title: Human flavin-containing monooxygenase 3 on graphene oxide for drug metabolism screening.
abstract: Human flavin-containing monooxygenase 3 (hFMO3), a membrane-bound hepatic protein, belonging to the second most important class of phase-1 drug-metabolizing enzymes, was immobilized in its active form on graphene oxide (GO) for enhanced electrochemical response. To improve protein stabilization and to ensure the electrocatalytic activity of the immobilized enzyme, didodecyldimethylammonium bromide (DDAB) was used to mimic lipid layers of biological membranes and acted as an interface between GO nanomaterial and the hFMO3 biocomponent. Grazing angle attenuated total reflectance Fourier transform infrared (GATR-FT-IR) experiments confirmed the preservation of the protein secondary structure and fold. Electrochemical characterization of the immobilized enzyme with GO and DDAB on glassy carbon electrodes was carried out by cyclic voltammetry, where several parameters including redox potential, electron transfer rate, and surface coverage were determined. This system's biotechnological application in drug screening was successfully demonstrated by the N-oxidation of two therapeutic drugs, benzydamine (nonsteroidal anti-inflammatory) and tamoxifen (antiestrogenic widely used in breast cancer therapy and chemoprevention), by the immobilized enzyme.
authors: Department of Life Sciences and Systems Biology, University of Torino , via Accademia Albertina 13, 10123 Torino, Italy.
authors: Silvia
authors: S
authors: Castrignanò
authors: Gianfranco
authors: G
authors: Gilardi
authors: Sheila J
authors: SJ
authors: Sadeghi
mesh_terms: D018931:Antineoplastic Agents, Hormonal / Q000737:chemistry; D001591:Benzydamine / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004563:Electrochemistry; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D049329:Nanostructures / Q000737:chemistry; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000737:chemistry / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D013629:Tamoxifen / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D018931:Antineoplastic Agents, Hormonal; D004800:Enzymes, Immobilized; D013629:Tamoxifen; D001591:Benzydamine; D006108:Graphite; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)