Optimization of cellulase production by flavobacteriumbolustinumunder submerged fermentation using response surfacemethodologyAuthor(s): Suman Malik, Rishi Pal Mandhan, Ajay Kansal
A 2 step Response Surface Methodology (RSM) employing central composite design (CCD) was applied for the optimization of cellulase production by a newly isolated strain Flavobacteriumbolustinum. Initially, Plackett Burman revealed the 4 significant factors (Temperature, Pineapple peel, Rice strawand NH4Cl) out of 9 on the basis of their F-value. Secondly, a 23 full factorial central composite design and RSMwere applied to determine the optimal concentration of each significant factor. The result showed that linear effect of temperature and pineapple peel and their interactions (as P<0.05) were more significant than the linear and interactive effect of the other variables. It showed the optimumcondition for cellulase production as Pineapple peel 2%, Rice straw 0.6%, NH4Cl 0.15%,MgSO40.4%, Glycerol 1%pH 9 Incubating at 40°C for 24 hrs at 200 rpm resulted in 2.26 fold increase in activity(601.08U/ml) as compared to initial level (265U/ml). Analysis of variance (ANOVA) showed a high coefficient of determination (R2) value 91.87 ensuring a satisfactory adjustment of quadratic model with the experimental data. UsingAgro waste, Pineapple and Rice strawas carbon source reduces the cost of enzyme production thus making the process economically sound, a major challenge in cellulase production.