A Regulator Based "Semi-Targeted" Approach to Activate Silent Biosynthetic Gene Clusters.

Autor: Mingyar E; Department of Microbiology and Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen Auf der Morgenstelle 28, 72076 Tübingen, Germany.; German Center for Infection Research (DZIF), Partner Site Tübingen, 72076 Tübingen, Germany., Mühling L; Department of Microbiology and Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen Auf der Morgenstelle 28, 72076 Tübingen, Germany., Kulik A; Department of Microbiology and Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen Auf der Morgenstelle 28, 72076 Tübingen, Germany.; Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany., Winkler A; Center for Biotechnology (CeBiTec), Universität Bielefeld, 33615 Bielefeld, Germany., Wibberg D; Center for Biotechnology (CeBiTec), Universität Bielefeld, 33615 Bielefeld, Germany., Kalinowski J; Center for Biotechnology (CeBiTec), Universität Bielefeld, 33615 Bielefeld, Germany., Blin K; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800 Kgs. Lyngby, Denmark., Weber T; Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800 Kgs. Lyngby, Denmark., Wohlleben W; Department of Microbiology and Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen Auf der Morgenstelle 28, 72076 Tübingen, Germany.; German Center for Infection Research (DZIF), Partner Site Tübingen, 72076 Tübingen, Germany.; Cluster of Excellence EXC 2124-Controlling Microbes to Fight Infections, 72076 Tübingen, Germany., Stegmann E; Department of Microbiology and Biotechnology, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen Auf der Morgenstelle 28, 72076 Tübingen, Germany.; German Center for Infection Research (DZIF), Partner Site Tübingen, 72076 Tübingen, Germany.; Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.; Cluster of Excellence EXC 2124-Controlling Microbes to Fight Infections, 72076 Tübingen, Germany.
Jazyk: angličtina
Zdroj: International journal of molecular sciences [Int J Mol Sci] 2021 Jul 15; Vol. 22 (14). Date of Electronic Publication: 2021 Jul 15.
DOI: 10.3390/ijms22147567
Abstrakt: 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.
Databáze: MEDLINE
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