Abstract

One of the lignocellulolytic and hydrolytic enzymes needed for a number of processes is cellulase. The biochemical characteristics of pure cellulase derived from Oryctes rhinoceros were documented in this work. Following the extraction of the precipitant from the crude cellulase generated by ammonium sulphate, the enzyme purity was ensured by gel filtration and affinity chromatography employing Reactive Blue 2-Agarose resins and Biogel P-100 resins, respectively. With a 15% yield and 3.59 U/mg of specific activity, the purified cellulase displayed a purification fold of 10.2. The isolated cellulase showed peak activity at pH 4.0 and 70°C, with a native molecular weight of 54,300daltons. Using glucose as the substrate, the Vmax and Km of the isolated enzyme were deduced as 50µmol/min/ml and 5.0mg/ml respectively. The enzyme's broad substrate specificity was demonstrated by the addition of other carbohydrates to the enzyme assay. Certain metal ions and substances like urea, 2-mercaptoethanol, and EDTA inhibited inhibit the enzymatic activity. The gut cellulase of Rhinoceros larvae has shown properties that are useful in biotechnological and industrial operations.

Key words: Characterization, metabisulfite, Oryctes rhinoceros, purification, tetraborate