海洋源纤溶酶高产菌株的诱变育种及液体培养基优化  

Mutation breeding of strains with high fibrinolytic activity derived from marine and optimization of its liquid fermentation medium

作　　者：刘延廖 漆光辉 李培玲[1] 梁智群[1,2,3] 陈桂光[1,2,3] 曾伟[1,2,3] LIU Yanliao QI Guanghui LI Peiling LIANG Zhiqun CHEN Guiguang ZENG Wei(State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, China Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, Guangxi University, Narming 530004, China College of Life Science and Technology, Guangxi University, Nanning 530004, China)

机构地区：[1]广西大学亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004 [2]广西大学微生物及植物遗传工程教育部重点实验室,广西南宁530004 [3]广西大学生命科学与技术学院,广西南宁530004

出　　处：《中国酿造》2016年第12期53-58,共6页China Brewing

基　　金：国家自然基金项目(21062001);广西自然科学基金项目(2015GXNSFBA139052);广西大学博士启动项目(XBZ160113);2016年度广西高等教育创优计划教学相关项目-优势特色专业项目(优质本科专业)

摘　　要：为获得纤溶酶高产菌株及其最优发酵条件,以海洋环境来源的枯草芽孢杆菌(Bacillus subtilis)H-A-03为出发菌株,经过^(60)Co辐射诱变获得了一株遗传稳定性好的纤溶酶高产菌株F8-C,其酶活力较出发菌株提高28.78%。采用统计学优化方法获得了组分更简单、用量更少、更利于菌株产酶的培养基:可溶性淀粉41.6g/L、黄豆粕粉26.2g/L、CaCl_2 1.0g/L,KH_2PO_42.0g/L。在此基础上,3L摇瓶放大试验结果显示,菌株F8-C的产酶量达到(3231±21)U/mL,较出发菌株H-A-3提高44.8%。结果表明利用^(60)Co辐射诱变育种,并采用统计学方法优化培养基,对于提高枯草芽孢杆菌纤溶酶的产酶量是一种有效策略。该研究结果也将为下一步发酵罐扩大试验奠定基础。A marine bacterium, Bacillus subtilis H-A-03 was used as initial strain to obtain mutants with higher fibrinolytic activity and optimal fermentation conditions. After mutagenesis by 60Co radiation, a mutant strain F8-C with good genetic stability was obtained, and the enzyme activity was 28.78% higher than the original strain. By using statistical optimization methods, an optimized fermentation medium which had simple components, less dosage, and easier for fibrinolytic enzyme production was determined as follows： soluble starch 41.2 g/L, soybean meal 26.2 g/L, CaCl2 1 g/L and KH2PO4 2 g/L. The fibrinolytic enzyme activity reached （3 231±21） U/ml in 3 L shake flask experiment under the optimal medium and fermentation conditions, which was 44.8% higher than the initial strain H-A-3. Results showed that 60Co radiation mutation breeding and statistical optimal methods were efficient strategies to improve the fibrinolytic enzyme production. The research results also established a foundation for industrial production of fibrinolytic enzyme in the future.

关 键 词：枯草芽孢杆菌 纤溶酶 发酵培养基 发酵条件 优化 

分 类 号：Q815[生物学—生物工程]