Supporting data for “Discovery and Engineering of Antibacterial Avi(Me)Cys-containing Cyclopeptides”

we first establish a rule-based omics mining pipeline to systematically uncover the landscape of Avi(Me)Cys-containing RiPP biosynthetic gene clusters (BGCs), followed by detailed experimental characterization and bioassay. We identify 1,172 RiPP BGCs responsible for Avi(Me)Cys-containing peptides formation from a vast pool of over 50,000 bacterial genomes. Subsequently, we successfully establish the connection between three identified BGCs and the biosynthesis of five peptide antibiotics via biosynthetic rule-guided metabolic analysis. we discover a class V lanthipeptide, massatide A, which displays excellent activity against gram-positive pathogens, with a relatively low risk of resistance and favorable safety profile, positions it as a promising candidate for antibiotic development. To further improve the druggability of massatide A, we successfully obtained twenty-six massatide A derivatives. Finally, we confirmed three candidates exhibited superior potential for antibiotic development compared to native massatide A.The dataset includes the biosynthetic gene cluster of potential ACyPs, mass analysis of three BGCs, MS data and NMR data of the newly identified compounds.