Purification and characterization of beta-1,4-glucosidase from Clostridium papyrosolvens.
Clostridium papyrosolvens CFR-1010 was selected for the anaerobic extracellular production of beta-1,4-glucosidase. The enzyme was purified by alcohol precipitation and DEAE ion-exchange chromatography. Its homogeneity 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 degrees C. The enzyme activity was inhibited by Ca2+, Co2+, Cu2+, Zn2+, Fe2+, Mg2+ and Na+ ions. However, the activity increased (158%) in the presence of MnCl2, whereas it decreased by 80% in the presence of N-bromosuccinimide, suggesting the presence of tryptophan residues at the active site of enzyme. The enzyme had a K(m) of 15 mg/ml and Vmax of 125 units/min per mg of protein.
Sharmila
T
T
Department of Food Microbiology, Central Food Technological Research Institute, Mysore, India.
Sreeramulu
G
G
Nand
K
K
D003013:Clostridium / Q000737:chemistry / Q000201:enzymology*; D004586:Electrophoresis / Q000379:methods; D008670:Metals / Q000494:pharmacology; D001617:beta-Glucosidase / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*
D008670:Metals; D001617:beta-Glucosidase