数字农科院2.0

Biodegradation of nitriles derived from glucosinolates in rapeseed meal by BnNIT2: a nitrilase from Brassica napus with wide substrate specificity

文献类型： 外文期刊

作者： Zhang, Heng;Zhang, Honghai;Qin, Xing;Wang, Xiaolu;Wang, Yuan;Tu, Tao;Wang, Yaru;Yao, Bin;Huang, Huoqing;Luo, Huiying

作者机构：

关键词： Brassica napus;Nitrilase;Rapeseed meal;Thermostability improvement;Detoxification

期刊名称： APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

ISSN： 0175-7598

年卷期： 2022 年

页码：

收录情况： SCIE（2022版） ; EI（2022版）

摘要： Nitriles derived from glucosinolates (GSLs) in rapeseed meal (RSM) can cause lesions on animal liver and kidneys. Nitrilase converts nitriles to carboxylic acids and NH3, eliminating their toxicity. Here we describe a nitrilase, BnNIT2, from Brassica napus (optimal temperature, 45 degrees C; pH, 7.0) that is stable at 40 degrees C and has a wide substrate specificity. Recombinant BnNIT2 converted the three main nitriles from GSLs (3-hydroxy-4-pentenenitrile, 3-butenenitrile, and 4-pentenenitrile), with the highest specific activity (58.6 U/mg) for 4-pentenenitrile. We used mutagenesis to improve the thermostability of BnNIT2; the resulting mutant BnNIT2-H90M had an similar to 14.5% increase in residual activity at 50 degrees C for 1 h. To verify the functionality of BnNIT2, GSLs were extracted from RSM and converted into nitriles at pH 5.0 in the presence of Fe2+. Then, BnNIT2 was used to degrade the nitriles from GSLs; ultimately, similar to 80% of nitriles were removed. Thus BnNIT2 is a potential enzyme for detoxification of RSM.

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