title: Enzymatic synthesis of an ezetimibe intermediate using carbonyl reductase coupled with glucose dehydrogenase in an aqueous-organic solvent system.
abstract: (4S)-3-[(5S)-5-(4-Fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one ((S)-ET-5) is an important chiral intermediate in the synthesis of chiral side chain of ezetimibe. Recombinant Escherichia coli expressing carbonyl reductase (CBR) was successfully constructed in this study. The total E. coli biomass and the specific activity of recombinant CBR in 5L fermenter culture were 10.9gDCWL-1and 14900.3Ug-1DCW, respectively. The dual-enzyme coupled biocatalytic process in an aqueous-organic biphasic solvent system was first constructed using p-xylene as the optimal organic phase under optimized reaction conditions, and 150gL-1(4S)-3-[5-(4-fluorophenyl)-1,5-dioxophentyl]-4-phenyl-1,3-oxazolidin-2-one (ET-4) was successfully converted to (S)-ET-5 with a conversion of 99.1% and diastereomeric excess of 99% after 24-h, which are the highest values reported to date for the production of (S)-ET-5.
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Liu
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Si-Chuan
authors.initials: SC
authors.lastname: Dong
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Huan-Huan
authors.initials: HH
authors.lastname: Yin
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Xue
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Tang
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
authors.forename: Xiao-Jian
authors.initials: XJ
authors.lastname: Zhang
authors.affiliation: School of Pharmaceutical Engineering, Zhejiang Pharmaceutical College, Ningbo 315100, PR China.
authors.forename: Jun-Yao
authors.initials: JY
authors.lastname: He
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China. Electronic address: zhengyg@zjut.edu.cn.
authors.forename: Yu-Guo
authors.initials: YG
authors.lastname: Zheng
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D055162:Biocatalysis / Q000187:drug effects; D019149:Bioreactors; D001711:Biotransformation; D002021:Buffers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D000069438:Ezetimibe / Q000378:metabolism*; D005285:Fermentation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009249:NADP / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D012997:Solvents / Q000737:chemistry*; D013696:Temperature; D014867:Water / Q000737:chemistry*
chemical_list: D002021:Buffers; D011994:Recombinant Proteins; D012997:Solvents; D014867:Water; D009249:NADP; D000429:Alcohol Oxidoreductases; D042843:Glucose 1-Dehydrogenase; D000069438:Ezetimibe; D005947:Glucose
keywords: Aqueous-organic solvent system; Carbonyl reductase; Chiral intermediate of ezetimibe; Dual-enzyme coupled system; Glucose dehydrogenase