Abstract
Recent efforts in genome mining of ribosomally synthesized and post-translationally modified peptides (RiPPs) have expanded the diversity of post-translational modification chemistries. However, RiPPs are rarely reported as hybrid molecules incorporating biosynthetic machinery from other natural product families. Here we report lipoavitides, a class of RiPP/fatty-acid hybrid lipopeptides that display a unique, putatively membrane-targeting 4-hydroxy-2,4-dimethylpentanoyl (HMP)-modified N terminus. The HMP is formed via condensation of isobutyryl-coenzyme A (isobutyryl-CoA) and methylmalonyl-CoA catalysed by a 3-ketoacyl-(acyl carrier protein) synthase III enzyme, followed by successive tailoring reactions in the fatty acid biosynthetic pathway. The HMP and RiPP substructures are then connected by an acyltransferase exhibiting promiscuous activity towards the fatty acyl and RiPP substrates. Overall, the discovery of lipoavitides contributes a prototype of RiPP/fatty-acid hybrids and provides possible enzymatic tools for lipopeptide bioengineering.
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Data availability
We declare that all data supporting the findings of this study are presented in the main text and Supplementary Information. NCBI (https://www.ncbi.nlm.nih.gov/), Protein Data Bank (PDB; https://www.rcsb.org/) and Uniprot (https://www.uniprot.org/) accessions are referenced in Supplementary Information, and these accessions are publicly accessible on the respective NCBI, RCSB PDB and Uniprot websites. Source data are provided with this paper.
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Acknowledgements
We thank J. Arrington from the Roy J. Carver Biotechnology Center for HR-MS/MS assistance. This work was supported by a grant from the National Institutes of Health (AI144967 to D.A.M. and H.Z.). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the paper.
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H.R. performed bioinformatics analysis, direct cloning, heterologous expression, HR-MS/MS analysis, stereochemistry determination, bioactivity assays, mutational analysis and enzymatic reactions. C.H. performed compound purification, the isotopic feeding study and intermediate characterization. C.H. also designed, performed and analysed NMR experiments. Y.P. assisted in compound purification and genetic manipulation. H.C. performed the AlphaFold2 and AutoDock analysis. D.A.M. contributed to the bioinformatics analysis of lpv genes. S.R.D. performed Marfey’s analysis, reductive desulfurization and interpretation of NMR data to assign stereochemistry with oversight from D.A.M.; M.L. designed and assisted in synthesizing the racemic and (S)-DPD standards. M.G.G. performed and analysed 1H–1H ROESY NMR experiments with oversight from D.A.M.; H.R. and C.H. wrote the paper with editorial oversight from H.Z., along with input from all other authors. H.Z. conceived of and supervised the overall project.
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Ren, H., Huang, C., Pan, Y. et al. Non-modular fatty acid synthases yield distinct N-terminal acylation in ribosomal peptides. Nat. Chem. (2024). https://doi.org/10.1038/s41557-024-01491-3
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DOI: https://doi.org/10.1038/s41557-024-01491-3