title: Overexpression and characterization of a novel (S)-specific extended short-chain dehydrogenase/reductase from Candida parapsilosis.
abstract: The gene encoding a putative protein from Candida parapsilosis CDC317 (CPE) was cloned and overexpressed in Escherichia coli. The protein was amenable to overexpression in E. coli and constituted 35% of the total cell protein content. The optimal activity was determined at pH 5.5 and 40°C with the substrate 4-chloro-3-oxobutanoate ethyl ester (COBE). The optical purity of the product was over 99% enantiomeric excess for the (S)-isomer, and the molar conversion yield of the product was 91.1%. The apparent k(m) value for COBE was 0.19±0.01mM, which is an order of magnitude lower than that of other enzymes in the literature.
authors.affiliation: Research Center for Biomedicine and Health, College of Life and Environmental Science, Hangzhou Normal University, Hangzhou 310012, China.
authors.forename: Qiuyan
authors.initials: Q
authors.lastname: Wang
authors.forename: Linghong
authors.initials: L
authors.lastname: Shen
authors.forename: Tingting
authors.initials: T
authors.lastname: Ye
authors.forename: Dan
authors.initials: D
authors.lastname: Cao
authors.forename: Rong
authors.initials: R
authors.lastname: Chen
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Pei
authors.forename: Tian
authors.initials: T
authors.lastname: Xie
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Weibo
authors.initials: W
authors.lastname: Gong
authors.forename: Xiaopu
authors.initials: X
authors.lastname: Yin
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D055162:Biocatalysis; D002175:Candida / Q000201:enzymology*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D064429:Fatty Acid Synthases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D010088:Oxidoreductases; C036543:short chain trans-2-enoyl-CoA reductase; D009247:NADH, NADPH Oxidoreductases; D064429:Fatty Acid Synthases