Analysis Note

Protein determined by biuret.

Application

Several publications cite use of the G2133 glucose oxidase in their protocols and in various applications, such as the following:a) Biosensor development: Diazoresin nanofilm coatings on alginate microspheres: Srivastava, R. et al., Biotechnol. Bioeng., 91(1), 124-131 (2005). Paper-based glucose biosensor: Lankelma, J. et al., Anal. Chem., 84(9), 417-4152 (2012) Microfluidic device with glucose oxidase immobilized on hydrogel for glucose analysis of blood: He, R.-Y. et al., RSC Adv., 9, 32367-32374 (2019). b) Single-molecule FRET study of human RAD51 filament formation: Subramanyam, S. et al., Methods Enzymol., 600, 201-232 (2018).c) Enzymatic fuel-cells with chitosan-based membranes: Bahar, T., and Yazici, M.S., Electroanalysis, 32(6), 1304-1314 (2020).

Glucose oxidase is widely used in the food and pharmaceutical industries as well as a major component of glucose biosensors.

Biochem/physiol Actions

Glucose oxidase catalyses the oxidation of β-d-glucose to d-glucono-β-lactone and hydrogen peroxide, with molecular oxygen as an electron acceptor.

General description

Molecular Weight: 160 kDa (gel filtration)pI: 4.2Extinction coefficient: E^1% = 16.7 (280 nm)Glucose oxidase from Aspergillus niger is a dimer consisting of 2 equal subunits with a molecular mass of 80 kDa each. Each subunit contains one flavin adenine dinulceotide moiety and one iron. The enzyme is a glycoprotein containing ~16% neutral sugar and 2% amino sugars. The enzyme also contains 3 cysteine residues and 8 potential sites for N-linked glycosylation.Glucose oxidase is capable of oxidizing D-aldohexoses, monodeoxy-D-glucoses, and methyl-D-glucoses at varying rates. The pH optimum for glucose oxidase is 5.5, while it has a broad activity range of pH 4-7. Glucose oxidase is specific for β-D-glucose with a K_M of 33-110 mM.Glucose oxidase does not require any activators, but it is inhibited by Ag⁺, Hg²⁺, Cu²⁺, phenylmercuric acetate, and p-chloromercuribenzoate. It is not inhibited by the nonmetallic SH reagents: N-ethylmaleimide, iodoacetate, and iodoacetamide.Glucose oxidase can be utilized in the enzymatic determination of D-glucose in solution. As glucose oxidase oxidizes β-D-glucose to D-gluconolactate and hydrogen peroxide, horseradish peroxidase is often used as the coupling enzyme for glucose determination. Although glucose oxidase is specific for β-D-glucose, solutions of D-glucose can be quantified as α-D-glucose will mutorotate to β-D-glucose as the β-D-glucose is consumed by the enzymatic reaction.

Physical form

Lyophilized powder containing phosphate buffer salts and sodium chloride

Quality

May contain traces of amylase, maltase, glycogenase, invertase, and galactose oxidase.

Unit Definition

One unit will oxidize 1.0 µmole of β-D-glucose to D-gluconolactone and H₂O₂ per min at pH 5.1 at 35 °C, equivalent to an O₂ uptake of 22.4 µl per min. If the reaction mixture is saturated with oxygen, the activity may increase by up to 100%.