Sunday, August 10, 2008
P21

Characterization of a cellulase from Bacillus sp. strain IC73 isolated from saline soil

Ashok K. Dubey, Manish Jain, Satyanarayana V. K., Poonam Mishra, and Deepika Pandey. Specialty and fine chemicals, Tata Chemicals Ltd Innovation Center, Anmol Pride, Baner Road, Pune, 411045, India

Cellulose, a renewable resource which is abundant in nature can be degraded by microorganisms. Its abundance in natural habitats e.g. soil, leaf litter, fallen wood, water courses and also at semi natural habitats such as compost heaps and anaerobic digests, makes it very important to look for suitable biodegradation strategies. Carboxymethyl cellulase (CMCase / endo-β-1,4-glucanase) is the key enzyme used  in textile, laundry, and pulp and paper industry. In this study, a strain with high cellulase activity was isolated from high saline soil and classified as Bacillus sp. strain IC73 based on 16S rRNA sequence analysis. Cellulase (endo-β-1,4-glucanase) was active on carboxymethyl cellulose (CMC). CMCase activities were assayed using CMC. The enzyme is active in alkaline conditions.  The endocellulase activity from culture broth was determined. Analysis of the enzyme by SDS-PAGE  and Native PAGE revealed a protein band showing cellulose hydrolyzing activity. The apparent molecular weight was about ~ 40 KDa as determined by SDS-PAGE. In situ activity determination was carried out by SDS-PAGE and native PAGE by incorporating CMC in the separation gel. Zymograms were prepared after a renaturation treatment and incubation by staining the gel with Congo red. This enzyme has a potential application in the biotechnology industry due to its activity in high saline conditions.