Isolation and Biochemical Characterization of Cellulase Produced by Bacterial Isolates from Sugarcane Waste Soil

Abstract

Cellulase is one of the most economically important enzyme, which aids in catalyzing cellulolysis, the decomposition of cellulose and other related polysaccharides. So the demand/importance of this enzyme in both domestic and commercial sectors cannot be over emphasized. In this research cellulase-producing bacteria were isolated from soil around sugarcane waste dumping area, which was identified to be P. fulorescens after numerous biochemical and microbiological analysis. The bacteria were then grown and used to ferment certain biomass, with the aim of using the organisms to produce the cellulase enzyme. The total protein/cellulase enzyme activity of the medium was ascertained. Optimization/characterization for maximum cellulase activity was done by varying the temperature, pH, enzyme concentration and substrate concentration, in which the optimum condition for cellulase production was ascertain to be at a temperature and pH of 40˚C and pH 7 respectively. SDS-PAGE electrophoresis was carried out to determine and reconfirm the presence and molecular weight of the isolated enzyme. The estimated extrapolated molecular weight of the enzyme was found to be 13.5KDa.