Enrichment of sialic acid-containing casein glycomacropeptide in protein hydrolysates using phenylboronic acid-functionalized mesoporous silica nanoparticles

Abstract

Glycomacropeptide (GMP) is a bioactive peptide of high value, rich in glycosylation sites and with physiological and dietary therapeutic value. The enrichment and detection of GMP facilitates the accurate quantification and the identification of adulteration of GMP in food products. In GMP, sialic acid is an abundant glycosyl group and is mainly located at the end of the sugar chain. Here, we propose a novel GMP enrichment strategy based on the affinity of sialic acid for phenylboronic acid groups that shift with environmental pH. As an enrichment material, mesoporous silica nanoparticles were progressively modified with aminopropyl and phenylboronic acid groups. The developed material showed excellent selectivity for sialic acid in the presence of galactose and fucose as interferents. The adsorption behavior of sialic acid-containing GMP fits the Langmuir adsorption model, offering a recovery of 71.72% (in terms of sialic acid content) and a GMP relative purity of 0.957. Results from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography confirm that the enriched GMP contains almost no other unexpected proteins and peptides, indicating that the developed strategy holds promise for purifying GMP in various dairy systems.

Keywords: Boronate affinity; Enrichment; Glycomacropeptide; Sialic acid; Solid-phase extraction.