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The key reaction for carbon dioxide fixation in reductive TCA cycle

Research Project

Project/Area Number 08660094
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionThe University of Tokyo

Principal Investigator

ISHII Masaharu  The University of Tokyo, Grad.Sch of Agei.Life Sci.Department of Biotechnology, Associate professor, 大学院・農学生命科学研究科, 助教授 (30193262)

Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordsreductive TCA / antotroph / cycle / フェレドキシン / 独立栄養細菌 / 炭酸固定 / 炭酸ガス
Research Abstract

In order to clarify that reductive TCA cycle is operative in a thermophilic hydrogenxidizing bacterium, Hydrogenobater thermophilus, the followings were demonstrated.
1.Purification and characterization of ferredoxin
Purification of ferredoxin was performed by applying cell-free extract successively. to Q-Sepharose, DEAE-5PW,and Superdex 75. Characterization of the prorein was also perfprmed.
2.Purification and characterization of pyruvate synthase
Purification of pyruvate synthase was performed by applying cell-free extract successively to DEAE-Sepharose CL-6B,PA-QA,Hydroxyapatite, and Superdex 200. Characterization of the protein was also performed.
3.Reactivity of ferredoxin toward 2-oxoglutarate synthase or pyruvate synthase
In the first step, reaction was performed toward CO_2 evolution direction ; both enzymes reacted with ferredoxin. In the next step, reduced ferredoxin was used as a reductant to show the carboxylation reaction. However, only a little carboxylation reaction occurred probably due to the instability of CoA-compounds. Finally, CoA-compounds were produced through enzymatic reactions ( ATP : citrate lyase and succinyl-CoA synthetase ) . By this system carboxylation reaction was detected and also the expected products were identified.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Masaharu Ishii: "Purification and characterization of ferredokin from Hydrogenobacter thermophilus strain TK-6" Biosci.Biotech.Biochem.60・9. 1513-1515 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ki-Seok Yoon: "Carboxylation reactions of pyruvate:ferredoxin oxidoveductase and 2-oxoglutarate:ferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6" Biosci.Biotech.Biochem.61・3. 510-513 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ki-Seok Yoon: "Purification and characterization of pyruvute:ferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6" Arch.Microbiol. 167・2. 275-279 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Masaharu Zshii, Yasufumi Ueda, Ki-Seok Yoon Yasuo Igarashi and Tohru Kodama: "Purification and characterization of ferredoxin from Hydyogenobacter thermophilus strain TK-6" Biosci. Biotech. Biochem.60-9. 1513-1515 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ki-Seok Yoon, Masaharu Ishii, Tohru Kodama and Yasuo Igarashi: "Corboxylation reactions of pyruvate : ferredoxin oxidoreductase and 2-oxoglutarate : ferredoxin oxido reductase from Hydrogenobacter thermophilus TK-6" Biosci. Biotech. Biochem.61-3. 510-513 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ki-Seok Yoon, Masaharu Ishii, Tohru Kodama and Yasuo Igarashi: "Purification and characterization of pyruvate : ferredoxin oxidoreductase from Hydrogenobacter theromphilus TK-6" Arch. Microbiol.167-2. 275-279 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ki-Seok Yoon ら: "Carboxylation Reactions of Pyruvate:Ferredoxin Oxidoreductase and 2-Oxogluterate Ferredoxin Oridoreductase from Hydrogenobacter thermophilus TK-6" Biosci.Biotech.Biochem.61・3. 510-513 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ki-Seok Yoon: "Purification and characterization of pyruvate:ferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6" Arch.Microbiol.167・2. 275-279 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Masaharu Ishii ら: "Purification and characterization of Ferredoxin from Hydrogenobacter thermophilus Strain TK-6" Biosci.Biotech.Biochem.60・9. 1513-1515 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Masaharu Ishii: "Purification and characterization of ferredoxin from Hydrogenobacter thermophilus strain TK-6" Biosci.Biotech,Biochem.60・9. 1513-1515 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ki-Seok Yoon: "Purification and characterization of pyruvate:ferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6" Arch.Microbiol.(in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Ki-Seok Yoon: "Carboxylation reactions of pyruvate:ferredoxin oxidoreductase and 2-oxoglutarate iferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6" Biosci.Biotech.Biochem.(in press).

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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