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Isolation and Characterization of Cellulose Degrading Bacteria from Saliva of‘Jaffrabadi’Breed of Buffalo

Ashok A. Shinde Faiyaz K. Shaikh Ketali P. Pisal Pratiksha S. Patil
Vol 10(9), 96-104

Microorganisms with cellulolytic activity are considered as potential candidates in the processing of cellulosic biomass for biotechnological applications. This study was conducted to isolate and characterize cellulose-degrading bacteria collected from the saliva of ‘Jaffrabadi’ breed of buffalo by morphological and biochemical analysis. The cellulolytic bacteria were isolated and purified on CMC (Carboxy methyl cellulose) agar. Bacterial isolate produced a clear transparent zone when stained with Congo red, confirming cellulase activity. Morphological examination revealed Gram-positive, rod or bacilli- shaped, non-motile, and spore forming nature of bacterial isolate. Different biochemical tests suggested that bacterial isolate was Bacillus subtilis. The cellulase enzyme was partially purified by (NH4)2SO4 and studied using CMCase assay. The activity of partially purified cellulose was higher (8µg/ml/min) as compared to that of crude supernatant (5.5µg/ml/min). The optimum pH and optimum temperature range were found to be 6.5 - 7.5 and 40 – 50 °C. Protein profiling using 10% SDS-PAGE further confirmed the purity of the partially purified sample.

Keywords : Bacillus subtilis Biotechnology Cellulase CMC agar Ruminant SDS-PAGE

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