title: Formate oxidase, an enzyme of the glucose-methanol-choline oxidoreductase family, has a His-Arg pair and 8-formyl-FAD at the catalytic site.
abstract: 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.
authors: Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui.
authors: Daiju
authors: D
authors: Doubayashi
authors: Takumi
authors: T
authors: Ootake
authors: Yosifumi
authors: Y
authors: Maeda
authors: Masaya
authors: M
authors: Oki
authors: Yuji
authors: Y
authors: Tokunaga
authors: Akihiko
authors: A
authors: Sakurai
authors: Yukio
authors: Y
authors: Nagaosa
authors: Bunzo
authors: B
authors: Mikami
authors: Hiroyuki
authors: H
authors: Uchida
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001236:Aspergillus oryzae / Q000737:chemistry / Q000201:enzymology*; D001665:Binding Sites; D020134:Catalytic Domain; D002794:Choline / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry / Q000378:metabolism*; D005561:Formates / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol / Q000378:metabolism; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011199:Potentiometry; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D005561:Formates; D005656:Fungal Proteins; D011994:Recombinant Proteins; D005182:Flavin-Adenine Dinucleotide; D000429:Alcohol Oxidoreductases; D002794:Choline; D000432:Methanol