文献类型：期刊文献

英文题名：Efficient production of ethyl (R)-2-hydroxy-4-phenylbutyrate using a cost-effective reductase expressed in Pichia pastoris

作者：Qian, Xiao-Long[1];Pan, Jiang[1];Shen, Nai-Dong[1];Ju, Xin[1];Zhang, Jie[1];Xu, Jian-He[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biocatalysis & Synthet Biotechnol, Shanghai 200237, Peoples R China

年份：2014

卷号：91

起止页码：72

外文期刊名：BIOCHEMICAL ENGINEERING JOURNAL

收录：;EI(收录号：20144300129213);WOS:【SCI-EXPANDED(收录号:WOS:000343950800010)】；

基金：This work was financially supported by the National Natural Science Foundation of China (Nos. 21276082 and 31200050), Ministry of Science and Technology, China (No. 2011CB710800, 2011AA02A210), and Shanghai Commission of Science and Technology (No. 11431921600).

语种：英文

外文关键词：Biocatalysis; (R)-HPBE; Enzyme production; Recombinant Pichia pastoris; Fermentation; Cost-effective reductase; Fed-batch culture

摘要：Recombinant strains of Pichia pastoris expressing carbonyl reductase CgKR2 from Candida glabrata were constructed for stereoselective reduction of ethyl-2-oxo-4-phenylbutyrate (OPBE) to ethyl (R)-2-hydroxy-4-phenylbutyrate [(R)-HPBE], an important building block for synthesis of angiotensin-converting enzyme (ACE) inhibitors. An intracellular expression level of 6.67 g L-1 and a secretion expression level of 3.0 g L-1 were obtained, respectively, by high cell density fermentation. By using whole cells of KM71/CgKR2 in aqueous system, the CgKR2-mediated bioreduction was performed, achieving a complete conversion of 1.0 M OPBE at 0.5 L scale, with a final yield of 77.9% and an enantiomeric excess (ee) of 97.3%. The secreted enzyme CgKR2 appeared to be a better catalyst with respect to the perfect ee of the product (R)-HPBE (>99%) when comparing with the recombinant cells of KM71/CgKR2. The cost of the resultant (R)-HPBE was reduced by 1/4 using the secreted CgKR2. (C) 2014 Elsevier B.V. All rights reserved.