title: A novel fungal enzyme, NADPH-dependent carbonyl reductase, showing high specificity to conjugated polyketones. Purification and characterization.
abstract: A novel enzyme which specifically catalyzes the reduction of conjugated polyketones was purified to homogeneity from cells of Mucor ambiguus AKU 3006. The enzyme has a strict requirement for NADPH and irreversibly reduces a number of quinones such as p-benzoquinone, alpha-naphthoquinone and acenaphthenequione. The enzyme also reduces polyketones such as isatin and ketopantoyl lactone, and their derivatives. The apparent Km values for isatin and ketopantoyl lactone are 49.9 microM and 714 microM, respectively. The reduction of ketopantoyl lactone proceeds stereospecifically to yield L-(+)-pantoyl lactone. The pro-S (A) hydrogen at C-4 of NADPH is transferred to the substrate. The enzyme is not a flavoprotein and consists of two polypeptide chains with an identical relative molecular mass of 27,500. Quercetin, dicoumarol and some SH reagents inhibit the enzyme activity. 3-Methyl-1,2-cyclopentanedione and 1,3-cyclohexanedione are uncompetitive inhibitors with Ki values of 80.9 microM and 64.5 microM, respectively, to ketopantoyl lactone.
authors.affiliation: Department of Agricultural Chemistry, Kyoto University, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Shimizu
authors.forename: S
authors.initials: S
authors.lastname: Hattori
authors.forename: H
authors.initials: H
authors.lastname: Hata
authors.forename: H
authors.initials: H
authors.lastname: Yamada
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives*; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007510:Isatin / Q000378:metabolism; D007700:Kinetics; D007783:Lactones / Q000378:metabolism; D008970:Molecular Weight; D009089:Mucor / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D007783:Lactones; C008232:keto-pantoyllactone; D007510:Isatin; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase; D015107:4-Butyrolactone