Abstract
Glycomacropeptide (GMP) was purified from sweet whey dialyzed in water by anion-exchange chromatography on DEAE-Sephacel at pH 2.0–4.5. The optimum pH range was 2.5–4.0. The yield of purified GMP increased and its sialic acid concentration decreased with increasing pH value. The GMP had an apparent isoelectric point < 3.8. Dialysis of sweet whey was shown to be important to maximize the yield of GMP adsorbed to the anion-exchanger. Only highly sialylated GMP, accounting for approximately 55% of total sialic acid content, was adsorbed on the anion-exchanger from non-dialyzed sweet whey.
Similar content being viewed by others
References
Abd El-Salam MH, El-Shibiny S, Buchheim W (1996) Characteristics and potential uses of the casein macropeptide. Int. Dairy J. 6: 327–341.
AOAC (1998) Official Methods of Analysis of AOAC International, 16th edn. Gaithersburg, MD: Association of Official Analytical Chemists.
Ayers JS, Coolbear KP, Elgar DF (1998) Process for isolating glycomacropeptide from dairy products with a phenylalanine impurity of 0.5% w/w. PCT/WO 98/14071.
Dische Z, Borenfreund E (1950) A spectrophotometric method for the microdetermination of hexosamines. J. Biol. Chem. 184: 517–522.
Dziuba J, Minkiewicz P (1996) Influence of glycosylation on micelle-stabilizing abilty and biological properties of C-terminal fragment of cow's _-casein. Int. Dairy J. 6: 1017–1044.
Eigel WN, Butler JE, Ernstrom CA, Farrel HM Jr, Harwalker VR, Jeness R, Whitney RMcL (1984) Nomenclature of proteins of cow's milk: fifth revision. J. Dairy Sci. 67: 1599–1631.
Erdmann P, Neumann F (1998) Procédé de traitement d'une matiére première lactosérique (in French). Demande de Brevet Europeen EP 0 880 902 A1.
Gottschalk A (1960) The Chemistry and Biology of Sialic Acid and Related Substances. London: Cambridge University Press.
Idota T, Kawakami H, Nakajima I (1994) Bifidus growth-promoting activity effect of N-acetyl-neuraminic acid-containing substances. Biosci. Biotechnol. Biochem. 58: 1720–1722.
Kawakami H, Kawasaki Y, Dosako S, Tanimoto M, Nakajima I (1992) Determination of _-casein glycomacropeptide by high performance liquid chromatography without trichloroacetic acid pretreatment. Milchwissenschaft 47: 688–693.
Kawasaki Y, Dosako S (1994) Process for producing kappa-casein glycomacropeptides. European Patent Specification 0 488 589 B1.
Kawasaki Y, Dosako S, Shimatani M, Idota T (1992) Production of kappa-casein glycomacropeptides using an anion-exchange resin. U.K. Patent Application GB 2 251 858 A.
Léonil J, Mollé D (1991) A method for determination of macropeptide by cation-exchange fast protein liquid chromatography and its use for following the action of chymosin in milk. J. Dairy Sci. 58: 321–328.
Nakano T, Ozimek L (1999) Purification of glycomacropeptide from non-dialyzable fraction of sweet whey by anion-exchange chromatography. Biotechnol. Tech. 13: 739–742.
Reid JR, Coolbear T, Ayers JS, Coolbear KP (1997) The action of chymosin on _-casein and its macropeptide: Effect of pH and analysis of products of secondary hydrolysis. Int. Dairy J. 7: 559–569.
Skudder PJ (1985) Evaluation of a porous silica-based ion-exchange medium for the production of protein fractions from rennet-and acid-whey. J.Dairy Res. 52: 167–181.
Tanimoto M, Kawasaki Y, Dosako S, Ahiko K (1992) Large scale preparation of _-casein glycomacropeptide from rennet casein whey. Biosci. Biotechnol. Biochem. 56: 140–141.
Trevelyan W, Harrison JS (1952) Studies on yeast metabolism. I. Fractionation and microdetermination of cell carbohydrates. Biochem. J. 50: 298–303.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nakano, T., Ozimek, L. Purification of glycomacropeptide from dialyzed and non-dialyzed sweet whey by anion-exchange chromatography at different pH values. Biotechnology Letters 22, 1081–1086 (2000). https://doi.org/10.1023/A:1005618619167
Issue Date:
DOI: https://doi.org/10.1023/A:1005618619167