Promoter Activation in Delta hfq Mutants as an Efficient Tool for Specialized 
Metabolite Production Enabling Direct Bioactivity Testing

Bode, Edna; Heinrich, Antje K.; Hirschmann, Merle; Abebew, Desalegne; Shi, Yan-Ni; Tien Vo, Duy; Wesche, Frank; Shi, Yi-Ming; Gruen, Peter; Simonyi, Svenja; Keller, Nadine; Engel, Yvonne; Wenski, Sebastian; Bennet, Reuel; Beyer, Sophie; Bischoff, Iris; Buaya, Anthony; Brandt, Sophie; Cakmak, Ibrahim; Cimen, Harun; Eckstein, Simone; Frank, Denia; Fuerst, Robert; Gand, Martin; Geisslinger, Gerd; Hazir, Selcuk; Henke, Marina; Heermann, Ralf; Lecaudey, Virginie; Schaefer, Wilhelm; Schiffmann, Susanne; Schueffler, Anja; Schwenk, Rebecca; Skaljac, Marisa; Thines, Eckhard; Thines, Marco; Ulshoefer, Thomas; Vilcinskas, Andreas; Wichelhaus, Thomas A.; Bode, Helge B.

doi:10.1002/anie.201910563

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Promoter Activation in Delta hfq Mutants as an Efficient Tool for Specialized Metabolite Production Enabling Direct Bioactivity Testing

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Citation

Bode, E., Heinrich, A. K., Hirschmann, M., Abebew, D., Shi, Y.-N., Tien Vo, D., et al. (2019). Promoter Activation in Delta hfq Mutants as an Efficient Tool for Specialized Metabolite Production Enabling Direct Bioactivity Testing. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 58(52), 18957-18963. doi:10.1002/anie.201910563.

Cite as: https://hdl.handle.net/21.11116/0000-000A-05A4-7

Abstract

Natural products (NPs) from microorganisms have been important sources for discovering new therapeutic and chemical entities. While their corresponding biosynthetic gene clusters (BGCs) can be easily identified by gene-sequencesimilarity-based bioinformatics strategies, the actual access to these NPs for structure elucidation and bioactivity testing remains difficult. Deletion of the gene encoding the RNA chaperone, Hfq, results in strains losing the production of most NPs. By exchanging the native promoter of a desired BGC against an inducible promoter in Dhfq mutants, almost exclusive production of the corresponding NP from the targeted BGC in Photorhabdus, Xenorhabdus and Pseudomonas was observed including the production of several new NPs derived from previously uncharacterized non-ribosomal peptide synthetases (NRPS). This easyPACId approach (easy Promoter Activated Compound Identification) facilitates NP identification due to low interference from other NPs. Moreover, it allows direct bioactivity testing of supernatants containing secreted NPs, without laborious purification.