Sequential optimization of xylanase production using seed waste in .

The production of xylanase from using reetha seed waste as substrate was studied using sequential optimization of fermentation parameters by response surface methodology. Five different lignocellulosic agricultural wastes as a substrate were studied to replace commercially available xylan, amongst which reetha seed waste was found to be the most suitable substrate for xylanase production. A sequential two-stage optimization strategy was used for the fermentation parameter optimization. The Plackett-Burman design was first employed for screening the 6 different physicochemical parameters affecting xylanase production (inoculum concentration, substrate concentration, temperature, pH, media volume, and agitation). The significant factors affecting the xylanase yield were further optimized by Box-Behnken Design in order to obtain the values contributing the highest enzyme yield. Three parameters, namely, temperature, inoculum concentration, and substrate concentration, can be interpreted as the most significant parameters based on the results of Plackett-Burman design. The optimum values by Box-Behnken Design (BBD) are 35 °C temperature, 3 g/L substrate concentration, and inoculum concentration of 4% (v/v) that resulted in maximum xylanase productivity of 5.75 IU/mL at 24 h of the incubation period. Sequential optimization strategy enhanced the xylanase yield by 4.8 fold to that of an unoptimized process.