antimicrobial activity of the indigenously microbial fermented fuzhuan brick-tea

1
S722 Abstracts / Journal of Biotechnology 136S (2008) S717–S742 The levels of triglycerides were also lower, due to uptake of mushroom mycelia. As a result of conducted studies it is possible to draw a conclusion that the additives of submerged mycelia of several basidiomycetes, to manage diabetes and leads to essential decrease in a risk of development of arteriosclerosis. doi:10.1016/j.jbiotec.2008.07.1716 VIII1-P-004 Thermal stability of immobilized lipase from Candida antarctica in 1-alcohols and glycerol Shuji Adachi , Takashi Kobayashi Division of Food Science and Biotechnology, Graduate School of Agri- culture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan E-mail address: [email protected] (S. Adachi). The immobilized lipase from Candida antarctica has been widely used for the synthesis of esters through condensation or transester- ification. 1-Alcohols and glycerol themselves are one of substrates for their condensation with a carbonic acid to produce their corresponding esters. The condensation at a high temperature accelerates the reaction rate, but inactivation of the enzyme would become more significant. In this context, we examined the thermal stability of the immobilized lipase in 1-alcohols with 4–8 carbons at 70 C to 115 C and in glycerols with various water contents at 80 C to 100 C. The plots of the residual activity versus the incu- bation time on a semi-logarithmic scale did not give a straight line for all the tested 1-alcohols and glycerol at any temperature, indi- cating that the inactivation did not obey the 1st-order kinetics. Assuming the heterogeneity in the susceptibility of the immobi- lized enzyme to the inactivation, the process was expressed by the following equation (Kawamura et al., 1981): C e C e0 = RT 2 −∞ exp R 2 T 2 (ln k d ln ¯ k d ) 2 2 2 exp(k d t )d(ln k d ) (1) where C e is the active enzyme concentration, C e0 is the initial C e , R is the gas constant, T is the absolute temperature, k d is the rate con- stant of thermal inactivation, ¯ k d is the k d corresponding to the mean value of the free energy of activation, G , and is the standard deviation of G . The inactivation processes of the immobilized lipase from Candida antarctica in all the tested 1-alcohols and glyc- erols could be well expressed by Eq. (1). The results indicated that alkyl or glyceryl esters could be synthesized using the enzyme at an abnormally high temperature. Acknowledgement This study was partially supported by Cooperative for Innovative Technology and Advanced Research in Evolutional Area (CITY AREA) program from the Ministry of Education, Culture, Sports, Science and Technology, Japan. Reference Kawamura, Y., Nakanishi, K., Matsuno, R., 1981. Stability of immobilized - chymotrypsin. Biotechnol. Bioeng. 23, 1219–1236. doi:10.1016/j.jbiotec.2008.07.1718 VIII1-P-009 Antimicrobial activity of the indigenously microbial fermented Fuzhuan brick-tea Haizhen Mo 1,, Hao Zhang 1 , Yingqiu Li 2 , Yang Zhu 3 1 School of Food Science, Henan Institute of Science and Technology, Xinxiang, 453003, China 2 College of Food and Bioengineering, Shandong Institute of Light Industry, Jinan 250353, China 3 Food and Bioprocess Engineering Group, Wageningen University, P.O. Box 8129, 6700 EV Wageningen, Netherlands E-mail address: [email protected] (H. Mo). Antimicrobial activity of extracts from an indigenously fermented tea, Fuzhuan, was tested in addition to the microbiological analysis. Microbial counting and identification revealed that Aspergillus spp., Penicillium spp. and Eurotium spp. were the main microorganisms isolated from the samples of fermentation and Eurotium spp. was the dominating fungus responsible for the fermentation. Antibac- terial tests of extracts of fermented tea showed inhibitory effect on several food borne bacteria, including spore forming bacteria Bacillus cereus, Bacillus subtilis, Clostridium perfringens and Clostrid- ium sporogenes. The antibacterial activity increased with the course of the fermentation. This implied that certain metabolites of the fungi growing on tea leaves had the feature of inhibiting certain food borne spoilage and pathogen microorganisms. These natural antimicrobial substances have the potential as innovative and mild food preservatives. Keywords: Antimicrobial activity; Aspergillus spp.; Penicillium spp.; Erothium spp.; Fuzhuan brick-tea; Solid-state fermentation; Food fermentation References Chou, C.C., Lin, L.L., Chung, K.T., 1999. Antimicrobial activity of tea as affected by the degree of fermentation and manufacturing season. Int. J. Food Microbiol. 48 (2), 125–130. Kawakami, M., Kobayashi, A., Yamanishi, T., Shoujaku, S., 1987. Flavour constituents of the microbially fermented teas Zhuan-cha and Koku-cha. Nippon Nogeika- gaku Kaishi. J. Agric. Chem. Soc. Jpn. 61 (4), 457–465. Mo, H.Z., Xu, X.Q., Yan, M.C., Zhu, Y., 2005. Microbiological analysis and antibacterial effects of the indigenous fermented Puer tea. Agro Food Ind. Hi-Tech. 16 (6), 16–18. Mo, H., Zhu, Y., Chen, Z., 2008. Microbial fermented tea – a potential source of natural food preservatives. Trends Food Sci. Technol. 19 (3), 124–130. Schillinger, U., Geisen, R., Holzapfel, W.H., 1996. Potential of antagonistic microor- ganisms and bacteriocins for the biological preservation of foods. Trends Food Sci. Technol. 7 (5), 158–164. Si, W.D., Gong, J., Tsao, R., Kalab, M., Yang, R., Yin, Y.L., 2006. Bioassay-guided purifica- tion and identification of antimicrobial components in Chinese green tea extract. J. Chromatogr. A 1125 (2), 204–210. Xu, X.Q., Mo, H.Z., Yan, M.C., Zhu, Y., 2007. Analysis of characteristic aroma of fungal fermented Fuzhuan brick-tea by gas chromatography/mass spectrophotometry. J. Sci. Food Agric. 87 (8), 1502–1504. doi:10.1016/j.jbiotec.2008.07.1719

