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Analysis of sorbitol metabolism in genus Gluconobacter.

Research Project

Project/Area Number 18580078
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionUniversity of the Ryukyus (2007)
Yamaguchi University (2006)

Principal Investigator

TOYAMA Hirohide  University of the Ryukyus, University of the Ryukyus, Agriculture, Professor (60240884)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,830,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥330,000)
Fiscal Year 2007: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000)
KeywordsGluconobacter / Acetic acid bacteria / Sorbitol / Oxidative fermentation
Research Abstract

It is revealed that D-sorbitol, not L-sorbose, is the inducer of sldSLC, the gene for FAD-sorbitol dehydrogenase (SLDH), yielding L-sorbose from D-sorbitol. The obtained results were published.
PQQ-glycerol dehydrogenase (GLDH), yielding also L-sorbose from D-sorbitol, connects efficiently to cytochrome bo 3 terminal oxidase and higher energy conservation ratio, thus it plays a major role in L-sorbose production. On the other hand, FAD-SLDH linked preferably to the cyanide-insensitive terminal oxidase, CIO. The obtained results were published.
The SboA enzyme expressed and purified from an Escherichia coli transformant showed NADPH-dependent L-sorbose reductase (NADPH-SR) activity. The obtained results are published. Now the enzyme is served for structural analysis by x-ray crystallography
Reverse transcription-PCR experiments indicated that sboRA comprises an operon. It is expected that SboR is a transcriptional regulator which binds with L-sorbose. SboR was expressed and purified from E. coli transformant and used for gel-shift assay with the DNA fragment containing the promoter-operator region for sboRA, however, we failed to demonstrate that SboR has an ability to bind the DNA fragment with or without L-sorbose. The obtained results were published.
The gene for FAD-gluconate dehydrogenase (GADH), yielding 2-ketogluconic acid from D-gluconate, was cloned and sequenced.
For easy measurement for 5-ketogluconate (5KGA) and 2-ketogluconate (2KGA), genes for 5KGA reductase and 2KGA reductase were cloned and expressed in E. coli, Both enzyme preparations were easily obtained from E. coli transformants after one column chromatography.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (12 results)

All 2008 2007

All Journal Article (12 results) (of which Peer Reviewed: 7 results)

  • [Journal Article] Distinct physiological roles of two membrane-bound dehydrogenases responsible for D-sorbitol oxidation in Gluconobacter frateurii2008

    • Author(s)
      W.Soemphol, et al.
    • Journal Title

      Biosci.Biotechnol.Biochem 72

      Pages: 842-850

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Distinct physiological roles of two membrane-bound dehydrogenases responsible for D-sorbitol oxidation in Gluconobacter frateurii2008

    • Author(s)
      W.Soemphol, et al.
    • Journal Title

      Bioscience, Biotechnology and Biochemistry 72

      Pages: 842-850

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Distinct physiological roles of two membrane-bound dehydrogenases responsible for D-sorbitol oxidation in Gluconobacter frateurii.2008

    • Author(s)
      W. Soemphol, et. al.
    • Journal Title

      Biosci. Biotechnol. Biochem 72

      Pages: 842-850

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] L-Sorbose reductase and its transcriptional regulator involved in L-sorbose utilization of Gluconobacter frateurii2007

    • Author(s)
      W.Soemphol, et al.
    • Journal Title

      J.Bacteriol. 189

      Pages: 4800-4808

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Membrane-bound, 2-keto D-gluconate-yielding D-gluconate dehydrogenase from Gluconobacter dioxyacetonicus IFO 3271 : Molecular properties and gene disruption2007

    • Author(s)
      H.Toyama, et al.
    • Journal Title

      Appl.Environ.Microbiol. 73

      Pages: 6551-6556

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Preparation of Enzymes Required for Enzymatic Quantification of 5-Keto-D-gluconate and 2-Keto-D-gluconate2007

    • Author(s)
      I.Saichana, et al.
    • Journal Title

      Biosci.Biotechnol.Biochem 71

      Pages: 2478-2486

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] L-Sorbose reductase and its transcriptional regulator involved in L-sorbose utilization of Gluconobacter frateurii2007

    • Author(s)
      W.Soemphol, et al.
    • Journal Title

      Journal of Bacteriology 189

      Pages: 4800-4808

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Membrane-bound, 2-keto D-gluconate-yielding D-gluconate dehydrogenase from Gluconobacter dioxyacetonicus IFO 3271 : Molecular properties and gene disruption2007

    • Author(s)
      H.Toyama, et al.
    • Journal Title

      Applied and Environmental Microbiology 73

      Pages: 6551-6556

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Preparation of Enzymes Required for Enzymatic Quantification of 5-Keto-D-gluconate and 2-Keto-D-gluconate2007

    • Author(s)
      I.Saichana, et al.
    • Journal Title

      Bioscience, Biotechnology and Biochemistry 71

      Pages: 2478-2486

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Membrane-bound, 2-keto D-gluconate-yielding D-gluconate dehydrogenase from Gluconobacter dioxyacetonicus IFO 3271: Molecular properties and gene disruption.2007

    • Author(s)
      H. Toyama, et. al.
    • Journal Title

      Appl Environ. Microbiol. 73

      Pages: 6551-6556

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Preparation of Enzymes Required for Enzymatic Quantification of 5-Keto-D-gluconate and 2-Keto-D-gluconate.2007

    • Author(s)
      I. Saichana, et. al.
    • Journal Title

      Biosci. Biotechnol. Biochem 71

      Pages: 2478-2486

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] L-Sorbose reductase and its transcriptional regulator involved in L-sorbose utilization of Gluconobacter frateurii2007

    • Author(s)
      W.Soemphol et al.
    • Journal Title

      J.Bacteriol. (掲載予定)

    • Related Report
      2006 Annual Research Report

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Published: 2006-04-01   Modified: 2016-04-21  

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