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在Streptomyces fungicidicus中应用多种基因组编辑技术,以提高恩杜拉西丁产量

Yanan Sun¹, Guoguo Wu¹, Yining Wang¹

¹Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, PR China.

Synthetic and systems biotechnology

|March 17, 2025

概括

研究人员优化了Streptomyces fungicidicus中的基因编辑,用于新合成. 增强的同源重组和CRISPR系统使得有效的基因群删除成为可能,改善了菌株生长特征.

科学领域:

微生物生物技术微生物生物技术
合成生物学合成生物学
遗传学遗传学是一个

背景情况:

菌株Streptomyces fungicidicus是Enduracidin生产的有价值的工业菌株,也是新型小合成的潜在底盘.
有效的基因编辑工具对于通过有针对性地删除二次代谢物基因集群来提高微生物菌株性能至关重要.
在Streptomyces中现有的基因编辑方法可能是低效的,阻碍了改进的工业菌株的开发.

研究的目的:

优化现有的基因编辑方法,并在Streptomyces fungicidicus中建立新的方法.
为了促进二次代谢物基因集群的删除,以改善菌株.
设计Streptomyces fungicidicus作为一个用于增强生物合成的细胞底盘.

主要方法:

通过整合选择和反选择标记,优化了传统的同源重组.
评估了CRISPR/cBEST系统的基础编辑,特别是用于停止编码子插入.
建立了一个CRISPR/Cas9系统,在S. fungicidicus中建立了有效基因编辑的反选择标记.

主要成果:

使用优化同源重组方法,成功删除了高达54.4kb的二次代谢物基因群.
证明了CRISPR/cBEST系统能够插入具有不同突变率的停止编码子的能力.
开发了一种CRISPR/Cas9系统,可以显著缩短基因编辑周期.
产生了单个和累积基因群删除的突变,表现出改善的生长特征 (延长的对数阶段,增加的生物量).

结论:

在Streptomyces fungicidicus中成功建立了优化和新型的基因编辑系统 (同类重组,CRISPR/cBEST,CRISPR/Cas9).
基因群删除导致了增长特征的改善,这表明了未来菌株优化潜力.
开发的基因编辑工具为设计其他Streptomyces物种提供了坚实的基础.

关键词:

这就是CRISPR/CasPR.恩杜拉西丁是一种耐药物.增长的特征增长的特征在NRPS/PKS基因集群中删除NRPS/PKS基因集群删除.Streptomyces fungicidicus 这种真菌的生长方式.

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