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Construction and Characterization of Composite Biocatalysts

Research Project

Project/Area Number 13125203
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Science and Engineering
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

ESAKI Nobuyoshi  Kyoto Univ., Institute for Chemical Research, Professor, 化学研究所, 教授 (50135597)

Co-Investigator(Kenkyū-buntansha) HIRATAKE Jun  HIRATAKE,Jun, 化学研究所, 助教授 (80199075)
FUJII Ikuo  Osaka Prefecture Univ., Research Institute for Advanced Science and Technology, Professor, 先端科学研究所, 教授 (70189984)
YOSHIMURA Tohru  Nagoya, Univ., Graduate School of Bioagricultural sciences, 大学院・生命農学研究科, 教授 (70182821)
藤井 郁夫  生物分子工学研究所, 機能創製プロジェクト部門, 部長
畑 安雄  京都大学, 化学研究所, 助教授 (10127277)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥79,700,000 (Direct Cost: ¥79,700,000)
Fiscal Year 2003: ¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2002: ¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2001: ¥54,900,000 (Direct Cost: ¥54,900,000)
Keywordscomposite biocatalyst / coenzyme / structure-activity relationship / pyridoxal phosphate / NADPH / iron-sulfur cluster / asymmetric synthesis / Suf / ジヒドロピリミジン / システインデスルフラーゼ / 鉄-硫黄クラスター / システインデルスフラーゼ / 抗体酵素
Research Abstract

We analyzed the mechanism of biosynthesis of iron-sulfur clusters, which are useful as components of composite biocatalysts. We also developed novel processes for the production of chiral compounds by the use of novel composite biocatalysts. In addition, reaction mechanisms of several composite biocatalysts such as amino acid racemases which require pyridoxal phosphate as a coenzyme were clarified.
1. Sulfur atoms of iron-sulfur clusters are supplied from L-cysteine by the sulfur-elimination reaction catalyzed by cysteine desulfurases. We studied the detailed mechanisms of iron-sulfur cluster assembly involving the following cysteine desulfurases : IscS and SufS from Escherichia coli and SsCsd3 from Synechocystis sp. PCC6803. Interactions between these cysteine desulfurases and proteins functioning as scaffold proteins for the assembly of iron-sulfur clusters were analyzed.
2. We isolated and characterized a novel NADPH enzyme, N-methyl-L-amino acid dehydrogenase. The gene coding for this enzyme was cloned from Pseudomonas putida ATCC12633, and its overexpression system was constructed. Using this enzyme, we developed an enzymatic process in which chiral N-methyl-L-amino acids are produced from α-keto acids and methylamine.
3. We isolated and characterized a novel NADPH enzyme that ctalyzes the asymmetric reduction of a carbon-carbon double bond in halogenated unsaturated compounds. We developed an enzymatic process in which (S)-2-CPA, which is used as a building block for the synthesis of aryloxyphenoxypropionic acid herbicide, is produced by asymmetric reduction of a carbon-carbon double bond in 2-chloroacrylate.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (123 results)

All Other

All Publications (123 results)

  • [Publications] A.Watanabe et al.: "Stereochemistry of hydrogen abstraction from pyridoxamine phosphate catalyzed by alanine racemase of Bacillus stearothermophilus"J.Mol.Catal.B.Enzymatic. 12. 145-150 (2001)

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  • [Publications] T.Uo et al.: "Purification and characterization of alanine racemase from hepatopancreas of black-tiger prawn, Penaeus monodon"J.Mol.Catal.B : Enzymatic. 12. 137-144 (2001)

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  • [Publications] T.Uo et al.: "Functional characterization of alanine racemase from Schizosaccharomyces pombe : a eucaryotic counterpart to bacterial alanine racemase"J.Bacteriol.. 183. 2226-2233 (2001)

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  • [Publications] T.Suzuki et al.: "Cold-active lipolytic activity of psychrotrophic Acinetobacter sp. strain no.6"J.Biosci.Bioeng.. 92. 144-148 (2001)

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  • [Publications] K.Soda et al.: "Stereospecificity for the hydrogen transfer of pyridoxal enzyme reactions"The Chemical Record. 1. 373-384 (2001)

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  • [Publications] S.Kato et al.: "Screening and identification of a protein interacting with IscU"微量栄養素研究. 18. 105-109 (2001)

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  • [Publications] M.Kashiwa et al.: "Removal of soluble selenium by a selenate-reducing bacterium Bacillus sp. SF-1"BioFactors. 14. 261-265 (2001)

