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Analysis of Structure-Function Relationship of the Signal Peptide-Independent Secretory Apparatus in the Gram-negative Bacterium

Research Project

Project/Area Number 09650878
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 生物・生体工学
Research InstitutionTottori University

Principal Investigator

YANASE Hideshi  Tottori University Department of Biotechnology Professor, 工学部, 教授 (20158033)

Co-Investigator(Kenkyū-buntansha) OKAMOTO Kenji  Tottori University Department of Biotechnology Associate Professor, 工学部, 助手 (80283969)
KITA Keiko  Tottori University Department of Biotechnology Associate Professor, 工学部, 助教授 (70234226)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsABC exporter / Gram-negative bacteria / GSP / Invertase / Levansucrase / Secretion / Signal peptide-independent pathway / Zymomonas mobilis / ABC exporter / General secretion pathway / Invertase / Levansucrase / Secretion / Signal peptide / Transcriptional activator / signal peptide-independent / secretion / secrettion channel / gram-negative bacteria / sec protein-independent / Fructosyltransferase
Research Abstract

Zymomonas mobilis is a facultative anaerobic Gram-negative bacterium known as an efficient ethanol producer on glucose, and its ethanol productivity is superior to that of commercial yeast. Our focus has been on the production of fuel ethanol from agricultural wastes and municipal solid waste using Z.mobilis. To breed a Z.mobilis strain capable of converting cellulose to et hanol directory, the bacterium was transformed with the endoglucanase and β-glucosidase genes. However, the recombinant strains were still unable to produce ethanol from cellulose, presumable because the cells did secrete necessary saccharifying enzymes. We have therefore begun investigating secretory mechanism in Z.mobilis, and found that levansucrase (SucZE2) and invertase(SucZE3) are translocated via a signal peptide-independent pathway in Z.mobilis. To clarify the secretory mechanism we cloned a 1.7 kb DNA fragment containing zliE and zliS genes complements the inability of secretive production of both enzymes in a Suc- mutant from Z.mobilis ATCC29191. By subcloning each genes into two different Suc mutants of Z.mobilis, it has been found that zliE activates the production of the extracellular levansucrase and invertase, and zliS stimulates the secretion of the enzymes. The results obtained from detailed analyses of zliS function suggested that the expression of zliS affects the cell membrane to stimulate the liberation of cell-associated proteins.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] H. Yanase, et al.: "Enzymatic One-Step Production of the Polysaccharide Levan"New, Technology Japan. 25 (4). 38-39 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 簗瀬英司: "フルクトシルトランスフェラーゼ(FTase)の触媒ドメイン解析と糖転移機能改変"応用糖質科学. 44(2). 203-211 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Tonouchi, et al.: "Increased Cellulose Production from Sucrose with Reduced Levan Accumulation by an Acetobacter Strain Harboring a Recombinant Plasmid"Biosci. Biotech. Biochem.. 62 (5). 833-836 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Yanase, et al.: "Expression of the Extracellular Levansucrase and Invertase Genes from Zymomonas mobilis in Escherichia coli Cells"Biosci. Biotech. Biochem.. 62 (9). 1802-1805 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y. Sakai, et al.: "A New carboxylesterase from Brevibacterium lines IFO12171 Responsible for the Conversion of 1,4-Butanediol Diacrylate to 4-hydroxybutyl Acrylate"Biosci. Biotech. Biochem.. 63 (4). 688-679 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y. Sakai, et al.: "Organization of the genes involved in the ribulose monophosphate pathway in an obligate methylotrophic bacterium, Methylomonas aminofaciens 77a"FEMS Microbiol. Letter. 176. 125-130 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Yanase: "Encyclopedia of Food Microbiology (Zymomonas)"ACADEMIC PRESS. 2372 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Yanase, et al.: "Enzymatic One-Step Production of the Polysaccharide Levan"New, Technology Japan. 25(4). 38-9 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Yanase: "Analysis of Structure-function Relationship in Fructosyltransferase of Zymomonas mobilis"J. Appl. Glycosci.. 44(2). 203-211 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N.Tonouchi, et al.: "Increased Cellulose Production from Sucrose with Reduced Levan Accumulation by an Acetobacter Strain Harboring a Recombinant Plasmid"Biosci. Biotech. Biochem.. 62(5). 833-836 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Yanase, et al.: "Expression of the Extracellular Levansucrase and Invertase Genes from Zymomonas mobilis in Escherichia coli Cells"Biosci. Biotech. Biochem.. 62(9). 1802-1805 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Yanase, et al.: "A New carboxylesterase from Brevibacterium linens IFO12171 Responsible for the Convertion of 1, 4-Butanediol Diacrylate to 4-hydroxybutyl Acrylate"Biosic. Biotechnol. Biochem.. 63(4). 688-679 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y.Sakai at.al.: "Organization of the genes involved in the ribulose monophosphate pathway in an obligate methylotrophic bacterium, Methylomonas aminofaciens 77a"FEMS Microbiol. Letter. 176. 125-130 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Yanase: "Encyclopedia of Food Microbiology (Zymomonas)"ACADEMIC PRESS. 2372 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y.Sakai,et al.: "A New carboxylesterase from Brevibacterium linens IFO12171 Responsible for the Convertion of 1,4-Butanediol Diacrylate to 4-hydroxybutyl Acrylate"Biosic.Biotechnol.Biochem.. 63・4. 688-679 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Y.Sakai,at al.: "Organization of the genes involved in the ribulose monophosphate pathway in an obligate methylotrophic bacterium,Methylomonas aminofaciens 77a"FEMS Microbiol.Letter. 176. 125-130 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] H.Yanase: "Encyclopedia of Food Microbiology(Zymomonas)"ACADEMIC PRESS. 2372 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Tonouchi,H.Yanase,Y.Kojima,T.Tsuchida,F.Yoshinaga,and S.Horinouchi: "Increased Cellulose Production from Sucrose with Reduced Levan Accumulation by an Acetobacter Strain Harboring a Recombinant Plasmid." Biosci.Biotechnol.Biochem.62・5. 833-836 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Yanase: "Enzyme to Efficient Decompose Formaldehyde" New Technology Japan. 26・3. 9-10 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Yanase, J.Fujimoto, M.Maeda, K.Okamoto, K.Kita, and K.Tonomura: "Expression of the Extracellular Levansucrase and Invertase Genes from Zymomonas mobilis in Escherichia coli Cells" Biosci.Biotechnol.Biochem.62・9. 1802-1805 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hideshi Yanase et.al.: "Enzymatic one-step production of the polysacharide levan" New Technology Japan. 25・4. 38-39 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 簗瀬 英司 他: "フルクトシルトランスフェラーゼ(FTase)の触媒ドメイン解析と糖転移機能改変" 応用糖質科学. 44・2. 203-211 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 簗瀬 英司 他: "Zymomonas属細菌におけるシグナルペプチド非依存型分泌酵素の分泌に関与する遺伝子" 日本生物工学大会講演要旨集. 20 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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