PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR CHITINASE FROM THE ENTOMOPATHOGEN METARHIZIUM-ANISOPLIAE

Chitinases are produced by Metarhizium anisopliae when it is grown in the presence of chitin. A chitinase from the culture filtrate of Metar hizium anisopliae was successively purified by precipitation with ammo nium sulphate, followed by anion-exchange chromatography on DEAE-Sepha cel. The purified enzyme, which has a molecular mass of approximately 30 kDa by sodium dodecyl sulphate - polyacrylamide gel electrophoresis , catalyses the hydrolysis of p-nitrophenyl beta-N-diacetylchitobiose with an apparent K-m of 0.537 mmol and V-max of 4.86 nmol.mL(-1).min(- 1). The optimum pH and temperature were 4.5-5.0 and 40-45 degrees C, r espectively.