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高效枯草芽孢杆菌168全细胞生物催化剂的构建及其在L-鸟氨酸生产中的应用。

Construction of a highly efficient Bacillus subtilis 168 whole-cell biocatalyst and its application in the production of L-ornithine.

作者信息

Wang Meizhou, Xu Meijuan, Rao Zhiming, Yang Taowei, Zhang Xian

机构信息

The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, Jiangsu, People's Republic of China.

出版信息

J Ind Microbiol Biotechnol. 2015 Nov;42(11):1427-37. doi: 10.1007/s10295-015-1672-z. Epub 2015 Aug 29.

Abstract

L-Ornithine, a non-protein amino acid, is usually extracted from hydrolyzed protein as well as produced by microbial fermentation. Here, we focus on a highly efficient whole-cell biocatalyst for the production of L-ornithine. The gene argI, encoding arginase, which catalyzes the hydrolysis of L-arginine to L-ornithine and urea, was cloned from Bacillus amyloliquefaciens B10-127 and expressed in GRAS strain Bacillus subtilis 168. The recombinant strain exhibited an arginase activity of 21.9 U/mg, which is 26.7 times that of wild B. subtilis 168. The optimal pH and temperature of the purified recombinant arginase were 10.0 and 40 °C, respectively. In addition, the recombinant arginase exhibited a strong Mn(2+) preference. When using whole-cell biocatalyst-based bioconversion, a hyper L-ornithine production of 356.9 g/L was achieved with a fed-batch strategy in a 5-L reactor within 12 h. This whole-cell bioconversion study demonstrates an environmentally friendly strategy for L-ornithine production in industry.

摘要

L-鸟氨酸是一种非蛋白质氨基酸,通常从水解蛋白中提取,也可通过微生物发酵生产。在此,我们聚焦于一种用于生产L-鸟氨酸的高效全细胞生物催化剂。编码精氨酸酶(催化L-精氨酸水解生成L-鸟氨酸和尿素)的基因argI,从解淀粉芽孢杆菌B10-127中克隆出来,并在公认安全(GRAS)菌株枯草芽孢杆菌168中表达。重组菌株表现出21.9 U/mg的精氨酸酶活性,是野生型枯草芽孢杆菌168的26.7倍。纯化后的重组精氨酸酶的最适pH和温度分别为10.0和40℃。此外,重组精氨酸酶对Mn(2+)有强烈偏好。在基于全细胞生物催化剂的生物转化中,采用分批补料策略,在5-L反应器中12小时内实现了356.9 g/L的超高L-鸟氨酸产量。这项全细胞生物转化研究展示了一种工业生产L-鸟氨酸的环境友好策略。

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