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Page 1: Antimicrobial activity of the indigenously microbial fermented Fuzhuan brick-tea

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References

Chou, C.C., Lin, L.L., Chung, K.T., 1999. Antimicrobial activity of tea as affected by thedegree of fermentation and manufacturing season. Int. J. Food Microbiol. 48 (2),125–130.

Kawakami, M., Kobayashi, A., Yamanishi, T., Shoujaku, S., 1987. Flavour constituentsof the microbially fermented teas Zhuan-cha and Koku-cha. Nippon Nogeika-gaku Kaishi. J. Agric. Chem. Soc. Jpn. 61 (4), 457–465.

Mo, H.Z., Xu, X.Q., Yan, M.C., Zhu, Y., 2005. Microbiological analysis and antibacterialeffects of the indigenous fermented Puer tea. Agro Food Ind. Hi-Tech. 16 (6),16–18.

Mo, H., Zhu, Y., Chen, Z., 2008. Microbial fermented tea – a potential source of natural

S722 Abstracts / Journal of Biotec

The levels of triglycerides were also lower, due to uptake ofmushroom mycelia.

As a result of conducted studies it is possible to draw aconclusion that the additives of submerged mycelia of severalbasidiomycetes, to manage diabetes and leads to essential decreasein a risk of development of arteriosclerosis.

doi:10.1016/j.jbiotec.2008.07.1716

VIII1-P-004

Thermal stability of immobilized lipase from Candida antarcticain 1-alcohols and glycerol

Shuji Adachi ∗, Takashi Kobayashi

Division of Food Science and Biotechnology, Graduate School of Agri-culture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan

E-mail address: [email protected] (S. Adachi).

