枯草芽孢杆菌甘油激酶编码基因定点突变提升甘油利用水平的研究  

Effect of site-specific mutagenesis of glycerol kinase coding gene on the glycerol utilization of Bacillus subtilis

作　　者：王光路 张帆 周忆菲 王梦园 赵金垒 李明笑 杨雪鹏 马歌丽 WANG Guanglu;ZHANG Fan;ZHOU Yifei;WANG Mengyuan;ZHAO Jinlei;LI Mingxiao;YANG Xuepeng;MA Geli(College of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450001,China;Food Inspection and Testing Institute of He′nan Province,Zhengzhou 450003,China)

机构地区：[1]郑州轻工业大学食品与生物工程学院,河南郑州450001 [2]河南省口岸食品检验检测所,河南郑州450003

出　　处：《轻工学报》2020年第6期1-8,共8页Journal of Light Industry

基　　金：国家自然科学基金联合基金项目(U1904101);河南省科技攻关重点研发与推广专项项目(202102310021,182102310607)。

摘　　要：采用无痕等位基因置换方法,将枯草芽孢杆菌基因组上的甘油激酶编码基因glpK第270位氨基酸残基M突变为I,构建了突变工程菌株枯草芽孢杆菌M270I,并分析了该突变菌株的生长特性.结果表明:对甘油激酶编码基因glpK进行定点突变可有效提升枯草芽孢杆菌对甘油的利用水平,与出发菌株枯草芽孢杆菌168Δupp相比,突变菌株枯草芽孢杆菌M270I在M9甘油基本盐液体培养基中的比生长速率提升了11%,延滞期缩短了2~4h,最大菌体生物量提升了16%.By employing a site-specific mutagenesis strategy for glycerol kinase(encoding by glpK),the active-site residue Met270 was replaced with Ile in the Bacillus subtilis genome.A mutant strain Bacillus subtilis M270I was constructed successfully by using the markerless gene replacement system.The results of growth analysis indicated that the mutant strain got a higher level of glycerol utilization.Compared with the control strain Bacillus subtilis 168Δupp,the mutant strain exhibited higher specific growth rate(increased by 11%)and higher biomass yield(improved by 16%)with a shorten lag phase(shorten 2~4 h)in the M9 glycerol minimal medium.

关 键 词：枯草芽孢杆菌 甘油激酶 定点突变 无痕等位基因置换 

分 类 号：TS201.3[轻工技术与工程—食品科学] Q789[轻工技术与工程—食品科学与工程]