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  • [Publications] H.Mihara et al.: "Role of cysteine desulfurase in the biosynthesis of formate dehydrogenase H containing Mo, molybdopterin, selenocysteine, and Fe_4S_4 cluster"Biomed.Res.Trace.Elem.. 12. 261-262 (2001)

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  • [Publications] A.Lewandowicz et al.: "Chlorine kinetic isotope effect on fluoroacetate dehalogenase reaction"J.Am.Chem.Soc.. 123. 9192-9193 (2001)

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  • [Publications] K.Yokoigawa et al.: "Structure and function of psychrophilic alanine racemase"J.Mot.Catal.B : enzymatic. 12. 27-35 (2001)

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  • [Publications] A.Watanabe et al.: "Reaction mechanism of alanine racemase from Bacillus stearothermophilus : X-ray crystallographic studies of the enzyme bound with N-(5'-phosphopyridoxyl)-alanine"J.Biol.Chem.. 277. 19166-19172 (2002)

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  • [Publications] T.Uo et al.: "Gene cloning, purification and characterization of 2,3-diaminopropionate ammonia-lyase from Escherichia col"Biosci.Biotechnol.Biochem.. 66. 2639-2644 (2002)

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  • [Publications] U.Tokumoto et al.: "Network of protein-protein interactions among iron-sulfur cluster assembly proteins in Eschrichia col"J.Biochem.. 131. 713-719 (2002)

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  • [Publications] T.Suzuki et al.: "A cold-active esterase with a substrate preference for vinyl esters from a psychrotroph, Acinetobacter sp.strain no.6 : gene cloning, purification, and characterization"J.Mol.Catal.B : Enzymatic. 16. 255-263 (2002)

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  • [Publications] T.Suzuki et al.: "Primary structure and catalytic properties of a cold-active esterase from a psychrotroph, Acinetobacter sp.strain no.6 isolated from Siberian soil"Biosci.Biotechnol.Biochem.. 66. 1682-1690 (2002)

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  • [Publications] S.Kato et al.: "Cys328 of IscS and Cys63 of IscU are the sites of disulfide bridge formation in a covalently-bound IscU/IscS complex : Implications for the mechanism of iron-sulfur cluster assembly"Proc.Natl.Acad.Sci.. 99. 5948-5952 (2002)

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  • [Publications] E.Pilon-Smits et al.: "Characterization of a NifS-like chloroplast protein from Arabidopsis. Implications for its role in sulfur and selenium metabolism"Plant Physiol.. 130. 1309-1318 (2002)

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  • [Publications] H.Bae et al.: "Application of a thermostable glutamate racemase from Bacillus sp. SK-1 for the production of D-phenylalanine in a multi-enzyme system"J Mol Catal B : Enzymatic. 17. 223-233 (2002)

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  • [Publications] S.Ichiyama et al.: "Catalysis-linked inactivation fluoroacetate dehalogenase by ammonia : A novel approach to the active-site environment"J.Biochem.. 131. 671-677 (2002)

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  • [Publications] T.A.Muranova et al.: "Crystallization and preliminary X-ray analysis of substrate complexes of leucine dehydrogenase from Thermoactinomyces intermedius"Acta Cryst.. D58. 1059-1062 (2002)

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  • [Publications] H.Mihara, N.Esaki: "Bacterial cysteine desulfurases : their function and mechanisms"Appl.Microbiol.Biotechnol.. 60. 12-23 (2002)

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  • [Publications] H.Mihara, N.Esaki: "Selenocysteine lyase from mouse liver"Methods in Enzymology. 347. 198-203 (2002)

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  • [Publications] H.Mihara et al.: "The iscS gene is essential for the biosynthesis of 2-selenouridine in tRNA and the selenocysteine containing formate dehydrogenase H"Proc.Natl.Acad.Sci.. 99. 6679-6683 (2002)

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  • [Publications] H.Mihara et al.: "Structure of external aldimine of Escherichia coli CsdB, an IscS/NifS homolog implications for its specificity toward selenocysteine"J.Biochem.. 131. 679-685 (2003)

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  • [Publications] K.Yoshimune et al.: "Hsc62, Hsc56, and GrpE, the third Hsp70 chaperone system of E.coli"Biochem.Biophys.Res.Commun.. 293. 1389-1395 (2002)

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  • [Publications] M.Kwak et al.: "Protein interaction between selenocysteine lyase and MUP-I"Biomedical Research on Trace Elements. 13. 238-239 (2002)

