多杀菌素产生菌复合诱变选育及发酵培养基优化  

Breeding of spinosad producing strains by combined mutation and optimization of fermentation medium

作　　者：扶教龙 徐敏强 张松 张瀚 鞠鑫 姚雪梅 胡翠英 李良智 Fu Jiao-long;Xu Min-qiang;Zhang Song;Zhang Han;Ju Xin;Yao Xue-mei;Hu Cui-ying;Li Liang-zhi(Suzhou University of Science and Technology,School of Chemistry,Biology and Materials Engineering,Suzhou 215009)

机构地区：[1]苏州科技大学化学生物与材料工程学院,苏州215009

出　　处：《中国抗生素杂志》2018年第7期824-830,共7页Chinese Journal of Antibiotics

基　　金：苏州市科技计划项目(No.SNG2017053);国家自然科学基金项目(No.21376156);苏州科技学院科研基金项目(No.XKZ201411)

摘　　要：目的利用诱变结合抗性筛选方法选育多杀菌素高产菌株,并通过发酵培养基优化进一步提高多杀菌素产量。方法分别确定链霉素、安普霉素和鼠李糖3种抗性的最小抑菌浓度(minimal inhibitory concentration,MIC),然后以S.s1-4为出发菌株,通过紫外(UV)结合链霉素、安普霉素、鼠李糖抗性因子诱变选育,在此基础上利用亚硝基胍(NTG)结合上述抗性因子诱变选育,并利用响应面实验设计对发酵培养基中葡萄糖、糊精、棉籽蛋白3种成分进行优化。结果出发菌株经过紫外照射30s,涂布于抗性平板上,筛选得到S.s2-21,S.s2-21再用NTG处理30min,涂布于抗性平板上,最终获得1株遗传性状稳定的菌株S.s3-37,产量为78.26mg/L,提高了45.71%;发酵培养基优化后,其产量达83.00mg/L。结论利用紫外和NTG结合抗性复合诱变选育获得多杀菌素高产菌株是有效的,通过发酵培养基优化,其产量较出发菌株提高了54.55%,获得良好的效果。Objective Mutation and resistance screening were used to breed high production strains of spinosad. Production of spinosad was further improved by optimizing fermentation medium. Methods The minimum inhibitory concentrations of streptomycin, apramycin, and rhamnose were determined, respectively. Breeding by employing UV combined with resistance factors of streptomycin, apramycin, and rhamnose was conducted and S.sl- 4 was used as the starting strain. On this basis, the mutants were induced by NTG combined with the above resistance factors. Fermentation medium components of glucose, dextrin, and cottonseed protein were optimized by the response surface design. Results The spores of the original strain treated by UV for 30s were spread onto the resistant plates, and the S.s2-21 mutants were obtained. The S.s2-21 mutants were then treated with NTG for 30min and spread onto the resistant plate. One genetically stable mutant S.s3-37 was obtained at last and its yield reached 78.26mg/L, which increased by 45.71%. The production reached 83.00mg/L after the optimization of fermentation medium. Conclusion Compound mutation of UV and NTG combined with resistance screening to obtain high production strain of spinosad was efficient. The yield of mutant increased by 54.55% compared with the original strain after the optimization of fermentation medium, which was a satisfactory result.

关 键 词：多杀菌素 刺糖多孢菌 抗性筛选 复合诱变育种 培养基优化 

分 类 号：R978.15[医药卫生—药品]