文献类型：期刊文献

英文题名：A high-throughput screening strategy for nitrile-hydrolyzing enzymes based on ferric hydroxamate spectrophotometry

作者：He, Yu-Cai[1,2,3];Ma, Cui-Luan[2,4];Xu, Jian-He[1,3];Zhou, Li[3]

机构：[1]Hubei Univ Technol, Key Lab Fermentat Engn, Minist Educ, Wuhan 430068, Peoples R China;[2]Changzhou Univ, Biochem Engn Lab, Coll Pharmaceut & Life Sci, Changzhou 213164, Peoples R China;[3]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[4]Changzhou Kangpu Pharmaceut Co Ltd, Changzhou 213172, Peoples R China

年份：2011

卷号：89

期号：3

起止页码：817

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20110513633657);WOS:【SCI-EXPANDED(收录号:WOS:000286429400035)】；

基金：This work was financially supported by the National Natural Science Foundation of China (No. 20506037), the Talent Introduction Foundation of Changzhou University (No. ZMF10020077), and the Students' Science & Technology Creation Foundation of Changzhou University (No. 10-Chem-A-11). This work was also partially supported by the Open Project Program of the State Key Laboratory of Bioreactor Engineering (No. 2008004) and the Key Laboratory of Fermentation Engineering (Ministry of Education).

语种：英文

外文关键词：Nitrilase; Carboxylic acids; High-throughput screening; Ferric hydroxamate spectrophotometry; Evaluation

摘要：Nitrile-hydrolyzing enzymes (nitrilase or nitrile hydratase/amidase) have been widely used in the pharmaceutical industry for the production of carboxylic acids and their derivatives, and it is important to build a method for screening for nitrile-hydrolyzing enzymes. In this paper, a simple, rapid, and high-throughput screening method based on the ferric hydroxamate spectrophotometry has been proposed. To validate the accuracy of this screening strategy, the nitrilases from Rhodococcus erythropolis CGMCC 1.2362 and Alcaligenes sp. ECU0401 were used for evaluating the method. As a result, the accuracy for assaying aliphatic and aromatic carboxylic acids was as high as the HPLC-based method. Therefore, the method may be potentially used in the selection of microorganisms or engineered proteins with nitrile-hydrolyzing enzymes.