Cells were thawed and resuspended in Buffer A (1 g cell pellet/ 5 mL) and lysed by sonication (1 s ON, 1 s OFF, 50 % amplitude, 10 min). Cell debris was removed by highspeed centrifugation at 18000 g for 20 min. The lysate supernatant was loaded onto 5 mL of Ni-NTA Agarose (Qiagen) and the column washed with 25 mL of wash Buffer B before finally eluting the purified protein with 10 mL of Buffer C. Pure fractions were concentrated by spin column (2000 rpm, Vivaspin 20, 30 kDA cutoff, Satorius) and buffer exchanged into 100 mM phosphate buffer, pH 8, 100 mM KPi using 10DG desalting columns (Bio-Rad). Purified protein was used for reaction assays or snap frozen in N2 (l) and stored at -80 °C. Active P450 protein concentrations were determined by UV-vis absorption at 419 nm.