高产那西肽活跃链霉菌的高通量选育

High-throughput screening of Streptomyces actuosus for high production of nosiheptide

为提高活跃链霉菌(Streptomyces actuosus)发酵生产那西肽的强度,对1株已应用于工业化生产那西肽的菌株11-27进行了恒温常压等离子体诱变(atmospheric and room temperature plasma,ARTP),并对最终获得的高产突变菌株进行了工业化生产应用。首先,根据那西肽的分子特征,建立了以FeCl_(3)为络合显色剂的多孔板高通量筛选方法。应用ARTP诱变方法对菌株11-27进行诱变处理,最终获得了1株高产突变菌株22-3B6,其那西肽的效价相对出发菌株11-27提高了13.7%。对高产菌株进行遗传稳定性分析后,在50 m^(3)工业生产罐上对最优菌株进行了16个批次的发酵验证,结果表明诱变菌株22-3B6生产那西肽的平均效价较对照菌株提高了10.1%。该文所建立的高通量筛选方法能简单、快速地获得高产突变菌株,从而有效地降低了那西肽的工业化生产成本。

In order to enhance the production of nosiheptide by Streptomyces actuosus,the atmospheric and room temperature plasma(ARTP)mutagenesis was carried out on an industrial strain 11-27,and then the obtained mutant was applied in industrial production.Firstly,a high throughput screening method was established according to the molecular characteristics of nosiheptide,which could form complex chromogenic reagent with FeCl_(3).Arter the treatment of ARTP on strain 11-27,a high-production mutant 22-3B6 was obtained,its production of nosiheptide was increased by 13.7%.Genetic stability analysis showed that the high-titer strain had a stable performance in nosiheptide production.Sixteen batches of fermentation of strain 22-3B6 in a 50 m^(3) fermenter showed that the average production of nosiheptide was 10.1%higher than that of the original strain 11-27.The high-throughput screening method established here could obtain excellent performance mutant simply and quickly,thus effectively reducing the industrial cost of nassipeptide production.