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

作　　者：张小朋 陈贵才 严发杰 郭李坤 曾伟主 周景文[3] ZHANG Xiaopeng;CHEN Guicai;YAN Fajie;GUO Likun;ZENG Weizhu;ZHOU Jingwen(Gansu Huineng Biological Engineering Co.Ltd.,Wuwei 733000,China;Zhejiang Huineng Biological Engineering Co.Ltd.,Haining 314422,China;Key Laboratory of Industrial Biotechnology of Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]甘肃汇能生物工程有限公司,甘肃武威733000 [2]浙江汇能生物股份有限公司,浙江海宁314422 [3]生物工程学院工业生物技术教育部重点实验室(江南大学),江苏无锡214122

出　　处：《食品与发酵工业》2021年第7期8-13,共6页Food and Fermentation Industries

基　　金：国家自然科学基金优秀青年基金项目(21822806)。

摘　　要：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.

关 键 词：那西肽 活跃链霉菌 高通量筛选 常压室温等离子体 诱变 

分 类 号：TQ927[轻工技术与工程—发酵工程]