分子微生物科学分野（協力講座：化学研究所） 教授：栗原達夫、助教：川本　純

化学を基盤にした微生物の機能解析と応用

酵素の精密反応機構解析と応用

微生物の環境適応機構の解析と応用

� 微生物の様々な生命現象を化学的なアプローチによって解き明かし、応用展開することを目指して研究を進めています。特に特殊環境微生物の環境適応を担う分子基盤の解析、ユニークな反応を触媒する新しい微生物酵素の探索と構造・機能解析、精密触媒機構の解析、生体膜の構築原理と機能発現機構の解析を行っています。また、それらの知見に基づき、物質生産や環境浄化に役立つ新しい微生物利用法の開発を行っています。

微生物酵素を利用した光学活性有機ハロゲン化合物（除草剤原料）の生産

C CH

H

Cl

COOHH3C C

HCOOH

Cl

京都大学農学研究科応用生命科学専攻　2014.7.10

生体膜における脂質とタンパク質の機能解析

　生体膜は細胞の内と外を隔て、細胞内外の物質輸送や情報伝達において必須の役割を担っています。私たちは、このような生体膜

の構築原理や機能発現機構の解明に取り組んでいます。特に、生体膜の主成分であるリン

脂質の機能に注目し、種々のリン脂質が生体膜でどのように分布しているか、膜タンパク質とどのように相互作用しているか、明らか

にするための研究を行っています。

H2NH2N OHPOOO

O

O

O

O

EPA-containing phospholipid Unfolded protein

Folded protein Membrane insertion (unfolded)

Synthetic !lipid probe!

　微生物の環境適応を担う分子基盤の解明と応用に取り組んでいます。地球上の生命圏の約8割は常時4℃以下に保たれた低温環境です。このような環境に生息する微生物の環境適応機構を重点的に解析

しています。また、これらの微生物を、低温で有用物質を生産するための微生物工場として開発する研究も行っています。

　微生物はきわめて多様な物質変換能を有しています。化学的に特殊な環境に生息する微生物は、ユニークな物質変換能をもつ

酵素を生産することで、環境中の化合物を資化・分解しています。それらの酵素には

物質生産や環境浄化への応用が期待されるものも多く、また、酵素化学的に興味深い反応メカニズムをもつものが多数あります。

このような微生物酵素の機能解析、精密反応機構解析、応用開発に取り組んでいます。

微生物、特殊環境微生物、低温菌、生体膜、タンパク質、リン脂質、酵素、補酵素、高度不飽和脂肪酸、反応機構、環境適応、物質生産

キーワード

最近の主な研究業績

Characterization of a Thermostable 2,4-Diaminopentanoate Dehydrogenase from Fervidobacterium nodosum Rt17-B1 Fukuyama S, Mihara H, Miyake R, Ueda M, Esaki N, Kurihara T (2014) J Biosci Bioeng 117 (5): 551-556  

Reciprocal Modulation of Surface Expression of Annexin A2 in a Human Umbilical Vein Endothelial Cell-derived Cell Line by Eicosapentaenoic Acid and Docosahexaenoic Acid Park J, Yamaura T, Kawamoto J, Kurihara T, Sato SB (2014) PLoS One. Jan 21; 9(1): e85045 Global Identification of Genes Affecting Iron-sulfur Cluster Biogenesis and Iron Homeostasis Hidese R, Mihara H, Kurihara T, Esaki N (2014) J Bacteriol 196 (6): 1238-1249   Selenocysteine Lyase: Delivering Selenium in Biosynthetic Pathway

Mihara H, Kurihara T, Esaki N (2014) Selenium in the Environment and Human Health - Proceedings of the 3rd International Conference on Selenium in the Environment and Human Health 181-182   化学的アプローチによる高度不飽和脂肪酸含有リン脂質の生理機能解析

栗原達夫, 川本純　(2014) 薬学雑誌 134 (4): 507-513   長鎖多価不飽和脂肪酸エイコサペンタエン酸の生理機能

栗原達夫, 川本純　(2014) 医学の歩み 248 (13): 1211-1227   Inhibition of Constitutive Akt (PKB) Phosphorylation by Docosahexaenoic Acid in the Human Breast Cancer Cell Line MDA-MB-453 Sato S B, Park J, Kawamoto J, Sato S, Kurihara,T (2013) Biochim Biophys Acta 1831: 306-313   Binding Modes of DL-2-Haloacid Dehalogenase Revealed by Crystallography, Modeling and Isotope Effects Studies Siwek A, Omi R, Hirotsu K, Jitsumori K, Esaki N, Kurihara T, Paneth P (2013) Arch Biochem Biophys 540: 26-32

