in the TM3 mouse Leydig cell line. These findings suggest that the hydrolysates stimulate the proliferation of TM3 cells via the AKT, ERK, and JNK pathways. The decline in the circulating testosterone levels in older men is associated with various adverse health effects. Following daily intake of hydrolysates for 12 weeks, the circulating level of testosterone and the sperm count in mice were measured. We found increased sperm motility (sperm count) as well as an increase in the testosterone level in male mice following a 12-week intake of hydrolysates derived from H. abdominalis. Hence, it can be suggested that seahorse-derived hydrolysates play an important role in improving male health by improving the serum testosterone level.

Dong-Ho Kim, Sung-Yong Hong, Mi-Hyeon Jeon, Jae-Min An, Sung-Youn Kim, Hyo-Young Kim, Bo Ryun Yoon, Soo Hyun Chung. Simultaneous determination of the levels of deoxynivalenol, 3-acetyldeoxynivalenol, and nivalenol in grain and feed samples from South Korea using a high-performance liquid chromatography–photodiode array detector. Appl. Biol. Chem. 59(6): 881–887An HPLC method combined with immunoaffinity clean-up was developed for the simultaneous analysis of deoxynivalenol (DON), nivalenol (NIV), and 3-acetyldeoxynivalenol (3AcDON) in grains and feeds. The limits of quantification of the method for DON, 3AcDON, and NIV were 11.0, 27.6, and 13.5 ng/g, respectively. HPLC analyses of 300 samples of grains and feeds collected in South Korea showed that the incidence of DON and NIV in corn was 22.5 and 7.5%, while that in rice was 4.0 and 54.0%, respectively. The incidence of DON and NIV in mixed grain powder was 62.0 and 24.0%, respectively. Our study also showed that animal feeds were mainly contaminated

with DON (98.1%) in the range of 32.8–950.25 ng/g with the mean concentration of 353.32 ng/g. The levels of DON in grains did not exceed the maximum allowable limit (1 mg/kg) set by the Korean Food and Drug Administration.