title: Extracting kinetic parameters for homogeneous [Os(bpy)2ClPyCOOH]+ mediated enzyme reactions from cyclic voltammetry and simulations.
abstract: The homogeneous reaction between glucose oxidase and osmium bipyridine-pyridine carboxylic acid in the presence of glucose has been studied in detail by cyclic voltammetry and digital simulation. Combination of the analytical equations that describe the dependence of the amperometric response on enzyme, substrate and co-substrate concentrations for the limiting cases with digital simulation of the coupled enzyme reaction diffusion problem allows us to extract kinetic parameters for the substrate-enzyme reaction: K(MS)=10.8 mM, k(cat)=254 s(-1) and for the redox mediator-enzyme reaction, k=2.2x10(5) M(-1) s(-1). The accurate determination of the kinetic parameters at low substrate concentrations (<7 mM) is limited by depletion of the substrate close to the electrode surface. At high substrate concentrations (>20 mM) inactivation of the reduced form of glucose oxidase in the bulk solution must be taken into account in the analysis of the results.
authors.affiliation: INQUIMAE, Departamento de Quimica Inorganica, Analitica y Quimica Fisica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EHA, Argentina.
authors.forename: V
authors.initials: V
authors.lastname: Flexer
authors.forename: M V
authors.initials: MV
authors.lastname: Ielmini
authors.forename: E J
authors.initials: EJ
authors.lastname: Calvo
authors.forename: P N
authors.initials: PN
authors.lastname: Bartlett
mesh_terms: D004563:Electrochemistry / Q000379:methods*; D004789:Enzyme Activation; D005949:Glucose Oxidase / Q000737:chemistry*; D007700:Kinetics; D009942:Organometallic Compounds / Q000737:chemistry*; D009992:Osmium / Q000737:chemistry*; D010084:Oxidation-Reduction
chemical_list: D009942:Organometallic Compounds; D009992:Osmium; D005949:Glucose Oxidase