Figure. 1. Comprehensive Bioactivity-Guided Workflow for Natural Product Discovery:
Figure. 1. Comprehensive Bioactivity-Guided Workflow for Natural Product Discovery:
(a) A schematic overview of the GNP discovery pipeline, including microbial genome analysis, gene detection, structure prediction, combinatorial library generation, and metabolite identification via LC-MS/MS.
(b) Gene cluster cal1-cal40 from Streptomyces calvus ATCC 13382 encoding specific modules (Cal17 to Cal23) involved in metabolite biosynthesis.
(c) LC-MS/MS chromatogram highlighting the isolated GNP hit with m/z = 1,023.88, identified at a retention time of ~40 minutes.
(d). Illustrates a bioactivity guided workflow for isolating bioactive natural products (NPs) from microbes, starting with solvent based extraction to maximize diversity, followed by bioactivity evaluation and multiple fractionation steps to isolate single compounds. Culturing challenges, such as organisms failing to grow or produce NPs outside their natural habitat, are addressed through innovations like in situ analysis, advanced culturing techniques, and biosynthetic gene expression. Crude extracts face limitations including known or nondrug like NPs and insufficient quantities, which are mitigated through de-replication, pre-fractionation, and optimized extraction methods. Bioactivity guided fractionation further refines extracts, enabling the isolation of novel compounds while overcoming rediscovery challenges. Finally, advanced methods for elucidating molecular modes of action address the difficulty in identifying molecular targets of bioactive compounds, significantly enhancing drug discovery efficiency.
