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In this study, cellulase enzyme production was assessed by pretreating cotton stalk with dilute sulphuric acid which was optimized through response surface methodology. Pretreatment conditions were optimized using three variable with three levels like sulfuric acid concentrations (0.6%, 0.8%, 1% v/v), biomass loading (5%, 10%, 15% w/v), and residence time (4, 6 and 8h). After pretreatment process, cellulase production was achieved by solid substrate which was conducted in 250mL capacity Erlenmeyer flask in which incubation of Bacillus subtilis was carried out for 24 h of fermentation period at temperature of 50o
C. Results showed that cellulase production was greatly affected by the thermochemical pretreatment as compared to the chemical pretreatment. At the pretreatment condition of 1% H2 SO4
conc, 6h residence time, 15% substrate concentration maximum CMCase production (0.858 IU/ml/ min) was obtained while at pretreatment conditions of 1% H2 SO4 concentration, 8h residence
time, 10% substrate concentration at room temperature followed by routine autoclaving, FPase production up to 0.876 IU/ml/min was recorded. The results of present study were found significant. The cellulase enzyme produced in this process affectively hydrolyzes the pretreated substrate at temperature of 50o C and 53 h of incubation period by releasing reducing sugars of 0.74 mg/ml. This study ensures the effective usage of lignocellulosic biomass at large scale biofuel production.
Noor Fatima, Muhammad Irfan, Hafiz Abdullah Shakir, Javed Iqbal Qazi. (2018) Dilute Sulphuric Acid Pretreatment Optimization of Cotton Stalk for Cellulase Production through Box-Bhenken Design, Punjab University Journal of Zoology, Volume 33, Issue 1.
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