surfactin高产菌株的等离子体诱变及其高通量筛选  

High-throughput screening of Bacillus subtilis mutants with high yield of surfactinby ARTP

作　　者：李光[1] 唐小玲[1] 韦璇[2] 朱学亮[1] 张勇 张乐乐 陈双喜[1] 

机构地区：[1]河南大学生命科学学院,河南开封475000 [2]沈阳师范大学生命科学学院,辽宁沈阳110000 [3]无锡源清天木生物科技有限公司,江苏无锡214000

出　　处：《食品与发酵工业》2017年第2期67-72,共6页Food and Fermentation Industries

摘　　要：利用常压室温等离子体(atmospheric pressure and room temperature plasma,ARTP)方法对枯草芽孢杆菌Bacillus subtilis E7进行快速诱变,筛选表面活性素(surfactin)高产菌株。采用表观初筛,溶血圈初筛,孔板复筛,多功能酶标仪检测,摇瓶验证相结合的方法提高了筛选通量,加快了筛选进程,构建了高通量诱变筛选surfactin高产菌株的体系,并使用高效液相(high performance liquid chromatography,HPLC)检测方法对产物进行检测。优化处理时间为220 s,获得了遗传稳定的突变菌株Bacillus subtillus SF90-5,其产surfactin能力提高到约0.8 g/L,是原始菌株的5倍。ARTP诱变技术结合高通量筛选体系极大程度减少了工作量,提高了筛选通量,加快了筛选进度,并快速获得了目的菌株,为脂肽类表面活性剂进一步的工艺研究提供了基础。In this study,ARTP（atmospheric pressure and room temperature plasma, ARTP） method was used to obtain rapid mutagenesis of Bacillus subtilis E7 in atmospheric pressure and room temperature, to screen strain with high yield of surfactin. Several methods such as appearance screening, determination of diameter of hemolytic circle, culture in Microflask, detection by muhifunctional enzyme standard detector, and shake flask test were combined to improve screening throughput and speed up screening process. A high-throughput screening system for mutant strain with high yield of surfactin was preliminarily built. High performance liquid chromatography （ HPLC ） was used to detect target product. After mutagenesis for 220 s, Bacillus subtillus strain SF90-5 with good genetic stability was obtained. Its surfactin yield increased to 0.8 g/L, which was 5 times of that of the original strain. Mutagenesis with AR- TP resulted inreduced workload,effectively increased screening flux, and accelerated screening process. Desired target strain was obtained fleetly. This experiment provided a foundation for study of strain with high yield of surfactin.

关 键 词：SURFACTIN 常压室温等离子体 枯草芽孢杆菌 高通量 

分 类 号：TS202.3[轻工技术与工程—食品科学]