Fig. 6: ETE of RgCYP87A3 for Prn production and mechanism analysis.

A The conformation of Cho in the catalytic pocket of wild-type (cyan stick) and Mutant (green stick) RgCYP87A3. V118A, A281D, and T285E were shown in red. B In the wild-type catalytic pocket, the distances of C20 and C22 of Cho from the FeO center are 3.9 Å each. In the mutant catalytic pocket, these distances are 4.2 Å for C20 and 2.6 Å for C22. C Free energy calculation for residues within 4 Å of Cho in wild type (WT-RgCYP87A3, red) and mutant (Mu-RgCYP87A3, gray) RgCYP87A3’s heme center. D Schematic drawings the three steps of side chain cleaving process by D281, E285, and heme center. E Distances between FeO (heme) of WTRgCYP87A3 and C20/22 of Cho were measured by MD simulation in 100 ns. F Distances between FeO (heme) of MuRgCYP87A3 and C20/22 of Cho were measured by MD simulation in 100 ns. G QM/MM calculation calculated energy profile (in kcal/mol) for RgCYP87A3 cleaving Cho side chain process in engineered catalytic pocket (red) and wild type pocket (black). (H) QM/MM-optimized the structures in the RgCYP87A3 and mutant RgCYP87A3 cleaving reaction. Key distances of TS1, TS2, TS3, TS1’, TS2’, and TS3’ are given in Å. Values are shown as mean±s. d. from three biological replicates. Source data are provided as a Source Data file.