Immobilization of enzymes on solid surfaces has become a fundamental technique for the preparation of enzyme-based biosensors. One of the most used oxide-reductase in this field is the Glucose Oxidase enzyme (GOD). Thin layers of Glucose Oxidase have been covalently and non-covalently bound to functionalized glass cover slips. A new method for the determination of the number of immobilized GOD molecules, based on the amperometric determination of Flavin Adenine Dinucleotide (FAD), was developed. Kinetic parameters (Km, kc, and kc/Km) were measured for the immobilized GOD and compared with the unbound enzyme. Atomic Force Microscopy (AFM) images were obtain in aqueous solution for the covalently and non-covalently bound enzyme, and revealed a homogeneous coverage of the glass support by the covalently bound GOD.

Kinetic and morphological characterization of Glucose Oxidase thin layers

DI PAOLO, MARIA LUISA;VIANELLO, FABIO;ZENNARO, LUCIO
1998

Abstract

Immobilization of enzymes on solid surfaces has become a fundamental technique for the preparation of enzyme-based biosensors. One of the most used oxide-reductase in this field is the Glucose Oxidase enzyme (GOD). Thin layers of Glucose Oxidase have been covalently and non-covalently bound to functionalized glass cover slips. A new method for the determination of the number of immobilized GOD molecules, based on the amperometric determination of Flavin Adenine Dinucleotide (FAD), was developed. Kinetic parameters (Km, kc, and kc/Km) were measured for the immobilized GOD and compared with the unbound enzyme. Atomic Force Microscopy (AFM) images were obtain in aqueous solution for the covalently and non-covalently bound enzyme, and revealed a homogeneous coverage of the glass support by the covalently bound GOD.
Scuola Nazionale di Biofisica, VI ciclo
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/177463
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