Production and Properties of Bacterial Cellulase from Arecanut Waste

Background: Cellulases are the second most popular industrial enzyme because of their many uses, especially in turning cellulosic biomass into sugars for further processing. Objectives: The current study focused on the isolation, screening, molecular characterization, optimization of culture conditions and partial purification of the cellulase producing bacteria. Materials and Methods: Cellulose degrading bacteria were isolated from soil surrounding arecanut waste. Single colonies were isolated using selective media and serial dilution. Biochemical tests, plate assays, spectrophotometric measures, and molecular methods such as PCR were used in the screening process and identification. The production of cellulase increased by optimizing the culture conditions for pH, temperature, incubation time and nutrition sources. The enzyme was partially purified using acetone, ammonium sulphate precipitation and dialysis. The molar mass was determined by SDS-PAGE. Results: Three bacterial strains B1, B2 and B3 produced clear transparent zone on CMC agar plate, were identified as cellulase producing bacteria. The B2 strain showed high potentiality for maximum cellulase production at pH 7.0 after 72 hr incubation at 37ºC 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 thuringiensis JN194. The specific activity, recovery and purification fold of enzyme were 2.64 mg/mL, 57.5% and 3.47 respectively. Conclusion: A novel strain from genus Bacillus has been isolated producing the enzyme cellulase which has application in both agriculture and industry.