Bielectrode Strategy for Determination of CYP2E1 Catalytic Activity: Electrodes with Bactosomes and Voltammetric Determination of 6-Hydroxychlorzoxazone.

ABSTRACT: We describe a bielectrode system for evaluation of the electrocatalytic activity of cytochrome P450 2E1 (CYP2E1) towards chlorzoxazone. One electrode of the system was employed to immobilize Bactosomes with human CYP2E1, cytochrome P450 reductase (CPR), and cytochrome b₅ (cyt b₅). The second electrode was used to quantify CYP2E1-produced 6-hydroxychlorzoxazone by its direct electrochemical oxidation, registered using square-wave voltammetry. Using this system, we determined the steady-state kinetic parameters of chlorzoxazone hydroxylation by CYP2E1 of Bactosomes immobilized on the electrode: the maximal reaction rate (V_max) was 1.64 ± 0.08 min^-1, and the Michaelis constant (K_M) was 78 ± 9 μM. We studied the

SUBMITTER: Kuzikov AV 

PROVIDER: S-EPMC10812958 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature