Abstract: A novel continuous preparation process of casein phosphopeptides (CPPs) was developed using enzymatic membrane reactor and anion-exchange chromatography. Alcalase was used to continuously hydrolyze 10% casein at pH 8.5 and 50 ℃ for 3 h in a tangential flow filter (TFF) enzymatic membrane reactor (EMR). The separation and purification of the resulting CPPs using anion-exchange resin D303 was investigated and the technological parameters were optimized by single factor design based on the recovery rate of phosphate and the molar ratio of nitrogen to phosphate. The results showed that the appropriate adorption/desorption conditions of D303 were as follows: loading permeate flow rate of 1 BV/h, sample loading quantity of 100 mL, elution temperature of 45 ℃, and hydrochloric acid concentration for elution of 0.15 mol/L, and the recovery rate of phosphate from the permeate was 93.11% and the molar ratio of nitrogen to phosphate was 7.21 under these conditions.

Key words: enzymatic membrane reactor, chromatography isolation, casein phosphopeptides, N/P molar ratio, purification