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Elucidation of expression/regulation of genes involved in microbial polyhydroxylalkanoate (PHA)

Research Project

Project/Area Number 11217207
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Science and Engineering
Research InstitutionNagoya University

Principal Investigator

YAMANE Tsuneo  Nagoya University, Graduate School of Bio & Agro Sciences, Professor, 大学院・生命農学研究科, 教授 (70026102)

Co-Investigator(Kenkyū-buntansha) UEDA Shunsaku  Utsunomiya University, Faculty of Agriculture, Professor, 農学部, 教授 (80160167)
NAKANO Hideo  Nagoya University, Bio-& Agro Sciences, Associate Professor, 大学院・生命農学研究科, 助教授 (00237348)
Project Period (FY) 1999 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥44,500,000 (Direct Cost: ¥44,500,000)
Fiscal Year 2002: ¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 2001: ¥10,500,000 (Direct Cost: ¥10,500,000)
Fiscal Year 2000: ¥11,000,000 (Direct Cost: ¥11,000,000)
Fiscal Year 1999: ¥16,000,000 (Direct Cost: ¥16,000,000)
KeywordsPHA / PHB / methylotroph / microbial denitrifyer / Paracoccus denitrificans / phaA / phaR / PHA depolymerase / P(3HB) / P(3HV) / phaP / PHB depolymerase / pha C / pha P / pha R
Research Abstract

With aim of elucidating regulation mechanism of PHA synthesis/degradation within denitrifyer and methylotroph, Paracoccus denitrificans, studies were carried out on genes/enzymes/metabolism levels, and the following have been achieved.
1. A gene named phaR, was discovered downstream of phaP that encoded PHA granule-associated protein, phasin. PhaR was a transcription factor which bound promoter region of phaP and also bound the PHA granule. It regulated the size of PHA granule.
2. Genes involved in PHA synthesis/degradation, i.e phaA, phaB, phaC, and phaZ, were expressed almost constitutively irrespective of PHA synthesis level.
A metabolite, Coenzyme A, was proposed as a global regulator of PHA biosynthesis.
3. Three enzymes involved in the intracellular degradation of PHA, i.e. two different kinds of PHA depolymerases, and PHB oligomer hydrolase, were purified and their biochemical characteristics were elucidated. One of the PHA depolymerases was a novel enzyme which yielded 3-hydroxylbutyrate monomer directly from PHB polymer.

