Milk protein allergy is of great concern when involves neonates and small children. The enzymatic hydrolysis of milk protein is used to modify proteins so changing the immunoreactivity of allergens. In this work the enzymatic hydrolysis of milk proteins was assessed using Alcalase and low-heat milk. The hydrolysis curve was determined following the release of -NH groups versus reaction time using the pH-stat technique. Progressive addition of NaOH 0.5 N was used to maintain a constant pH. The added volume of NaOH was used to calculate the released -NH groups. The substrate was prepared by dispersion of milk powder in mineral water under agitation. The final concentration was 3.1% (w/w) of protein corresponding to 33.1 mmol of peptidic bonds per 100 g of dispersion. Two sets of experiments were carried on at pH 6.5. The first one varying enzyme activity between 19-231 mAU (Anson Units) at 55°C and the second one between 47.9°C-62.1°C at 125 mAU. Results indicated that the degrees of hydrolysis (DH) after 60 min of reaction varied from 16% to 33% for the first set and from 12% to 31% for the second set. The higher DH (33.1%) was obtained in the experiment carried out at 62.1°C and 125 mAU. In conclusion Alcalase is a suitable protease for the hydrolysis of this kind of milk due to the high hydrolysis level achieved in just one hour of reaction with a low amount of enzyme.