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Improvement of Functions of Novel Enzymes in the Desulfurization Metaoblism of Petroleum by Protein Engineering and Molecular Genetics

Research Project

Project/Area Number 15580063
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionNational University Corporation Tottori University

Principal Investigator

IZUMI Yoshikazu  Tottori University, Faculty of Engineering, Professor, 工学部, 教授 (40026555)

Co-Investigator(Kenkyū-buntansha) TANOKURA Masaru  the University of Tokyo, Graduate School of Agricultural and Life Science, Professor, 大学院・農学生命化学研究科・応用生命化学専攻, 教授 (60136786)
OHSHIRO Takashi  Tottori University, Faculty of Engineering, Lecturer, 工学部, 講師 (00233106)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordsdesulfurization / dibenzothiophene / Rhodococcus erythropolis / desulfinase / DszB / DszC / Bds / Bfl / BdsA / desuofurization
Research Abstract

It is difficult to remove polycyclic sulfur compounds in fossil fuels. Dibenzothiophene(DBT) is considered as a model polycyclic sulfur compound contained in fossil fuels. Some bacteria such as Rhodococcus erythropolis D-1 utilize DBT as a sole source of sulfur without breaking its carbon-carbon backbone through the sulfur-specific pathway. In this pathway, DBT is oxidized to DBT sulfone via DBT sulfoxide by DszC,DBT sulfone is converted to 2-hydroxybiphenyl 2-sulfinic acid (HBPSi) by DszA, and HBPSi is desulfurized to 2-hydroxybiphenyl by DszB. Flavin reductase is necessary for monooxygenase reactions by DszC and DszA.
The research results of this study are summarized as follows.
1.DszB of R.erythropolis KA2-5-1 and its C27S mutant complexed with biphenyl-2-sulfinic acid were crystallized and preliminary X-ray crystallographic analyses were conducted.
2.The gene of thermostable falvin reductase (Bfl) from Bacillus sp.DSM411, which coupled with DszC, was overproduced in Escherichia coli, and the recombinant enzymes were purified and characterized.
3.Following the above 1, the mutant Q65H of DszB showed higher thermostability and optimal temperature than the parent DszB enzyme.
4.Three enzymes (BdsC,A,B) involved in the DBT desulfurization were purified from the moderately thermophilic bacterium, Bacillus subtilis WU-S2B and characterized.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report

Research Products

(12 results)

All 2005 2004 2003 Other

All Journal Article (9 results) Publications (3 results)

  • [Journal Article] Dibenzothiophene Desulfurizing Enzymes from a Moderately Thermophilic Bacterium Bacillus subtilis WU-S2B-Purification, Characterization and Overexpression-2005

    • Author(s)
      T.Ohshiro, Y.Ishii, K.Ueda, Y.Izumi, K.Kino, K.Kirimura
    • Journal Title

      J.Biosci.and Bioeng. (in press)

    • NAID

      210000130368

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Crystallization and preliminary X-ray analyses of desulfurization enzyme DszB and its C27S mutant complexed with biphenyl-2-sulfinic acid2004

    • Author(s)
      W.C.Lee, T.Ohshiro, T.Matsubara, Y.Izumi, M.Tanokura
    • Journal Title

      Acta.Crystallogr D60

      Pages: 1636-1638

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Thermostable falvin reductase that couples with dibenzothiophene monooxygenase, from Bacillus sp.DSM411 : Purification, characterization, and gene cloning2004

    • Author(s)
      T.Ohshiro, H.Yamada, Y.Shimoda, T.Matsubara, Y.Izumi
    • Journal Title

      Biosci.Biotechnol.Biochem. 68(8)

      Pages: 1712-1721

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Crystallization and preliminary X-ray analyses of desulfurization enzyme DszB and its C27S mutant complexed with biphenyl-2-sulfinic acid2004

    • Author(s)
      W.C.Lee, T.Ohshiro, T.Matsubara, Y.Izumi, M.Tanokura
    • Journal Title

      Acta.Crystallogr D 60

      Pages: 1636-1638

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Crystallization and preliminary X-ray analyses of desulfurization enzyme DszB and its C27S mutant complexed with biphenyl-2-sulfinic acid2004

    • Author(s)
      W.C.Lee, T.Ohshiro, T.Matsubara, Y.Izumi, M.Tanokura
    • Journal Title

      Acta.Cryst.allogr D60

      Pages: 1-3

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Thermostable falvin reductase that couples with dibenzothiophene monooxygenase, from Bacillus sp. DSM411: Purification, characterization, and gene cloning2004

    • Author(s)
      T.Ohshiro, H.Yamada, Y.Shimoda, T.Matsubara, Y.Izumi
    • Journal Title

      Biosci.Biotechnol.Biochem. 68(8)

      Pages: 1712-1721

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Biosynthesis of ε-poly-L-lysine in a cell-free system of Streptomyces albulus2003

    • Author(s)
      T.Kawai, T.Kubota, J.Hiraki, Y.Izumi
    • Journal Title

      Biochem.Biophys.Res.Commun. 311

      Pages: 635-640

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] 石油の硫黄・窒素・金属の除去2003

    • Author(s)
      大城 隆, 和泉好計
    • Journal Title

      農芸化学の事典

      Pages: 757-760

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] 電解生成水によるズブチリシン,α-アミラーゼおよびリパーゼの活性化2003

    • Author(s)
      原 安夫, 小林健治, 和泉好計
    • Journal Title

      日本食品科学工学会誌 50

      Pages: 563-569

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] T.Kawai, T.Kubota, J.Hiraki, Y.Izunmi: "Biosynthesis of ε-poly-L-lysine in a cell-free system of Streptomyces albulus"Biochem.Biophys.Res.Commun. 311. 635-640 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 大城 隆, 和泉好計: "石油の硫黄・窒素・金属の除去"農芸化学の事典. 757-760 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 原 安夫, 小林健治, 和泉好計: "電解生成水によるズブチリシン,α-アミラーゼおよびリパーゼの活性化"日本食品科学工学会誌. 50. 563-569 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-03-31   Modified: 2016-04-21  

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