Optimization of Casein Hydrolysis for Obtaining High Contents of Small Peptides: Use of Subtilisin and Trypsin

With the aim of producing casein hydrolysates for dietary supplement, the optimization of hydrolysis conditions using two enzymes was studied in order to obtain high di- and tripeptide contents. Trypsin and subtilisin were used, separately and in association (simultaneously or successively) and the hydrolysates were then analysed. After fractionating by size-exclusion chromatography, a rapid quantitation of peptides was performed using the correct fraction area method. The hydrolysate with the highest content of small peptides was obtained using only subtilisin with a 4% E:S ratio. Among the enzymatic associations studied, the successive one employing firstly subtilisin for 5 min was the most advantageous due to the lower level of large peptides produced.