Identification of a gene fromStreptomyces rimosusM527 negatively affecting rimocidin biosynthesis and morphological differentiation
Identification of a gene fromStreptomyces rimosusM527 negatively affecting rimocidin biosynthesis and morphological differentiation
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鉴定出对抗生素生物合成和形态分化产生负面影响的链霉菌 M527 基因
DOI：
10.1007/s00253-020-10955-8
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发表时间：
2020-10-15
影响因子：
5
通讯作者：
Yu, Xiaoping
中科院分区：
文献类型：
--
作者：
Liao, Zhijun;Song, Zhangqing;Yu, Xiaoping
由龟裂链霉菌M527产生的多烯大环内酯类龟裂杀菌素被发现对广泛的植物真菌病原体非常有效。目前对裂膜链霉菌M527中裂膜霉素生物合成和形态分化的调控机制了解有限。NsdA被认为是一种负调控因子，参与了一些链霉菌的形态分化和次生代谢产物的生物合成。本研究从S. rimosus M527.通过基因缺失、互补和过表达研究nsdA（sr）在rimocidin生物合成和形态分化中的作用。使用CRISPR/Cas9获得Delta nsdA（sr）突变体。突变体产生更多的rimocidin（46%）和加速形态分化比野生型菌株。nsdA（sr）的过度表达导致rimocidin产量减少和形态分化受损。定量RT-PCR分析显示，在Delta nsdA（sr）突变株中，rimocidin生物合成相关基因的转录水平上调，而在thensdA（sr）过表达株中，rimocidin生物合成相关基因的转录水平下调。类似的效果已经被描述为天蓝色链霉菌M145和工业丰卡霉素生产菌株Streptomycesacatochromogenes 1628。
The polyene macrolide rimocidin, produced byStreptomyces rimosusM527, was found to be highly effective against a broad range of fungal plant pathogens. Current understanding of the regulatory mechanism of rimocidin biosynthesis and morphological differentiation inS. rimosusM527 is limited. NsdA is considered a negative regulator involved in morphological differentiation and biosynthesis of secondary metabolites in someStreptomycesspecies. In this study,nsdA(sr)was cloned fromS. rimosusM527. The role ofnsdA(sr)in rimocidin biosynthesis and morphological differentiation was investigated by gene deletion, complementation, and over-expression. A Delta nsdA(sr)mutant was obtained using CRISPR/Cas9. The mutant produced more rimocidin (46%) and accelerated morphological differentiation than the wild-type strain. Over-expression ofnsdA(sr)led to a decrease in rimocidin production and impairment of morphological differentiation. Quantitative RT-PCR analysis revealed that transcription ofrimgenes responsible for rimocidin biosynthesis was upregulated in the Delta nsdA(sr)mutant but downregulated in thensdA(sr)over-expression strain. Similar effects have been described forStreptomyces coelicolorM145 and the industrial toyocamycin-producing strainStreptomyces diastatochromogenes1628.