Glucose oxidase preparation

Enzymes themselves can be analyzed by measuring the amount of substrate transformed in a given time or the product that is produced in a given time. The substrate, houldJ2eJn ccess so that the reaction rate depends oiily on the enzyme concentration. The results are expressed as international units of enzyme. For example, the activity of a glucose oxidase preparation can be determined by measuring manometrically or amperometrically the number of micromoles of oxygen consumed per minute. On the other hand, the use of enzymes to develop specific procedures for the determination of substrates, particularly in clinical chemistry, has proved to be extremely useful. In this case, the enzyme concentration is in excess so the reaction rate is dependent on the substrate concentration. 

The permanent inclusion of solution constituents during the electropolymerization of pyrrole was also used to prepare modified electrodes with cobalt phthalocya-nine and glucose oxidase 122423) examples. Another technique of perhaps... 

A colloidal suspension of conductive vanadium pentoxide [130] can be used to perform intercalation, adsorption or encapsulation of electroactive molecules or biomolecules for electrodes or biosensor realization [131]. Encapsulation of glucose oxidase in nanocomposite films made with polyvinyl alcohol and V205 sol-gel matrix or in ferrocene intercalated V2Os sol-gel [132] were envisaged to prepare glucose biosensors. 

Enzyme containing Nation membranes prepared according to the proposed protocol have shown high specific activity and stability of immobilized glucose oxidase. As expected, the simplicity of preparation provided high reproducibility. When the same casting solution is used, the maximum deviation in membrane activity is <2%. This, however, is also the precision limit for kinetic investigations. 

J. Pei and X. Li, Amperometric glucose enzyme sensor prepared by immobilizing glucose oxidase on CuPtC16 chemically modified electrode. Electroanalysis 11, 1266-1272 (1999). 

A. Salimi, R.G. Compton, and R. Hallaj, Glucose biosensor prepared by glucose oxidase encapsulated sol-gel and carbon-nanotube-modified basal plane pyrolytic graphite electrode. Anal. Biochem. 333, 49— 56 (2004). 

Ikariyama [2] described a unique method for the preparation of a glucose oxidase (GOD) electrode in their work. The method is based on two electrochemical processes, i.e. electrochemical adsorption of GOD molecules and electrochemical growth of porous electrode. GOD immobilized in the growing matrix of platinum black particles is employed for the microfabrication of the enzyme electrode. It demonstrated high performance with high sensitivity and fast responsiveness. 

Glucose oxidase (GOD) is a typical flavin enzyme with flavin adenine dinucleotide (FAD) as redox prosthetic group. Its biological function is to catalyze glucose to form gluconolaction, while the enzyme itself is turned from GOD(FAD) to GOD(FADH2). GOD was used to prepare biosensors in extensive fields. Many materials that can be used to immobilize other proteins can be suitable for GOD. GOD adsorbed on CdS nanoparticles maintained its bioactivity and structure, and could electrocatalyze... 

In contrast to the molecular wire of molecular interface, electron mediators are covalently bound to a redox enzyme in such a manner as an electron tunneling pathway is formed within the enzyme molecule. Therefore, enzyme-bound mediators work as molecular interface between an enzyme and an electrode. Degani et al. proposed the intramolecular electron pathway of ferrocene molecules which were covalently bound to glucose oxidase [ 4 ]. However, few fabrication methods have been developed to form a monolayer of mediator-modified enzymes on the electrode surface. We have succeeded in development of a novel preparation of the electron transfer system of mediator-modified enzyme by self-assembly in a porous gold-black electrode as schematically shown in Fig.12 [14]. 

Alonso et al. prepared ferrocenyl silicon dendrimers [67], which could be used as mediators in glucose biosensors, based on glucose oxidase [68, 69], The ferrocenyl units are located at the end of long, flexible, silicon containing branches and serve to electrically connect the enzyme to the electrode. The flexibility of the dendrimer is proposed to play an important role in the interaction with the redox center of glucose oxidase. 

Glucose oxidase, glycoproteins from plasma membranes of rat liver Analytical and semi preparative separations Lectin Affinity (Concanavalin A) disks [76]... 

Zhao et al. prepared magnetite (FesO nanoparticles modified with electroactive Prussian Blue [44]. These modified NPs were drop-cast onto glassy-carbon electrodes. They observed the redox processes commonly observed for PB (similar to that seen in Figure 4.8), and also demonstrated that the Prussian White material produced by PB reduction at 0.2 V served as an electrocatalyst for Fi202 reduction. They also prepared LbL films in which PB NPs and glucose oxidase were alternated between PD DA layers [99]. These were demonstrated to act as electrocatalysts for Fi202 reduction. Based on the ability to sense the product of the enzymatic reaction, these structures were shown to act as glucose sensors. 

The preparation and application of SAM systems patterned by STM and their use in catalysis was demonstrated by Wittstock and Schuhmann [123]. The patterning (local desorption) of SAMs from alkane thiols on gold was performed by scanning electrochemical microscopy (SECM), followed by the assembly of an amino-deriva-tized disulfide and coupling of glucose oxidase to form a catalytically active pattern of the enzyme. The enzymatic activity could be monitored/imaged by SECM. 

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