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  • [Publications] E.Kuwana et al.: "Effects of selenium deficiency on the expression of selenoprotein mRNAs in mouse brain"BiOMEUICAL RESEARCH ON TRACE ELEMETS. 14. 293-296 (2003)

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  • [Publications] Y.Wei et al.: "A novel esterase from a psychrotrophic bacterium, Acinetobacter sp. strain no.6, that belongs to the amidase signature family"J.Mol.Catal.B : Enzymatic. 23. 357-365 (2003)

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  • [Publications] T.Kurihara et al.: "Purification, characterization, and gene cloning of a novel fluoroacetate dehalogenase from Burkholderia sp. FA1"J.Mol.Catal.B : Enzymatic. 23. 347-355 (2003)

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  • [Publications] T.Kurihara et al.: "Assembly of iron-sulfur clusters mediated by cysteine desulfurases, IscS, CsdB and CSD, from Escherichia coil"Biochimica et Biophysica Acta. 1647. 303-309 (2003)

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  • [Publications] L.Kulakova et al.: "Improvement of thermostability of cold-active serine alkaline protease from the psychrotrophic bacterium Shewanella sp.strain Ac10 by rational mutagenesis"Journal of Molecular Catalysis B : Enzymatic. 22. 113-117 (2003)

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  • [Publications] T.Suzuki et al.: "Cloning, heterologous expression, renaturation, and characterization of a cold-adapted esterase with unique primary structure from a psychrotroph Pseudomonas sp.strain B11-1"Protein Expression and Purification. 30. 171-178 (2003)

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  • [Publications] S.Kato et al.: "Purification and characterization of alanine dehydrogenase from a marine bacterium, Vibrio proteolyticus"J.Mol.Catal.B : Enzymatic. 23. 373-378 (2003)

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  • [Publications] M.Pilon et al.: "Enhanced selenium tolerance and accumulation in transgenic Arabidopsis expressing a mouse selenocysteine lyase"Plant Physiol.. 131. 1250-1257 (2003)

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  • [Publications] C.Park et al.: "A new DL-2-haloacid dehalogenase acting on 2-holoacid amides : Purification, characterization, and mechanism"J.Mol.Catal.B : Enzymatic. 23. 329-336 (2003)

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  • [Publications] A.Galkin et al.: "Cold-active enzymes from cold-adapted microorganisms : Their possible applications and structural relationships with mesophilic and thermophilic counterparts"Recent Advances in Marine Biotechnology. 8. 1-27 (2003)

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  • [Publications] G.Yow et al.: "Conversion of the catalytic specificity of alanine racemase to a D-amino acid aminotransferase activity by a double active-site mutation"J.Mol.Catal.B : Enzymatic. 23. 311-319 (2003)

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  • [Publications] T.Yoshimura, N.Esaki: "Amino acid racemases : Functions and mechanisms"Journal of Bioscience and Bioengineering. 96. 103-109 (2003)

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  • [Publications] Y.Li et al.: "Mass spectrometric analysis of the reactions catalyzed by L-2-haloacid dehalogenase mutants and implications for the roles of the catalytic amino acid residues"J.Mol.Catal.B : Enzymatic. 23. 337-345 (2003)

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  • [Publications] Y.Fuchikami et al.: "D-Amino acid aminotransferase: Fragmentation at a flexible loop is an efficient metond to generate mutant enzymes with new substrate specificities and elevated activities"J.Mol.Catal.B : Enzymatic. 23. 321-328 (2003)

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  • [Publications] M.Kwak et al.: "A novel regulatory function of selenocysteine lyase, a unique catalyst to modulate major urinary protein"J.Mol.Catal.B : Enzymatic. 23. 367-372 (2003)

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  • [Publications] H.Yamamoto et al.: "A novel NADH-dependent carbonyl reductase from Kluyveromyces aestuarii and comparison of NADH-regeneration system for the synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate"Biosci.Biotechnol.Biochem.. 68. 638-649 (2004)

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  • [Publications] L.Kulakova et al.: "Cold-active esterase from Psychrobacter sp. Ant300: gene cloning, characterization and the effects of Gly->Pro substitution near the active site on its catalytic activity and stability"Biochimica et Biophysica Acta. 1696. 59-65 (2004)

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  • [Publications] T.Takakura et al.: "Assay method for antitumor L-methionine γ-lyase : comprehensive kinetic analysis of the complex reaction with L-methionine"Analytical Biochemistry. 327. 233-340 (2004)

