文献类型：期刊文献

英文题名：A novel D-mandelate dehydrogenase used in three-enzyme cascade reaction for highly efficient synthesis of non-natural chiral amino acids

作者：Fan, Chang-Wei[1];Xu, Guo-Chao[1];Ma, Bao-Di[1];Bai, Yun-Peng[1];Zhang, Jie[1];Xu, Jian-He[1]

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

年份：2015

卷号：195

起止页码：67

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20150300422937);WOS:【SCI-EXPANDED(收录号:WOS:000348839500010)】；

基金：Mandelate racemase used in this study was kindly provided by the group of Prof. Hong-Wei Yu from Zhejiang University. Financial support from NSFC (No. 21276082) is gratefully acknowledged.

语种：英文

外文关键词：D-Mandelate dehydrogenase; L-Phenylglycine; Asymmetric synthesis; Cascade reaction; Process optimization

摘要：A novel NAD(+)-dependent D-mandelate dehydrogenase was identified from Lactobacillus brevis (LbDMDH). After purified to homogeneity, the optimum pH and temperature for oxidation of D-mandelate were pH 10.0 and 40 degrees C, and the K-m and k(cat) were 1.1 mM and 355 s(-1) respectively. Employing the LbDMDH together with a mandelate racemase from Pseudomonas putida and a leucine dehydrogenase (EsLeuDH) from Exiguobacterium sibiricum, we established a three-step one-pot domino reaction system for preparing chiral L-phenylglycine from racemic mandelic acid with internal cofactor recycling. Under the optimum conditions, 30.4 g rac-mandelic acid (0.2 M) at 1 L scale had been converted into chiral L-phenylglycine, with 96.4% conversion, 86.5% isolation yield, >99% ee(p) and 50.4 g L-1 d(-1) space-time yield. (C) 2014 Elsevier B. V. All rights reserved.