The group of bacterial non-ribosomally produced peptides (NRPs) forms a rich source of antibiotics, such as daptomycin, vancomycin, and teixobactin. The difficulty of functionally expressing and engineering the corresponding large biosynthetic complexes is a bottleneck in developing variants of such peptides. Here, we apply a strategy to synthesize mimics of the recently discovered antimicrobial NRP brevicidine. We mimicked the molecular structure of brevicidine by ribosomally synthesized, post-translationally modified peptide (RiPP) synthesis, introducing several relevant modifications, such as dehydration and thioether ring formation. Following this strategy, in two rounds peptides were engineered in vivo, which showed antibacterial activity against Gram-negative pathogenic bacteria susceptible to wild-type brevicidine. This study demonstrates the feasibility of a strategy to structurally and functionally mimic NRPs by employing the synthesis and post-translational modifications typical for RiPPs. This enables the future generation of large genetically encoded peptide libraries of NRP-mimicking structures to screen for antimicrobial activity against relevant pathogens.

细菌非核糖体产生的肽（NRP）组形成了丰富的抗生素来源，例如达托霉素，万古霉素和替ixobactin。在功能上表达和工程化相应的大型生物合成复合物的困难是开发此类肽的变体的瓶颈。在这里，我们应用一种策略来合成最近发现的抗菌NRP brevicidine的模拟物。我们通过核糖体合成，翻译后修饰的肽（RiPP）合成模仿了短vic啶的分子结构，引入了几种相关的修饰，例如脱水和硫醚环形成。遵循这一策略，在体内设计了两轮多肽表现出对易受野生型布雷维定影响的革兰氏阴性致病菌的抗菌活性。这项研究通过采用RiPPs的典型合成和翻译后修饰，证明了在结构和功能上模仿NRP的策略的可行性。这使得将来可以生成大型的NRP模拟结构的基因编码肽库，以筛选针对相关病原体的抗菌活性。