Oxidation, glucose, immobilized enzyme reaction

The first enzyme biosensor was a glucose sensor reported by Clark in 1962 [194], This biosensor measured the product of glucose oxidation by GOD using an electrode which was a remarkable achievement even though the enzyme was not immobilized on the electrode. Updark and Hicks have developed an improved enzyme sensor using enzyme immobilization [194], The sensor combined the membrane-immobilized GOD with an oxygen electrode, and oxygen measurements were carried out before and after the enzyme reaction. Their report showed the importance of biomaterial immobilization to enhance the stability of a biosensor. 

In this test, oxygen from air is used for the enzymatic oxidation of (3-D-glucose in the presence of immobilized GOD to gluconic acid and hydrogen peroxide. Hydrogen peroxide can be determined by a second enzyme reaction. With horse radish peroxidase as catalyst o-phenylendiamine is oxidized by H2O2 to 2,3-diaminophenazine, which can be photometrically determined at 490 nm, thus establishing a quantitative relationship between active GOD sites and the intensity of the absorption band. 

The bioelectrocatalytic oxidation or reduction of substrates at biocatalyst electrodes can be accelerated by the presence of a small molecule which functions as an electron transfer mediator between the electrode and the prosthetic group of the immobilized enzyme " . Two types of the enzyme-modified electrode with entrapped mediator have been designed" , a carbon paste electrode and a porous electrode, both modified with enzyme and a reservoir of mediator. In the former type of electrode , where glucose oxidase (GOD) was immobilized on the surface of a carbon paste electrode along with p-benzoquinone (BQ) by coating the enzyme-loaded surface with a semipermeable membrane, BQ was dissolved into the immobilized enzyme layer and retained there effectively to serve as an electron transfer mediator between the carbon paste electrode and the immobilized enzyme. It has been shown " that the kinetics of the bioelectrocatalytic oxidation of D-glucose (Glc) at the membrane-coated GOD-modified carbon paste electrode with mixed-in BQ can be explained by theoretical equations in which the diffusion enzyme reaction of the substrate and mediator in the immobilized-enzyme layer and the diffusion of the substrate (and mediators) in the coating membrane are taken into account - . In this study, a number of quinone derivatives and a few ferrocene derivatives were examined as electron transfer mediators in the GOD electrode, and the mediator activity of the compounds was evaluated on the basis of theoretical equations. 

The glucose oxidation reaction in the enzyme immobilized membrane of the glucose sensors is performed according to the following reaction. 

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