This article describes that electrochemical polymerization of 4-methoxyphenol in the presence of enzyme glucose oxidase produces adherent polymeric films containing the active enzyme onto the surface of platinum electrodes. Polymeric electrodes prepared in this one-step procedure can be operated for the glucose determination. The effects of the electrochemical polymerization parameters (for example, concentrations of monomer, electrolyte, and enzyme; film thickness; and polymerization potential) on the electrode preparation and the effects of amperometric measurement parameters (for example, pH, temperature) on the amperometric responses to the glucose of the prepared electrodes were systematically investigated, and optimal values were determined. Furthermore, glucose specificity and storage stability of the enzyme electrode were investigated.