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

Abstract

Casein glycomacropeptide (CMP) is a 64- amino acid peptide found in cheese whey, which is released after κ-casein specific cleavage by chymosin. CMP lacks aromatic amino acids, 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 its different glycosylated forms (gCMP A and gCMP B). The most predominant carbohydrate of gCMP is N-acetylneuraminic acid (sialic acid). Here, we developed a CMP purification process based 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. Maximum adsorption capacities were calculated according to the Langmuir model from adsorption 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% (specifically 33.7% of gCMP A and B, 75.8% of aCMP A, and 93.9% of aCMP B). The CMP was recovered as an aggregate with an overall yield of 64%. Therefore, the matrices developed are promising for CMP purification from cheese whey. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:171-180, 2017.

Advances in separation and identification of biologically important milk proteins and peptides

“…Further, the separation of milk proteins/peptides has been based on PAGE [90] and on chromatographic separations [84], such as SEC [13], IEC [13,91], and affinity chromatography [92]. However, analytical separation methods, such as capillary HPLC (especially RP-HPLC) and CE, are very efficient and sensitive for milk proteins/peptides purification, and various techniques have been developed over years [84].…”

Detection of Glucomacropeptide in Raw Milk Adulterated with Cheese Whey in Ecuador

“…GMP is therefore a 64-amino acid peptide, present exclusively in whey cheese, released after the specific cleavage of κ-casein by chymosin; consists of two non-glycosylated isoforms (aGMP A and aGMP B) and their different glycosylated forms (gGMP A and gGMP B), the most predominant carbohydrate of gGMP being N-acetylneuraminic acid (sialic acid) [9]. It should be noted that some psychotropical microorganisms (especially Pseudomonas fluorescens)…”

“…Studies have shown that GMP helps the production of cytokines, regulating immune system interactions [11], is an immunomodulator that has an intestinal anti-inflammatory effect in rats with colitis type I [12] [13] [14], has gastric inhibitory activity [15] and is used for the nutritional management of phenylketonuria in humans [16] [17] [18], has a gastric inhibitory activity, because it lacks aromatic amino acids, making it usable as a nutritional supplement for these people [9]. Studies also show that GMP has anti-inflammatory, prebiotic and immunomodulatory effects, including for atopic dermatitis in humans [19].…”