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腈水解酶在枯草芽胞杆菌中的表达及在烟酸合成中的应用

发表时间:2020-10-10  浏览量:635  下载量:106
全部作者: 王顺治,汪子凯,龚劲松,李恒,李会,许正宏,史劲松
作者单位: 江南大学生物工程学院;江南大学药学院
摘 要: 研究将腈水解酶基因NITR与大肠杆菌−枯草芽孢杆菌穿梭质粒pMA5连接,并导入到枯草芽孢杆菌WB600中进行重组表达,重组菌在发酵培养36 h后酶活达到38 U·mL-1. 本文进一步研究了重组枯草芽孢杆菌腈水解酶的酶学性质,以及批次补料模式下细胞的转化能力。在底物补料浓度为200 mmol·L-1时,可以在450 min内完全转化12批底物3-氰基吡啶,反应体系中累积的烟酸浓度达到295 g·L-1. 本研究对枯草芽孢杆菌中腈水解酶重组表达的报道尚属首次。
关 键 词: 微生物学;枯草芽孢杆菌;腈水解酶;烟酸;批次转化;3-氰基吡啶
Title: Expression of nitrilase in Bacillus subtilis and its application in nicotinic synthesis
Author: WANG Shunzhi, WANG Zikai, GONG Jinsong, LI Heng, LI Hui, XU Zhenghong, SHI Jinsong
Organization: School of Biotechnology, Jiangnan University; School of Pharmaceutical Sciences, Jiangnan University
Abstract: In this study, the nitrilase gene NITR was ligated with the Escherichia coli-Bacillus subtilis shuttle plasmid pMA5 and introduced into B. subtilis WB600 for recombinant expression. The enzyme activity reached as high as 38 U·mL-1 when the recombinant bacteria was fermentation cultured for 36 h. The enzymatic properties of the recombinant B. subtilis nitrilase were studied and the cell transformation ability under batch feeding mode was investigated. 12 batches of substrate 3-cyanopyridine could be completely transformed within 450 min with final substrate feeding concentration of 200 mmol·L-1 and the accumulated nicotinic acid concentration was 295 g·L-1 in the reaction system with no residual substrate detected. This is the first report about the recombinant expression of nitrilase in B. subtilis.
Key words: microbiology; Bacillus subtilis; nitrilase; nicotinic acid; batch conversion; 3-cyanopyridine
发表期数: 2020年9月第3期
引用格式: 王顺治,汪子凯,龚劲松,等. 腈水解酶在枯草芽胞杆菌中的表达及在烟酸合成中的应用[J]. 中国科技论文在线精品论文,2020,13(3):272-280.
 
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