Abstract
A consecutive optimization based on statistical approach was applied for a-glucosidase production by both wild and mutant T. dupontii. Plackett Burman design (PBD) with two levels was employed in order to screen the significant effect of different nutritional and physical parameters through submerged fermentation. Among all nine variables tested in PBD, incubation time, inoculum size and ammonium sulphate concentration were selected. The Box-Behnken approach was further applied for process optimization. The a-glucosidase production for both wild and mutant T.dupontii was obtained at 72 h of incubation, 1.25 mL inoculum size and 0.25% ammonium sulphate concentration with relatively 95% correlation between the experimentally predicted and observed values. The duration of maximum enzyme production in RSM was cost-saving and fast. The quadratic model was in satisfactory adjustment with the experimental data with high R2 value which describes 98.90% of response variability of the model. Moreover, the novel approach of this present work is that, consecutive optimization were applied for maximum a-glucosidase production using response surface methodology by both wild and mutant thermophillic T. dupontii. Results revealed that thermophillic mutant T. dupontii could be potential candidate for industrial applications.
Highlights
Cellulases that belongs to hydrolytic group is used for the conversion of lignocellulose into simple sugars
Optimization of physical and nutritional parameters by response surface methodology (RSM) In present work Plackett Burman design (PBD) was used to screen the factors that significantly affect the β-glucosidase production by both wild and mutant T. dupontii. β-glucosidase production of nine variables were screened in PBD (Table 1)
The data from Pareto chart revealed that out of all nine factors, three variables such as (X1) incubation time, (X4) inoculum size and (X8) ammonium sulphate concentration had a positive effect on β-glucosidase production, while the remaining factors did not show any significant effect on β-glucosidase production
Summary
Cellulases that belongs to hydrolytic group is used for the conversion of lignocellulose into simple sugars. The Cellulases comprises of three major hydrolases including cellobiohydrolase, endoglucanase and β-glucosidases that act synergistically on cellulose and break into glucose units [1]. Statistical methods offers many advantages over classical method and considered more reliable and rapid, short lists the significant variables and help to understand the interactions between the variables at different concentrations. Statistical tool such as response surface methodology (RSM) is used for optimization and modeling of numerous variables [4]. The aim of present work is to determine the effect of several variables on beta-glucosidase production by both wild and mutant thermophilic T. dupontii using two steps procedure. In the first step Plackett Burman design (PBD) was applied to optimize all
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