采用全基因组改组技术选育蛋白酶高产菌株  

Whole Genome Shuffling of Bacillus subtilis for Enhanced Protease Production

作　　者：王景会[1,2] 刘景圣[1] 高岩[1] 马颖辉[3] 李玉秋[2] 牛春华[2] 

机构地区：[1]吉林农业大学食品科学与工程学院,吉林长春130118 [2]吉林省农业科学院农产品加工研究中心,吉林长春130033 [3]黑龙江八一农垦大学食品学院,黑龙江大庆163319

出　　处：《食品科学》2012年第19期211-215,共5页Food Science

基　　金：国家"863"计划项目(2008AA100802);吉林省自然科学基金项目(201215196)

摘　　要：为了提高枯草芽孢杆菌B4的蛋白酶产量,采用全基因组改组技术进行菌株选育,先通过紫外诱变构建了B4菌的突变体库,在优化其原生质体制备和再生条件的基础上,以其中4株诱变菌株(BRS1、BRS2、BRS5、BRS6)作为亲本,采用PEG介导的方法进行两次多亲本的全基因组改组,同时,结合双亲灭活的筛选方法,共筛选出3株酶活力大幅度提高并能稳定遗传的优良菌株,为BRS1、BRS5、BRS6,酶活力最高达2175.81U/mL,达到原始菌株的3.01倍。Whole genome shuffling technique was used to enhance protease production by Bacillus subtilis B4. A candidate mutant library was constructed by UV irradiation of Bacillus subtilis B4. Based on the optimum conditions for protoplast preparation and regeneration, multi-parental whole genome shuffling was carded out with 4 mutant strains （BRS1, BRS2, BRS5 and BRS6） by PEG mediated protoplasts fusion. Then BRS 1, BRS5 and BRS6 were identified by biparental inactivation method as 3 excellent strains with greatly improved protease activity and good genetic stability. The maximum activity was as high as 2175.81 U/mL, which was increased 3.01 fold compared with the initial strain B4.

关 键 词：枯草芽孢杆菌 蛋白酶 基因组改组 

分 类 号：Q819[生物学—生物工程]