Technical note: Comparison of chromatographic profile of glycomacropeptide from cheese whey isolated using different methods.
Glycomacropeptide (GMP) has heterogeneous carbohydrates, and this attributes to its various biological activities. In this study, we compared the chromatographic profiles of GMP isolated by three methods (trichloracetic acid fractionation, ethanol precipitation, and ultrafiltration) from whey protein isolate (WPI). Seven sharp heterogeneous GMP peaks were eluted from GMP prepared by ethanol precipitation and ultrafiltration using Mono Q anionic chromatography, while only 5 peaks were seen in TCA treated sample. The TCA pretreatment recovered only sialo-GMP (glycosylated) and eliminated all contaminated proteins; however, the recovery rate was the lowest (6.7% of the initial WPI). Ethanol precipitation recovered 20.4% of GMP from WPI and 75.7% was glycosylated, but the heating process might lead to degradation of glycosidic residues. Ultrafiltration was found to be the most effective in recovering GMP. The recovery rate was 33.9% with 81.6% sialo-GMP. We concluded that carbohydrate profile of GMP varied widely and depended on the isolation method. Based on the high recovery of sialo-GMP, the combination of ultrafiltration and anionic chromatography might be a suitable and practical approach on an industrial scale.
Li
Esther W Y
EW
Department of Food Science, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Mine
Yoshinori
Y
D002364:Caseins / Q000737:chemistry; D002611:Cheese / Q000032:analysis*; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008894:Milk Proteins / Q000032:analysis*; D008970:Molecular Weight; D014238:Trichloroacetic Acid; D000067816:Whey Proteins
D002364:Caseins; D006020:Glycopeptides; D008894:Milk Proteins; D000067816:Whey Proteins; D000431:Ethanol; D014238:Trichloroacetic Acid