Purification and characterization of an endocellulase from the thermophilic fungus Chaetomium thermophilum CT2

Chaetomium thermophilum CT2 produced endocellulases at 50degreesC, when grown on 2% microcrystalline cellulose, 1% soluble starch, and 0.4% yeast extract medium. A major endocellulase component was purified to homogeneity by fractional ammonium sulphate precipitation, ion-exchange chromatography on DEAE-Sepharose, Phenyl-Sepharose hydrophobic interaction chromatography and gel filtration on Sephacryl S-100. The molecular weight of the enzyme was estimated to be 67.8 kDa and the enzyme was found to be a glycoprotein containing 18.9% carbohydrate. The K-m of the purified enzyme for carboxymethyl cellulose, sodium salt (CMC), was 4.6 mg ml(-1). The enzyme displayed highest activity towards CMC and significantly lower activities towards phosphoric acid swollen cellulose and filter paper. The activity was enhanced in the presence of Na+, K+ and Ca2+ but inhibited by Hg2+, Zn2+, Ag+, Mn2+, Ba2+, Fe2+, Cu2+, Mg2+ and NH4+. Optimum activity was at 60degreesC and pH 4.0. The enzyme was stable over 60 min incubation at 60degreesC and half-life at 70, 80 and 90degreesC was approximately 45, 24 and 7 min, respectively. (C) 2003 Elsevier Inc. All rights reserved.