Chemoenzymatic Late‐Stage Modifications Enable Downstream Click‐Mediated Fluorescent Tagging of Peptides

Aromatic prenyltransferases from cyanobactin biosynthetic pathways catalyse the chemoselective and regioselective intramolecular transfer of prenyl/geranyl groups isoprene donors to an electron-rich position in these macrocyclic linear peptides. These enzymes often demonstrate relaxed substrate specificity are considered useful biocatalysts for structural diversification Herein, we assess donor N1-tryptophan prenyltransferase AcyF anacyclamide A8P pathway using a library 22 synthetic alkyl pyrophosphate analogues, which many display reactive that amenable additional functionalization. We further used introduce moiety into tryptophan-containing cyclic peptide subsequently click chemistry fluorescently label enzymatically modified peptide. This chemoenzymatic strategy allows late-stage modification peptides is applications.