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The electroanalytical characteristics of recombinant cytochrome P450 3A4 (P450 3A4) immobilized  on the surface of screen-printed graphite electrodes modified with multi-walled carbon nanotubes have been studied.  The role and the influence of graphite working electrode modification with carbon nanotubes on electroanalytical  characteristics of cytochrome P450 3A4 have been demonstrated. The conditions for the immobilization of cytochrome  P450 3A4 on the obtained screen-printed graphite electrodes modified with carbon multi-walled nanotubes have been  optimized. The electrochemical parameters of the oxidation and reduction of the heme iron of the enzyme have  been estimated. The midpoint potential E0′ was -0.35±0.01 V vs Ag/AgCl; the calculated heterogeneous electron  transfer rate constant ks, was 0.57±0.04 s-1; the amount of electroactive cytochrome P450 3A4 on the electrode Г0,  was determined as (2.6±0.6)⋅10-10 mol/cm2. The functioning mechanism of P450 3A4-based electrochemical sensor  followed the “protein film voltammetry”. In order to develop electrochemical analysis of drugs being substrates  of that hemoprotein and respective medical biosensors the voltammetric study of catalytic activity of immobilized  cytochrome P450 3A4 was carried out. Electrocatalytic properties of cytochrome P450 3A4, immobilized on modified  screen-printed graphite electrodes, has been investigated using erythromycin (macrolide antibiotics). It has been shown  that the modification of electrodes plays a decisive role for the study of the properties of cytochromes P450  in electrochemical investigations. Smart electrodes can serve as sensors for analytical purposes, as well  as electrocatalysts for the study of biotransformation processes and metabolic processes. Electrodes modified  with carbon nanomaterials are applicable for analytical purposes in the registration of hemoproteins. Electrodes  modified with synthetic membrane-like compounds (e.g. didodecyldimethylammonium bromide) are effective  in enzyme-dependent electrocatalysis.

Electroanalytical and electrocatalytical characteristics of cytochrome P450 3A4 using electrodes modified with nanocomposite carbon nanomaterials