This study investigated the angiotensin I converting enzyme (ACE) inhibitory activity and the anti-hypertensive effect of Enzyme-Modified Cheese (EMC). EMC was synthesized from the hydrolysis of Danish skim milk cheese with the combination of commercial protease, endo-type protease, and exo-type protease. EMC-No. 9 made with Protease N amano, Umamizyme, and Flavourzyme showed the highest ACE inhibitory effect (IC50=36.3μg/ml). Single oral administration of EMC-No. 9 (100, 500 and 100mg/kg) dose-dependently and significantly lowered the systolic blood pressure in spontaneously hypertensive rats for up to 8 hours. Thus, EMC-No. 9 could be useful for the treatment of hypertension.