title: Purification and characterization of an NADH-dependent alcohol dehydrogenase from Candida maris for the synthesis of optically active 1-(pyridyl)ethanol derivatives.
abstract: A novel (R)-specific alcohol dehydrogenase (AFPDH) produced by Candida maris IFO10003 was purified to homogeneity by ammonium sulfate fractionation, DEAE-Toyopearl, and Phenyl-Toyopearl, and characterized. The relative molecular mass of the native enzyme was found to be 59,900 by gel filtration, and that of the subunit was estimated to be 28,900 on SDS-polyacrylamide gel electrophoresis. These results suggest that the enzyme is a homodimer. It required NADH as a cofactor and reduced various kinds of carbonyl compounds, including ketones and aldehydes. AFPDH reduced acetylpyridine derivatives, β-keto esters, and some ketone compounds with high enantioselectivity. This is the first report of an NADH-dependent, highly enantioselective (R)-specific alcohol dehydrogenase isolated from a yeast. AFPDH is a very useful enzyme for the preparation of various kinds of chiral alcohols.
authors.affiliation: Frontier Biochemical and Medicinal Research Laboratories, Kaneka Corporation, Takasago, Hyogo, Japan. Shigery.Kawano@kaneka.co.jp
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kawano
authors.forename: Miho
authors.initials: M
authors.lastname: Yano
authors.forename: Junzo
authors.initials: J
authors.lastname: Hasegawa
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Yasohara
mesh_terms: D000426:Alcohol Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism*; D001709:Biotechnology / Q000379:methods*; D002175:Candida / Q000737:chemistry / Q000201:enzymology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007659:Ketones / Q000378:metabolism*; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism*; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000447:Aldehydes; D007659:Ketones; D009243:NAD; D000426:Alcohol Dehydrogenase