Fermentation of sugarcane baggase for the production and characterization of exoglucanase by Phaeolus spadiceus

Abstract

The current study is designed having aims i.e., to check cellulase production potential of P. spadiceus, optimization of different culture conditions for the production of exoglucanase using sugarcane bagasse as substrate and characterization of exoglucanase. P. spadiceus, a mushroom was used to enhance the production of exoglucanase enzyme by Solid State Fermentation (SSF). Effect of different conditions was observed including, fermentation period, temperature, pH, moisture level and different carbon and nitrogen sources to obtain maximum yield of exoglucanase. Crude-enzyme was then centrifuged and characterized for temperature, pH, effect of metal ions and effect of different substrate concentration for the determination of Km and Vmax. Maximum activity of exoglucanase was observed after day 4 (57.64 IU/mL/min), at 25-30oC and 50% moisture level (31.5 IU/mL/min), at pH 4.5 (47.03 IU/mL/min) and also with carbon source (5% sucrose) and nitrogen source (2.5% urea) which was 39.2 IU/mL/min. The enzyme was subjected to characterization to find out its optimum temperature, pH, effect of different metal ions and finding Km and Vmax. Exoglucanase activity at different conditions of each parameter showed that the optimum temperature was 30oC and pH 4.5. The Km and Vmax were determined by the Line-weaver-Burk-plot between 1/[VO] on the Y-axis and 1/[S] on the X-axis that results 66.66 mM and 48.73 µM/mL/min respectively The study of exoglucanase enzyme production from this fungus and to optimize different conditions will aid in more enzyme production, decrease environmental pollution and produce valued things from the resulting fermentable-sugars.