Electrochemical Assessment of CYP3A4 Catalytic Activity in Bactosomes

In this work, we present investigation on the behavior of bactosomes containing human cytochrome CYP3A4 in comparison with recombinant enzyme. The bactosomes were adsorbed on the surface of screen-printed electrodes modified with didodecyldimethylammonium bromide. We have shown that bactosomes are effective bioreactors in the case of drug metabolic electroenzymatic reactions. The electrocatalytic reaction was conducted at a working potential of - 0.50 divided by - 0.55 V. Drug conversion was registered by means of measuring of formaldehyde as product of N-demethylation reaction of erythromycin and time-dependent electrochemical oxidation of diclofenac during electrolysis at controlled working potential. The efficiencies of electrocatalysis of erythromycin N-demethylation as well-known cytochrome P450 3A4 substrate in the case of bactosomes containing human cytochrome P450 3A4 was 3.5 +/- 0.4 times higher in comparison with highly purified cytochrome P450 3A4-electrode as catalyst. The efficiencies of electrocatalysis of diclofenac 5-hydroxylation in the case of bactosomes were 3.1 +/- 0.3 times higher than for purified enzyme. Bactosomes are effective electrochemical enzymatic nanobioreactors with a high electroenzymatic efficiency for metabolite production.