Process scale chromatographic isolation, characterization and identification of tryptic bioactive casein phosphopeptides

A process scale isolation of 40 kg tryptic Casein Phosphopeptides (CPP) was implemented using tryptic hydrolysis, acid precipitation, diafiltration and anion exchange chromatography. The obtained yield corresponded to 20% (w/w) CPPs from caseinate. The CPP-product was obtained either as Ca-CPP (6.97% Ca, 0.23% Na) or Na-CPP (0.01% Ca, 8.53% Na). These products were completely soluble at pH as low as 0.5 and displayed no particular taste. All produced CPPs were phosphorylated peptides which exhibited calcium binding ability and inhibited calcium phosphate crystallization. The peptides were identified by Mass Spectrometry and N-terminal sequencing as genuine truncated tryptic peptides. A high purity CPP product, available as Na-CPP and Ca-CPP, was obtained by this high yield process scale isolation of tryptic CPPs. (C) 1999 Elsevier Science Ltd. All rights reserved.