ISOLATION AND PURIFICATION OF Β-CN FROM SHEEP MILK AND MEASURING THE EFFECTIVENESS OF ITS ENZYMATIC HYDROLYSIS IN INHIBITING ACE1

Milk is a biological important fluid that contains minerals, vitamins, fat, and proteins as crucial elements inside the human body that are used to support the body with natural supplements and might be used for therapeutic purposes. Purification of casein using salt and urea was used and it was partially purified using DEAE-Cellulose and then Sephadex-G-75. The resulting hydrolysate was estimated after incubation with pepsin, trypsin, and a mixture of them to study the inhibition rate of the Angiotensin-converting enzyme (ACE 1). β-CN was partially purified with one peak after DEAE-Cellulose ion exchange and there is also one peak that appeared after the Sephadex G-25 gel filtration technique. Maximum hydrolysis was gained after 8 hours of incubation using a mixture of pepsin and trypsin that inhibit ACE1 at an optimal level. The inhibition rate reached about 71% after 8 hours of incubation with a mixture of pepsin and trypsin with a hydrolysis concentration of 0.083μmol. Results showed that casein extracted from sheep milk can be highly purified using both ion exchange and gel filtration chromatography. Hydrolysis of α and β-CN produces low molecular weight protein using pepsin and trypsin and a mixture of them.