Functional analysis of cytochrome P450 monooxygenases from the white-rot basidiomycete Trametes versicolor: CYP502B15 decorates cadinane-type sesquiterpenoids

Cytochrome P450 monooxygenases (P450) play crucial roles in the diversification of secondary metabolic systems in living organisms. Basidiomycetes produce a wide variety of natural products as secondary metabolites, with their relevance to pharmaceutical and agricultural applications garnering significant interest. Thus, the functional characterization of fungal P450s is essential for basic and applied research of basidiomycetes. We herein describe the identification and functional characterization of P450 in the white-rot basidiomycete Trametes versicolor (TvCYP). A total of 182 TvCYP genes were identified in the T. versicolor genome. Moreover, 145 cDNAs encoding possible mature open reading frames were isolated/generated. Furthermore, we constructed a library of TvCYPs using a yeast expression system that allowed for the semi-comprehensive screening of latent TvCYP functions. Notably, we used this library to identify TvCYPs responsible for the metabolism of cadinane-type sesquiterpenoids. CYP502B15 was revealed to have significant activities against γ-cadinene and epicubenol. On the basis of NMR and X-ray crystal structure experiments, CYP502B15 was observed to catalyze the 3α-hydroxylation of γ-cadinene and epicubenol. To the best of our knowledge, this is the first report of the identification of basidiomycetous P450s responsible for the metabolism of cadinane-type sesquiterpenoids.