PURIFICATION AND CHARACTERIZATION OF BETA-1,4-GLUCOSIDASE FROM CLOSTRIDIUM PAPYROSOLVENS

Clostridium papyrosolvens CFR-1010 was selected for the anaerobic extr acellular production of beta-1,4-glucosidase, The enzyme was purified by alcohol precipitation and DEAE ion-exchange chromatography. Its hom ogeneity was confirmed by SDS/PAGE. The enzyme had a molecular mass of 85 kDa, The maximum enzyme activity was observed at pH 5.0 and 50 deg rees C. The enzyme activity was inhibited by Ga2+, Co2+, Cu2+, Zn2+, F e2+, Mg2+ and Na+ ions. However, the activity increased (158%) in the presence of MnCl2, whereas it decreased by 80% in the presence of N-br omosuccinimide, suggesting the presence of tryptophan residues at the active site of enzyme, The enzyme had a K-m of 15 mg/ml and Y-max of 1 25 units/min per mg of protein.