Gene Cloning and Characterization of Mycobacterium phlei Flavin Reductase Involved in Dibenzothiophene Desulfurization(Enzymology, Protein Engineering, and Enzyme Technology)

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概要

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• Mycobacterium phlei WU-F1 possesses the ability to convert dibenzothiophene (DBT) to 2-hydroxybiphenyl with the release of inorganic sulfur over a wide temperature range from 20℃ to 50℃. The conversion is initiated by consecutive sulfur atom-specific oxidations by two monooxygenases, and a flavin reductase is essential in combination with these flavin-dependent monooxygenases. The flavin reductase gene (frm) of M. phlei WU-F1, which encodes a protein of 162 amino acid residues with a molecular weight of 17,177, was cloned and the deduced amino acid sequence shares approximately 30% identity with those of several flavin reductases in two protein-component monooxygenases. It was confirmed that the coexpression of frm with the DBT-desulfurization genes (bdsABC) from M. phlei WU-F1 was critical for high DBT-desulfurizing ability over a wide temperature range from 20℃ to 55℃. The frm gene was overexpressed in Escherichia coli cells, and the enzyme (Frm) was purified to homogeneity from the recombinant cells. The purified Frm was found to be a 34-kDa homodimeric protein with a monomeric molecular mass of 17kDa. Frm exhibited high flavin reductase activity over a wide temperature range, and in particular, the turnover rate for FMN reduction with NADH as the electron donor reached 564 s^<-1> at 50℃, which is one of the highest activities among all of the flavin reductases previously reported. Intriguingly, Frm also exhibited a high ferric reductase activity.

• 社団法人日本生物工学会の論文
• 2005-06-25

著者

• Kino Kuniki

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

• 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

• Takahashi Shusuke

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

• Furuya Toshiki

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

• Ishii Yoshitaka

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

• IWASAKI YUICHIRO

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

• Iwasaki Yuichiro

  Department Of Applied Chemistry School Of Science And Engineering Waseda University

• Kirimura Kohtaro

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Furuya Toshiki

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Takahashi Shusuke

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Ishii Yoshitaka

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

• Ishii Yoshitaka

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

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