Dibenzothiophene Desulfurizing Enzymes from Moderately Thermophilic Bacterium Bacillus subtilis WU-S2B : Purification, Characterization and Overexpression(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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概要

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• The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized diben-zothiophene (DBT) at 50℃ through the selective cleavage of carbon-sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2'-hydroxybiphenyl 2-sulfinic acid des-ulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail.

• 社団法人日本生物工学会の論文
• 2005-09-25

著者

• Kino Kuniki

  Department of Applied Chemistry, Faculty of Science and Engineering, Waseda University

• 大城 隆

  鳥取大・工・生応工

• Izumi Yoshikazu

  Dep. Of Biotechnology Tottori Univ.

• KIRIMURA Kohtaro

  Department of Applied Chemistry, School of Science and Engineering, Waseda University

• Kino K

  Department Of Applied Chemistry School Of Science And Engineering Waseda University

• Kino Kuniki

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Kino Kuniki

  Department Of Applied Chemistry School Of Science And Engineering Waseda University

• Ohshiro Takashi

  Department of Biotechnology, Tottori University

• Ishii Yoshitaka

  Department of Applied Chemistry, School of Science and Engineering, Waseda University

• IZUMI Yoshikazu

  Department of Biotechnology, Tottori University

• Matsubara Toshiyuki

  Department of Biotechnology, Tottori University

• Ueda Koichi

  Department of Biotechnology, Tottori University

• Kino Kuniki

  Dep. Of Applied Chemistry Fac. Of Sci. And Engineering Waseda Univ. 3-4-1 Ohkubo Shinjuku-ku Tokyo 1

• Ishii Yoshitaka

  Institute For Biomedical Engineering Consolidated Research Institute For Advanced Science And Medica

• Kirimura Kohtaro

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Ohshiro Takashi

  Department Of Biotechnology Faculty Of Engineering Tottori University

• Ishii Yoshitaka

  Bio-refining Process Laboratory Advanced Technology And Research Institute Petroleum Energy Center

• Izumi Yoshikazu

  Department Of Agricultural Chemistry Kyoto University

• Ishii Yoshitaka

  Department Of Applied Chemistry School Of Science And Engineering Waseda University

• 大城 隆

  Department Of Biotechnology Tottori University

• Matsubara Toshiyuki

  Department Of Biotechnology Tottori University

• Ishii Yoshitaka

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Ueda Koichi

  Department Of Biotechnology Tottori University

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