Affinity chromatography matrices for depletion and purification of casein glycomacropeptide from bovine whey

MARTINEZ, MARÍA JULIA; URTASUN, N.; BAIELI, M. FERNANDA; MIRANDA, M. V.; PILOSOF, ANA M. R.; HIRSCH, DANIELA B.; CASCONE, O.; WOLMAN, F. J.

BIOTECHNOLOGY PROGRESS

AMER CHEMICAL SOC

Lugar: Washington; Año: 2017 vol. 33 p. 171 - 180

8756-7938

Casein glycomacropeptide (CMP) is a 64- amino acid peptide found in cheese whey,which is released after j-casein specific cleavage by chymosin. CMP lacks aromatic aminoacids, a characteristic that makes it usable as a nutritional supplement for people with phenylketonuria.CMP consists of two nonglycosylated isoforms (aCMP A and aCMP B) and itsdifferent glycosylated forms (gCMP A and gCMP B). The most predominant carbohydrate ofgCMP is N-acetylneuraminic acid (sialic acid). Here, we developed a CMP purification processbased on the affinity of sialic acid for wheat germ agglutinin (WGA). After formation of chitosan beads and adsorption of WGA, the agglutinin was covalently attached with glutaraldehyde.Two matrices with different WGA density were assayed for CMP adsorption. Maximumadsorption capacities were calculated according to the Langmuir model fromadsorption isotherms developed at pH 7.0, being 137.0 mg/g for the matrix with the best performance.In CMP reduction from whey, maximum removal percentage was 79% (specifically33.7% of gCMP A and B, 75.8% of aCMP A, and 93.9% of aCMP B). The CMP wasrecovered as an aggregate with an overall yield of 64%. Therefore, the matrices developed are promising for CMP purification from cheese whey.