Cloning and expression of three formate oxidase genes from Debaryomyces vanrijiae MH201.
Debaryomyces vanrijiae MH201 produces formate oxidase (FOD) at estimated pI values by native isoelectric focusing of 5.1, 5.4, and 5.9. We cloned and expressed three formate oxidase cDNAs, FOD1, FOD2, and FDO3, of the yeast using Escherichia coli. The open reading frames of FOD1, FOD2, and FDO3 were 1,731 bp long, and encoded 576-amino acid polypeptides with molecular masses of 64,142, 63,794, and 63,836 Da respectively. Expression of FOD1, FOD2, and FOD3 resulted in the production of three isozymes, with pI values of 5.1, 5.9, and 5.9 respectively. Co-expression of FOD1 and FOD2 and of FOD1 and FOD3 resulted in the production of additional isozymes with pI values, of 5.4. The three amino acid sequences of FOD1, FOD2, and FOD3 contained a consensus motif of a flavin adenine dinucleotide binding site in their N-terminal parts and a glucose-methanol-choline oxidoreductase signature pattern, suggesting that formate oxidase ought to be classified in the glucose-methanol-choline oxidoreductase family.
Maeda
Yoshifumi
Y
Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, Japan.
Oki
Masaya
M
Fujii
Yutaka
Y
Hatanaka
Akira
A
Hojo
Masayuki
M
Hirano
Kouzou
K
Uchida
Hiroyuki
H
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005561:Formates / Q000378:metabolism*; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010802:Phylogeny; D004718:Saccharomycetales / Q000201:enzymology* / Q000235:genetics
D018076:DNA, Complementary; D005561:Formates; C030544:formic acid; D010088:Oxidoreductases