Optimization of sulphuric acid pre-treatment of Acacia saw dust through box-bhenken design for cellulase production by B. Subtilis

Background: Cellulases are enzymes which are capable of degrading lignocellulosic biomass. The current study is centred on optimization of dilute sulphuric acid pre-treatment of Acacia saw dust for maximizing cellulase production (CMCase and FPase). Hydrolysis or saccharification of lignocellulosic biomass is brought about by cellulases and the sugar thus released can be used for further bioethanol production.Methods: Box- Bhenken design (BBD) was employed for optimization of pre-treatment conditions for Acacia saw dust. Three variables i.e. sulphuric acid concentration (0.6%, 0.8% and 1.0% v/v), substrate concentration (5%,10% and15%) and reaction time (4h,6h and 8h) was optimized. The pre-treated saw dust was used in the study as a substrate for producing cellulase enzyme through submerged fermentation by Bacillus subtilis (K-18).Results: An optimum conditions i.e. (0.8% H2SO4 conc., 15% substrate conc. and 4h of reaction time) yielded highest filter paper activity (1.3617 IU/ml/min) and CMCase activity (0.7783 IU/ml/min). The suggested model was significant as revealed by F-value, coefficient of determination (R2) andP-value.Conclusion: Results concluded that pre-treated substrate (Acacia sawdust) significantly increased cellulase production as compared to untreated substrate that could be utilized for further biofuel production