Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization

Abstract

CDH, an enzyme produced by wood-decomposing fungi, has diverse applications in biosensor production, bioremediation, and biomedical industries. In this study, CDH from Schizophyllum commune BCC26414 was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and Sephadex G-200 chromatography. The purification fold achieved was 65.81 with a specific activity of 1612.34 U/mg. The purity and molecular weight of CDH was confirmed using native and SDS PAGE. Optimal temperature and pH were found to be 30°C and 5, respectively. The purified CDH exhibited stability over a wide pH range (3.5 to 6.5) for 24 hrs and retained complete activity at 40°C, with reduced activity at 50°C when observed for 150 min. KCl, MgSO4, ZnSO4, and NiCl2 at a concentration of 5 mM enhanced CDH activity and HgCl2 and CuSO4 inhibited the enzyme activity. The kinetic constants, Km and Vmax of CDH for lactose were observed to be 125 mM and 13.26 U/ml, respectively. The purified CDH may be utilized commercially in various applications.