The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus

ABSTRACT Daptomycin (DAP) is a cyclic lipopeptide antibiotic produced by Streptomyces roseosporus, and its biosynthesis is precisely regulated by a complex regulatory network. Although the biosynthetic pathway of DAP has been elucidated, the regulatory mechanism governing its biosynthesis at the tra...

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Main Authors: Fuqiang He, Xinpeng Liu, Haiyi Wang, Xu Li, Yun Wu, Dan Zhang, Shufang Liang
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Microbial Biotechnology
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Online Access:https://doi.org/10.1111/1751-7915.70110
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author Fuqiang He
Xinpeng Liu
Haiyi Wang
Xu Li
Yun Wu
Dan Zhang
Shufang Liang
author_facet Fuqiang He
Xinpeng Liu
Haiyi Wang
Xu Li
Yun Wu
Dan Zhang
Shufang Liang
author_sort Fuqiang He
collection DOAJ
description ABSTRACT Daptomycin (DAP) is a cyclic lipopeptide antibiotic produced by Streptomyces roseosporus, and its biosynthesis is precisely regulated by a complex regulatory network. Although the biosynthetic pathway of DAP has been elucidated, the regulatory mechanism governing its biosynthesis at the transcriptional level is not yet fully understood. In the present study, a new transcriptional regulator, DhyR, was identified. A deletion mutant of dhyR was constructed using the CRISPR‐Cas9 tool to elucidate the biological role of DhyR thanks to functional and transcriptomic analyses. The results demonstrated that DhyR positively regulates DAP biosynthesis in S. roseosporus. The in‐frame deletion of the dhyR gene resulted in a significant downregulation of the transcription levels of all structural genes within the DAP biosynthetic gene cluster and a significant decrease in DAP yield. In contrast, overexpression of dhyR enhanced the transcription levels of the DAP biosynthetic gene cluster, leading to a 23% increase in DAP yield. Deletion of dhyR caused significant changes in the expression of multiple genes involved in carbohydrate metabolism, energy metabolism and amino acid metabolic pathways through transcriptome analysis. Especially, deletion of dhyR led to a significant downregulation of transcription levels of three DAP biosynthesis‐associated genes, including atrA, depR1 and ssig‐05090. In summary, DhyR positively regulates DAP biosynthesis in S. roseosporus by influencing the expression of the DAP gene cluster and modulating precursor flux. It functions as a pleiotropic regulator of primary and secondary metabolism in S. roseosporus.
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spelling doaj-art-ac96f1c0e99d47f5ac5c7b66247877d22025-08-20T02:11:14ZengWileyMicrobial Biotechnology1751-79152025-03-01183n/an/a10.1111/1751-7915.70110The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporusFuqiang He0Xinpeng Liu1Haiyi Wang2Xu Li3Yun Wu4Dan Zhang5Shufang Liang6Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital Sichuan University Chengdu Sichuan P.R. ChinaDepartment of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital Sichuan University Chengdu Sichuan P.R. ChinaDepartment of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital Sichuan University Chengdu Sichuan P.R. ChinaDepartment of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital Sichuan University Chengdu Sichuan P.R. ChinaDepartment of Cell Biology, College of Life Science Sichuan Normal University Chengdu Sichuan P.R. ChinaDepartment of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital Sichuan University Chengdu Sichuan P.R. ChinaDepartment of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital Sichuan University Chengdu Sichuan P.R. ChinaABSTRACT Daptomycin (DAP) is a cyclic lipopeptide antibiotic produced by Streptomyces roseosporus, and its biosynthesis is precisely regulated by a complex regulatory network. Although the biosynthetic pathway of DAP has been elucidated, the regulatory mechanism governing its biosynthesis at the transcriptional level is not yet fully understood. In the present study, a new transcriptional regulator, DhyR, was identified. A deletion mutant of dhyR was constructed using the CRISPR‐Cas9 tool to elucidate the biological role of DhyR thanks to functional and transcriptomic analyses. The results demonstrated that DhyR positively regulates DAP biosynthesis in S. roseosporus. The in‐frame deletion of the dhyR gene resulted in a significant downregulation of the transcription levels of all structural genes within the DAP biosynthetic gene cluster and a significant decrease in DAP yield. In contrast, overexpression of dhyR enhanced the transcription levels of the DAP biosynthetic gene cluster, leading to a 23% increase in DAP yield. Deletion of dhyR caused significant changes in the expression of multiple genes involved in carbohydrate metabolism, energy metabolism and amino acid metabolic pathways through transcriptome analysis. Especially, deletion of dhyR led to a significant downregulation of transcription levels of three DAP biosynthesis‐associated genes, including atrA, depR1 and ssig‐05090. In summary, DhyR positively regulates DAP biosynthesis in S. roseosporus by influencing the expression of the DAP gene cluster and modulating precursor flux. It functions as a pleiotropic regulator of primary and secondary metabolism in S. roseosporus.https://doi.org/10.1111/1751-7915.70110CRISPR‐Cas9daptomycinDhyRStreptomyces roseosporustranscriptional regulatortranscriptome
spellingShingle Fuqiang He
Xinpeng Liu
Haiyi Wang
Xu Li
Yun Wu
Dan Zhang
Shufang Liang
The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus
Microbial Biotechnology
CRISPR‐Cas9
daptomycin
DhyR
Streptomyces roseosporus
transcriptional regulator
transcriptome
title The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus
title_full The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus
title_fullStr The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus
title_full_unstemmed The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus
title_short The Transcriptional Regulator DhyR Positively Modulates Daptomycin Biosynthesis in Streptomyces roseosporus
title_sort transcriptional regulator dhyr positively modulates daptomycin biosynthesis in streptomyces roseosporus
topic CRISPR‐Cas9
daptomycin
DhyR
Streptomyces roseosporus
transcriptional regulator
transcriptome
url https://doi.org/10.1111/1751-7915.70110
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