Stabilization of Glucose Oxidase Enzyme in Liquid Reagent

Aktarun Nahar, Abdul Alim, Nusrat Zahan, Mohammad Taufiq Alam

Abstract

Attempts were made to improve the stability of glucose oxidase (GOD) enzyme to expand its feasibility of application using sodium gluconate (1%, 2%, 3% and so on), disaccharide trehalose (1%, 2%, 3% etc.), glycerol (1%, 2%, 3% etc.) and sorbitol (1%, 2%, 3% and so on) in liquid form, having a pH of 7.4 and containing phosphate buffer saline at a concentration of 50 mM. Different combinations of these stabilizers were applied to the glucose oxidase preparation and the stabilizing effects of these combinations were verified and discussed. The stability of the enzyme preparation were checked with respect to concentration of the stabilizing agent for 7 days at 37°C. For all stabilizers, an increasing activity of the enzyme was found with increasing percentage of stabilizer in the stabilizing solution up to a certain limit. Among the formulations top three were selected for the ‘degradation of enzyme activity with respect to time’ tests to find the most effective formulation. Results showed that, formulation containing 5% sodium gluconate with 5% trehalose could retain at least 99% initial activity of glucose oxidase enzyme during 28 days of assay. So, it was concluded that, sodium gluconate together with trehalose is the most appropriate for stabilization of glucose oxidase in liquid reagent at a certain concentration.

Conclusion

The enzyme undergoes different kinds of denaturation processes during production, storage and application. It can be reversed if the denaturing influence is removed. This study evaluates the stabilization efficiency of stabilizing agents at various concentrations on the biocatalyst activity of glucose oxidase. Sodium gluconate with trehalose appeared to be appropriate for the stabilization of glucose oxidase subjected to extreme environmental condition. The experiments revealed that the stabilizing agents provided much better long-term storage stability at pH 7.4, by stabilizing enzyme activity. The application of stabilizing agents improved the temperature profile of enzyme activity and shifted the activity optimum toward the region of higher temperature. The noticeable increase in enzyme activity in the presence of sodium gluconate with trehalose could be of great biotechnological importance while performing enzymatic conversions at low concentrations.

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