Fig. 2

HPLC chromatograms of the in vivo and in vitro conversion of medrane with bovine CYP21A2 and CPR. a Medrane was converted with resting cells of C43(DE3) containing the bovine CYP21A2 isoform and CPR encoding vector p21b_bRED. Samples were taken after 24 h and extracted for HPLC analysis. The steroids were separated by an acetonitril:water gradient. b Substrate conversions of medrane were performed with purified bovine CYP21A2 and its redox partner CPR for 30 min. Steroids were extracted and analyzed by HPLC to verify a selective conversion of medrane to premedrol. The steroids were separated by an acetonitril:water gradient, showing the 21-hydroxylated product premedrol