The plant is a rich source of protein, especially enzymes. In this study, a milk clotting enzyme (MCE) was produced from Portulaca oleracea to be a substitute for commercial MCE. The first steps of partial purification were carried out by precipitation with ammonium sulfate (AS) and were the concentration of ammonium sulfate at 40% gave high the enzyme activity, followed by using sequential chromatographic technique of the most active fraction on sephadex G100 by rate purification to 3.77, Yield with 6.59 % recovery. To the MCE of optimum temperature was 40 °C and the enzyme activity was stable at 20 to 70 °C. The MCE enzyme showed the optima of pH 6 and was more stable in broad from pH 4.0 to 7.0. Effect of calcium chloride at a concentration of 25 mM gave the highest relative activity of the purified MCE. Sodium chloride at a concentration of 2 % gave the highest relative activity of the purified MCE 139.2. The metal ions at 1 and 5 mM gave activators and inhibitors of the purified MCE. On the other hand, soft Cheese was made from pasteurized milk with rennet from microbial and animal sources. tow cheese treatments were made from raw milk and pasteurized milk using purified MCE, the results showed and it was found that the soft cheese making by MCE from plant developed lower Moisture, pH and titratable acidity, while TN, SN and TVFA were high in cheeses manufactured by MCE from Portulaca oleracea plant and displayed better flavor and greater acceptability than other treatments cheese. Increased rate of proteolysis in cheese manufactured by MCE from plant extract had a direct relation to accelerated ripening soft cheese.