Purification and some properties of a beta-glucosidase from Trichoderma harzianum type C-4.
Type C-4 strain of Trichoderma harzianum was isolated as a microorganism with high cellulolytic activity. Beta-glucosidase is involved in the last step of cellulose saccharification by degrading cellobiose to glucose, and plays an important role in the cellulase enzyme system with a synergic action with endoglucanase and cellobiohydrolase for cellulose degradation. Beta-glucosidase from T. harzianum type C-4 was purified to homogeneity through Sephacryl S-300, DEAE-Sephadex A-50, and Mono P column chromatographies. It was a single polypeptide with the molecular mass of 75,000 by SDS-PAGE. The enzyme was very active at pH 5.0 and 45 degrees C. No significant inhibition was observed in the presence of metal ions, thiol reagents, or EDTA. The enzyme was stable in the presence of 5% ox gall and digestive enzymes. p-Nitrophenyl-beta-D-cellobioside worked as a substrate for the enzyme as much as p-nitrophenyl-beta-glucopyranoside. Glucose and gluconolactone showed competitive inhibition with a Ki of 1 mM and 1.8 microM, respectively, while galactose, mannose, and xylose did not inhibit the enzyme significantly.
Yun
S I
SI
Department of Biological Sciences, University of Ulsan, Korea.
Jeong
C S
CS
Chung
D K
DK
Choi
H S
HS
D002480:Cellulase / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D005942:Gluconates / Q000378:metabolism / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D005960:Glucosides / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007783:Lactones; D008670:Metals / Q000378:metabolism / Q000494:pharmacology; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology*; D001617:beta-Glucosidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*
D004791:Enzyme Inhibitors; D005942:Gluconates; D005960:Glucosides; D007783:Lactones; D008670:Metals; C025193:4-nitrophenyl beta-D-glucoside; C034170:4-nitrophenyl beta-cellobioside; D002482:Cellulose; D004492:Edetic Acid; D001617:beta-Glucosidase; D002480:Cellulase; D005947:Glucose; C010730:beta-glucono-1,5-lactone