Isolation, purification and characterization of carboxymethyl cellulase (CMCase) from endophytic Fusarium oxysporum producing podophyllotoxin

Refaz Ahmad Dar, Iram Saba, Mohd Shahnawaz, Manisha Kashinath Sangale, Avinash Bapurao Ade, Shabir Ahmad Rather, Parvaiz Hassan Qazi

Department of Botany, University of Pune, Pune, Maharashtra, India.
Microbial Biotechnology Division, Indian Institute of Integrative Medicine (CSIR), Sanatnagar, Srinagar, Jammu and Kashmir, India.

DOI: 10.4236/aer.2013.14010                

Abstract

Endophytic fungus Fusarium oxysporum is a rich source of cellulases. In the present study, the highest activity was reported at 28 ° C, pH 5.6 with 2% Carboxymethyl cellulose (CMC) as carbon source. CMC was purified using Sephadex G and DEAE cellulose chromatography to 15.9 folds and the molecular weight was determined to be 84 kDa by SDS-PAGE analysis and was subsequently characterized. The purified enzyme was stable over the pH range from 4.0 to 8.0 and at temperatures below 50 ° C. The enzyme was highly active on CMC and reduced or no activity on Avicel, cellobiose and it was suggested to be CMCase/endoglucanase. The activity of endoglucanase was enhanced in the presence of MgCl₂, CoCl₂, FeCl₃, CaCl₂, FeCl₂ and intensive to HgCl₂. The purified enzyme showed its optimum activity at pH 5.0 - 6.0 and was quite stable at 50 ° C for 30 min and retained 45% of original activity.

Keywords

Endophyte; Fusarium oxysporum; Juniperus recurva; Podophyllotoxin; CMCase

Conflicts of Interest

The authors declare no conflicts of interest.

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