6533b86efe1ef96bd12cb51f
RESEARCH PRODUCT
Promoter Activation in Dhfq Mutants as an Efficient Tool for Specialized Metabolite Production Enabling Direct Bioactivity Testing
Andreas VilcinskasSimone EcksteinVirginie LecaudeyTien Duy VoSebastian L WenskiReuel BennetSvenja SimonyiMarco ThinesHarun CimenWilhelm SchäferRalf HeermannEckhard ThinesYi-ming ShiAntje K. HeinrichPeter GrünGerd GeisslingerNadine KellerDesalegne AbebewSusanne SchiffmannYvonne EngelThomas A. WichelhausMartin GandRobert FürstMerle HirschmannAnthony BuayaMarina HenkeIbrahim CakmakYan-ni ShiRebecca SchwenkMarisa SkaljacEdna BodeSophie BeyerThomas UlshöferFrank WescheHelge B. BodeAnja SchüfflerSelcuk HazirSophie C. BrandtIris BischoffDenia Franksubject
bioactivity testingnatural productsMetaboliteMutantPeptide SynthetasesXenorhabdus010402 general chemistry01 natural sciencesCatalysisBiosynthesis | Very Important Paperchemistry.chemical_compoundddc:570RNA chaperoneHumansMetabolomicsGeneResearch ArticlesBiological Productsbiology010405 organic chemistryPseudomonastechnology industry and agricultureGeneral MedicineGeneral Chemistrybiology.organism_classification0104 chemical sciencesBiosynthetic PathwayseasyPACIdchemistryBiochemistryddc:540proteobacteriaPhotorhabdussimplified productionResearch Articledescription
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‐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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2019-12-12 |