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Structure and function of hexulose phosphate synthase

Research Project

Project/Area Number 02660120
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 応用微生物学・発酵学
Research InstitutionTottori University

Principal Investigator

KATO Nobuo  Tottori University ; Faculty of Engineering ; Professor, 工学部, 教授 (50026556)

Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsHexulose phosphate synthase / Phosphohexuloisomerase / Methylotroph / ^<13>C-labeled compounds / Fructose-6-phospahte / Formaldehyde / ホスホヘキシュロイソメラ-ゼ / フルクト-スー6-リン酸 / アルド-ル縮合 / リブロ-スモノリン酸経路
Research Abstract

Hexulose phosphate synthase catalyzes aldol condensation of formaldehyde and ribulose 5-phosphate, which is a key enzyme in the fixation of C_1 compounds in methylotrophic bacteria. And it is suggested to be correlated to the CO_2-fixing enzyme, ribulose bisphospahte caboxylase, from the view point of the biological evolution.
Base on the amino acid sequences of the N-terminal region, three DNA probes were synthesized and used to detect the hexulose phosphate synthase gene. 5-kb EcoRl and 4-kb PstI fragments of Methylomonas aminofaciens 77a that is a facultative methylotroph was detected in Southern hybridization with the probes. Three fragments were cloned into pUC118 by colony hybridization. Further Southern hybridization revealed that the fragments contained the hexulose phosphate synthase gene. The sequencing of the positive clones are in proceeding.
The formaldehyde-fixing system of a methonol-using bacterium, Methylomonas aminofaciens 77a was used for the production of ^<13>C-labeled D-fructose-6-phosphate from ^<13>C-formaldehyde and D-ribose-5-phosphate. The specific and high yield production of[1- ^<13>C]D-fructose-6-phosphate was performed using an enzyme system that contained partially purified hexulose phosphate synthase and phosphohexuloisomerase from the bacterium, and spinach phosphoriboisomerase. A fed-batch reaction, in which 50 mM formaldehyde and 75 mM D-ribose-5-phosphate were added at 15-min intervals, afforded 203 mM D-fructose-6-phosphate for 90 min.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] 加藤 暢夫: "C_1微生物の機能と物質変換" バイオサイエンスとインダストリ-. 49. 20-24 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Yanase: "Production of D-〔1- ^<13>C〕Fructose 6-phosphate by a Formaldehydefixing System in Methylomonas aminofaciens 77a" Biosci.Biotech.Biochem.56. 541-542 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Yanase: "Enzymatic preparation of 〔1-13C〕D-fructose-6-phosphate from 13C-fromaldehyde and D-ribose-5-phosphate using formaldehydefixing systme of Methylomonas aminofaciens 77a" Appl.Microbiol.Biotechnol.(1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] N. Kato: "Methylotrophs, metabolic function and application to bioconversion" Biosciens and Industry. 49. 20-24 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H. Yanase et al.: "Production of D-[1- ^<13>C] Fructose 6-phosphate by a Formaldehyde-fixing System in Methylomonas aminofaciens 77a" Biosh. Biotech. Biochem.56. 541-542 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H. Yanase et al.: "Enzymatic preparation of [1- ^<13>C] D-fructose-6-phospahte from ^<13>C-formaldehyde and D-ribose-5-phosphate using formaldehyde-fixing system of Methylomonas aminofacidens 77a" Appl Microbiol. Biotechnol.(1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 加藤 暢夫: "C_1微生物の機能と物質変換" バイオサイエンスとインダストリ-. 49. 20-24 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] H.Yanase: "Production of Dー〔1ー^<13>C〕Fructose 6ーphosphate by a Formaldehydeーfixing System in Methylomonas aminofaciens 77a" Biosci.Biotech.Biochem.56. 541-542 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] H.Yanase: "Enzymatic preparation of〔1ー13C〕Dーfructoseー6ーphosphate from 13Cーformaldehyde and Dーriboseー5ーphosphate using formaldehydeーfixing system of Methylomonas aminofaciens 77a" Appl.Microbiol.Biotechinol.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 加藤 暢夫: "C_1微生物の機能と物質変換" バイオサイエンスとバイオインダストリ-. 49. 20-24 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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