EXTRACTION OF MILK – CLOTTING ENZYME FROM Cartamus tinctorius SEEDS AND PARTIAL CHARACTERIZATION

Abstract

The partial characterization and purification of milk clotting enzyme from defatted meal of safflower seeds was done . The enzyme was extracted using four different types of buffers ,the most efficient extraction solution was found to be 3% NaCl in phosphate buffer . The enzyme was partially purified by precipitation with ammonium sulphate at 60% saturation , giving specific milk clotting of 9.33 unit/mg . The study of some specifies of partially purified enzyme pointed out that the maximum milk clotting activity observed at pH value 6 . At determination the optimum temperature for activity the enzyme from safflower seeds exhibited the maximum coagulation activity at 75 C. Upon examining the effect of the addition of CaCl2 on coagulation activity at range of 10 – 100 mM ,the enzymatic activity increases as the concentration of CaCl 2 increases ,and the high activity at a salt conc. of 70 mM ,after that the activity decrease . To know the effects of some inhibitors on activity like Iodo acetic acid , EDTA and β-mercaptoethanol ,the enzyme was incubated with these compounds , it was clear that they had no significant inhibitory effect on enzyme coagulation activity .