Preparation of casein phosphopeptides using a novel continuous process of combining an enzymatic membrane reactor with anion-exchange chromatography Original Research Article

A novel process for continuous preparation of casein phosphopeptides (CPPs) was developed using an enzymatic membrane reactor (EMR) and anion-exchange chromatography (AEC). Alcalase was selected to hydrolyse casein (with an initial concentration of 6% (w/w)) at pH 8.5 and temperature 50 °C in an enzymatic membrane reactor. CPPs were then separated from the enzymatic hydrolysates through a weakly basic macroporous anion-exchanger. The yields of CPPs and phosphorus from casein were 8.11%, 15.66% and 21.41%, and 19.10%, 52.70% and 90.01% for EMR fitted with 3 kDa, 5 kDa and 10 kDa membranes, respectively. The yield and quality of CPPs improved with increase in the nominal molecular weight cut-off (NMWCO) of the membrane. However, use of a high NMWCO membrane resulted in substantial enzyme leakage in the permeate.