Fig. 11
The conversion and major hydroxylated products of 11-deoxycorticosterone (a) and cortodoxone (b) catalyzed by CYP260B1 and CYP260B1-T224A. Reaction conditions: purified enzymes of CYP260B1 and its T224A (0.5 μM), AdR (1.5 µM), Adx4–108 (10 µM), MgCl2 (1 mM), glucose-6-phosphate (5 mM) and glucose-6-phosphate dehydrogenase (1 U) in a final volume of 250 µl of potassium phosphate buffer (20 mM, pH 7.4) was used. The steroids (10 mM stock solution in EtOH) were added to a final concentration of 200 µM. The reactions were initiated by adding 500 µM NADPH, 30 min, 30 °C