Regulation of Cytochrome c- and Quinol Oxidases, and Piezotolerance of Their Activities in the Deep-Sea Piezophile Shewanella violacea DSS12 in Response to Growth Conditions Ohke Y, Sakoda A, Kato C, Sambongi Y, Kawamoto J, Kurihara T, Tamegai H (2013) Biosci Biotechnol Biochem 77: 1522-1528  

Proteins Involved in Cold Adaptation Yoshimune K, Kawamoto J, Kurihara T (2013) Cold-Adapted Microorganisms 97-110   細菌細胞膜の新しいマイクロドメイン 栗原達夫, 川本純 (2013) 化学と生物 51: 358-360   細菌における高度不飽和脂肪酸含有リン脂質の生理機能 川本純, 栗原達夫 (2013) オレオサイエンス 13: 221-229

  地球環境と微生物 栗原達夫 (2013) 海洋化学研究 26: 28-30  

Occurrence of a Bacterial Membrane Microdomain at the Cell Division Site Enriched in Phospholipids with Polyunsaturated Hydrocarbon Chains Sato S, Kawamoto J, Sato S B, Watanabe B, Hiratake J, Esaki N, Kurihara T (2012) J Biol Chem 287: 24113-24121   Eicosapentaenoic Acid Facilitates the Folding of an Outer Membrane Protein of the Psychrotrophic Bacterium, Shewanella livingstonensis Ac10 Dai X-Z, Kawamoto J, Sato S B, Esaki N, Kurihara T (2012) Biochem Biophys Res Commun. 425: 363-367

  Identification of Cold-Inducible Inner Membrane Proteins of the Psychrotrophic Bacterium, Shewanella livingstonensis Ac10, by Proteomic Analysis Park J, Kawamoto J, Esaki N, Kurihara T (2012) Extremophiles 16: 227-236

  Characterization of 1-Acyl-sn-Glycerol-3-Phosphate Acyltransferase from a Polyunsaturated Fatty Acid-Producing Bacterium, Shewanella livingstonensis Ac10 Cho H-N, Kasai W, Kawamoto J, Esaki N, Kurihara T (2012) Trace Nutrients Research 29: 92-99

  Functional Analysis of an Eicosapentaenoic Acid Biosynthesis Protein Orf2 from a Psychrotrophic Bacterium, Shewanella livingstonensis Ac10 Gong C, Kawamoto J, Esaki N, Kurihara T (2012) Trace Nutrients Research 29: 84-91   Pseudomonas putida PydR, a RutR-Like Transcriptional Regulator, Represses the Dihydropyrimidine Dehydrogenase Gene in the Pyrimidine Reductive Catabolic Pathway Hidese R, Mihara H, Kurihara T, Esaki N (2012) J Biochem 152: 341-346

Substrate Specificity of Fluoroacetate Dehalogenase: An Insight from Crystallographic Analysis, Fluorescence Spectroscopy, and Theoretical Computations Nakayama T, Kamachi T, Jitsumori K, Omi R, Hirotsu K, Esaki N, Kurihara T, Yoshizawa K (2012) Chemistry 18: 8392-8402  

Eicosapentaenoic Acid Plays a Role in Stabilizing Dynamic Membrane Structure in the Deep-Sea Piezophile Shewanella violacea: A Study Employing High-Pressure Time-Resolved Fluorescence Anisotropy Measurement Usui K, Hiraki T, Kawamoto J, Kurihara T, Nogi Y, Kato C, Abe F (2012) Biochim Biophys Acta 1818: 574-583  

好冷性微生物の活用 栗原達夫 (2012) 極限環境微生物の産業展開: 131-136   遺伝子工学で用いる酵素

栗原達夫(2012) 基礎生物学テキストシリーズ１０　遺伝子工学 15-30   Favourable effects of Eicosapentaenoic Acid on the Late Step of the Cell Division in a Piezophilic Bacterium, Shewanella violacea DSS12, at High-hydrostatic Pressures

Kawamoto J, Sato T, Nakasone K, Kato C, Mihara H, Esaki N, Kurihara T (2011) Environ Microbiol 13: 2293–2298   In vitro Refolding of an OmpA Homolog, a Major Cold-Inducible Outer Membrane Protein, from a Psychrotrophic Bacterium, Shewanella livingstonensis Ac10 Dai X-Z, Kawamoto J, Esaki N, Kurihara T (2011) Journal of Japanese Society for Extremophiles 10: 90-96   Differential Roles of Internal and Terminal Double Bonds in Docosahexaenoic Acid: Comparative Study of Cytotoxicity of Polyunsaturated Fatty Acids to HT-29 Human Colorectal Tumor Cell Line Sato S B, Sato S, Kawamoto J, Kurihara T (2011) Prostaglandins Leukot Essent Fatty Acids 84: 31-37  

