UGT93s, Plant UGTs Generating Prenylated Phenolic Glycosides

Abstract • Prenylated phenolic glycosides, such as nodakenin, represent a class of natural products with diverse bioactivities. The metabolic engineering production of these compounds remains largely unexplored, primarily due to the scarcity of efficient UDP-glycosyltransferases (UGTs) capable of catalyzing prenylated phenolic substrates.• Through UGT classification and functional characterization, 14 UGT93s from Angelica decursiva were identified to generate nodakenin utilizing nodakenetin.• Six UGT93s showed catalytic preference toward prenylated phenolics. Through functional characterization of more UGT93s and ancestral sequence reconstruction, this catalytic characteristic was found in most UGT93s.• Two hydrophobic clusters wrapping around the substrates are probably the structural basis for the catalytic activities toward prenylated phenolics.• Functionally characterizing the novel UGT93s provided valuable biosynthetic modules for biomanufacturing. And finding the catalytic preference highlight the UGT93 family as a promising source of biocatalysts for the biosynthesis of prenylated phenolic glycosides, offering new opportunities for their scalable production.