食品与发酵工业 >

2021 , Vol. 47 >Issue 7: 8 - 13

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.025310

研究报告

高产那西肽活跃链霉菌的高通量选育

  • 张小朋 ,
  • 陈贵才 ,
  • 严发杰 ,
  • 郭李坤 ,
  • 曾伟主 ,
  • 周景文
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  • 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)
硕士,高级工程师(周景文教授为通讯作者,E-mail:zhoujw1982@jiangnan.edu.cn)

收稿日期: 2020-08-08

  修回日期: 2020-09-08

  网络出版日期: 2021-05-20

基金资助

国家自然科学基金优秀青年基金项目(21822806)

收起

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

  • ZHANG Xiaopeng ,
  • CHEN Guicai ,
  • YAN Fajie ,
  • GUO Likun ,
  • ZENG Weizhu ,
  • ZHOU Jingwen
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  • 1(Gansu Huineng Biological Engineering Co.Ltd.,Wuwei 733000,China)
    2(Zhejiang Huineng Biological Engineering Co.Ltd.,Haining 314422,China)
    3(Key Laboratory of Industrial Biotechnology of Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China)

Received date: 2020-08-08

  Revised date: 2020-09-08

  Online published: 2021-05-20

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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FeCl3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 m3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。

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

本文引用格式

张小朋 , 陈贵才 , 严发杰 , 郭李坤 , 曾伟主 , 周景文 . 高产那西肽活跃链霉菌的高通量选育[J]. 食品与发酵工业, 2021 , 47(7) : 8 -13 . DOI: 10.13995/j.cnki.11-1802/ts.025310

Abstract

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 FeCl3. 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 m3 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.

Key words： nosiheptide; Streptomyces actuosus; high-throughput screening; atmospheric and room temperature plasma(ARTP); mutagenesis

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