biostudies-literatureUnknownShukla RNIAID NIH HHSDutch Research Council (NWO)Antimicrobial resistance is a leading mortality factor worldwide. Here we report the discovery of clovibactin, a new antibiotic, isolated from uncultured soil bacteria. Clovibactin efficiently kills drug-resistant bacterial pathogens without detectable resistance. Using biochemical assays, solid-state NMR, and atomic force microscopy, we dissect its mode of action. Clovibactin blocks cell wall synthesis by targeting pyrophosphate of multiple essential peptidoglycan precursors (C ₅₅ PP, Lipid II, Lipid _WTA ). Clovibactin uses an unusual hydrophobic interface to tightly wrap around pyrophosphate, but bypasses the variable structural elements of precursors, accounting for the lack of resistance. Selective and efficient target binding is achieved by the irreversible sequebioRxiv : the preprint server for biology2023.05.15.540765https://www.ebi.ac.uk/biostudies/studies/S-EPMC10245560biostudies-literatureA new antibiotic from an uncultured bacterium binds to an immutable target.PMC10245560711.018.001R01 AI170962R43 AI091224184.032.207R44 AI091224R43 AI136137P01 AI118687R44 AI136137723.014.003Honorato RVLing LLKubitscheck ULelli MKrueger AMVermeulen BJASpoering ALAchorn Cde Benedetti SHughes DSchwalen CJLewis KLavore FNitti ASchneider TGrein FDerks MGNMaity SRoos WHBreukink ELudwig KCWeingarth MShukla RPeoples AJBonvin AfalseA new antibiotic from an uncultured bacterium binds to an immutable target.Antimicrobial resistance is a leading mortality factor worldwide. Here we report the discovery of clovibactin, a new antibiotic, isolated from uncultured soil bacteria. Clovibactin efficiently kills drug-resistant bacterial pathogens without detectable resistance. Using biochemical assays, solid-state NMR, and atomic force microscopy, we dissect its mode of action. Clovibactin blocks cell wall synthesis by targeting pyrophosphate of multiple essential peptidoglycan precursors (C ₅₅ PP, Lipid II, Lipid _WTA ). Clovibactin uses an unusual hydrophobic interface to tightly wrap around pyrophosphate, but bypasses the variable structural elements of precursors, accounting for the lack of resistance. Selective and efficient target binding is achieved by the irreversible seque2023-01-01T00:00:00Z2023 May2025-04-21T14:40:26.971Z2025-04-21T14:40:26.971ZS-EPMC102455603729262410.1101/2023.05.15.540765