产多杀菌素刺糖多孢菌的诱变选育  

Mutagenesis and breeding of spinosad-producing strains of Saccharopolyspora spinosa

作　　者：王欣荣 张爱雪 唐智超 翟龙飞 张新宜 林家富 黄挺 Wang Xin-rong;Zhang Ai-xue;Tang Zhi-chao;Zhai Long-fei;Zhang Xin-yi;Lin Jia-fu;Huang Ting(Antibiotics Research and Re-evaluation Key Laboratory of Sichuan Province,Sichuan Industrial Institute of Antibiotics,Chengdu University,Chengdu 610052)

机构地区：[1]抗生素研究与再评价四川省重点实验室,成都大学四川抗菌素工业研究所,成都610052

出　　处：《中国抗生素杂志》2020年第9期873-877,共5页Chinese Journal of Antibiotics

基　　金：“重大新药创制”化学结构导向的分子设计育种体系建设(No.2018ZX09711001-007-006)。

摘　　要：目的采用甲基磺酸乙酯(EMS)、核糖体工程育种和常压室温等离子体(ARTP)方法处理产多杀菌素刺糖多孢菌(Saccharopolyspora spinosa)SIIA-1802,采用含蛋氨酸培养基筛选得到高产菌株。方法首轮采用EMS诱变;第二轮采用链霉素抗性育种;第三轮采用ARTP诱变育种进一步巩固育种成效;采用对照和添加蛋氨酸的发酵培养基考察突变菌株的发酵水平。结果出发株刺糖多孢菌SIIA-1802经过EMS诱变、链霉素抗性筛选和ARTP诱变得到的突变株ESA-611,发酵水平提高了671.8%,采用含有蛋氨酸的发酵培养基进行筛选,发酵水平进一步提高了57.6%。在ARTP诱变过程中,筛选到一株多杀菌素A显著下降,但产生较高水平多杀菌素J的菌株ESA-598。结论本方法简单经济,突变效率高,能够快速获得传代稳定的多杀菌素高产突变株。另外,通过诱变拓宽了代谢产物谱,得到了具有更高潜在价值的组分。Objective To obtain the hyperproducing Saccharopolyspora spinosa strains,S.spinosa SIIA-1802 was treated with ethyl methane sulfonate(EMS),ribosomel engineering breeding and atmospheric and room temperature plasma(ARTP),or cultured with methionine.Methods The original strain S.spinosa SIIA-1802 was treated with EMS in the first round mutagenesis.And then the S.spinosa SIIA-1802 was treated with streptomycin in the second round mutagenesis.Finally,ARTP breeding was performed in the third mutagenesis round to further consolidate the breeding effects.The fermentation level of mutant strains was investigated by adding methionine to fermentation medium.Results The mutant strain ESA-611 was obtained by EMS mutagenesis,streptomycin resistance screening and ARTP induction.The fermentation level was 671.8%higher than original strain SIIA-1802.The mutant strains were screened in the fermentation medium containing methionine and the fermentation level was further increased by 57.6%.During the ARTP mutagenesis process,the production of spinosad A was significantly decreased in one mutant strain,but the mutant strain ESA-598 that produced higher levels of spinosad J was obtained.Conclusion This method is simple,economical,and has a high mutation efficiency.The stable and hyperproducing S.spinosa mutants for spinosad can be quickly obtained.In addition,the spectrum of metabolites in S.spinosa was improved by mutagenesis and components with higher potential value were obtained.

关 键 词：多杀菌素A 诱变育种 发酵 多杀菌素J 刺糖多孢菌 

分 类 号：R978.1[医药卫生—药品]