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Characterization and functional improvement of novel enzymes involved in the desulfi dibenzothiophene in microorganisms

Research Project

Project/Area Number 13650857
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

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

Co-Investigator(Kenkyū-buntansha) OHSHIRO Takashi  Tottori University, Faculty of Engineering, Lecturer, 工学部, 講師 (00233106)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywordsdibenzothiophen (DBT) / DBT-desulfurizing microorganism / DszB / HBSPiデスルフィナーゼ / HBPSiデスルフィナーゼ / DszC / ジベンゾチオフェン / 脱硫菌 / DszA / フラビンレダクターゼ / Rhodococcus erythropolis / 好熱性Bacillus / モノオキシゲナーゼ
Research Abstract

Organic sulfur compounds are found in fossil fuels, the combustion of which causes serious environmental problems, such as acid rain. At the refinery, hydrodesulfurization is currently performed to remove sulfur compounds form fossil fuels. However, it is difficult to remove polycyclic sulfur compounds. As legislative limits on sulfur emissions have become tighter, the need to remove polycyclic sulfur compounds form fuel has become more pressing. Dibenzothiophene (DBT) is considered as a model polycyclic sulfur compound contained in fossil fuels. We have reported that 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 monooxygen … More ase reactions by DszC and DszA.
The research results of this study are summarized as follows.
(1) We purified and characterized the flavin reductase from R. erythropolis D-1 grown in a medium containing DBT as the sole source of sulfur. The purified flavin reductase contains no chromogenic cofactors and was found to have a molecular mass of 86 kDa and four identical 22 kDa subunits. The purified enzyme was characterized: 1) NADH-dependent reduction of flavin mononucleotide (FMN), the Km values for NADH and FMN (208 and 10.8 μM. respectively), the optimal temperature and optimal pH (35^。C and 6.0, respectively), and the heat stability (30% activity retaining at 80^。C for 30 minutes). The flavin reductase gene was amplified with primers designed by using a DszD of R. erythropolis IGTS8, and the enzyme was overexpressed in Escherichiacoli. The specific activity in crude extracts of the overexpressed strain was about 275-fold that of the wild-type strain.
(2) DszB of a DBT-desulfurizing bacterium Rhodococcus erythropolis KA2-5-1 was overproduced in Escherick coexpression with chaperonin genes, groEL/groES. The recombinant DszB was purified to homogeneity and character was thought to be a novel enzyme (HBPSi desulfinase) in that it could specifically cleave the carbon-sulfurbond of H2-HBP and sulfite ion without the aid of any other proteinic components and coenzymes. Less

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Yoshikazu Izumi: "Purification, characterization, and overexpression of flavin reductase involoved in dibenzothiophene desulfurization by Rhodococcus erythropolis D-1"Applied and Environmental Microbiology. 67,2. 1179-1184 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshikazu Izumi: "Purification and characterization of enzymes involved in desulfurization of dibenzothiophene in fossil fuels"Journal of Molecular Catatysis B. Enzymatic. 11,2. 1061-1064 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshikazu Izumi: "A Novel enzyme, 2-hydroxybiphenyl-2'-sulfinate desulfinase (DszB) from dibenzothiophene desulfurizing bacterium, Rhodococcus erythropolis KA2-5-1: Gene overexpression and enzyme characterization"Biochemica et Biophysica Acta. 1598. 122-130 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Matsubara, T. Ohshiro, Y. Nishina and Y. Izumi: "Purification, characterization, and overexpression of flavin reductase involved in dibenzothiophene desulfurization by Rhodococcus erythropolis D-1"Applied and Environmental Microbiology. 67(2). 1179-1184 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Ohshiro and Y. Izumi: "Purification and characterization of enzymes involved in desulfurization of dibenzothiophene in fossil fuels"Journal of Molecular Catatysis B. Enzymatic. 11(2). 1061-1064 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Nakayama, T. Matsubara, T.Ohshiro, Y. Moroto, Y. Kawata, K. Koizumi, Y. Hirakawa, M. Suzuki, K. Maruhashi, Y. Izumi, and R. Kurane: "A novel enzyme, 2-hydroxybiphenyl-2-sulfinate desulfinase (DszB) from dibenzothiophene desulfurizing bacterium, Rhodococcus erythropolis KA2-5-1: Gene overexpression and enzyme characterization"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshikazu Izumi: "A novel enzyme, 2-hydroxybiphenyl-2'-sulfinate desulfinase (DszB) from dibenzothiophene desulfurizing bacterium, Rhodococcus erythropolis KA2-5-1 : Gene overexpression and enzyme characterization"Biochimica et Biophysica Acta. 1598. 122-130 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yoshikazu Izumi: "Purification and characterization of enzymes involved in desutfurization of dibenzothiophene in fossil fuels"J. Mol. Cat. B. Enzymatic. 11.3. 1061-1064 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yoshikazu Izumi: "Purification, characterization, and overexpression of flavin reductase involved in dibenzothiophene desulfurization by Rhodococcus erythropolis D-1"Appl. Environ. Microbiol.. 67.1. 1179-1184 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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