CELLULOSE HYDROLYSIS BY THE CELLULASES FROM TRICHODERMA-REESEI - A NEW MODEL FOR SYNERGISTIC INTERACTION

The hydrolysis of Whatman no. 1 filter paper by purified cellulolytic components from Trichoderma reesei and the synergistic action of binary combinations of these enzymes on the same substrate were investigated. At 20 g/l filter paper, enzyme concentrations needed to obtain half-maximal hydrolysis rates (K(E) values) were in the 3-4 muM range for the cellobiohydrolases (CBHs) and 0.05-0.10 muM for the endoglucanases (EGs). Catalytic-core proteins of CBH I and EG III, lacking the cellulose-binding domain, exhibit K(E) values 2.3 and 5.1 times higher than those of the intact enzymes. In synergistic combinations of two cellulases, the K(E) value of at least one enzyme was 3-10-fold reduced. CBH I/CBH II and CBH I/EG III combinations showed the most powerful synergism, and optimal ratios were a function of the total protein concentration. Results obtained in activity and adsorption assays using filter paper pretreated with one component, followed by inactivation and subsequent hydrolysis with the same or another cellulase component, point to a sequential enzymic attack of the cellulose and seems consistent with the mathematical model presented.