Purification and biochemical properties of a new thermostable xylanase from symbiotic fungus, Termitomyces sp.
A xylanase was purified from symbotic fungus, Termitomyces sp. by chromatography on columns of DEAE-Sepharose, CM-Sepharose, gel filtration and Phenyl-Sepharose. The preparation was shown to be homogenous by polyacrylamide gel electrophoresis. The purified enzyme displayed two protein bands on SDS-polyacrylamide gel electrophoresis and its molecular mass was estimated to 80-87 kDa. The xylanase exhibited maximum activity at 65-70°C and at pH 5.6, but it retained more than 80% of its activity in the pH range 5.0-6.0. The enzyme was stable for a long time-period up to 50°C and for 1 h at 60°C. Although the xylanase had a lower carboxymethylcellulase activity, it lacked activity towards substituted xylan, xylobiose, inulin, starch, polygalacturonic acid or pNP glycosides. Kinetic parameters indicated higher efficiency in the hydrolysis of beechwood xylan and birchwood xylan. The xylanase activity was stimulated by K+, Mn2+ and dithiol-reducing agents and was sensitive to Cu2+, Fe2+, Zn2+ and detergent agents. The enzymatic activity was observed in presence of urea up to a 1% (w/v) concentration. The enzyme could also be used in the presence of organic solvents such as acetone or dioxane (5%, v/v) without loss of activity.
Keywords: Xylanase, Thermostable Termitomyces sp., Macrotermes subhyalinus, Termitidae