PARTIAL PURIFICATION AND SOME PROPERTIES OF PROTEASE FROM MICROSPORUMCANIS

Clinical strain Microsporum canis was isolated from specimen of patient with Tinea corporis in arm and it has ability to produce extra cellular keratinolytic protease in a broth containing human hair. The enzyme was partial purified 35.8- fold from culture filtrate by sequential steps through salting out with ammonium sulfate precipitation (80% saturation), ion exchange Chromatography by batch wise ion exchange by CM- Cellulose cation resin and anion resin by DEAEcellulose column. The partially purified enzyme had an optimum activity at pH 9 and maximum activity at pH11. The activity was stable in the alkaline pH 9 for 30 min at 25°C. Enzyme activity toward casein increased when temperature raised more than 20°C and maximal activity attained at 55° C. The enzyme was stable at temperature under 25˚C and approximately 80% of its activity abolished by incubation of the enzyme at 60 ° C for 30 min. Protease had activation energy equal 3.314 Kcal/Mole that to be able to transform casein to product. On the other hand, the activation energy for denaturation was equal to 49.675 Kcal/Mole. The result of this experiment demonstrated that the enzyme is heat labile.