title: Purification and properties of a carbonyl reductase involved in stereoselective reduction of ethyl 4-chloro-3-oxobutanoate from Cylindrocarpon sclerotigenum IFO 31855.
abstract: A NADPH-dependent carbonyl reductase (CSCR1) was purified to homogeneity from Cylindrocarpon sclerotigenum IFO 31855. The enzyme catalyzed the stereoselective reduction of ethyl 4-chloro-3-oxobutanoate to the corresponding (S)-alcohol with a >99% enantiomer excess. The relative molecular mass of the enzyme was estimated to be 68,000 by gel filtration chromatography and 24,800 on SDS polyacrylamide gel electrophoresis. The enzyme had an extremely narrow substrate specificity and it highly reduced conjugated diketone, 2,3-butanedion, in addition to ethyl 4-chloro-3-oxobutanoate. The enzyme activity was inhibited by HgCl(2) (100%), 5,5'-dithiobis(2-nitrobenzoic acid) (56%), dicoumarol (42%), and CuSO(4) (46%). The N-terminal amino acid sequence of the enzyme (P-Q-G-I-P-T-A-S-R-L) showed no apparent similarity with those of other oxidoreductases.
authors.affiliation: Research Institute for Advanced Science and Technology, Osaka Prefecture University, 1-2 Gakuen-cho, Sakai, Osaka 599-8570, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Saratani
authors.forename: Eiji
authors.initials: E
authors.lastname: Uheda
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Atsuo
authors.initials: A
authors.lastname: Nishimura
authors.forename: Fumiki
authors.initials: F
authors.lastname: Yoshizako
mesh_terms: D000090:Acetoacetates / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000438:Alcohols; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D003931:Diacetyl / Q000378:metabolism; D004791:Enzyme Inhibitors; D006865:Hydrogenation; D003904:Mitosporic Fungi / Q000201:enzymology*; D008970:Molecular Weight; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000090:Acetoacetates; D000438:Alcohols; D004791:Enzyme Inhibitors; C111982:ethyl 4-chloro-3-oxobutanoate; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase; D003931:Diacetyl