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  • [Publications] S.Ichiyama et al.: "Reactivity of asparagine residue at the active site of the D105N mutant of fluoroacetate dehalogenase from Moraxella sp.B"Biochimica et Biophysica Acta. 1698. 27-36 (2004)

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  • [Publications] W.Guo, J.Hiratake, K.Ogawa, M.Yamamoto, S.-J.Ma, K.Sakata: "β-Glycosylamidines : potent, selective, and easily accessible β-glycosidase inhibitors"Bioorg.Med.Chem.Lett.. 11. 467-470 (2001)

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  • [Publications] S.K.Boehlein, T.Nakatsu, J.Hiratake, R.Thirumoorthy, J.D.Stewart, N.G.J.Richards, S.M.Schuster: "Characterization of inhibitors acting at the synthetase site of Escherichia coli asparagine synthetase B"Biochemistry. 40. 11168-11175 (2001)

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  • [Publications] 藤井亮太, 平竹 潤: "リパーゼの進化分子工学"酵素工学ニユース (Enzyme Engineering News). 46. 22-27 (2001)

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  • [Publications] S.-J.Ma, J.Hiratake, Y.Nakai, M.Izumi, K.Fukase, S.Kusumoto, K.Sakata: "A direct continuous spectrophotometric assay for glycosidases with 3-nitro2-pyridyl glycosides by tautomerization of 2-hydroxy-3-nitropyridine"Anal.Biochem.. 302. 291-297 (2002)

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  • [Publications] J.Hiratake, M.Inoue, K.Sakata: "γ-Glutamyltranspeptidase and γ-glutamyl peptide ligases : Use of fluorophosphonate and phosphonodifluoromethyl ketone analogues as probes of the tetrahedral transition state and the γ-glutamyl-P intermediate"Methods in Enzymology. 354. 272-295 (2002)

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  • [Publications] J.Hiratake, T.Irie, N.Tokutake, J.Oda: "Recognition of a cysteine substrate by E.coli γ-glutamylcysteine synthetase probed by sulfoximine-based transition-state analogue inhibitors"Biosci.Biotechnol.Biochem.. 66. 1500-1514 (2002)

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  • [Publications] J.Hiratake: "Rational design and synthesis of enzyme inhibitors : Significance in probing the function of enzymes"J.of Synthetic Organic Chemistry, Special Issue in English. 60. 1095-1103 (2002)

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  • [Publications] R.Fujii, Y.Utsunomiya, J.Hiratake, A.Sogabe, K.Sakata: "Highly sensitive active-site titration of lipase in microscale culture media using fluorescent organophosphorus ester"Biochim.Biophys.Acta (Molecular and Cell Biology of Lipids). 1631. 197-205 (2003)

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  • [Publications] J.Hiratake, K.Sakata: "Glycosylamidines as potent selective and easily accessible glycosidase inhibitors and their application to affinity chromatography"Methods in Enzymology. 363. 421-444 (2003)

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  • [Publications] K.Inoue, J.Hiratake, M.Mizutani, M.Takada, M.Yamamoto, K.Sakata: "β-Glycosylamidine as ligand for affinity chromatography tailored to glycon substrate specificity of β-glycosidases"Carbohydr.Res.. 338. 1477-1491 (2003)

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  • [Publications] 平竹 潤: "遷移状態アナログを用いて酵素の構造と機能をさぐる"生化学. 74. 38-42 (2002)

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  • [Publications] 平竹 潤, 坂田完三: "選択的β-グリコシダーゼ阻害剤の分子設計と応用"化学と生物. 40. 352-354 (2002)

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  • [Publications] 平竹 潤: "研究ツールとしてのグリコシダーゼ阻害剤-その設計と応用-"日本農芸化学会誌. 77. 409-412 (2003)

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  • [Publications] 平竹 潤, 坂田完三: "新規グリコシダーゼ阻害剤 β-グリコシルアミジン誘導体の開発ど応用"BIOINDUSTRY. 8. 44-51 (2003)

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  • [Publications] 江崎信芳 他: "生化学基礎の基礎"化学同人. 326 (2002)

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  • [Publications] N.Esaki et al.: "Isomerizations. Enzyme Catalysis in Organic Synthesis second edition (Edited by K Drauz and H.Waldmann)"VCH. 53 (2002)

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  • [Publications] J.Hiratake, H.Suzuki, H.Kumagai: "Glutamyl transpeptidase and its precursor. in Handbook of Proteolytic Enzymes, 2nd edition"Academic Press (In press). (2004)

