Abstract
Fidaxomicin, an 18-membered macrolide antibiotic, is highly active against Clostridium difficile, the most common cause of diarrhea in hospitalized patients. Though the biosynthetic mechanism of fidaxomicin has been well studied, little is known about its regulatory mechanism. Here, we reported that FadR1, a LAL family transcriptional regulator in the fidaxomicin cluster of Actinoplanes deccanensis Yp-1, acts as an activator for fidaxomicin biosynthesis. The disruption of fadR1 abolished the ability to synthesize fidaxomicin, and production could be restored by reintegrating a single copy of fadR1. Overexpression of fadR1 resulted in an approximately 400 % improvement in fidaxomicin production. Electrophoretic mobility shift assays indicated that fidaxomicin biosynthesis is under the control of FadR1 through its binding to the promoter regions of fadM, fadA1-fadP2, fadS2-fadC, and fadE-fadF, respectively. And the conserved binding sites of FadR1 within the four promoter regions were determined by footprinting experiment. All results indicated that fadR1 encodes a pathway-specific positive regulator of fidaxomicin biosynthesis and upregulates the transcription levels of most of genes by binding to the four above intergenic regions. In summary, we not only clearly elucidate the regulatory mechanism of FadR1 but also provide strategies for the construction of industrial high-yield strain of fidaxomicin.
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Acknowledgments
We gratefully thank Dr. Chris Wood, a native English biologist, for his critical reading of this manuscript.
Funding
This work was supported by the National Natural Science Foundation of China (No 31730002 and No 31520103901) and the National Key Research and Development Program (2016YFD0400805).
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Yue-Ping Li and Pin Yu performed the experiments and wrote the paper, and these authors contributed equally to this work; Ji-Feng Li, Yi-Li Tang, and Qing-Ting Bu assisted with the primary data analysis; Yong-Quan Li and Xu-Ming Mao conceived and designed the project; Yong-Quan Li supervised the project and revised the manuscript. All authors reviewed the manuscript.
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Li, YP., Yu, P., Li, JF. et al. FadR1, a pathway-specific activator of fidaxomicin biosynthesis in Actinoplanes deccanensis Yp-1. Appl Microbiol Biotechnol 103, 7583–7596 (2019). https://doi.org/10.1007/s00253-019-09949-y
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DOI: https://doi.org/10.1007/s00253-019-09949-y