文献类型：期刊文献

英文题名：Enhancement of ethyl (S)-4-chloro-3-hydroxybutanoate production at high substrate concentration by in situ resin adsorption

作者：Chen, Li-Feng[1];Fan, Hai-Yang[1];Zhang, Yi-Ping[1];Wei, Wei[1];Lin, Jin-Ping[1];Wei, Dong-Zhi[1];Wang, Hua-Lei[1]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2017

卷号：251

起止页码：68

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20171703601090);WOS:【SCI-EXPANDED(收录号:WOS:000402470300009)】；

基金：This work was supported by the National Natural Science Foundation of China (Grant No. 21406068/B060804), the Fundamental Research Funds for the Central Universities.

语种：英文

外文关键词：Ethyl (S)-4-chloro-3-hydroxybutanoate; Carbonyl reductase; Asymmetric reduction; Product inhibition; Resin adsorption

摘要：Asymmetric reduction of ethyl 4-chloro-3-oxobutyrate (COBE) by carbonyl reductases presents an efficient way to produce Ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE), an important chiral intermediate for the synthesis of hydroxymethylglutaryl-CoA reductase inhibitors such as Lipitor (R). In this study, an NADPH-dependent carbonyl reductase (SrCR) from Synechocystis sp. was characterized to demonstrate a broad substrate spectrum, and the highest activity (53.1 U/mg protein) with COBE. To regenerate the cofactor NADPH, Bacillus subtilis glucose dehydrogenase was successfully coexpressed with SrCR. Owing to the product inhibition, no more than 400 mM of COBE could be completely reduced to (S)-CHBE using the recombinant Escherichia coli/pET-SrCR-GDH. The macroporous adsorption resin HZ 814 was applied to adsorb (S)-CHBE in situ to alleviate the product inhibitio. Consequently, 3000 mM (494 g/L) of COBE was bioconverted within 8 h, resulting in a (S)-CHBE yield of 98.2%, with 99.4% ee and total turnover number of 15,000, revealed great industrial potential of (S)-CHBE production.