Isolation, Screening and Partial Purification of Cellulase from Cellulase Producing Bacteria

The cellulase producing bacteria was isolated from cow dung and characterized by morphological and biochemical analysis. 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. Optimized conditions of CMC and Coir waste as substrates were Glucose, Yeast extract of pH-6 at 40oC for 48 hour but saw dust as substrate has optimized pH-7 with Xylose, Yeast extract at 50oC for 48 hours. Among three substrates used, Carboxy Methyl Cellulose was found to be the best substrate for cellulase production, when compared to coir waste and saw dust as substrates. This is due to the less complexity and hence easy assimilation of Carboxy Methyl Cellulose by the isolated microbe. Ammonium sulphate precipitation followed by dialysis was performed to partially purify the cellulase enzyme. The molecular weight was found to be 32.5 kilodaltons by SDS-PAGE method. On performing zymogram staining, the cellulolytic property was confirmed. 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.