1,4-Benzoquinone-based electrophoretic assay for glucose oxidase

Area of Science:

• Biochemistry
• Analytical Chemistry
• Enzyme Assays

Background:

• Glucose oxidase is a crucial enzyme in various biological and industrial processes.
• Accurate and efficient measurement of glucose oxidase activity is essential for research and applications.
• Existing assays may involve multiple steps or require additional reagents, impacting efficiency.

Purpose of the Study:

• To demonstrate the feasibility of a novel enzyme microassay for glucose oxidase.
• To develop a simplified assay protocol utilizing 1,4-benzoquinone as an electron acceptor.
• To establish a sensitive method for quantifying glucose oxidase concentration.

Main Methods:

• Utilized electrophoretically mediated microanalysis in a plug-plug mode with nanolitre volumes.
• Employed 1,4-benzoquinone as the electron acceptor for glucose oxidase.
• Separated reactant (1,4-benzoquinone) and product (hydroquinone) using differential binding to sulfated-beta-cyclodextrin in phosphate buffer (pH 7).
• Monitored absorbance in UV spectra for quantification.

Main Results:

• Successfully demonstrated the feasibility of the enzyme microassay for glucose oxidase.
• The assay effectively covers glucose oxidase concentrations ranging from 0.01 to 0.1 mg/mL.
• Eliminated the need for a secondary enzyme (peroxidase) due to strong UV absorbance of reactant and product.

Conclusions:

• The developed microassay offers a feasible and efficient method for glucose oxidase determination.
• The protocol simplifies enzyme quantification by eliminating the need for peroxidase.
• This approach provides a sensitive and direct measurement of glucose oxidase activity.