Partial purification and characterization of β-glucanase produced by Bacillus sphaericus CE-3.

β-glucanase produced by Bacillus sphaericus CE-3 was purified to homogeneity by dialysis in 4M sucrose solution, ion-exchange chromatography on Q-sepharose FF and by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B. The relative molecular mass of the enzyme was estimated to be 24,000 by SDS-polyacrylamide gel electrophoresis. The enzyme was optimally active at pH 9.0 and 40^oC, stable at pH 9.0 and unusually retained over 90% activity between 50-100^oC (30 min). It was strongly activated by Mn²⁺ but inhibited by Ba²⁺, Co²⁺, Hg²⁺, Pb²⁺, Cu²⁺, Sr²⁺, Fe²⁺, Ca²⁺ and Zn²⁺. The β-glucanase displayed high catalytic activity with untreated sawdust, followed by Avicel, while sodium hydroxide treated sawdust was the least hydrolyzed.