THE PURIFICATION OF PROTEASE FROM COWSLIP (PRIMULA VERIS) AND ITS USE IN FOOD PROCESSING

Proteases perform very important functions and are used in different objectives as in vitro and in vivo. In recent years, proteases have been used for clinical, pharmaceutical and industrial applications. Most of these proteases are obtained from animal sources. News of diseases passing over from animals to human (severe acute respiratory syndrome SARS], mad cow) has created suspicion of contamination in animal food and its product. Therefore, a new vegetal protease source for use in the food industry was studied. Ammonium sulfate fractionation and CM‐sephadex ion exchange chromatography were used in the purification of the enzyme. The purified enzyme has an optimum pH of 7.0 and its optimum temperature was 70C. The V_max and K_m values determined by Lineweaver–Burk graphics were 0.44 mg/mL.min and 40 mM, respectively. The purification degree was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE ). The native molecular weight of the enzyme was 46 kDa using a gel filtration chromatograph. SDS‐PAGE results show that the enzyme has two subunits, which have protease activity at 32 and 14 kD. It was investigated whether the purified and characterized protease enzyme would cause to congeal milk or could produce digestion of chicken and cow meat. The results showed that the protease enzyme can be used for industrial production.