High-throughput screening methods for nitrilases

被引:22
作者
Xue, Ya-Ping [1 ,2 ]
Yang, Yue-Kai [1 ,2 ]
Lv, Sheng-Zhi [1 ,2 ]
Liu, Zhi-Qiang [1 ,2 ]
Zheng, Yu-Guo [1 ,2 ]
机构
[1] Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China
[2] Zhejiang Univ Technol, Minist Educ, Engn Res Ctr Bioconvers & Biopurificat, Hangzhou 310014, Zhejiang, Peoples R China
基金
中国国家自然科学基金;
关键词
Nitrilase; High-throughput screening; Nitrile; Carboxylic acid; Biocatalysis; ALCALIGENES-FAECALIS NITRILASE; SITE SATURATION MUTAGENESIS; (R)-(-)-MANDELIC ACID; RACEMIC MANDELONITRILE; HYDROLASE LIBRARIES; DIRECTED EVOLUTION; PH INDICATORS; GLYCOLIC ACID; ASSAY; BIOTRANSFORMATIONS;
D O I
10.1007/s00253-016-7381-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Nitrilases have been widely acknowledged as important alternatives to chemical catalysts, as they have been proved to transform an immense variety of nitriles under mild conditions and often in a stereoselective or regioselective manner. In the discovery of new nitrilases to establish viable industrial processes, screening plays an important role in identifying which subset of candidates contains a nitrilase of interest from a collection of organisms, clone banks, or enzyme libraries. However, the traditional methods for evaluating the nitrilases are a time-consuming, laborious, and costly process and have been regarded as a bottleneck in developing these nitrilases as industrial biocatalysts. In the past few years, a number of high-throughput screening methods have been developed for rapid evaluation and identification of nitrilases. Here, we review the various methodologies developed for high-throughput screening of nitrilases and focus on their advantages and limitations.
引用
收藏
页码:3421 / 3432
页数:12
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