Rapid Glucose Determination by Mutarotase

Aldose 1-epimerase or mutarotase is the enzyme that responsible for carbohydrate metabolism and converts the alpha anomer into beta anomer of glucose. This enzyme was extracted from bovine kidney cortex. Crude enzyme exhibited the activity of 14.92 U mL-1 with specific activity of 0.153 U mg-1 proteins. The enzyme activity and specific activity was increased to 53.75 UmL-1 4.981 Umg-1 respectively after 38-60% ammonium sulfate precipitation and it was further increased to 73.27 UmL-1 and 11.67 Umg-1 when subjected to diethylaminoethyl (DEAE) cellulose chromatography. Further purification was carried out by passing it through Sephadex G-150 column and observed increase in activity 79.26 UmL-1 with 19.55 Umg-1 specific activity. The optimum pH and temperature were recorded as 8.5 and 37 oC respectively. Different stabilizers (glycerol, sodium benzoate, sodium citrate) were used to study their effect on stability of enzyme.