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

Mingyar E, Muhling L, Kulik A, et al. A Regulator Based "Semi-Targeted" Approach to Activate Silent Biosynthetic Gene Clusters. International journal of molecular sciences. 2021;22(14): 7567.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 Tubingen 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. TU17, Streptomyces sp. TU10 and Streptomyces sp. TU102. Introduction of the CSRs-plasmid into strain S. sp. TU17 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. TU102, the introduction of the SARP-plasmid triggered the production of a chartreusin-like compound. Insertion of the CSRs-plasmid into strain S. sp. TU10 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. TU102 and aur1P and pntR in of S. sp. TU10