Isolation and Characterization of Cellulase Producing Bacteria from Rumen Fluid

The rumen is known as a paunch, forms the larger part of the reticulorumen. The rumen is the first chamber in the alimentary canal of ruminant animals. The rumen is huge amount of different microbes, including bacteria, fungi, protozoa, and others. These microbes assist the bovine to digest ingested food. Bovines can eat a ample range of feeds as they have many different kinds of microbes to help them in digestion. In this study efforts were made to isolate the bacteria occur in the rumen of buffalo. Twelve cellulase producing bacteria (CPB) were isolated from eight different buffalo rumen fluid by enriching rumen specific medium which contains CMC(Carboxy Methyl Cellulose) as a substrate for cellulose degradation. To indicate the cellulase activity of the organisms, diameter of clear zone around the colony on Congo-red agar media were measured. Isolate CPB 1 &CPB4 exhibited the maximum zone of clearance around the colony. Whereas for all isolates zone of clearance and Hydrolysis Capacity (HC) value were found between 5 to 33mm and 1.3 to 3.7 respectively. Celluase enzyme assay was carried out using Miller’s. The extracellular cellulase activities ranged from 0.6 to 1.3µmol/min/ml. All the cultures were further tested for their capacity to degrade CMC at various pH, temperature and NaCl concentration. The maximum CPB was estimated to be CPB 2. For the isolates CPB 1-4Growth kinetics, partial protein purification and that protein estimation were done. Cellulase activity from that partially purified enzyme is ranged from 1.2 to 1.4µmol/min/ml.