Amperometric Sensor and Immobilized Enzyme Electrode for the Determination of Enzymatic Activities

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Abstract
A sensitive method for the rapid determination of activities of soluble or immobilized enzymes, based on the electrochemical detection of H2O2 is described. Kinetic studies (Vmax and Km determinations) can be performed for all H2O2 generating enzymes (i.e., most of the oxidases) using an amperometric probe with a Pt anode at a fixed potential. When associated with an immobilized glucose oxidase membrane, this sensor constitutes a glucose electrode and the activity of any hydrolase which releases glucose can be measured. There is no need for other auxiliary enzymes and no preincubation step is required. The possibility to carry out continuous analysis constitutes the main advantage of the described method.