title: Characterization and high-level production of D-amino acid oxidase in Candida boidinii.
abstract: D-Amino acid oxidase (DAO, EC 1.4.3.3) from a methylotrophic yeast, Candida boidinii, was produced at a high level under the control of the alcohol oxidase gene promoter in the original host. The enzyme was a peroxisomal and monomeric enzyme, and contained noncovalently-bound FAD as a cofactor. The enzyme was active toward several D-amino acids such as D-Ala, D-Met, and D-Ser. An alcohol oxidase-depleted strain (aod1delta) was found to be a more suitable host for DAO production than the wild-type strain. Several post-translational effects may be responsible for the improvement of the DAO productivity by the aod1delta strain. Finally, an aod1delta strain transformant having multi-copies of an expression plasmid on its chromosome could produce DAO amounting up to 30% of the total soluble proteins.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yurimoto
authors.forename: T
authors.initials: T
authors.lastname: Hasegawa
authors.forename: Y
authors.initials: Y
authors.lastname: Sakai
authors.forename: N
authors.initials: N
authors.lastname: Kato
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000502:physiology; D002175:Candida / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D003605:D-Amino-Acid Oxidase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015870:Gene Expression; D014170:Transformation, Genetic
chemical_list: D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D003605:D-Amino-Acid Oxidase