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.本研究首先构建基于规则的组学挖掘流程，系统性地揭示含Avi(Me)Cys的核糖体合成并经翻译后修饰的肽（Ribosomally synthesized and post-translationally modified peptides，RiPP）生物合成基因簇（BGCs）的全貌，随后开展细致的实验表征与生物活性测定。我们从逾5万株细菌基因组的庞大数据库中，鉴定出1172个负责合成含Avi(Me)Cys肽类的RiPP BGCs。后续通过基于生物合成规则的代谢组分析，成功建立了3个鉴定得到的BGCs与5种肽类抗生素生物合成之间的关联。本研究发现了一种V类羊毛硫肽（lanthipeptide）massatide A，其对革兰氏阳性病原菌展现出优异的抑菌活性，且耐药风险较低、安全性良好，是极具开发前景的抗生素候选化合物。为进一步提升massatide A的成药性，我们成功获得了26个massatide A衍生物。最终证实，相较于天然型massatide A，3个候选衍生物展现出更优的抗生素开发潜力。本数据集涵盖潜在ACyPs的生物合成基因簇、3个BGCs的质谱分析数据，以及新鉴定化合物的质谱（MS）与核磁共振波谱（NMR）数据。