Yuxia Ma, Jinming Di, Jin-Long Lu, Weixu Zhai, Guang-Cheng Lou, Xinrong Li, Jiao‐Jiao Cui, Jiang Xiong, Hongping Han, Kun Gao, Xinxiang Lei, Shangwen Luo, Shi‐Hui Dong
Lasso peptides and lanthipeptides are two major classes of ribosomally synthesized and post-translationally modified peptides (RiPPs), each defined by distinct post-translational modification (PTM) enzymes. We identify a hybrid biosynthetic gene cluster that integrates PTM enzymes from both pathways to produce a unique lasso peptide featuring a dehydroalanine residue and C-terminal Asp side-chain methylation. Elucidation of its biosynthesis uncovers unprecedented combinatorial RiPP maturation. This study establishes a paradigm for hybrid RiPP biosynthesis and enables discovery and engineering of novel RiPPs.