title: Highly efficient bioreduction of 2-hydroxyacetophenone to (S)- and (R)-1-phenyl-1,2-ethanediol by two substrate tolerance carbonyl reductases with cofactor regeneration.
abstract: Optically pure 1-phenyl-1,2-ethanediol is a very important chiral building block and intermediate in fine chemical and pharmaceutical industries. Reduction of 2-hydroxyacetophenone provides a straightforward approach to access these important compounds. In this study, two enantiocomplementary carbonyl reductases, BDHA (2,3-butanediol dehydrogenase from Bacillus subtilis) and GoSCR (polyol dehydrogenase from Gluconobacter oxydans) were discovered for the first time to convert 2-hydroxyacetophenone (2-HAP) to (R)-1-phenyl-1,2-ethanediol ((R)-PED) and (S)-1-phenyl-1,2-ethanediol ((S)-PED) with excellent stereochemical selectivity, respectively. The two enzymes were purified and characterized. In vitro bioreduction of 2-HAP catalyzed by BDHA and GoSCR coupled with glucose dehydrogenase (GDH) from Bacillus subtilis for cofactor regeneration were demonstrated, affording both (R)-PED and (S)-PED in>99% ee and 99% conversion. Recombinant Escherichia coli whole cells co-expressing both GDH and BDHA or GoSCR genes were used to asymmetric reduction of 2-HAP to (R)-PED or (S)-PED. Under the optimized conditions, the bioreduction of 400mM (54g/L) substrate was proceeded smoothly without the external addition of cofactor, and the product (R)-PED and (S)-PED were obtained with 99% yield, >99% ee and 18.0g/L/h volumetric productivity. These results offer a practical biocatalytic method for the preparation of both (R)-PED and (S)-PED with high volumetric productivity.
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, Shanxi, 030024, PR China.
authors.forename: Zhi-Mei
authors.initials: ZM
authors.lastname: Cui
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, Shanxi, 030024, PR China. Electronic address: zhangjiandong@tyut.edu.cn.
authors.forename: Jian-Dong
authors.initials: JD
authors.lastname: Zhang
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, Shanxi, 030024, PR China.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacture, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, PR China.
authors.forename: Gao-Wei
authors.initials: GW
authors.lastname: Zheng
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, Shanxi, 030024, PR China. Electronic address: changhonghong@tyut.edu.cn.
authors.forename: Hong-Hong
authors.initials: HH
authors.lastname: Chang
authors.affiliation: Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, Shanxi, 030024, PR China.
authors.forename: Wen-Long
authors.initials: WL
authors.lastname: Wei
mesh_terms: D000098:Acetophenones / Q000737:chemistry / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D001412:Bacillus subtilis / Q000201:enzymology; D001711:Biotransformation; D002072:Butylene Glycols / Q000378:metabolism; D003001:Cloning, Molecular; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005026:Ethylene Glycols / Q000737:chemistry / Q000378:metabolism*; D020600:Gluconobacter oxydans / Q000201:enzymology / Q000235:genetics; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism; D007064:L-Iditol 2-Dehydrogenase / Q000378:metabolism; D018832:Molecular Chaperones; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D000098:Acetophenones; D002072:Butylene Glycols; D029968:Escherichia coli Proteins; D005026:Ethylene Glycols; D018832:Molecular Chaperones; C031474:styrene glycol; C087908:2'-hydroxyacetophenone; C026978:2,3-butylene glycol; D000429:Alcohol Oxidoreductases; D007064:L-Iditol 2-Dehydrogenase; D042843:Glucose 1-Dehydrogenase
keywords: 2-Hydroxyacetophenone; Carbonyl reductase; Chiral 1-phenyl-1,2-ethanediol; Cofactor regeneration; Highly enantioselective