生物加工过程2026，Vol.24Issue(2)：135-143,9.DOI:10.3969/j.issn.1672-3678.2026.02.003
特异性生成(S)-3-氯-1-苯基-1-丙醇的酮还原酶的异源表达及酶学性质
Heterologous expression and enzymatic characterization of a ketoreductase for specifical generation of (S)-3-chloro-1-phenyl-1-propanol
摘要
Abstract
A ketoreductase(KRED1),isolated from Lentilactobacillus kefiri,can specifically catalyze the reduction of 3-chloro-1-phenylpropanone.Upon the heterologous expression in Escherichia coli BL21(DE3)and subsequent purification of the recombinant KRED1,its enzymatic properties were characterized.As a result,KRED1 proved to be a novel NADPH-preference ketoreductase,with an optimal temperature of 35 ℃ and an optimal pH of 7.0.The Michaelis constant(Km)of KRED1 for 3-chloro-1-phenylpropanone was determined to be 0.6 mmol/L.Additionally,metal ions including Mn2+,Cu2+,and Zn2+showed strong inhibitory effects on the enzymatic activity of KRED1.Preliminary exploration was also conducted for the KRED1-catalyzed synthesis of(S)-3-chloro-1-phenylpropanol through a glucose dehydrogenase(GDH)-based coenzyme regeneration system.When the reaction was carried out at 35 ℃ and pH 8.0,(SS)-3-chloro-1-phenylpropanol could be generated at significantly higher rate,along with 99％enantiomeric excess(e.e.)Overall,our research demonstrated the potential application of KRED1 in the enzymatic synthesis of(S)-3-chloro-1-phenylpropanol.关键词
酮还原酶/(S)-3-氯-1-苯基-丙醇/异源表达/酶学性质Key words
ketoreductase/(S)-3-chloro-1-phenyl-propanol/heterologous expression/enzymatic properties分类
生物科学引用本文复制引用
安杨,刘飞,张积强,刘延新,李溢真,张小刚,张秀华..特异性生成(S)-3-氯-1-苯基-1-丙醇的酮还原酶的异源表达及酶学性质[J].生物加工过程,2026,24(2):135-143,9.基金项目
山东省重点研发计划(2022SFGC0105) （2022SFGC0105）
济南市合成生物学与绿色生物制造技术研发中心课题(202534034) （202534034）
