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Identification of a small molecule signaling factor that regulates the biosynthesis of the antifungal polycyclic tetramate macrolactam HSAF in Lysobacter enzymogenes

  • Applied genetics and molecular biotechnology
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Abstract

Lysobacter species are emerging as new sources of antibiotics. The regulation of these antibiotics is not well understood. Here, we identified a small molecule metabolite (LeDSF3) that regulates the biosynthesis of the antifungal antibiotic heat-stable antifungal factor (HSAF), a polycyclic tetramate macrolactam with a structure and mode of action distinct from the existing antifungal drugs. LeDSF3 was isolated from the culture broth of Lysobacter enzymogenes, and its chemical structure was established by NMR and MS. The purified compound induced green fluorescence in a reporter strain of Xanthomonas campestris, which contained a gfp gene under the control of a diffusible signaling factor (DSF)-inducible promoter. Exogenous addition of LeDSF3 in L. enzymogenes cultures significantly increased the HSAF yield, the transcription of HSAF biosynthetic genes, and the antifungal activity of the organism. The LeDSF3-regulated HSAF production is dependent on the two-component regulatory system RpfC/RpfG. Moreover, LeDSF3 upregulated the expression of the global regulator cAMP receptor-like protein (Clp). The disruption of clp led to no HSAF production. Together, the results show that LeDSF3 is a fatty acid-derived, diffusible signaling factor positively regulating HSAF biosynthesis and that the signaling is mediated by the RfpC/RpfG-Clp pathway. These findings may facilitate the antibiotic production through applied genetics and molecular biotechnology in Lysobacter, a group of ubiquitous yet underexplored microorganisms.

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Acknowledgments

This work was supported in part by the 973 Project (2013CB734002, 2015CB150602), the NIH (R01AI097260), and Program for Changjiang Scholars and Innovative Research Team in University (IRT13028). We thank Prof. Lindow for the generous gift of DSF reporter strain X. campestris 8523/pKLN55. Ron Cerny, Martha Morton, Kurt Wulser, and Javier Seravalli are thanked for technical assistance.

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Authors and Affiliations

  1. Department of Chemistry, University of Nebraska—Lincoln, Lincoln, NE, 68588, USA

    Yong Han, Yan Wang, Simon Tombosa, Stephen Wright, Justin Huffman & Liangcheng Du

  2. Key Laboratory of Chemical Biology, School of Pharmaceutical Sciences, Shandong University, 250100, Jinan, China

    Yong Han & Yuemao Shen

  3. Department of Plant Pathology, University of Nebraska—Lincoln, Lincoln, NE, 68583, USA

    Gary Yuen

  4. College of Plant Protection, Nanjing Agricultural University, 210095, Nanjing, China

    Guoliang Qian & Fengquan Liu

  5. College of Marine Life Sciences, Ocean University of China, 266003, Qingdao, China

    Yan Wang

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  1. Yong Han
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  2. Yan Wang
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Correspondence to Yuemao Shen or Liangcheng Du.

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Yong Han and Yan Wang contributed equally to this work.

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Han, Y., Wang, Y., Tombosa, S. et al. Identification of a small molecule signaling factor that regulates the biosynthesis of the antifungal polycyclic tetramate macrolactam HSAF in Lysobacter enzymogenes . Appl Microbiol Biotechnol 99, 801–811 (2015). https://doi.org/10.1007/s00253-014-6120-x

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