Training old dogs to do new tricks: A general multi-pronged activation approach for natural product discovery in Actinomycetes

Abstract

Abstract Natural products are a family of diverse compounds with multiple impactful applications, especially in therapeutics. Recent advances in genomics and bioinformatics have also hinted at vast untapped chemical potential within Nature. However, despite the many strategies available for activation and upregulation of natural product biosyntheses in native and heterologous microbial strains, there is yet to be a generalizable and efficient approach for interrogating diverse native strain collections. Here, we describe and demonstrate a flexible and robust one-step integrase-mediated genetic- and cultivation-based approach to perturb and activate antibiotics production in a set of 54 actinobacterial strains. Our multi-pronged strategy significantly increases accessible metabolite space by two-fold, resulting in the discovery of the first example of Gram-negative bioactivity in new tetramic acid analogs. We envision these results to serve as the first step toward a more streamlined, accelerated, and scalable strategy to unlock the full potential of Nature’s chemical repertoire.