Purification of κ-Casien Glycomacropeptide from Sweet Whey with Undetectable Level of Phenylalanine

• Published in: Biotechnology progress 2002, Vol.18 (2), p.409-412
• Main Authors: Nakano, Takuo, R. Silva-Hernandez, Eryck, Ikawa, Noriaki, Ozimek, Lech
• Format: Article
• Language: English
• Subjects: Amino Acids - analysis; Animals; Carbohydrates - analysis; Caseins - analysis; Caseins - chemistry; Caseins - isolation & purification; Caseins - metabolism; Cattle; Chromatography, Gel - methods; Chymosin - metabolism; Glycopeptides - analysis; Glycopeptides - chemistry; Glycopeptides - isolation & purification; Milk - chemistry; Milk - metabolism; N-Acetylneuraminic Acid - analysis; Phenylalanine - analysis; Phosphorus - analysis; Trichloroacetic Acid - chemistry
• ISSN: 8756-7938; 1520-6033
• DOI: 10.1021/bp010195u

Glycomacropeptide (GMP) found in sweet whey is a biologically active compound released from κ‐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 μg, 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.