Production of cellulases and β-glucosidase in Trichoderma reesei mutated by proton beam irradiation

To obtain mutant strains producing high levels of cellulases (FPase and CMCase) and beta-glucosidase, Trichoderma reesei KCTC 6950 was mutated by proton beam irradiation. Five mutants were selected out of 1,000 mutants of T.reesei treated with proton beam irradiation, based on their ability for enzyme production on a plate screening medium. In submerged cultures containing Mandel's fermentation medium, the mutant strain T-2 (MT-2) demonstrated a 165% increase in the activity of FPase, a 146% increase in the activity of CMCase, and a 313% increase in the activity of beta-glucosidase, compared with the wild type strain. Additionally, the properties of high level beta-glucosidase produced by MT-2 were the same as those of the wild type strain, e.g., an optimum pH of 4.8, and an optimum temperature of 65 A degrees C. Moreover, the protein concentrations of beta-glucosidase produced by the wild type strain and MT-2 were measured by SDS-PAGE, and then beta-glucosidase activities were detected by the MUG-zymogram assay.