Cellobiose dehydrogenase(CDH) as a hemoflavoenzyme is secreted out of cell in the cellulose degradation. As CDH strongly bound to amorphous cellulose, it helps cellulose hydrolysis by cellulase. CDH may have an important role of saccharification process for bioethanol production. In this study, Phanerochaete chrysosporium ATCC 32629 was selected for the production of CDH among other strains tested. The optimal temperature and pH of CDH produced by P. chrysosporium ATCC 32629 were 55 ℃ and 4, respectively. To improve the activity of CDH, the mutation of P. chrysosporium was performed using proton beam that has high energy level partially. As a result, P. chrysosporium mutant with the high activity was selected at 1.2 kGy in a range of 99.9% lethal rate. The CDH and β-glucosidase activities of mutant were 1.4 fold and 20 fold higher than those of wild strain. Therefore, P. chrysosporium mutant with the high activities of CDH and β-glucosidase was obtained from mutation by proton beam irradiation.