Biosynthesis of Collinodin Unveils Iterative Oxidative and Prenylation Modifications
Jing Shi, Yi Zhang, Wei Qin Ren, Yang Shi, Yuan Yuan Wei, Bo Zhang, Rui Jiao, Hui Ming Ge
Abstract
Lasso peptides are a unique family belonging to ribosomally synthesized and post-translationally modified peptides (RiPPs), characterized by unique isopeptide bonds and knotted structures. In recent years, genome mining for RiPPs has significantly expanded the diversity of post-translational modification chemistries. However, lasso peptides with multiple modifications are rarely reported. In this study, we used genome mining tools to identify a lasso peptide biosynthetic gene cluster that encodes unusual tailoring enzymes, including an α-KG-dependent dioxygenase and a prenyltransferase. Through heterologous expression, we successfully produced a lasso peptide with prenylation modification and elucidated its biosynthetic pathway in detail through in vivo and in vitro experiments. We identified a rare α-KG-dependent dioxygenase that acts twice during the maturation of the lasso peptide. Additionally, we verified the function of the prenyltransferase and its substrate broad specificity through in vitro experiments.