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Development of New Methods on Isotope-Labeling of Amino Acid and Vitamin with Microbial Enzymes

Research Project

Project/Area Number 63880021
Research Category

Grant-in-Aid for Developmental Scientific Research (B).

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionKyoto University

Principal Investigator

SODA Kenji  Kyoto University, Institute for Chemical Research, Professor, 化学研究所, 教授 (30027023)

Co-Investigator(Kenkyū-buntansha) TOMITA Kousuke  Unitica Co. Ltd., Director, 中央研究所, 部長
YOSHIMURA Tohru  Kyoto Univ., Inst. for Chem. Res., Assistant Prof., 化学研究所, 助手 (70182821)
TANIZAWA Katsuyuki  Osaka Univ., Inst. Sci. Indus. Res., Associate Prof., 産業科学研究所, 助教授 (20133134)
ESAKI Nobuyoshi  Kyoto Univ., Inst. for Chem. Res., Associate Prof., 化学研究所, 助教授 (50135597)
TANAKA Hidehiko  Okayama Univ., Fac. of Agric., Prof., 農学部, 教授 (90065912)
平沢 敏子  京都大学, 化学研究所, 教務職員
冨田 耕右  ユニチカ株式会社, 中央研究所, 部長
Project Period (FY) 1988 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1989: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1988: ¥6,000,000 (Direct Cost: ¥6,000,000)
KeywordsAmino acid / Vitamin / Microbial enzyme / Isotope-labeling / 同位元素標識法 / 同位体元素標識法
Research Abstract

New methods on isotope-labeling of amino acid and vitamin were established with microbial enzymes, which have high stereo-specificity of reaction. First, or ^<35>S- labeling ofS-alkylcysteine was established by beta -replacement reaction of S-alkylcysleine alpha, beta -lyase, which was newly purified from a thermophile (Bacillus sp. 41A). Second, deuterium or tritium-labeling of D-amino acid was developed Stereoselectively with D-amino acid aminotransferase. Stereoselectively-deulerated NADH and NADPH were also prepared by coupling of glutamate racemase and glutamate dehydrogenase. Further, double isotope-labeling of Lysine was carried out by L-lysine epsilon -dehydrogenase and amino acid racemase. Finally. Substitution of S- (beta -aminoethyl) - cysteine for active-site lysine of thermostable aspartate aminotransferase was established by gene technology and chemical modification. Thus, many effective isotope-labeling of amino acid, vitamin and protein were developed.

Report

(4 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • 1988 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] K.ヒヨウシキホウ17784-17789

