FLUORESCENCE-BASED DETERMINATION OF ENZYME ACTIVITY OF RECOMBINANT CYP51bl (STEROL 14α-DEMETHYLASE) WITH COUMARIN DERIVATIVES
The current investigation was undertaken with the aim to carry out an <i>in vitro </i>evaluation of the ability of coumarin derivatives as probe substrates to predict the activity of CYP51b1. The results obtained indicate that 7-aminocoumarin-4-acetic acid (ACAC) can be used to determine the recombinant CYP51bl activity. Determination of CYP51bl activity with ACAC is based on the direct registration of fluorescence increasing at 30°C. It was found also that BMR in a simple soluble model system can be used as an electron donor for CYP51B1. Fluorescence-based assay is highly sensitive and can be used for the screening of sterol 14alpha-demethylase inhibitors.