基于深孔板培养高通量筛选rakicidin B1高产菌的研究

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。

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

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 490mg/L compared to 144mg/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