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Cloning and structure analyzes of genes from Paracoccus denitrificans related to denitrification

Research Project

Project/Area Number 05454036
Research Category

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

Allocation TypeSingle-year Grants
Research Field 生物・生体工学
Research InstitutionNagoya University

Principal Investigator

IIJIMA Shinji  Nagoya University, Department of Biotechnology, Professor, 工学部, 教授 (00168056)

Co-Investigator(Kenkyū-buntansha) MIYAKE Katsuhide  Nagoya University, Department of Biotechnology, Research associate, 工学部, 助手 (90252254)
KOBAYASHI Takeshi  Nagoya University, Department of Biotechnology, Professor, 工学部, 助手 (10043324)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1994: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥4,600,000 (Direct Cost: ¥4,600,000)
KeywordsEnvironment / Waste water treatment / Denitrification / Nitrous oxide / 窒素化合物
Research Abstract

In order to bread denitrifying bacterium which acts under aerobic conditions by recombinant DNA technology, we cloned genes for denitrification and determined the DNA sequence of NirS locus. In this year, we determined the DNA sequence of NirS downstream flanking region (6.7 kbp SmaI-BamHI fragment). For this, DNA was cloned as 3 pieces in pUC 118 and 119 vectors and DNA sequences of each fragment were determined.
We also tried to determin the sequence of NirS upstream flanking region. Although we have not yet completely determined the entire sequence, a part of the lucus was homologous to norB gene of Ps. aeruginosa which encodes a subunit of nitrous oxide reductase.
We found 2 additional open reading frames designated as ORFs 4 and 6. ORF6 which located 4 kbp downstream of NirS gene was partly homologous to pyrolquinoline quinone synthetase in amino acid level (30.2%). Furthermore, ORF4 which located 5 kbp downstream from NirS gene (just adiacent to ORF6) showed partial homology to NirC (40%) and NirM (25%) of Ps. aeruginosa in amino acid level. NirC and Nir M genes of Ps. aeruginosa encoded cytochrome C and cytochrome C-551, respectively. Therefore, it is possible that the locus located 4-5kbp downstream of NirS gene of P.denitrificans encoded a proteins which are essential for electron transfer.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] 飯島信司: "Cloning and sequencing of a gene encoding nitrite reductase from paracoccus denitrificans and expression of the gene in Escherichia coli" Journal of Fermentation and Bioengineering. 76. 83-88 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 飯島信司: "膜型バイオリアクターを用いたphoプロモーターからの遺伝子発現の誘導と物質生産の効率化" 生物工学会誌. 71. 317-323 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 飯島信司: "Structure of denitrifying enzyme gene cluster of Paracoccus denitrificans" Journal of Fermentation and Bioengineering. (発表予定).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Iijima: "Cloning and sequencing of a gene encoding nitrite reductase from Paracoccus denitrificans and expression of the gene in Escherichia coli" Journal of Fermentation and Bioengineering. 76. 82-88 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Iijima: "Effective production of recombinant protein by using inducible pho promoter" Seibutukougakukaishi. 71. 317-323 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 飯島信司: "Cloning and sequencing of a gene encoding nitrite reductase from Paracoccus denitrificans and expression of the gene in Escherichia coli" Journal of Fermentation and Bioengineering. 76. 83-88 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] 飯島信司: "膜型バイオリアクターを用いたphoプロモーターからの遺伝子発現の誘導と物質生産の効率化" 生物工学会誌. 71. 317-323 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] 飯島信司: "Structure of denitrifying enzyme gene cluster of Paracoccus denitrificans" Journal of Fermentation and Bioengineering. (発表予定).

    • Related Report
      1994 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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