基于深孔板培养高通量筛选rakicidin B1高产菌的研究  

Studies on high throughput screening for high rakicidin B1-producing strains with deep-well plates

作　　者：周剑[1] 江红[1] 林风[1] Zhou Jian;Jiang Hong;Lin Feng(Fujian Key Laboratory of Screening for Novel Microbial Products,Fujian Institute of Microbiology,Fuzhou 350007)

机构地区：[1]福建省微生物研究所福建省新药(微生物)筛选重点实验室

出　　处：《中国抗生素杂志》2019年第12期1352-1355,共4页Chinese Journal of Antibiotics

基　　金：国家重点研发计划(No.2018YFC0311004);国家“十三五”重大新药创制项目(No.2018ZX09711001-007-007);福建省属公益类科研院所基本科研专项(No.2019R11020006-6)

摘　　要：目的获得rakicidin B1的高产突变菌株。方法建立深孔板发酵rakicidin B1的方法,结合常压室温等离子体技术(ARTP)及核糖体工程对rakicidin B1的产生菌进行高通量诱变选育。结果获得一株高产突变菌株R36-27,其摇瓶发酵产量达到490mg/L左右,较出发菌株提高了237.9%;经遗传稳定性考察验证了该菌株稳定性较好。结论采用ARTP为诱变源,以庆大霉素抗性为选择压力,结合深孔板高通量筛选,可以快速有效地获得高产菌株。Objective To obtain high-yield and genetic stable mutants of Rakicidin B1 production.Methods The atmospheric and room temperature plasma(ARTP),the ribosome engineering and high-throughput screening(HTS)were comprehensively applied in the screening of high-yield strains of rakicidin B1.Results Finally,a mutant R36-27 with gentamicin resistance was selected by high-throughput screening.In the shaking flask culture,the mutant R36-27 was hereditarily stabilized and its yield of rakicidin B1 increased to 490 mg/L compared to 144 mg/L produced by the original strain,namely,increased by 237.9%.Conclusion This study shows that the screening high-yield strains can be enhanced effectively by combining the atmospheric and room temperature plasma(ARTP),the ribosome engineering and high-throughput screen methods(HTS).

关 键 词：常温等离子体 核糖体工程 高通量筛选 深孔板 Rakicidin 

分 类 号：R978.1[医药卫生—药品]