Formate Oxidase, an Enzyme of the Glucose-Methanol-Choline Oxidoreductase Family, Has a His-Arg Pair and 8-Formyl-FAD at the Catalytic Site

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概要

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• Formate oxidase of Aspergillus oryzae RIB40 contains an 8-replaced FAD with molecular mass of 799 as cofactor. The 1H-NMR spectrum of the cofactor fraction obtained from the enzyme indicated that the 8-replaced FAD in the fraction was 8-formyl-FAD, present in open form and hemiacetal form. The oxidation-reduction potentials of the open and hemiacetal forms were estimated by cyclic voltammetry to be −47 and −177 mV vs. Normal Hydrogen Electrode respectively. The structure of the enzyme was constructed using diffraction data to 2.24 Å resolution collected from a crystal of the enzyme. His511 and Arg554 were situated close to the pyrimidine part of the isoalloxazine ring of 8-formyl-FAD in open form. The enzyme had 8-formyl-FAD, the oxidation potential of which was approximately 160 mV more positive than that of FAD, and the His-Arg pair at the catalytic site, unlike the other enzymes belonging to the glucose-methanol-choline oxidoreductase family.

著者

• Mikami Bunzo

  Division Of Applied Life Science Graduate School Of Agriculture Kyoto University

• Uchida Hiroyuki

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

• 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 Fukui

• Tokunaga Yuji

  Department Of Materials Science And Engineering Faculty 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

• Nagaosa Yukio

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

• OOTAKE Takumi

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

• MAEDA Yosifumi

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

• 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

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