文献类型：期刊文献

英文题名：Cloning, Overexpression, and Characterization of a High Enantioselective Nitrilase from Sphingomonas wittichii RW1 for Asymmetric Synthesis of (R)-Phenylglycine

作者：Qiu, Jian[1];Su, Er-Zheng[2];Wang, Hua-Lei[1];Cai, Wen-Wen[1];Wang, Wei[1];Wei, Dong-Zhi[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, Shanghai 200237, Peoples R China;[2]Nanjing Forestry Univ, Coll Light Ind Sci & Engn, Enzyme & Fermentat Technol Lab, Nanjing 210037, Jiangsu, Peoples R China

年份：2014

卷号：173

期号：2

起止页码：365

外文期刊名：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录：;EI(收录号：20142017712460);WOS:【SCI-EXPANDED(收录号:WOS:000336492000003)】；

基金：This work was supported by the National Basic Research Program of China (No. 2012CB721003) and the Fundamental Research Funds for the Central Universities.

语种：英文

外文关键词：Nitrilase; Sphingomonas wittichiiRW1; Kinetic resolution; (R)-phenylglycine

摘要：In this study, a high (R)-enantioselective nitrilase gene from Sphingomonas wittichii RW1 was cloned and overexpressed in Escherichia coli BL21 (DE3). The recombinant nitrilase was purified to homogeneity with a molecular weight of 40 kDa. The pH and temperature optima were shown to be pH 8.0 and 40 A degrees C, respectively. The purified nitrilase was most active toward succinonitrile, approximately 30-fold higher than that for phenylglycinonitrile. Using the E. coli BL21/ReSWRW1 whole cells as biocatalysts, the kinetic resolution for asymmetric synthesis of (R)-phenylglycine was investigated at pH 6.0. A yield of 46 % was obtained with 95 % enantiomeric excess (ee), which made it a promising biocatalyst for synthesis of (R)-phenylglycine.