Isolation, Characterization and Molecular weight determination of Cellulase from Trichoderma viride

Abstract

Cellulose hydrolyzing enzyme from fungus Trichoderma viride was purified and characterized. The cellulase production was variable depending upon the type of cellulose the fungus grew on; it was higher when grown on cellulose or whatmann filter paper than on other carbon source viz carboxy methyl cellulose. Enzyme purification to homogeneity was carried out by anion exchange chromatography on DEAE-Sepharose. SDS-PAGE revealed molecular mass of 87 kDa. Maximal activity of the enzymes was observed at 50°C at pH 4 and was stimulated by Ca²⁺, Co²⁺, Mg²⁺ (test at 10 Mm each) and inhibited by Fe²⁺. Ethanol at an optimum concentration of 2% stimulated the initial enzyme activity. The end product of cellulase action was glucose and cellobiose. The enzyme therefore qualifies as an exo-β 1, 4-glucanase. Thermostability, pH and stability in the presence of surface active agents make this enzyme potentially useful in industry particularly for ethanol production.

Keywords: Enzyme, cellulose, cellulase

African Journal of Biotechnology Vol. 12(28), pp. 4512-4518