Abstract
Enzymes containing domain of unknown function (DUF) 3328 are proposed to catalyze varied reactions in fungal natural product biosynthesis, including halogenation, hydroxylation, oxidative cyclization, etc. However, only two DUF3328 enzymes have been very recently characterized: halogenase ApnU and macrocyclase AprY. We report the biochemical characterization of CctR, a DUF3328 enzyme that catalyzes copper- and oxygen-dependent C(sp(3))-H hydroxylation in hydroxycyclochlorotine biosynthesis. This work expands the catalytic repertoire of this widely distributed enzyme family.