Enzymatic Hydrolysis of Cellulose Using Mixes of Mutant Forms of Cellulases from Penicillium verruculosum

Abstract

Cellulases are the major components of multienzyme systems applied in processes of bioconversion of renewable lignocellulosic feedstocks to various useful products. The hydrolytic efficiency of enzyme mixes based on recombinant wild-type endoglucanase II, cellobiohydrolases I and II from the Penicillium verruculosum fungus (in the presence of Aspergillus niger β-glucosidase) with mixes of mutant forms of these enzymes in the hydrolysis of cellulosic materials is compared, and the influence of temperature and substrate concentration on the glucose yield is studied. The mutant cellulases represented proteins, in which N-linked glycans were partially removed using site-directed mutagenesis. In the hydrolysis of microcrystalline cellulose and milled aspen wood by mixes of mutant cellulases, the yields of glucose after 24–72 h of an enzymatic reaction were higher by 31–38% and 11–27%, respectively, than those for the compositions based on the wild-type enzymes. The highest product concentrations, using mutant enzyme compositions, are achieved at 50°С when the hydrolysis temperature is varied in the range of 40 to 60°С. Increasing the substrate concentration in the reaction system from 5 to 50 g/L (while maintaining the enzyme dosage at the same level) led to a 2.6–2.8-fold increase in the glucose yield, accompanied by a decrease in the cellulose conversion degree.