Expression in Escherichia coli of an Unnamed Protein Gene from Aspergillus oryzae RIB40 and Cofactor Analyses of the Gene Product as Formate Oxidase

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概要

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• An unnamed protein of Aspergillus oryzae RIB40 (accession no. XP_001727378), the amino acid sequence of which shows high similarity to those of formate oxidase isoforms produced by Debaryomyces vanjiriae MH201, was produced in Escherichia coli in C-His6-tagged form. The gene product, purified by affinity column chromatography, catalyzed the oxidation of formate to yield hydrogen peroxide but showed no evidence of activity on the other substrates tested. The Km and Vmax values at 30 °C at pH 4.5 were 7.9 mM and 26.3 μmole/min mg respectively. The purified enzyme showed UV-visible spectra atypical of ordinary flavoproteins. The UV-visible spectra of the enzyme and the UV-visible spectra, fluorescence spectra, and mass spectrometry of the extract obtained by boiling the purified enzyme suggested that the enzyme has a non-covalently bound FAD analog, which is expected to be 8-formyl-FAD.

• 社団法人 日本農芸化学会の論文
• 2009-12-23

著者

• FUJII Yutaka

  Department of Applied Physics, University of Fukui

• UCHIDA Hiroyuki

  Department of Earth and Space Science, Osaka University

• Fujii Yutaka

  Department Of Applied Physics University Of Fukui

• Fujii Yutaka

  Dep. Of Molecular Biology And Chemistry Fac. Of Medicine Univ. Of Fukui

• ISA Kimio

  Department of Natural Science Education, Faculty of Education and Regional Studies, Fukui University

• Uchida Hiroyuki

  Department Of Applied Chemistry And Biotechnology Graduate School Of Engineering University Of Fukui

• Maeda Yoshifumi

  Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fuk

• Doubayashi Daiju

  Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fuk

• Oki Masaya

  Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fuk

• Nose Hiroaki

  Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fuk

• Isa Kimio

  Department Of Chemistry Faculty Of Education And Regional Studies Fukui University

• Oki Masaya

  Department Of Applied Chemistry And Biotechnology Graduate School Of Engineering University Of Fukui

• Nose Hiroaki

  Department Of Applied Chemistry And Biotechnology Graduate School Of Engineering University Of Fukui

• SAKURAI AKIHIKO

  Department of chemical Process Engineering, Faculty of Engineering, Hokkaido University

• Maeda Yoshifumi

  Department Of Applied Chemistry And Biotechnology Graduate School Of Engineering University Of Fukui

• Isa Kimio

  Department Of Science And Mathematics Education Faculty Of Education And Regional Studies University

• Sakurai Akihiko

  Department Of Applied Chemistry And Biotechnology Graduate School Of Engineering University Of Fukui

• Sakurai Akihiko

  Department Of Applied Chemistry And Biotechnology Faculty Of Engineering Fukui University

• Uchida Hiroyuki

  Department Of Applied Chemistry And Biotechnology Faculty Of Engineering University Of Fukui

• Uchida Hiroyuki

  Department Of Applied Chemistry And Biotechnology Faculty Of Engineering Fukui University

• Oki Masaya

  Department Of Applied Chemistry & Biotechnology Graduate School Of Engineering University Of Fuk

• Fujii Yutaka

  Deparment Of Bacteriology Hiroshima University School Of Medicine

• Doubayashi Daiju

  Department Of Applied Chemistry And Biotechnology Graduate School Of Engineering University Of Fukui

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