重离子诱变枯草芽孢杆菌高产复合酶菌种的筛选  

Screening of Bacillus subtilis Mutants of a High-yielding Compound Enzyme by Heavy-ion Beam Irradiation

作　　者：刘艳新[1,2] 胡建华 崔金娜[1,3] 张耀胜 李永丽 石雅丽 朱明达[1,3] 刘占英 LIU Yanxin;HU Jianhua;CUI Jinna;ZHANG Yaosheng;LI Yongli;SHI Yali;ZHU Mingda;LIU Zhanying(College of Chemical Engineering, Inner Mongolia University of Technology, Hohhot 010051, Inner Mongolia, China;Shenzhou Biology & Technology Co., Ltd. Hohhot 010206, Inner Mongolia, China;Inner Mongolia Engineering Center of Energy Conservation and Emission Reduction in Fermentation Industry, Hohhot 010051, Inner Mongolia, China)

机构地区：[1]内蒙古工业大学化工学院,内蒙古呼和浩特010051 [2]神舟生物科技有限责任公司,内蒙古呼和浩特010206 [3]内蒙古自治区发酵产业节能减排工程技术研究中心,内蒙古呼和浩特010051

出　　处：《激光生物学报》2019年第2期123-130,共8页Acta Laser Biology Sinica

基　　金：兰州重离子加速器国家实验室课题项目;内蒙古科技计划项目;中国科学院"西部之光"人才培养计划项目;国家自然科学基金项目(61361016);内蒙古自治区高等学校青年科技英才支持计划项目;内蒙古"草原英才"工程及"草原英才"滚动支持计划项目(CYYC20130034);内蒙古自治区自然科学基金项目(2018MS02019);内蒙古自治区科技成果转化项目(CGZH2018133)

摘　　要：重离子诱变作为一种新型高效的辐照诱变方式,具有突变率高、突变谱广、突变体稳定等特点。枯草芽孢杆菌(Bacillus subtilis)因具有多种水解酶活力而广泛应用于工业生产,虽然经过多次诱变和改造,其酶活力不断提高,但其复合酶活力仍有提升空间。同时,多次同种诱变剂的使用容易导致菌种产生"抗性",酶活力提高不大。为此,本研究首次利用不同剂量^(12)C^(6+)重离子束对枯草芽孢杆菌20076进行辐照诱变。诱变后通过透明圈初筛和酶活力验证复筛方式选育出一株复合酶活力较高的菌株B. subtilis KC-1。与原始菌株相比,其发酵液滤纸酶活力、内切葡聚糖酶活力、β-葡萄糖苷酶活力、外切葡聚糖酶活力、蛋白酶活力和α-淀粉酶活力分别提高了20. 2%、79. 6%、98. 2%、7. 4%、10. 4%和27. 4%。11代遗传稳定性实验结果表明该突变株具有良好的稳定性。As a new high efficiency irradiation mutation source, heavy ion mutagenesis has advantages of high mutation rate, broad mutation spectrum, and good genetic stability for mutants. Bacillus subtilis , which has a broad hydrolase system, is one of the most important bacteria for industrial application. In this research, Bacillus subtilis 20076 was irradiated by various doses of 12 C 6+ heavy ion beam to increase its enzyme production for industrial application. Based on transparent circle and enzyme activities, a target strain was obtained and named as KC-1. The strain showed an increase of 20.2%,79.6%, 98.2%, 7.4%,10.4%and 27.4%respectively in filter paper enzyme, endo-1,4-glucanase,β-glucosidase, exo-1,4-glucanase, protease and α-amylase when compared to the original strain. After the 11th passages, the mutant also had good stability in compound enzyme production.

关 键 词：枯草芽孢杆菌 复合酶 重离子诱变 遗传稳定性 

分 类 号：Q931[生物学—微生物学]