Abstract
Recent advancements in biorefinery processes necessitate search for cost effective and thermostable cellulases. This study was designed to characterize the cellulase obtained from a thermophilic bacterium, Neobacillus sedimentimangrovi UE25. A combined pretreatment of NaOH and methyltrioctylammonium chloride was given to sugarcane bagasse (SB) for lignin removal and the pretreated SB was utilized as a carbon source for the cellulase production. The thermostable cellulase thus obtained was characterized by adopting central composite design which has not been reported earlier for this purpose. Cellulase showed its maximum activity at pH 7 and temperature 60 degrees C and it remained active in the presence of many salts and detergents. Endoglucanase (EG) was found to be stable for 30 min at 80 degrees C. The purification of EG by using DEAE column yielded specific activity and purification fold of 365.866 IU mg(-1) and 4.264, respectively. The purified EG had a molecular weight of similar to 45 kDa. End product tolerance of EG was also evident, as an activity of 228.57 IU mL(-1) was observed in the presence of 60 mM glucose which revealed that it does not lose its activity upon accumulation of end-product when the reaction is prolonged. The purified EG exhibited V-max and K-m of 294 U mL(-1) min(-1) and 36 mu M, respectively, in the presence of 60 mM glucose. This novel thermostable cellulase can finds its applications in industrial sector.