Partial Purification and Characterization of Two Endo-β-1, 4-glucanase from Trichoderma sp. (Shmosa tri)

Two endoglucanases (EC 3.2.1.4) from Trichoderma sp. (shmosaTri) FJ937359 were purified to homogeneity using ammonium sulfate precipitation and gel filtration. Purity was confirmed by SDS/PAGE. Enzymatic properties and molecular weights were determined. Molecular weights of CMCase I and II were 58 and 34 KDa, respectively. The effect of temperature on the 2 endoglucanase activity was studied and results showed that optimum activity obtained at 50°C for both CMCase I and II. The enzymes withstand 60 min at 50°C without loss of enzymatic activity. CMCase I and II retained 14.0 and 26.5 % of their original activities at 70°C after 90 min. The optimum pH for CMCase I and II was 5.0. Results also show that CMCase I was active at room temperature after 24 hrs over a broad pH range (3.0-9.0) while CMCase II was relatively stable in pH range (4.0-6.0). Among different kinds of substrates, both enzymes showed a high preference for carboxymethyl cellulose while both CMCase I and II did not show any hydrolytic activity against chitin, starch and cellobiose. On the other hand both CMCase I and II have relatively low hydrolytic activity towards P glucan and xylan. All metallic ions used as well as EDTA and SDS at a concentration of 20 ug/ml of reaction mixture have an inhibitory effect on both CMCase I and II