Purification and characterization of carboxymethyl cellulase from Bacillus sp. isolated from a paddy field |
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Authors: | Vijayaraghavan Ponnuswamy Vincent S G Prakash |
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Affiliation: | Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam 629 502, Kanyakumari District, Tamilnadu, India. venzymes@gmail.com |
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Abstract: | A microorganism hydrolyzing carboxymethyl cellulose was isolated from a paddy field and identified as Bacillus sp. Production of cellulase by this bacterium was found to be optimal at pH 6.5, 37 degrees C and 150 rpm of shaking. This cellulase was purified to homogeneity by the combination of ammonium sulphate precipitation, DEAE cellulose, and sephadex G-75 gel filtration chromatography. The cellulase was purified up to 14.5 fold and had a specific activity of 246 U/mg protein. The enzyme was a monomeric cellulase with a relative molecular mass of 58 kDa, as determined by SDS-PAGE. The enzyme exhibited its optimal activity at 50 degrees C and pH 6.0. The enzyme was stable in the pH range of 5.0 to 7.0 and its stability was maintained for 30 min at 50 degrees C and its activity got inhibited by Hg2+, Cu2+, Zn2+, Mg2+, Na2+, and Ca2+. |
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