Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization

Vinita Yadav¹ and Sadhana Nighojkar^2*

¹School of Biotechnology, Devi Ahilya University Indore (M.P) India.

²Mata Gujri College of Professional Studies, Indore (M.P) India.

Corresponding Author E-mail:sadhana0301@gmail.com

DOI : http://dx.doi.org/10.13005/bbra/3141

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, K_m and V_max 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.

KEYWORDS: DEAE-cellulose anion exchange chromatography; Purification fold; Schizophyllum commune BCC26414; Sephadex G-200 gel filtration chromatography; Specific activity

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