Figure 1.

Characterization of recombinant RD20 activity in yeast. A, Mechanistic scheme for peroxygenase activity. Enz represents the peroxygenase, and Fe represents the heme of the protein; Sub and SubO represent the substrate and its oxidized form, respectively; R-O-OH and R-OH represent the hydroperoxide cosubstrate and its corresponding alcohol, respectively. B, Cooxidation of radiolabeled polyunsaturated fatty acids in the presence of cumene hydroperoxide (CuOOH), H₂O₂, or 13-HPOD. ¹⁴C-labeled substrates metabolized by microsomal fractions prepared from RD20-expressing yeast were separated by thin-layer chromatography and analyzed by radiodetection. 18:1, Oleic acid; 18:2, linoleic acid; 18:3, linolenic acid. Incubations of ¹⁴C-labeled substrates in the presence of microsomal fractions prepared from untransformed yeast and from yeast transformed with empty vector did not exhibit any activity.