Purification of kappa-casien glycomacropeptide from sweet whey with undetectable level of phenylalanine.
Glycomacropeptide (GMP) found in sweet whey is a biologically active compound released from kappa-casein by the action of chymosin during cheese making. This study was undertaken to purify GMP from sweet whey as a research chemical on a laboratory scale. Glycomacropeptide was isolated from proteins and other non-GMP compounds by deproteinization with trichloroacetic acid and gel chromatography on Sephacryl S-200. The purified GMP accounted for 0.12% of dry sweet whey powder and contained 107.0, 50.9, 61.2 and 4.3 microg, respectively, of sialic acid, galactose, galactosamine and phosphorus per mg dry weight. The GMP was of high purity, with its amino acid composition showing undetectable levels of phenylalanine, tyrosine and arginine, the amino acids that do not occur in bovine GMP. On gel electrophoresis, the GMP showed a single broad band with an average mobility faster than that of carbonic anhydrase (molecular weight = 31 kDa). The purified GMP may be used as a standard glycopeptide in chromatography and electrophoresis and may also be used to test various known or unknown properties and biological activities of this compound.
Nakano
Takuo
T
Alberta Dairy Association Research Unit, c/o Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, T6G 2P5, Canada. tnakano@ualberta.ca
Silva-Hernandez
Eryck R
ER
Ikawa
Noriaki
N
Ozimek
Lech
L
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002364:Caseins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods*; D012085:Chymosin / Q000378:metabolism; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry* / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D010649:Phenylalanine / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D014238:Trichloroacetic Acid / Q000737:chemistry
D000596:Amino Acids; D002241:Carbohydrates; D002364:Caseins; D006020:Glycopeptides; C020251:kappa-casein glycomacropeptide; D010758:Phosphorus; D010649:Phenylalanine; D014238:Trichloroacetic Acid; D012085:Chymosin; D019158:N-Acetylneuraminic Acid