caa a22 4500 605504296 CHVBK 20210128100615.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00253-014-6245-y doi (NATIONALLICENCE)springer-10.1007/s00253-014-6245-y Upscale production of ethyl ( S )-4-chloro-3-hydroxybutanoate by using carbonyl reductase coupled with glucose dehydrogenase in aqueous-organic solvent system [Elektronische Daten] [Zhi-Qiang Liu, Jing-Jing Ye, Zhen-Yang Shen, Hua-Bin Hong, Jian-Bo Yan, Yi Lin, Zheng-Xu Chen, Yu-Guo Zheng, Yin-Chu Shen] (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE) is an important chiral intermediate to synthesize the side chain of cholesterol-lowering drug atorvastatin. To biosynthesize the (S)-CHBE, a recombinant Escherichia coli harboring the carbonyl reductase and glucose dehydrogenase was successfully constructed. The recombinant E. coli was cultured in a 500-L fermentor; after induction and expression, the enzyme activity and cell biomass were increased to 23,661.65U/L and 13.90g DCW/L which was 3.24 and 2.60-folds compared with those in the 50L fermentor. The biocatalytic process for the synthesis of (S)-CHBE in an aqueous-organic solvent system was constructed and optimized with a substrate fed-batch strategy. The ethyl 4-chloro-3-oxobutanoate concentration reached to 1.7M, and the (S)-CHBE with yield of 97.2% and enantiomeric excess (e.e.) of 99% was obtained after 4-h reaction in a 50-L reactor. In this study, the space-time yield and space-time yield per gram of biomass (dry cell weight, DCW) were 413.17mM/h and 27.55mM/h/g DCWfor(S)-CHBE production, respectively, which were the highest values as compared to previous reports. Finally, (S)-CHBE was extracted from the reaction mixture with 82% of yield and 95% of purity. This study paved the foundation for the upscale production of (S)-CHBE by biocatalysis method. Springer-Verlag Berlin Heidelberg, 2014 Co-expression nationallicence Carbonyl reductase nationallicence Glucose dehydrogenase nationallicence Ethyl ( S )-4-chloro-3-hydroxybutanoate nationallicence Liu Zhi-Qiang Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut Ye Jing-Jing Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut Shen Zhen-Yang Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut Hong Hua-Bin Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut Yan Jian-Bo Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut Lin Yi Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut Chen Zheng-Xu Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut Zheng Yu-Guo Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut Shen Yin-Chu Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2119-2129 0175-7598 99:5<2119 2015 99 253 https://doi.org/10.1007/s00253-014-6245-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6245-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liu Zhi-Qiang Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Ye Jing-Jing Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Shen Zhen-Yang Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Hong Hua-Bin Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Yan Jian-Bo Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Lin Yi Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Chen Zheng-Xu Zhejiang Neo-DanKong Pharmaceutical Co., Ltd, 318000, Taizhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Zheng Yu-Guo Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 700 1- Shen Yin-Chu Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, Zhejiang, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/5(2015-03-01), 2119-2129 0175-7598 99:5<2119 2015 99 253