P450‐Modified Multicyclic Cyclophane‐Containing Ribosomally Synthesized and Post‐Translationally Modified Peptides

作者：Cheng-Li Liu, Zi Jie Wang, Jing Shi, Jing Shi, Zhang Yuan Yan, Guo Dong Zhang, Rui Jiao, Ren Xiang Tan, Hui Ming Ge · 发表于：Angewandte Chemie International Edition · 年份：2023 · DOI：10.1002/anie.202314046 · 被引用次数：22 · 研究领域：Chemical Synthesis and Analysis、Microbial Natural Products and Biosynthesis、RNA and protein synthesis mechanisms

Cyclic peptides with cyclophane linkers are an attractive compound type owing to the fine-tuned rigid three-dimensional structures and unusual biophysical features. Cytochrome P450 enzymes are capable of catalyzing not only the C-C and C-O oxidative coupling reactions found in vancomycin and other nonribosomal peptides (NRPs), but they also exhibit novel catalytic activities to generate cyclic ribosomally synthesized and post-translationally modified peptides (RiPPs) through cyclophane linkage. To discover more P450-modified multicyclic RiPPs, we set out to find cryptic and unknown P450-modified RiPP biosynthetic gene clusters (BGCs) through genome mining. Synergized bioinformatic analysis reveals that P450-modified RiPP BGCs are broadly distributed in bacteria and can be classified into 11 classes. Focusing on two classes of P450-modified RiPP BGCs where precursor peptides contain multiple conserved aromatic amino acid residues, we characterized 11 novel P450-modified multicyclic RiPPs with different cyclophane linkers through heterologous expression. Further mutation of the key ring-forming residues and combinatorial biosynthesis study revealed the order of bond formation and the specificity of P450s. This study reveals the functional diversity of P450 enzymes involved in the cyclophane-containing RiPPs and indicates that P450 enzymes are promising tools for rapidly obtaining structurally diverse cyclic peptide derivatives.