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  • [Publications] A.Watanabe, T.Yoshimura, Y.H.Lim, Y.Kurokawa, K.Soda, N.Esaki: "Stereochemistry of hydrogen abstraction from pyridoxamine phosphate catalyzed by alanine racemase of Bacillus stearothermophilus."J. Mol. Catal. B : Enzymatic. 12. 145-150 (2001)

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      2003 Final Research Report Summary
  • [Publications] T.Uo, T.Ueda, T.Nishiyama, T.Yoshimura, N.Esaki: "Purification and characterization of alanine racemase from hepatopancreas of black-tiger prawn, Penaeus monodon"J. Mol. Catal. B : Enzymatic. 12. 137-144 (2001)

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    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Uo, T.Yoshimura, N.Tanaka, K.Takegawa, N.Esaki: "Functional characterization of alanine racemase from Schizosaccharomyces pombe : a eucaryotic counterpart to bacterial alanine racemase."J. Bacteriol.. 183. 2226-2233 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Suzuki, T.Nakayama, T.Kurihara, T.Nishino, N.Esaki: "Cold-active lipolytic activiy of psychrotrophic Acinetobacter sp.strain no. 6."J. Biosci. Bioeng.. 92. 144-148 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] K.Soda, T.Yoshimura, N.Esaki: "Stereospecificity for the hydrogen transfer of pyridoxal enzyme reactions."The Chemical Record. 1. 373-384 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] M.Kashiwa, M.Ike, H.Mihara, N.Esaki, M.Fujita: "Removal of soluble selenium by a selenate-reducing bacterium Bacillus sp. SF-1."BioFactors. 14. 261-265 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] H.Mihara, S.Kato, T.Kurihara, T.Yoshimura, N.Esaki, G.M.Lacourdiere, T.C.Stadtman: "Role of cysteine desulfurase in the biosynthesis of formate dehydrogenase H containing Mo, molybdopterin, selenocysteine, and Fe_4S_4 cluster."Biomed. Res. Trace. Elem.. 12. 261-262 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] A.Lewandowicz, D.Sicinska, J.Rudzinski, S.Ichiyama, T.Kurihara, N.Esaki, P.Paneth: "Chlorine kinetic isotope effect on fluoroacetate dehalogenase reaction."J. Am. Chem. Soc.. 123. 9192-9193 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] K.Yokoigawa, Y.Okubo, H.Kawai, N.Esaki, K.Soda: "Structure and function of psychrophilic alanine racemase."J. Mol. Catal. B : Enzymatic. 12. 27-35 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] A.Watanabe, T.Yoshimura, B.Mikami, H.Hayashi, H.Kagamiyama, N.Esaki: "Reaction mechanism of alanine racemase from Bacillus stearothermophilus : X-ray crystallographic studies of the enzyme bound with N-(5'-phosphopyridoxyl)-alanine."J. Biol. Chem.. 277. 19166-19172 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Uo, T.Yoshimura, T.Nishiyama, N.Esaki: "Gene cloning, purification and characterization of 2,3-diaminopropionate ammonia-B from Escherichia coli."Biosci. Biotechnol. Biochem.. 66. 2639-2644 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] U.Tokumoto, S.Nomura, Y.Minami, H.Mihara, S.Kato, T.Kurihara, N.Esaki, H.Kanazawa, H.Matsubara, Y.Takahashi: "Network of protein-protein interactions among iron-sulfur cluster assembly proteins in Eschrichia coli."J. Biochem.. 131. 713-719 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Suzuki, T.Nakayama, T.Kurihara, T.Nishino.N.Esaki: "A cold-active esterase with a substrate preference for vinyl esters from a psychrotroph, Acinetobacter sp. strain no. 6 : gene cloning, purification, and characterization."J. Mol. Catal. B : Enzymatic. 16. 255-263 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Suzuki, T.Nakayama, T.Kurihara, T.Nishino.N.Esaki: "Primary structure and catalytic properties of a coldactive esterase from a psychrotroph, Acinetobacter sp. strain no. 6 isolated from Siberian soil."Biosci. Biotechnol. Biochem.. 66. 1682-1690 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] S.Kato, H.Mihara, T.Kurihara, Y.Takahashi, U.Tokumoto, T.Yoshimura N.Esaki: "Cys328 of IscS and Cys63 of IscU are the sites of disulfide bridge formation in a covalently-bound IscU/IscS complex : Implications for the mechanism of iron-sulfur cluster assembly."Proc. Natl. Acad. Sci.. 99. 5948-5952 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] E.Pilon-Smits, G.Garifullina, S.Abdel-Ghany, S.Kato, H.Mihara, K.Hale, J.Burkhead, N.Esaki, T.Kurihara, M.Pilon: "Characterization of a NifS-like chloroplast protein from Arabidopsis. Implications for its role in sulfur and selenium metabolism."Plant Physiol.. 130. 1309-1318 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] H.Bae, S.Hong, S.Lee, M.Kwak, N.Esaki, M.Sung: "Application of a thermostable glutamate racemase from Bacillus sp. SK-1 for the production of D-phenylalanine in a multi-enzyme system."J. Mol. Catal B : Enzymatic. 17. 223-233 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] S.Ichiyama, T.Kurihara, M.Miyagi, A.Galkin, S.Tsunasawa, H.Kawasaki, N.Esaki: "Catalysis-linked inactivation fluoroacetate dehalogenase by ammonia A novel approach to the active-site enviroment."J. Biochem.. 