title: Effective biosynthesis of ethyl (R)-4-chloro-3-hydroxybutanoate by supplementation of l-glutamine, d-xylose and β-cyclodextrin in n-butyl acetate-water media.
abstract: To avoid adding NAD(+) and effectively transform ethyl 4-chloro-3-oxobutanoate, the mixture of l-glutamine (200mM) and d-xylose (250mM) was added into in n-butyl acetate-water (10:90, v/v) biphasic system instead of NAD(+) for increasing the biocatalytic efficiency. To further improve the synthesis of optically pure ethyl (R)-4-chloro-3-hydroxybutanoate (>99% ee), β-cyclodextrin was also added into this reaction media, and ethyl (R)-4-chloro-3-hydroxybutanoate (>99% ee) could be effectively synthesized from 800mM ethyl 4-chloro-3-oxobutanoate in the yield of 100% by whole-cells of recombinant E. coli CCZU-A13. Finally, the possible mechanism for improving the reductase activity by supplementation of l-glutamine, d-xylose and β-CD was proposed. In conclusion, this strategy has high potential for the effective biosynthesis of ethyl (R)-4-chloro-3-hydroxybutanoate (>99% ee).
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China. Electronic address: heyucai2001@163.com.
authors: Yu-Cai
authors: YC
authors: He
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Zhi-Cheng
authors: ZC
authors: Tao
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Yun
authors: Y
authors: Ding
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Dan-Ping
authors: DP
authors: Zhang
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Yin-Qin
authors: YQ
authors: Wu
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Yun
authors: Y
authors: Lu
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Feng
authors: F
authors: Liu
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Yu-Feng
authors: YF
authors: Xue
authors: Laboratory of Biochemical Engineering, College of Pharmaceutical Engineering and Life Sciences, Changzhou University, Changzhou, China.
authors: Cheng
authors: C
authors: Wang
authors: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors: Jian-He
authors: JH
authors: Xu
mesh_terms: D000085:Acetates / Q000378:metabolism; D000090:Acetoacetates / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D002087:Butyrates / Q000378:metabolism*; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005973:Glutamine / Q000494:pharmacology*; D010088:Oxidoreductases / Q000378:metabolism; D014867:Water; D014994:Xylose / Q000494:pharmacology*; D047392:beta-Cyclodextrins / Q000494:pharmacology*
chemical_list: D000085:Acetates; D000090:Acetoacetates; D001426:Bacterial Proteins; D002087:Butyrates; D003470:Culture Media; D047392:beta-Cyclodextrins; C121244:ethyl 4-chloro-3-hydroxybutanoate; D014867:Water; D005973:Glutamine; C006848:butyl acetate; C111982:ethyl 4-chloro-3-oxobutanoate; D014994:Xylose; D010088:Oxidoreductases; C031215:betadex
keywords: Ethyl (R)-4-chloro-3-hydroxybutanoate; Ethyl 4-chloro-3-oxobutanoate; d-Xylose; l-Glutamine; β-Cyclodextrin, E. coli CCZU-A13