Bacterial Cellulase: Isolation, Characterization, And Production Optimization

Abstract Cellulose degrading bacteria were isolated from the soil. The colonies were isolated using CMC media and serial dilution. Biochemical tests, plate assays, and molecular methods such as PCR were used in the screening process and identification. The production of cellulase increased by ooptimizing the culture conditions for pH, temperature, incubation time and nutrition sources. The enzyme was partially purified using acetone, ammonium sulphate precipitation and dialysis. The bacterial strain that showed clear transparent zone on CMC agar plate, was identified as cellulase producing bacteria. The strain showed high potentiality for maximum cellulase production at pH 7.0 after 72 hours incubation at 37°C in a medium containing 3% CMC and yeast extract as nitrogen source. Through morphological and biochemical characteristics followed by 16S rRNA, the strain was identified as Bacillus cereus. The specific activity, recovery and purification fold of enzyme were found to be 2.64 U/mg, a yield of 71.9%, and a 3.51 respectively by ammonium sulphate precipitation. Keywords Cellulase, Bacteria, Enzyme isolation, optimization.