131. 671-677 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.A.Muranova, S.N.Ruzheinikov, S.E.Sedelnikova, P.J.Baker, A.Pasquo, A.Galkin, N.Esaki, T.Ohshima, K.Soda, D.W.Rice: "Crystallization and preliminary X-ray analysis of substrate complexes of leucine dehydrogenase from Thermoactinomyces intermedius."Acta Cryst.. D58. 1059-1062 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] H.Mihara, N.Esaki: "Bacterial cysteine desulfurases : their function and mechanisms."Appl. Microbiol Biotechnol.. 60. 12-23 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] H.Mihara, N.Esaki: "Selenocysteine lyase from mouse liver."Methods in Enzymology. 347. 198-203 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] H.Mihara, S.Kato, G.M.Lacourciere, T.C.Standman, R.A.J.D.Kennedy, T.Kurihara, U.Tokumoto, Y.Takahashi, N.Esaki: "The iscS gene is essential for the biosynthesis of 2-selenouridine in tRNA and the selenocysteine containing formate dehydrogenase H."Proc. Natl. Acad. Sci.. 99. 6679-6683 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] H.Mihara, T.Fujii, S.Kato, T.Kurihara, Y.Hata, N.Esaki: "Structure of external aldimine of Escherichia coli CsdB, an IscS/NifS homolog implications for its specificity toward selenocysteine."J. Biochem.. 131. 679-685 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] K.Yoshimune, T.Yoshimura, T.Nakayama, T.Nishino, N.Esaki: "Hsc62, Hsc56, and GrpE, the third Hsp70 chaperone system of E. coli."Biochem. Biophys. Res. Commune.. 293. 1389-1395 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] M.Kwak, H.Mihara, N.Esaki: "Protein interaction between selenocysteine lyase and MUP-1."Biomedical Research on Trace Elements. 13. 238-239 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] E.Kuwana, S.Kurokawa, H.Mihara, T.Kurihara, T.Yoshimura, N.Esaki: "Effects of selenium deficiency on the expression of selenoprotein mRNAs in mouse brain."BiOMEDICAL RESEARCH ON TRACE ELEMENTS. 14. 293-296 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Wei.T.Kurihara, T.Suzuki, N.Esaki: "A novel esterase from a psychrotrophic bacterium, Acinetobacter sp. strain no. 6, that belongs to the arnidase signature family."J. Mol. Catal. B : Enzymatic. 23. 357-365 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Kurihara, T.Yamauchi, S.Ichiyama, H.Takahata, N.Esaki: "Purification, characterization, and gene cloning of a novel fluoroacetate dehalogenase from Burkholderia sp. FA1."J. Mol. Catal. B : Enzymatic. 23. 347-355 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Kurihara, H.Mihara, S.Kato, T.Yoshimura, N.Esaki: "Assembly of iron-sulfur clusters mediated by cysteine desulfurases, IscS, CscS, CsdB and CSD, from Escherichia coil."Biochimica et Biophysica Acta. 1647. 303-309 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] L.Kulakova, A.Galkin, T.Nakayama, T.Nishino, N.Esaki: "Improvement of thermostability of cold-active serine alkaline protease from the psychrotrophic bacterium Shewanella sp. strain Ac10 by rational mutagenesis."Journal of Molecular Catalysis B : Enzymatic. 22. 113-117 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Suzuki, T.Nakayama, D.W.Choo, Y.Hirano, T.Kurihara, T.Nishino, N.Esaki: "Cloning, heterologous expression, renaturation, and characterization of a cold-adapted esterase with unique primary structure from a psychrotroph Pseudomonas sp. strain B11-1."Protein Expression and Purification. 30. 171-178 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] S.Kato, T.Ohshima, A.Galkin, L.Kulakova, T.Yoshimura, N.Esaki: "Purification and characterization of alanine dehydrogenase from a marine bacterium, Vibrio proteolyticus."J. Mol. Catal. B : Enzymatic. 23. 373-378 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Pilon, J.D.Owen, G.F.Garifullina, T.Kurihara, H.Mihara, N.Esaki, E.A.H.Pilon-Smits: "Enhanced selenium tolerance and accumulation in transgenic Arabidopsis expressing a mouse selenocysteine lyase."Plant Physiol.. 131. 1250-1257 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] C.Park, T.Kurihara, T.Yoshimura, K.Soda, N.Esaki: "A new DL-2-haloacid dehalogenase acting on 2-holoacid amides : Purification, characterization, and mechanism."J. Mol. Catal. B : Enzymatic. 23. 329-336 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] A.Galkin, L.Kulakova, T.Nakayama, T.Nishino, N.Esaki: "Cold-active enzymes from cold-adapted microorganisms : Their possible applications and structural relationships with mesophilic and thermophilic counterparts."Recent Advances in Marine Biotechnology. 8. 1-27 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] G.Yow, A.Watanabe, T.Yoshimura, N.Esaki: "Conversion of the catalytic specificity of alanine racemase to a D-amino acid aminotransferase activity by a double active-site mutation."J. Mol. Catal. B : Enzymatic. 23. 311-319 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Yoshimura, N.Esaki: "Amino acid racemases : Functions and mechanisms."Journal of Bioscience and Bioengineering. 