常压室温等离子体-紫外复合诱变选育新硫肽类抗生素166A高产菌株  

Breeding of new thiopeptide antibiotic 166A high-yield producing strain by MPMS composite mutagenesis with plasma and UV

作　　者：李艳青 戴剑漉[3] 蒋忠科[2] 罗辉 孙承航[2] Li Yan-qing;Dai Jian-lu;Jiang Zhong-ke;Luo Hui;Sun Cheng-hang(Southern Marine Science and Engineering Guangdong Laboratory,Guangdong Medical University,Zhanjiang 524023;Department of Microbial Chemistry,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050;Key Lab of Antibiotic Biotechnology,National Health Commission,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050)

机构地区：[1]广东医科大学,南方海洋科学与工程广东省实验室,湛江524023 [2]中国医学科学院&北京协和医学院医药生物技术研究所,微生物化学研究室,北京100050 [3]中国医学科学院&北京协和医学院医药生物技术研究所,国家卫健委抗生素生物工程重点实验室,北京100050

出　　处：《中国抗生素杂志》2022年第5期514-520,共7页Chinese Journal of Antibiotics

基　　金：中国医学科学院医学与健康科技创新工程项目(No.2021-1-I2M-028)。

摘　　要：目的通过对新硫肽类抗生素166A产生菌小单孢菌TMD166进行诱变选育研究,以期获得产166A的高产菌株。方法使用多功能等离子体诱变系统(multifunctional plasma mutagenesis system,MPMS)对出发菌株TMD166的孢子进行等离子体-紫外(MPMS-UV)复合诱变,单孢子悬液经照射处理、涂布培养后获得单菌落,并以牛津杯固体发酵高通量筛选方法对菌株进行初筛,之后对高产菌株进行摇瓶复筛,利用突变株摇瓶发酵的化学效价筛选出正突变菌株。结果对比MPMS-UV不同诱变剂量发现,MPMS105s-UV60s复合诱变剂量获得的正突变率最高,达到41.43%,相应致死率为99.88%。最终筛选出一株166A高产突变株TMD166-MU1,在培养温度为28℃、210 r/min的条件下,经过96~120 h培养后,166A产量相比出发菌株提高了7.47倍。结论采用MPMS-UV复合诱变方式,再结合牛津杯固体发酵高通量筛选方法和摇瓶复筛,可高效筛选获得166A高产菌株。Objective To obtain the new thiopeptide antibiotics 166A high-yield producing strain by means of mutation breeding of TMD166 strain.Methods The spores from the original strain were treated by Plasma and UV composite mutation in multifunctional plasma mutagenesis system(MPMS).The strain spore suspension of TMD166 treated under different mutagenesis conditions was spread on the cultural medium plates to obtain a single colony.Through the first high throughput screening with the oxford cup method and the following second screening with the flask fermentation culture,positive mutant strains were screened out by comparison of the fermentation titer.Results MPMS105s-UV60 composite mutagenesis can obtain the highest positive mutation rate among different MPMS-UV mutation conditions.Its positive mutation rate and lethality rate were 41.43%and 99.88%,respectively.After multiple conditions of MPMS or/and UV mutagenesis,a mutant strain coded as TMD166-MU1 was finally screened out and the yield of 166A was 7.47 times higher than that of the original strain after being cultured in the shaker at 210 r/min under 28℃for 96~120 h.Conclusion The MPMS-UV composite mutagenesis with oxford cup high throughput screening can significantly improve efficiency to screen out high-yield strains.

关 键 词：新硫肽类抗生素166A 多功能等离子体诱变系统 复合诱变 选育 高产菌株 

分 类 号：R978.1[医药卫生—药品]