title: Immobilized enzyme reactors based upon the flavoenzymes monoamine oxidase A and B.
abstract: Monoamine oxidase (MAO) catalyzes the oxidative deamination of amines. The enzyme exists in two forms, MAO-A and MAO-B, which differ in substrate specificity and sensitivity to various inhibitors. Membrane fractions containing either expressed MAO-A or MAO-B have been non-covalently immobilized in the hydrophobic interface of an immobilized artificial membrane (IAM) liquid chromatographic stationary phase. The MAO-containing stationary phases were packed into glass columns to create on-line immobilized enzyme reactors (IMERs) that retained the enzymatic activity of the MAO. The resulting MAO-IMERs were coupled through a switching valve to analytical high performance liquid chromatographic columns. The multi-dimensional chromatographic system was used to characterize the MAO-A (MAO-A-IMER) and MAO-B (MAO-B-IMER) forms of the enzyme including the enzyme kinetic constants associated with enzyme/substrate and enzyme/inhibitor interactions as well as the determination of IC(50) values. The results of the study demonstrate that the MAO-A-IMER and the MAO-B-IMER can be used for the on-line screening of substances for MAO-A and MAO-B substrate/inhibitor properties.
authors.affiliation: McGill University Health Centre, Montreal General Hospital, Montreal, Que., Canada.
authors.forename: Nektaria
authors.initials: N
authors.lastname: Markoglou
authors.forename: Ruth
authors.initials: R
authors.lastname: Hsuesh
authors.forename: Irving W
authors.initials: IW
authors.lastname: Wainer
mesh_terms: D019149:Bioreactors; D002851:Chromatography, High Pressure Liquid; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D008995:Monoamine Oxidase / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D004800:Enzymes, Immobilized; D008995:Monoamine Oxidase