Bioelectrochemical platform with human monooxygenases: FMO1 and CYP3A4 tandem reactions with phorate.

• Immobilisation of human FMO1 on glassy carbon electrode with cation surfactant DDAB. • Contact angle and AFM surface measurements of graphene oxide-DDAB. • Enhanced electrochemical signal of FMO1 in presence of graphene oxide. • FMO1 bioelectrode activity towards pesticides; phorate, methiocarb and fenthion. • Successful tandem oxidation of phorate with FMO1 and CYP3A4 bioelectrodes. It is highly advantageous to devise an in vitro platform that can predict the complexity of an in vivo system. The first step of this process is the identification of a xenobiotic whose monooxygenation is carried out by two sequential enzymatic reactions. Pesticides are a good model for this type of tandem reactions since in specific cases they are initially metabolised by human flavin-containing monooxygenase 1 (hFMO1), followed by cytochrome P450 (CYP). To assess the feasibility of such an in vitro platform, hFMO1 is immobilised on glassy carbon electrodes modified with graphene oxide (GO) and cationic surfactant didecyldimethylammonium bromide (DDAB). UV–vis, contact angle and AFM measurements support the effective decoration of the GO sheets by DDAB which appear as 3 nm thick structures. hFMO1 activity on the bioelectrode versus three pesticides; fenthion, methiocarb and phorate, lead to the expected sulfoxide products with K M values of 29.5 ± 5.1, 38.4 ± 7.5, 29.6 ± 4.1 µM, respectively. Moreover, phorate is subsequently tested in a tandem system with hFMO1 and CYP3A4 resulting in both phorate sulfoxide as well as phoratoxon sulfoxide. The data demonstrate the feasibility of using bioelectrochemical platforms to mimic the complex metabolic reactions of xenobiotics within the human body. [ABSTRACT FROM AUTHOR]