Function of FADH2-dependent 2-Haloacrylate Hydratase from a 2-Chloroacrylate-utilizing Bacterium, Burkholderia sp. WS Vasudevan A, Fujita M, Kurata A, Kawamoto J, Esaki N, Kurihara T (2011) Trace Nutri Res 28: 58-64  

Thermal Stability of Cytochrome c(5) of Pressure-Sensitive Shewanella livingstonensis Masanari M, Wakai S, Tamegai H, Kurihara T, Kato C, Sambongi Y (2011) Biosci Biotechnol Biochem 75: 1859-1861   Piezotolerance of the Respiratory Terminal Oxidase Activityof the Piezophilic Shewanella violacea DSS12 as Compared with Non-Piezophilic Shewanella Species Tamegai H, Ota Y, Haga M, Fujimori H, Kato C, Nogi Y, Kawamoto J, Kurihara T, Sambongi Y (2011) Biosci Biotechnol Biochem 75: 919-924  

Identification of Novel Mammalian Phospholipids Containing Threonine, Aspartate, and Glutamate as the Base Moiety Omori T, Honda A, Mihara H, Kurihara T, Esaki N (2011) J Chromatogr B 879: 3296-3302  

Mammalian Selenocysteine Lyase Is Involved in Selenoprotein Biosynthesis Kurokawa S, Takehashi M, Tanaka H, Mihara H, Kurihara T, Tanaka S, Hill K, Burk R, Esaki N (2011) J Nutr Sci Vitaminol 57: 298-305   Escherichia coli Dihydropyrimidine Dehydrogenase is a Novel NAD-dependent Heterotetramer Essential for the Production of 5,6-Dihydrouracil Hidese R, Mihara H, Kurihara T, Esaki N (2011) J Bacteriol 193: 989-993   A Mechanistic Analysis of Enzymatic Degradation of Organohalogen Compounds

Kurihara T (2011) Biosci Biotechnol Biochem 75: 189-198   好冷性細菌が生産する高度不飽和脂肪酸含有リン脂質の生理機能 川本純, 栗原達夫 (2011) バイオサイエンスとインダストリー 69: 305-307   IscS Functions as a Primary Sulfur-donating Enzyme by Interacting Specifically with MoeB and MoaD in the Biosynthesis of Molybdopterin in Escherichia coli Zhang W, Urban A, Mihara H, Leimkühler S, Kurihara T, Esaki N (2010) J Biol Chem 285: 2302-2308   Crystallization and Preliminary X-ray Analysis of L-Azetidine-2-Carboxylate Hydrolase from Pseudomonas sp. strain A2C

Toyoda M, Jitsumori K, Mikami B, P.Wackett L, Kurihara T, Esaki N (2010) Acta Crystallogr Sect F Struct Biol Cryst Commun 66: 801-804   The Distribution of Phosphatidyl-D-serine in the Rat Omori T, Mihara H, Kurihara T, Esaki N (2010) Biosci Biotechnol Biochem 74: 1953-1955   Reaction Mechanism and Molecular Basis for Selenium/Sulfur Discrimination of Selenocysteine Lyase Omi R, Kurokawa S, Mihara H, Hayashi H, Goto M, Miyahara I, Kurihara T, Hirotsu K, Esaki N (2010) J Biol Chem 285: 12133-12139   2-Haloacrylate Hydratase, a New Class of Flavoenzyme that Catalyzes the Addition of Water to the Substrate for Dehalogenation

Mowafy A M, Kurihara T, Kurata A, Uemura T, Esaki N (2010) Appl Environ Microbiol 76: 6032-6037  

2-Haloacrylate Hydratase Is a Bifunctional Enzyme with NADH-Dependent FAD Reductase Activity Mowafy A M, Kurihara T, Esaki N (2010) Trace Nutri Res 27: 52-55  

低温細菌における長鎖高度不飽和脂肪酸の生合成と機能 栗原達夫, 川本純, 江崎信芳 (2010) 蛋白質核酸酵素 55: 94-99   好冷酵素の特性と利用法 栗原達夫 (2010) 酵素利用技術大系: 239-243   有機ハロゲン化合物代謝関連酵素 栗原達夫 (2010) 酵素利用技術大系: 88-92

  Crystal Structure of a Homolog of Mammalian Serine Racemase from Schizosaccharomyces pombe Goto M, Yamauchi T, Kamiya N, Miyahara I, Yoshimura T, Mihara H, Kurihara T, Hirotsu K, Esaki N (2009) J Biol Chem 284: 25944-25952   Selenocysteine Is Selectively Taken Up by Red Blood Cells Imai T, Mihara H, Kurihara K, Esaki N (2009) Biosci Biotechnol Biochem 73: 2746-2748.   Possible Role of Red Blood Cells in Selenocysteine Metabolism Imai T, Mihara H, Kurihara T, Esaki N (2009) Trace Nutri Res 26: 22-25

  The Catalytic Mechanism of Fluoroacetate Dehalogenase: A Computational Exploration of Biological Dehalogenation Kamachi T, Nakayama T, Shitamichi O, Jitsumori K, Kurihara T, Esaki N, Yoshizawa K (2009) Chemistry 15: 7394-7403

  Identification of Proteins Interacting with Selenocysteine Lyase Tobe R, Mihara H, Kurihara T, Esaki N (2009) Biosci Biotechnol Biochem 73:1230-1232  

Roles of K151 and D180 in L-2-Haloacid Dehalogenase from Pseudomonas sp. YL: Analysis by Molecular Dynamics and Ab Initio Fragment Molecular Orbital Calculations Nakamura T, Yamaguchi A, Kondo H, Watanabe H, Kurihara T, Esaki N, Hirono S, Tanaka S (2009) J Comput Chem 30: 2625-2634 Serine Racemase with Catalytically Active Lysinoalanyl Residue Yamauchi T, Goto M, Wu H-Y, Uo T, Yoshimura T, Mihara H, Kurihara T, Miyahara I, Hirotsu K, Esaki N (2009) J Biochem 145: 421-424

 

Occurrence of Phosphatidyl-D-serine in the Rat Cerebrum Omori T, Mihara H, Kurihara T, Esaki N (2009) Biochem Biophys Res Commun 382: 415-418   Eicosapentaenoic Acid Plays a Beneficial Role in Membrane Organization and Cell Division of a Cold-Adapted Bacterium, Shewanella livingstonensis Ac10 Kawamoto J, Kurihara T, Yamamoto K, Nagayasu M, Tani Y, Mihara H, Hosokawa M, Baba T, Sato S B, Esaki N (2009) J Bacteriol 191: 632-640   X-ray Crystallographic and Mutational Studies of Fluoroacetate Dehalogenase from Burkholderia sp. FA1 Jitsumori K, Omi R, Kurihara T, Mihara H, Miyahara I, Hirotsu K, Esaki N (2009) J Bacteriol 191: 2630-2637   低温細菌における高度不飽和脂肪酸含有リン脂質の機能

栗原達夫 (2009) 生化学 81: 716-719   好冷性細菌の低温適応に関わるタンパク質とリン脂質 栗原達夫, 川本純, 江崎信芳 (2009) 生化学 81: 1072-1079   低温下で機能する酵素 栗原達夫, 江崎信芳 (2009) 化学工学 73: 324-327   新規不斉還元酵素によるキラル有機ハロゲン化合物の生産 栗原達夫 (2009) バイオサイエンスとインダストリー67: 161-163

  細胞がセレンをタンパク質に取り込む機構の解析とその応用 栗原達夫, 三原久明, 江崎信芳 (2009) 環境・資源保全のためのメタルバイオテクノロジー  

Investigation of the Roles of Cysteine Desulfurases in the Molybdopterin Synthesis in Escherichia coli Zhang W, Mihara H, Kurihara T, Esaki N (2008) Trace Nutri Res 25: 152-157  

Role of Cysteine Desulfurase IscS in the Biosynthesis of Molybdopterin Zhang W, Mihara H, Kurihara T, Esaki N (2008) Vitamins (Japan) 82: 645-650 Cold Adaptation of Eicosapentaenoic Acid-less Mutant of Shewanella livingstonensis Ac10 Involving Uptake and Remodeling of Synthetic Phospholipids Containing Various Polyunsaturated Fatty Acids Sato S, Kurihara T, Kawamoto J, Hosokawa M, Sato SB, Esaki N (2008) Extremophiles 12: 753-761  

The iscS Gene Deficiency Affects the Expression of Pyrimidine Metabolism Genes Mihara H, Hidese R, Yamane M, Kurihara T, Esaki N (2008) Biochem Biophys Res Commun 372: 407-411  

Thioredoxin Reductase 1 Is Important for Selenoprotein Biosynthesis in HeLa Cells Kurokawa S, Mihara H, Yokoyama I, Mochizuki M, Yodoi J, Tamura T, Kurihara T, Esaki N (2008) Biomed Res Trace Elem 19: 84-87   Proteomic Studies of Psychorophilic Microorganisms

Kurihara T, Esaki N (2008) Psychrophiles: From Biodiversity to Biotechnology 333-343   Bacterial Hydrolytic Dehalogenases and Related Enzymes: Occurrences, Reaction Mechanisms, and Applications Kurihara T, Esaki N (2008) Chem Rec 8: 67-74   Production of (S)-2-Chloropropionate by Asymmetric Reduction of 2-Chloroacrylate with 2-Haloacrylate Reductase Coupled with Glucose Dehydrogenase Kurata A, Fujita M, Mowafy A M, Kamachi H, Kurihara T, Esaki N (2008) J Biosci Bioeng 105: 429-431