Report

(5 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Akira Maehara et al.: "Analyses of a Polyhydroxyalkanoic Acid Granule-Associated 16-Kilodalton Protein and Its Putative Regulator in the pha Locus of Paracoccus denitrificans"Journal of Bacteriology. 181・9. 2914-2921 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yasuo Takagi et al.: "Biosynthesis of Polyhydroxyalkanoate with a Thiophenoxy Side Group Obtained from Pseudomonas putida"Macromolecules. 32. 8315-8318 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Dai Gao et al.: "Identification of the intracellular polyhydroxyalkanoate depolymerase gene of Paracoccus denitrificans and some properties of the gene product"FEMS Microbiology Letters. 196. 159-164 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Akira Maehara et al.: "PhaR, a protein of unknown function conserved among short-chain-length polyhydroxyalkanoic acids producing bacteria, is a DNA-binding protein and represses Paracoccus denitrificans phaP expression in vitro"FEMS Microbiology Letters. 200. 9-15 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Syunsaku Ueda et al.: "Purification and properties of D-(-)-3-hydroxybutyrate oligomer hydrolase of Paracoccus denitrificans"FEMS Microbiology Letters. 206. 179-184 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Akira Maehara et al.: "A Repressor Protein, PhaR, Regulates Polyhydroxyalkanoate (PHA) Synthesis via Its Direct Interacion with PHA"Journal of Bacteriology. 184. 3992-4002 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 山根恒夫: "グリーンプラスチック最新技術(井上義夫監修)、第6章"(株)シーエムシー. 13 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A.Maehara et al: "Analysis of polyhydroxyalkanoic acid granule-associated 16-kirodalton protein and its putative regulator in the pha locus of Paracoccus denitrificans"Journal of Bacteriology. 181. 2914-2921 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Takagi et al: "Biosyntesis of polyhydroxyalkanoate with a thiophenoxy side group obtained from Pseudomonas putida"Macromolecules. 32. 8315-8318 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] D.Gao et al: "Identification of the intracellular polyxydroxyalkanoate depolymerase gene of Paracoccus denitriificans and some properties of the gene product"FEMS Microbiology Letters. 196. 159-164 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A.Maehara et al: "PhaR, a protein of unknown function conserved among short-chain-length polyhydroxyalkanoic acids oroducing bacteria, is a DNA-binding protein and represses Paracoccus denitrificans expression in vitro"FEMS Microbiology Letters. 200. 9-5 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] S.Ueda et al: "Purification and properties of d- (-) -3 -hydroxybutyrate oligomer hydrolase of Paracoccus denitrificans"FEMS Microbiology Letters. 206. 179-184 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A.Maehara et al: "A represser protein, PhaR, regulates polyhydroxyalkanoate (PHA) synthesis via its direct interaction with PHA"Journal of Bacteriology. 184. 3992-4002 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Yamane, S.Ueda, and A. Maehara, ed. by Yoshio Inoue: "Synthesis of Biodegradable Plastics by Microorganisms, Recent Technology of Green Plastics"CMC.Co. Ltd.. Chap.6(71-83) (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Syunsaku Ueda et al.: "Purification and properties of D(-)-3-hydroxybutyrate oligomer hydrolase of Paracoccus denitrificans"FEMS Microbiology Letters. 206・2. 179-184 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Akira Maehara et al.: "A Repressor Protein, PhaR, Regulates Polyhydroxyalkanoate(PHA) Synthesis via Its Direct Interaction with PHA"Journal of Bacteriology. 184・14. 3992-4002 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 山根恒夫: "「グリーンプラスチック最新技術」(井上義夫監修)、第6章"(株)シーエムシー. 13 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yasuo Takagi: "Biosynthesis of polyhydroxyalkanoate with thiophenoxy side group obtained from Pseudomonas Putida"Macromolecules. 32・25. 8315-8318 (1999)

    • Related Report
      2001 Annual Research Report
  • [Publications] Dai Gao: "Identification of the intracellular polyhydroxyalkanoate depolymerase gene of paracoccus denitrificans and some properties of the gene product"FEMS Microbiology Letters. 196・2. 159-164 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Akira Maehara: "PhaR, a protein of unknown function conserved among short-chain-length polyhydroxyalkanoic acids producing bacteria, is a DNA-binding protein"FEMS Microbiology Letters. 200・1. 9-15 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Chanprateep: "Mulltivariable control of alcohol concentrations in the production of polyhydroxyalkanoates (PHAs) by Paracoccus denitrificans"Biotechnology and Bioengineering. 74・2. 116-124 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shunsaku Ueda: "Purification and properties of D(-)-3-hydroxybutyrate oligomer hydrolase of Paracoccus denitrificans"FEMS Microbiology Letters. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Yamane et al.: "Biosynthesis of Polyhydroxyalkanoate with a Thiophenoxy Side Group Obtained from Pseudomonas putida"Macromolecules. 32. 8315-8318 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] S.Ueda et al.: "Identification of the intracellular polyhydroxyalkanoate depolymeruse gene of Paracoccus denitrificans and the region essential for its expression"FEMS Microbiology Letters. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Akira Maehara: "Analysis of a Polyhydroxylkanoic Acid Granule-Associated 16-Kilodalton Protein and Its Putative Regulator in the pha Locus of Paracoccus denitrificans"Journal of Bacteriology. 181・9. 2914-2921 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yasuo Takagi: "Biosynthesis of Polyhydroxyalkanoate with a Thiophenoxy Side Group Obtained from Pseudomonas putida"Macromolecules. 32・25. 8315-8318 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Akira Maehara: "Paracoccus denitrificans Pha R, a regulatory protein, binds to a short DNA region located upstream of the phasin gene and regulates its expression in vitro"Journal of Bacteriology. (発表予定).

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2018-03-28  

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