title: Assessing the stereoselectivity of carbonyl reductases toward the reduction of OPBE and docking analysis.
abstract: The asymmetric reduction of prochiral carbonyl compounds by NAD(P)H-dependent carbonyl reductases represents a powerful method for the production of optically active alcohols. The stereoselectivity of a series of carbonyl reductases were evaluated toward the reduction of ethyl 2-oxo-4-phenylbutyrate (OPBE). A majority of reductases produced the ethyl (R)-2-hydroxy-4-phenylbutyrate ((R)-HPBE) with low to excellent enantiomeric excess (e.e.), whereas about 30% reductases catalyzed OPBE to form (S)-HPBE. Among them, the carbonyl reductase from Saccharomyces cerevisiae (SeCR) and short-chain dehydrogenase from Gluconobacter oxydans (GoKR) exhibited 100% e.e., yielding the corresponding (R) and (S)-HPBE, respectively. However, the SeCR showed relative higher activity (29.0 U/mg) and affinity (Km of 0.22 mM) than those of GoKR. Docking analysis found that the interaction of OPBE with enzyme-NADPH complex determined the NADPH-provided hydrogen transfer and the configuration of reductive product. These results indicated that the three-dimensional (3D) structure of enzymes controlled the stereoselectivity of the reductive product based on the geometry of the substrate and cofactor.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, People's Republic of China.|Center for Biomedicine and Health, Division of Basical Medicine, Hangzhou Normal University, Hangzhou, People's Republic of China.
authors.forename: Rong
authors.initials: R
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, People's Republic of China.
authors.forename: Jian
authors.initials: J
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, People's Republic of China.
authors.forename: Jinping
authors.initials: J
authors.lastname: Lin
authors.affiliation: Center for Biomedicine and Health, Division of Basical Medicine, Hangzhou Normal University, Hangzhou, People's Republic of China.
authors.forename: Xiaopu
authors.initials: X
authors.lastname: Yin
authors.affiliation: Center for Biomedicine and Health, Division of Basical Medicine, Hangzhou Normal University, Hangzhou, People's Republic of China.
authors.forename: Tian
authors.initials: T
authors.lastname: Xie
authors.affiliation: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, People's Republic of China.
authors.forename: Shengli
authors.initials: S
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, People's Republic of China.
authors.forename: Dongzhi
authors.initials: D
authors.lastname: Wei
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D055162:Biocatalysis; D007700:Kinetics; D062105:Molecular Docking Simulation; D010084:Oxidation-Reduction; D010654:Phenylbutyrates / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D010654:Phenylbutyrates; C572921:ethyl 2-oxo-4-phenylbutyrate; D000429:Alcohol Oxidoreductases
keywords: Prelog's rule; asymmetric reduction; carbonyl reductase; chiral alcohol; docking analysis