Partial purification and characterization of beta-glucosidase from Armillaria mellea

In this study, ß-glucosidase from Armillaria mellea mushroom harvested at Hıdırnebi Plateau in Trabzon were partially purified by using ion-exchange chromatography and characterized. ß-Glucosidase activity was determined in the crude extracts prepared by using buffers having different pH values and the suitable buffer was chosen for the extraction. The presence of ß-glucosidase in the crude enyzme extract and solution obtained after ion-exchange choromatography were put forward by native polyacrylamide gel electrophoresis and substrate staining. While three bands were observed in the crude extract, two bands was observed after ion-exchange choromatography. Partially purified enzyme was biochemically characterized. The optimum pH and optimum temperature of the enzyme were found to be 4.0 and 50 °C, respectively. After 24 h of incubation at 4 °C, the A mellea ß-glucosidase extremely stable in the range of pH 3.0-6.0 and 8.0. When the termal stability profile of the enzyme was analyzed, it was determined that the enzyme was exremely stable in the range of 20-50 °C after 1 h incubation. Vmax and Km values of ß-glucosidase in peresence of pNPG were found to be 3.646 U/mg and 0.295 mM, respectively. It was also determined that some metal ions and chemicals were effected the enzyme activity in different ratio.