High Production of beta-Glucosidase in Schizophyllum commune: Isolation of the Enzyme and Effect of the Culture Filtrate on Cellulose Hydrolysis

裂褶菌中β-葡萄糖苷酶的高产:酶的分离及培养滤液对纤维素水解的影响

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Abstract

Optimization experiments with response surface statistical analysis were performed with Schizophyllum commune to obtain high beta-glucosidase yields. The factors in the optimization experiment were the concentrations of cellulose, peptone, and KH(2)PO(4). Their optimal values were 3.2, 3.0, and 0.2 g/100 ml, respectively. Enzyme assays revealed very high beta-glucosidase (22.2 U/ml) and cellobiase (68.9 U/ml) yields. The avicelase yield was low as compared with that from Trichoderma reesei. Mixtures of S. commune and T. reesei culture filtrates caused faster and more extensive saccharification of Avicel than could be achieved by either filtrate alone. A beta-glucosidase was isolated and purified from the optimized culture filtrate of S. commune. The electrophoretic mobility of the purified beta-glucosidase indicated a molecular weight of 97,000. The amino acid composition was similar to that of beta-glucosidase from T. reesei. The acidic (aspartate and glutamate) residues or their amides or both made up approximately 20% of the protein. The NH(2)-terminal amino acid of the enzyme was histidine.

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