Figure 4
From: Steroid conversion with CYP106A2 – production of pharmaceutically interesting DHEA metabolites
Conversion of 400 μM DHEA. 50 ml B. megaterium ATCC13368 cultures were incubated 16 hours at 30°C and 150 rpm. DHEA (400 μM dissolved in ethanol) was added and cultures were incubated additional 180 min. 500 μl samples were taken at 0, 15, 30, 45, 60, 120 and 180 minutes after substrate addition. Samples were quenched and extracted with 500 μl ethyl acetate, evaporated to dryness and chromatographed using ethyl acetate as mobile phase. After anisaldehyde staining the relative substrate consumption in % was calculated using the spot intensity with the Image-Lab™ (BIO-RAD, Germany) software. The graph shows the mean values and standard deviation from three independent conversions.