A regulator based “semi-targeted” approach to activate silent biosynthetic gene clusters

Erik Mingyar, Lucas Mühling, Andreas Kulik, Anika Winkler, Daniel Wibberg, Jörn Kalinowski, Kai Blin, Tilmann Weber, Wolfgang Wohlleben, Evi Stegmann*

*Corresponding author for this work

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Abstract

By culturing microorganisms under standard laboratory conditions, most biosynthetic gene clusters (BGCs) are not expressed, and thus, the products are not produced. To explore this biosynthetic potential, we developed a novel “semi-targeted” approach focusing on activating “silent” BGCs by concurrently introducing a group of regulator genes into streptomycetes of the Tübingen strain collection. We constructed integrative plasmids containing two classes of regulatory genes under the control of the constitutive promoter ermE*p (cluster situated regulators (CSR) and Streptomyces antibiotic regulatory proteins (SARPs)). These plasmids were introduced into Streptomyces sp. TÜ17, Streptomyces sp. TÜ10 and Streptomyces sp. TÜ102. Introduction of the CSRs-plasmid into strain S. sp. TÜ17 activated the production of mayamycin A. By using the individual regulator genes, we proved that Aur1P, was responsible for the activation. In strain S. sp. TÜ102, the introduction of the SARP-plasmid triggered the production of a chartreusin-like compound. Insertion of the CSRs-plasmid into strain S. sp. TÜ10 resulted in activating the warkmycin-BGC. In both recombinants, activation of the BGCs was only possible through the simultaneous expression of aur1PR3 and griR in S. sp. TÜ102 and aur1P and pntR in of S. sp. TÜ10.

Original languageEnglish
Article number7567
JournalInternational Journal of Molecular Sciences
Volume22
Issue number14
Number of pages14
ISSN1661-6596
DOIs
Publication statusPublished - 2 Jul 2021

Bibliographical note

Funding Information:
Acknowledgments: The authors thank J. Wiese (Helmholz-Center for Ocean Research Kiel) for providing mayamycin. E.S. and W.W. were supported by the DZIF, the position of E.M. was funded by DZIF. The position of T.W. and K.B. were funded by the Nordisk Foundation.

Funding Information:
Funding: This research was funded by German Center for Infection Research (DZIF), grant number 9.818 (01.01.2017). T.W. would like to acknowledge funding from the Novo Nordisk Foundation, grant numbers NNF16OC0021746 (01.03.2017) and NNF20CC0035580 (1.1.2021).

Funding Information:
This research was funded by German Center for Infection Research (DZIF), grant number 9.818 (01.01.2017). T.W. would like to acknowledge funding from the Novo Nordisk Foundation, grant numbers NNF16OC0021746 (01.03.2017) and NNF20CC0035580 (1.1.2021). The authors thank J. Wiese (Helmholz-Center for Ocean Research Kiel) for providing mayamycin. E.S. and W.W. were supported by the DZIF, the position of E.M. was funded by DZIF. The position of T.W. and K.B. were funded by the Nordisk Foundation.

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Regulation
  • Secondary metabolites
  • Silent biosynthetic gene cluster
  • Streptomyces

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