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Isolation, screening, production, media optimization, partial purification and Characterisation of Cellulase producing Bacillus species isolated from Rhizosphere soil

Author: 
Kiran Kumar, S. J.
Subject Area: 
Life Sciences
Abstract: 

The cellulase producing Bacillus species was isolated from rhizosphere soil and characterized by various staining procedures, biochemical analysis and partial purification. Further partial purification of cellulase was carried out by dialysis and ammonium phosphate precipitation and also to determine molecular weight by SDS-PAGE. Bacterial isolated were grown on Carboxy Methyl Cellulose (CMC) agar at various optimum conditions such as parameters like pH, temperature, incubation period, carbon and nitrogen sources prior to examined and identified primarily as genus, Bacillus species for cellulase production by morphological and biochemical analysis. The cellular enzyme was completely active in a large pH range (5 -8) and presented an optimum activity of 195U/mL. The cellulase production was found to be more when incubated for two days (48 hrs). Ammonium sulphate precipitation followed by dialysis was performed to partially purify the cellulase enzyme. The molecular weight was found to be 32 kilodaltons by SDS-PAGE method. The specific activity of the enzyme was found to be 0.3 mg/mL. The use of microorganisms for the production of enzymes offers a promising approach for its large scale production and as a possible food supplement or in pharmaceutical industry.

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