Preparation, characterization, and potential application of an immobilized glucose oxidase.

A simple, one-step process, using 0.25 M p-benzoquinone dissolved in 20% dioxane at 50 degrees C for 24 h was applied to the activation of polyacrylamide beads. The activated beads were reacted with glucose oxidase isolated from Aspergillus niger. The coupling reaction was performed in 0.1 M potassium phosphate at pH 8.5 and 0-4 degrees C for 24 h. The protein concentration was 50 mg/mL. In such conditions, the highest activity achieved was about 100 U/g solid. The optimum pH for the catalytic activity was shifted by about 1 pH unit in the acidic direction to pH 5.5. Between 35 and 50 degrees C, the activity of the immobilized form depends on the temperature to a smaller extent than that of the soluble form. Above 50 degrees C, the activity of immobilized glucose oxidase shows a sharper heat dependence. The enzyme-substrate interaction was not profoundly altered by the immobilization of the enzyme. The heat resistance of the immobilized enzyme was enhanced. The immobilized glucose oxidase is most stable at pH 5.5. The practical use of the immobilized glucose oxidase was tested in preliminary experiments for determination of the glucose concentration in blood sera.