Identification of a gene from Streptomyces rimosus M527 negatively affecting rimocidin biosynthesis and morphological di
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APPLIED MICROBIAL AND CELL PHYSIOLOGY
Identification of a gene from Streptomyces rimosus M527 negatively affecting rimocidin biosynthesis and morphological differentiation Zhijun Liao 1 & Zhangqing Song 1 & Jie Xu 1 & Zheng Ma 1 & Andreas Bechthold 2 & Xiaoping Yu 1 Received: 2 August 2020 / Revised: 1 October 2020 / Accepted: 7 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract The polyene macrolide rimocidin, produced by Streptomyces rimosus M527, 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 in S. rimosus M527 is limited. NsdA is considered a negative regulator involved in morphological differentiation and biosynthesis of secondary metabolites in some Streptomyces species. In this study, nsdAsr was cloned from S. rimosus M527. The role of nsdAsr in rimocidin biosynthesis and morphological differentiation was investigated by gene deletion, complementation, and over-expression. A ΔnsdAsr mutant was obtained using CRISPR/Cas9. The mutant produced more rimocidin (46%) and accelerated morphological differentiation than the wild-type strain. Over-expression of nsdAsr led to a decrease in rimocidin production and impairment of morphological differentiation. Quantitative RT-PCR analysis revealed that transcription of rim genes responsible for rimocidin biosynthesis was upregulated in the ΔnsdAsr mutant but downregulated in the nsdAsr over-expression strain. Similar effects have been described for Streptomyces coelicolor M145 and the industrial toyocamycin-producing strain Streptomyces diastatochromogenes 1628. Key points • A negative regulator for sporulation and rimocidin production was identified. • The CRISPR/Cas9 system was used for gene deletion in S. rimosus M527. Keywords NsdAsr . Streptomyces rimosus . Rimocidin . CRISPR/Cas9
Introduction Streptomyces are Gram-positive bacteria with complex morphological differentiation and secondary metabolism. They are well-known to produce a wide variety of valuable natural Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10955-8) contains supplementary material, which is available to authorized users. * Zheng Ma [email protected] * Xiaoping Yu [email protected] 1
Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, College of Life Sciences, China Jiliang University, Xueyuan Street, Xiasha Higher Education District, Hangzhou 310018, Zhejiang Province, China
2
Institute for Pharmaceutical Sciences, Pharmaceutical Biology and Biotechnology, University of Freiburg, 79104 Freiburg, Germany
products with diverse biological activities (Kemung et al. 2018; Liu et al. 2018; Olanrewaju and Babalola 2019). In Streptomyces, the processes of morphological differentiation and antibiotic biosynthesis are tightly controlled via multiple levels of regulation that includes cluster-situated, pleiotropic, and global regul
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