文献类型：期刊文献

英文题名：Switching the secondary and natural activity of Nitrilase from Acidovorax facilis 72 W for the efficient production of 2-picolinamide

作者：Wang, Liuzhu[1];Jiang, Shuiqin[1];Sun, Yangyang[1];Yang, Zeyu[1];Chen, Zhi[1];Wang, Hualei[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, Shanghai 200237, Peoples R China

年份：2021

卷号：43

期号：8

起止页码：1617

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20211910329903);WOS:【SCI-EXPANDED(收录号:WOS:000648305100001)】；

基金：This work was supported by the China Postdoctoral Science Foundation (No. 2017M621390), the Fundamental Research Funds for the Central Universities (No. 222201814037), and National Natural Science Foundation of China (No. 21676090).

语种：英文

外文关键词：Biocatalyst; Catalytic Promiscuity; Nitrilase; Protein Engineering; 2-Picolinamide

摘要：Objectives Catalytic promiscuity, or the ability to catalyze a secondary reaction, provides new opportunities for industrial biocatalysis by expanding the range of biocatalytic reactions. Some nitrilases converting nitriles to amides, referred to as the secondary activity, show great potential for amides production. And our goal was exploiting the amide-forming potential of nitrilases. Results In this study, we characterized and altered the secondary activity of nitrilase from Acidovorax facilis 72 W (Nit72W) towards different substrates. We increased the secondary activity of Nit72W towards 2-cyanopyridine by 196-fold and created activity toward benzonitrile and p-nitrophenylacetonitrile by modifying the active pocket. Surprisingly, the best mutant, W188M, completely converted 250 mM 2-cyanopyridine to more than 98% 2-picolinamide in 12 h with a specific activity of 90 U/mg and showed potential for industrial applications. Conclusions Nit72W was modified to increase its secondary activity for the amides production, especially 2-picolinamide.