Purification And Characterization Of Beta-Glucosidase From Aspergillus Niger

Aspergillus niger, an isolate of soil contaminated with effluents from cotton ginning mill was grown in Czapek-Dox medium containing sawdust, Triton-X 100 and urea for production of an extracellular -glucosidase. -Glucosidase enzyme was purified (86-fold) from culture filtrate of A. niger by employing ammonium sulphate precipitation and gel filtration on sephadex G-75. The molecular mass of the purified enzyme was estimated to be 95 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The enzyme had an optimal activity on p-nitrophenyl -D-glucopyranoside at 50 degrees C and pH 5.0. The K-m and V-max of the enzyme on p-nitrophenyl -D-glucopyranoside at 50 degrees C and pH 5 were 8.0 mM and 166 mu mol/min/mg of protein, respectively. The enzyme could hydrolyze cellobiose and lactose but not sucrose. Heavy metals like Hg2+, Al3+, and Ag+ inhibited the activity, whereas Zn2+ and detergents such as Triton-X 100 and Tween-80 increased the activity at 0.01%. The enzyme activity increased in the presence of methanol and ethanol.