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Journal: Communications biology
Article Title: Exploring a general multi-pronged activation strategy for natural product discovery in Actinomycetes.
doi: 10.1038/s42003-023-05648-7
Figure Lengend Snippet: Fig. 1 Schematics of a multi-pronged general activation strategy for native microbial strains toward natural product discovery. A Microbial strains are genetically edited via one-step phiC31 integrase protocol for heterologous expression of genes for activation and/or upregulation of secondary metabolites. B Genetic-based activation is complemented with cultivation-based (one strain many compounds, OSMAC) activation where a range of fermentation media (with varied nutrient compositions) is employed to probe the full biosynthetic potential of the genetically edited mutants. C Extracts are profiled both chemically and with bioassays to characterize and discover new bioactive metabolite space. Bioactivity assays include antibacterial assays (Acinetobacter baumannii, Klebsiella aerogenes, Pseudomonas aeruginosa and Staphylococcus aureus Rosenbach), antifungal assays (Aspergillus fumigatus) and mammalian cytotoxicity assays (A549 human lung carcinoma cells). D In depth analysis of LCMS data allowed us to interrogate the effects of media and activators where we found the combination of 3 regulators and 3 media were sufficient to produce maximal metabolites production. E Bioactivity profile comparison between mutants and native strains revealed that the expanded chemical space was accompanied by increased or novel bioactivities, which were used for new NP discovery.
Article Snippet: 1 and 2 were active against
Techniques: Activation Assay, Expressing, Comparison