96. 103-109 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] Y.Li, T.Kurihara, S.Ichiyama, M.Miyagi, S.Tsunasawa, N.Esaki: "Mass spectrometric analysis of the reactions catalyzed by L-2-haloacid dehalogenase mutants and implications for the roles of the catalytic amino acid residues."J. Mol. Catal. B : Enzymatic. 23. 337-345 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] Y.Fuchikami, T.Yoshimura, N.Esaki: "D-Amino acid aminotransferase : Fragmentation at a flexible loop is an efficient metond to generate mutant enzymes with new substrate specificities and elevated activities."J. Mol. Catal. B : Enzymatic. 23. 321-328 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Kwak, H.Mihara, N.Esaki: "A novel regulatory function of selenocysteine lyase, a unique catalyst to modulate major urinary protein."J. Mol. Catal. B : Enzymatic. 23. 367-372 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] H.Yamamoto, K.Mitsuhashi, N.Kimoto, A.Matsuyama, N.Esaki, Y.Kobayashi: "A novel NADH-dependent carbonyl reductase from Kluyveromyces aestuarii and comparison of NADH-regeneration system for the synthesis of ethyl (S)-4-chloro-3-hydroxybutanoate."Biosci. Biotechnol. Biochem.. 68. 638-649 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] L.Kulakova, A.Galkin, T.Nakayama, T.Nishino, N.Esaki: "Cold-active esterase from Psychrobacter sp. Ant300 : gene cloning, characterization and the effects of Gly->Pro substitution near the active site on its catalytic activity and stability."Biochimica et Biophysica Acta. 1696. 59-65 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] T.Takakura, K.Mitsushima, S.Yagi, K.Inagaki, H.Tanaka, N.Esaki, K.Soda, A.Takimoto: "Assay method for antitumor L-methionine g-lyase : comprehensive kinetic analysis of the complex reaction with L-methionine."Analytical Biochemistry. 327. 233-240 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] S.Ichiyama, T.Kurihara, Y.Kogure, S.Tsunasawa, H.Kawasaki, N.Esaki: "Reactivity of asparagine residue at the active site of the D105N mutant of fluoroacetate dehalogenase from Moraxella sp. B."Biochimica et Biophysica Acta. 1698. 27-36 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] W.Guo, J.Hiratake, K.Ogawa, M.Yamamoto, S.-J.Ma, K.Sakata: "β-Glycosylamidines : potent, selective, and easily accessible-β-glycosidase inhibitors."Bioorg. Med. Chem. Lett.. 11. 467-470 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] S.K.Boehlein, T.Nakatsu, J.Hiratake, R.Thirumoorthy, J.D.Stewart, N.G.J.Richards, S.M.Schuster: "Characterization of ihibitors acting at the synthetase site of Escherichia coli asparagine synthetase B."Biochemistry. 40. 11168-11175 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] S.-J.Ma, J.Hiratake, Y.Nakai, M.Izumi, K.Fukase, S.Kusumoto, K.Sakata: "A direct continuous spectrophotometric assay for glycosidases with 3-nitro2-pyridyl glycosides by tautomerization of 2-hydroxy-3-nitropyridine."Anal. Biochem.. 302. 291-297 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] J.Hiratake, M.Inoue, K.Sakata: "γ-Glutamyltranspeptidase and γ-glutarnyl peptide ligases : Use of fluorophosphonate and phosphonodifluoromethyl ketone analogues as probes of the tetrahedral transition state and the γ-glutamyl-P intermediate."Methods in Enzymology. 354. 272-295 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] J.Hiratake, T.Irie, N.Tokutake, J.Oda: "Recognition of a cysteine substrate by E. coli γ-glutamylcysteine synthetase probed by sulfoximine-based transition-state analogue inhibitors."Biosci. Biotechnol. Biochem.. 66. 1500-1514 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] J.Hiratake: "Rational design and synthesis of enzyme inhibitors : Significance in probing the function of enzymes."J. of Synthetic Organic Chemistry, Special Issue in English. 60. 1095-1103 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] R.Fujii, Y.Utsunomiya, J.Hiratake, A.Sogabe, K.Sakata: "Highly sensitive active-site titration of lipase in microscale culture media using fluorescent organophosphorus ester."Biochim. Biophys. Acta (Molecular and Cell Biology of Lipids). 1631. 197-205 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] J.Hiratake, K.Sakata: "Glycosylamidines as potent selective and easily accessible glycosidase inhibitors and their application to affinity chromatography."Methods in Enzymology. 363. 421-444 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] K.Inoue, J.Hiratake, M.Mizutani, M.Takada, M.Yamamoto, K.Sakata: "β-Glycosylamidine as ligand for affinity chromatography tailored to glycon substrate specificity of β-glycosidases."Carbohydr. Res.. 338. 1477-1491 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
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      2003 Final Research Report Summary
  • [Publications] Tatsuo Kurihara et al.: "Assembly of iron-sulfur clusters mediated by cysteine desulfurases, JscS, CsdB and CSD, from Escherichia coil"Biochimica et Biophysica Acta. 1647. 113-117 (2003)

