Application of Box-Behnken Design for Optimization of Different Pretreatment Conditions for Cellulase Production

摘要

Effects of dilute acid and acid steam pretreatments were inspected for cellulose production of Eucalyptus leaves through Box-Behenken design,a three variable factors for response surface methodology by Bacillus subtilus K^(-1)8.Maximum cellulose production performed in 250 mL erlenmeyer flask with submerged fermentation attained at 50℃,p H 5,140 r·min^(-1) for 24 h.Results showed the efficient cellulose production from acid steam pretreatment(being autoclaved at 15 Psi for 15 min)than acid pretreatment.The optimum condition for maximum carboxymethyl cellulas(CMCase)was 1.811 IU·mL^(-1)·min^(-1)(0.8%acid conc.,10 g biomass loading,6 h reaction time)and filter paper activity(FPase)was 2.255 IU·mL^(-1)·min^(-1)(1%acid conc.,10 g biomass loading,8 h reaction time).Whereas,the acid steam maximum CMCase activity recorded was 2.585 IU·mL^(-1)·min^(-1)(0.8%acid conc.,15 g substrate loading and 8 h reaction time)and the highest FPase activity was 2.055 IU·mL^(-1)·min^(-1)(0.8%conc.,10 g biomass,6 h reaction time then autoclaved).Results revealed that acid pretreated Eucalyptus leaves were better lignocellulosic biomass for cellulose production by submerged fermentation.