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
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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168 Δupp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2~4 h，最大菌体生物量提升了16%.Abstract: 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 168 Δupp, 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.
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