Ion exchange centrifugal partition chromotography: an innovative tool for bioactive peptide capture in complex protein hydrolyzate
Résumé
Alfalfa (Medicago sativa) is a common feed, with a high protein contain of good nutritional value like RuBisCO (Ribulose-I ,5-biphosphatase). RuBisCO hydrolysis using Delvolase® leads to a complex mixture ofpeptides (average MW= 600 g.mol-1), which exhibits an opioid effect. Among this mixture, one dipeptide VW is an angiotensin-converting enzyme inhibitor. In order to develop nutraceutical anti-hypertensive product, VW content of 0.3% must be improved. Centrifugal Partition Chromatography (CPC) is a support free liquid/liquid chromatography process, which has demonstrated high interest for analyte capture inside complex matrix, typically natural product extracts or crude hydrolyzates, especially in ion exchange mode. Here is described a new ion exchange CPC process designed for peptide capture and applied to RuBisCO hydrolyzate. Enriched fractions were obtained, containing 11% of VW, with a high recovery rate of 97%.