LimF is a versatile prenyltransferase for histidine-C-geranylation on diverse non-natural substrates

Prenylation plays an important role in diversifying the structure and function of secondary metabolites. Although several cyanobactin prenyltransferases have been characterized, their chemistries are mainly limited to modification electron-rich heteroatoms. Here we report a prenyltransferase, LimF, from Limnothrix sp. CACIAM 69d, geranylating electron-deficient C2 atom His imidazole. Interestingly, addition its native substrate, LimF also modifies diverse exotic peptides, including thioether-closed macrocycles. We serendipitously uncovered Tyr-O-geranylating activity as providing evolutional insight into divergent repertoire prenylated peptides produced by PTases. Crystallographic analysis complexed with pentapeptide substrate prenyl donor analogue provides structural basis for recognition bifunctionality. show prenylation ability on various bioactive molecules containing imidazole group, non-amino acid small molecules, highlighting potential versatile biocatalyst chemically challenging C-geranylation. His-C2-directed is rarely occurs nature due low reactivity this position. Now, prenlytransferase has discovered applied geranylation histidine-containing imidazole-containing showcasing versatility biocatalyst.