The immobilized lipase from Candida antarctica has been widelyused for the synthesis of esters through condensation or transester-ification. 1-Alcohols and glycerol themselves are one of substratesfor their condensation with a carbonic acid to produce theircorresponding esters. The condensation at a high temperatureaccelerates the reaction rate, but inactivation of the enzyme wouldbecome more significant. In this context, we examined the thermalstability of the immobilized lipase in 1-alcohols with 4–8 carbonsat 70 ◦C to 115 ◦C and in glycerols with various water contents at80 ◦C to 100 ◦C. The plots of the residual activity versus the incu-bation time on a semi-logarithmic scale did not give a straight linefor all the tested 1-alcohols and glycerol at any temperature, indi-cating that the inactivation did not obey the 1st-order kinetics.Assuming the heterogeneity in the susceptibility of the immobi-lized enzyme to the inactivation, the process was expressed by thefollowing equation (Kawamura et al., 1981):

Ce

Ce0= RT√

2��

∫ ∞

−∞exp

(−R2T2(ln kd− ln k̄d)

2

2�2

)exp(−kdt)d(ln kd)

(1)

where Ce is the active enzyme concentration, Ce0 is the initial Ce, Ris the gas constant, T is the absolute temperature, kd is the rate con-stant of thermal inactivation, k̄ is the k corresponding to the mean

d dvalue of the free energy of activation, �G‡, and � is the standarddeviation of �G‡. The inactivation processes of the immobilizedlipase from Candida antarctica in all the tested 1-alcohols and glyc-erols could be well expressed by Eq. (1). The results indicated thatalkyl or glyceryl esters could be synthesized using the enzyme atan abnormally high temperature.

Acknowledgement

This study was partially supported by Cooperative for InnovativeTechnology and Advanced Research in Evolutional Area (CITY AREA)program from the Ministry of Education, Culture, Sports, Scienceand Technology, Japan.

Reference

Kawamura, Y., Nakanishi, K., Matsuno, R., 1981. Stability of immobilized �-chymotrypsin. Biotechnol. Bioeng. 23, 1219–1236.

doi:10.1016/j.jbiotec.2008.07.1718

gy 136S (2008) S717–S742

VIII1-P-009

Antimicrobial activity of the indigenously microbial fermentedFuzhuan brick-tea

Haizhen Mo 1,∗, Hao Zhang 1, Yingqiu Li 2, Yang Zhu 3

1 School of Food Science, Henan Institute of Science and Technology,Xinxiang, 453003, China2 College of Food and Bioengineering, Shandong Institute of LightIndustry, Jinan 250353, China3 Food and Bioprocess Engineering Group, Wageningen University, P.O.Box 8129, 6700 EV Wageningen, Netherlands

E-mail address: [email protected] (H. Mo).

Antimicrobial activity of extracts from an indigenously fermentedtea, Fuzhuan, was tested in addition to the microbiological analysis.Microbial counting and identification revealed that Aspergillus spp.,Penicillium spp. and Eurotium spp. were the main microorganismsisolated from the samples of fermentation and Eurotium spp. wasthe dominating fungus responsible for the fermentation. Antibac-terial tests of extracts of fermented tea showed inhibitory effecton several food borne bacteria, including spore forming bacteriaBacillus cereus, Bacillus subtilis, Clostridium perfringens and Clostrid-ium sporogenes. The antibacterial activity increased with the courseof the fermentation. This implied that certain metabolites of thefungi growing on tea leaves had the feature of inhibiting certainfood borne spoilage and pathogen microorganisms. These naturalantimicrobial substances have the potential as innovative and mildfood preservatives.

Keywords: Antimicrobial activity; Aspergillus spp.; Penicillium spp.;Erothium spp.; Fuzhuan brick-tea; Solid-state fermentation; Foodfermentation

food preservatives. Trends Food Sci. Technol. 19 (3), 124–130.Schillinger, U., Geisen, R., Holzapfel, W.H., 1996. Potential of antagonistic microor-

ganisms and bacteriocins for the biological preservation of foods. Trends FoodSci. Technol. 7 (5), 158–164.

Si, W.D., Gong, J., Tsao, R., Kalab, M., Yang, R., Yin, Y.L., 2006. Bioassay-guided purifica-tion and identification of antimicrobial components in Chinese green tea extract.J. Chromatogr. A 1125 (2), 204–210.

Xu, X.Q., Mo, H.Z., Yan, M.C., Zhu, Y., 2007. Analysis of characteristic aroma of fungalfermented Fuzhuan brick-tea by gas chromatography/mass spectrophotometry.J. Sci. Food Agric. 87 (8), 1502–1504.

doi:10.1016/j.jbiotec.2008.07.1719