Promoter activation in Δhfq mutants as an efficient tool for specialized metabolite production enabling direct bioactivity testing

  • 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-sequence-similarity-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 Δhfq 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.

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Author:Edna Bode, Antje Katharina Heinrich, Merle HirschmannGND, Desalegne Abebew, Yan-Ni Shi, Tien Duy Vo, Frank Wesche, Yi-Ming Shi, Peter Grün, Svenja Simonyi, Nadine Keller, Yvonne Engel, Sebastian Wenski, Reuel Bennet, Sophie Beyer, Iris Bischoff, Anthony Buaya, Sophie Brandt, Ibrahim Cakmak, Harun Çimen, Simone Eckstein, Denia Frank, Robert FürstORCiDGND, Martin Gand, Gerd GeisslingerORCiDGND, Selcuk Hazir, Marina Henke, Ralf Heermann, Virginie LecaudeyORCiDGND, Wilhelm Schäfer, Susanne SchiffmannORCiDGND, Anja Schüffler, Rebecca Schwenk, Marisa Skaljac, Eckhard Thines, Marco Thines, Thomas Ulshöfer, Andreas Vilcinskas, Thomas A. Wichelhaus, Helge Björn BodeORCiDGND
URN:urn:nbn:de:hebis:30:3-638202
DOI:https://doi.org/10.1002/anie.201910563
ISSN:1521-3773
Parent Title (English):Angewandte Chemie
Publisher:Wiley-VCH
Place of publication:Weinheim
Document Type:Article
Language:English
Date of Publication (online):2019/11/06
Date of first Publication:2019/11/06
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2022/01/25
Tag:bioactivity testing; easyPACId; natural products; proteobacteria; simplified production
Volume:58
Issue:52
Page Number:7
First Page:18957
Last Page:18963
Note:
Research Funding: LOEWE-Centre Translational Biodiversity Genomics. Grant Number: LOEWE TBG; Scientific and Technological Research Council of Turkey (TUBITAK). Grant Number: TOVAG-117O172; LOEWE-Centre Translational Medicine and Pharmacology. Grant Number: LOEWE TMP; LOEWE-Centre Insect Biotechnology and Bioressources. Grant Number: LOEWE Insect biotechnology; LOEWE research cluster MegaSyn. Grant Number: LOEWE MegaSyn; DFG. Grant Number: INST 161/810-1.
HeBIS-PPN:490966950
Institutes:Biowissenschaften
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften
5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung 4.0