Purification and characterization of carboxymethyl cellulase from Bacillus sp. isolated from a paddy field.
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+.
Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam 629 502, Kanyakumari District, Tamilnadu, India. venzymes@gmail.com
Ponnuswamy
P
Vijayaraghavan
S G Prakash
SG
Vincent
D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development; D002480:Cellulase / Q000302:isolation & purification* / Q000378:metabolism; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D012988:Soil Microbiology; D013379:Substrate Specificity; D013696:Temperature
D002480:Cellulase; C019826:carboxymethylcellulase