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Global and pathway-specific transcriptional regulations of pactamycin biosynthesis in Streptomyces pactum

  • Applied genetics and molecular biotechnology
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Abstract

Pactamycin, a structurally unique aminocyclitol natural product isolated from Streptomyces pactum, has potent antibacterial, antitumor, and anti-protozoa activities. However, its production yields under currently used culture conditions are generally low. To understand how pactamycin biosynthesis is regulated and explore the possibility of improving pactamycin production in S. pactum, we investigated the transcription regulations of pactamycin biosynthesis. In vivo inactivation of two putative pathway-specific regulatory genes, ptmE and ptmF, resulted in mutant strains that are not able to produce pactamycin. Genetic complementation using a cassette containing ptmE and ptmF integrated into the S. pactum chromosome rescued the production of pactamycin. Transcriptional analysis of the ΔptmE and ΔptmF strains suggests that both genes control the expression of the whole pactamycin biosynthetic gene cluster. However, attempts to overexpress these regulatory genes by introducing a second copy of the genes in S. pactum did not improve the production yield of pactamycin. We discovered that pactamycin biosynthesis is sensitive to phosphate regulation. Concentration of inorganic phosphate higher than 2 mM abolished both the transcription of the biosynthetic genes and the production of the antibiotic. Draft genome sequencing of S. pactum and bioinformatics studies revealed the existence of global regulatory genes, e.g., genes that encode a two-component PhoR-PhoP system, which are commonly involved in secondary metabolism. Inactivation of phoP did not show any significant effect to pactamycin production. However, in the phoP::aac(3)IV mutant, pactamycin biosynthesis is not affected by external inorganic phosphate concentration.

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Acknowledgements

The authors thank Benjamin Philmus for a critical reading of this manuscript and Maureen J. Bibb and John Innes Centre for providing plasmid pIJ6902.

Funding

This work was supported by grant AI129957 from the National Institute of Allergy And Infectious Diseases. The content is solely the responsibility of the authors and does not represent the official views of the National Institute of Allergy And Infectious Diseases, or the National Institutes of Health (NIH).

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  1. Wanli Lu and Abdullah R. Alanzi contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Oregon State University, Corvallis, OR, 97331-3507, USA

    Wanli Lu, Abdullah R. Alanzi, Mostafa E. Abugrain & Taifo Mahmud

  2. Faculty of Pharmacy, Osaka-Ohtani University, 3-11-1 Nisikiorikita, Tondabayashi, Osaka, 584-8540, Japan

    Takuya Ito

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  1. Wanli Lu
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  2. Abdullah R. Alanzi
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  3. Mostafa E. Abugrain
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Correspondence to Taifo Mahmud.

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Lu, W., Alanzi, A.R., Abugrain, M.E. et al. Global and pathway-specific transcriptional regulations of pactamycin biosynthesis in Streptomyces pactum. Appl Microbiol Biotechnol 102, 10589–10601 (2018). https://doi.org/10.1007/s00253-018-9375-9

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  • DOI: https://doi.org/10.1007/s00253-018-9375-9

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