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      2003 Annual Research Report
  • [Publications] Eriko Kuwana et al.: "Effects of Selenium Deficiency on the Expression of Selenoprotein mRNAs in Mouse Brain"BiOMEDICAL RESEARCH ON TRACE ELEMENTS. 14・4. 293-296 (2003)

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      2003 Annual Research Report
  • [Publications] Marinus Pilon et al.: "Enhanced Selenium Tolerance and Accumulation in Transgenic Arabidopsis Expressing a Mouse Selenocysteine Lyase."Plant Physiol"Plant Physiol.. 131. 1250-1257 (2003)

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      2003 Annual Research Report
  • [Publications] Mi-Sun Kwak et al.: "A novel regulatory function of selenocysteine lyase, a unique catalyst to modulate major urinary protein"Mol.Catal.B : Enzymatic. 23. 367-372 (2003)

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      2003 Annual Research Report
  • [Publications] Mihara, H.: "Bacterial cysteine desulfurases : their function and mechanisms"Appl.Microbiol.Biotechnol.. 60. 12-23 (2002)

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      2002 Annual Research Report
  • [Publications] Mihara, H.: "The iseS gene is essential for the biosynthesis of 2-selenouridine in tRNA and the selenocysteine-containing formatedehydrogenase H"Proc.Natl.Acad.Sci., USA. 99. 6679-6683 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Mihara, H.: "Structure of external aldimine of Escherichia coli CsdB, an IscS/NifS homolog : implications for its specificity towardselenocysteine"J.Biochem.. 131. 679-685

    • Related Report
      2002 Annual Research Report
  • [Publications] Kato, S.: "Cys328 of IscS and Cys63 of IscU are the Sites of DisulfideBridge Formation in a Covalently-bound IscS/IscU Complex : Implication for the Mecbanism of lron-Sulfur Cluster Assembly"Proc. Natl. Acad. Sci., USA. (印刷中).

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      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2018-03-28  

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