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kamitani Hiroshi: "Thermostabla SーAlkylcysteine α,βーLyase from a Thermophile:Purification and Properties" Agicultural Biological Chemistry. 54. 2069-2076 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Alvaro Martinez del Pozo: "Stereospecificity of Reactions Calalyzed hy Bacterial Dーamino Acid Transaminase" The Journal of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Esaki Nobuyoshi: "Enantioselective Synthesis of DーSelenoーmethionine with Dーamino acid Aminotransfese" Biotechnology and Bioengineering. 34. 1231-1233 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Esaki Nobuyoshi: "Preparation of Steroselectivelyーdeuterated NADH and NADOH by Coupling of Glutamate Racemase and Glutamate Dehydrogenase" Biotechnology Letters. 12. 105-110 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Misono Haruo: "Sterespecific Abstraction of εーproーRーHydrogen of LーLysine by LーLysine εーDehydrogenase from Agrobaclerium tumefaciens" Journal of Biochemistry. 107. 169-172 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishiwata Kenーichi: "Enzymatic Production of LーTryptophan from DLーSerine and Indole by a Coupled Reaction of Tryptophan Synthase and Amino Acid Rasemase" Biotechnology and Applied Biochemistry. 12. 141-149 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tohru Yoshimura: "Substitution of Sー(βーAminoethyl)ーCysteine for ActiveーSite Lysine of Thermostable Aspartate Aminotransferase" Journal of Biochemistry. 108. 699-700 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nakajima Nobuyoshi: "Enzymatic in Situ Analysis by ^< 1>HーNMR of the Hydrogen Transfer Slecreospecificity of NADP^+ーDependent Dehydrogenase" Journal of Biochemistry. 106. 515-517 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Esaki Nobuyoshi: "Enzymatic in Situ Determination of Stereospecificity of NADーdependent Dehydrogenase" The Journal of Biological Chemistry. 264. 9750-9752 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] MoonーHee Sung: "Purification and Characterization of Thermostable Aspartate Aminotransferase from a Thermophilic Bacillus Species" Journal of Bacteriology. 172. 1345-1351 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hiroshi Kamitani, Nobuyoshi Esaki, Hidehiko Tanaka and Kenji Soda: "Thermostable S-Alkylcysteine alpha, beta-Lyase from a Thermophile : Purification and Properties." Agricultural Biological Chemistry. 54. 2069-2076 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Alvaro Martinez del Pozo, Marcello Merola, Hiroshi Ueno, James M. Manning, Katuyuki Tanizawa, Kenji Soda and Dagmar Ringe: "Stereospecificity of Reaction Catalyzed by Bacterial D-amino Acid Transaminase." The Journal of Biological Chemistry. 264. 17784-17789 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nobuyoshi Esaki, Hiroko Shimoi, Hidehiko Tanaka and Kenji Soda: "Enantioseleclive Synthesis of D-Seleno- Methionine with D-amino acid Aminotransferase." Biotechnology and Bioengineering. 34. 1231-1233 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nobuyoshi Esaki, Nobuyoshi Nakajima, Kaoru Nakamura, Kazuo Yonaha, Hidehiko Tanaka and Kenji Soda: "Preparation of Stereoselectively-deuterated NADH and NADOH by Coupling of glutamate Racemase and Glutamate Dehydrogenase." Biotechnology Letters. 12. 105-110 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Haruo Misono, Tohru Yoshimura, Susumu Nagasaki, and Kenji Soda: "Stereospecific Abstraction of epsilon-pro-R-Hydrogen of L-Lysine by LーLysine epsilon-Dehydrogenase from Agrobacterium tumefaciens." Journal of Biochemistry. 107. 169-172 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ken-ichi Ishiwata, Nobuhiro Fukuhara, Masao Shimada, Nobuyoshi Makiguchi, and Kenji Soda: "Enzymatic Production of L-Tryptophan from DL-Serinc and lidole by a Coupled Reaction of Tryptophan Synthase and Amino Acid Racemase." Biotechnology and Applied Biochemistry. 12. 141-149 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tohru Yoshimura, Yutaka Matushima Katsuyuki Tanizawa, Moon-Hee Sung. Takayoshi Yamauchi, Mamoru Wakayama, Nobuyoshi Esaki, and Kenji Soda: "Substitution of S-(beta-Aminoethyl)-Cysteine for Active-Site Lysine of Thermostable Aspartate Aminotransferase." Journal of Biochemistry. 108. 699-700 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Moon-Hee Sung, Katsuyuki Tanizawa, Hidehiko Tanaka, Seiki Kuramitu, Hiroyuki Kagamiyama and Kenji Soda: "Purification and Characterization of Thermostable Aspartate Aminotransferase from a Thermophilic Bacillus Species." Journal of Bacteriology. 108. 699-700 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nobuyoshi Nakajima, Kaoru Nakayama, Nobuyoshi Esaki, Hidehiko Tanaka, and Kenji Soda: "Enzymatic In Situ Analysis by of the Hydrogen Transfer Stereospecificity of NADP^+-Dependent Dehydrogenase." Journal of Biochemistry. 106. 515-517 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nobuyoshi Esaki, Hiroko Shimoi, Nobuyoshi Nakajima, Toshihisa Ohshima, Hidehiko Tanaka and Kenji Soda: "Enzymatic in Situ Determination of Stereospecificity of NAD-dependent Dehydrogenasc" The Journal of Biological Chemistry. 264. 9750-9752 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tohru Yoshimura: "Substitution of Sー(βーAminoethyl)ーCysteine for ActiveーSite Lysine of Thermostable Aspartate Aminotransferase" Journal of Biochemistry. 108. 699-700 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] MoonーHee Sung: "Purification and Characterization of Thermostable Aspartate Aminotransferase from a Thermophilic Bacillus Species" Journal of Bacteriology. 172. 1345-1351 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Nobuyoshi Nakajima: "Enzymatic In Situ Analysis by 1-H-NMR of the Hydrogen Transfer Stereospecificity of NAD(P)-Dependent Dehydrogenases." Jounal of Biochemistry. 106. 515-517 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Katsuyuki Tanizawa: "Thermostable D-Amino Acid Aminotransferase from a Termophilic Bacillus Species" Journal of Biological Chemistry. 264. 2445-2449 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Katsuyuki Tanizawa: "The Primary Structure of Thermostable D-Amino Acid Aminotransferase from a Termophilic Bacillus Species and Its Correlation with L-Amino Acid Aminotransferase" Journal of Biological Chemistry. 264. 2450-2454 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Nobuyoshi Esaki: Journal of Biological Chemistry. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Nobuyoshi Esaki: Biotechnology and Applied Biochemistry. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Nobuyoshi Esaki: Biotechnology and Bioengineering. (1989)

    • Related Report
      1988 Annual Research Report

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Published: 1988-04-01   Modified: 2016-04-21  

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