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Production of sialic acid rich glycopeptide from bovine κ-casein glycomacropeptide by hydrolyzing with papain

Published online by Cambridge University Press:  04 September 2020

Takuo Nakano  and
Mirko Betti
Takuo Nakano*
Affiliation:
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, T6G 2P5, Canada
Mirko Betti
Affiliation:
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, T6G 2P5, Canada
*
Author for correspondence: Takuo Nakano, Email: takuonakano@hotmail.com
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Abstract

Bovine κ-casein glycomacropeptide (GMP) is a sialic acid containing glycopeptide having many biological activities. The study described in this research communication was undertaken to determine whether sialic acid rich glycopeptide can be produced from GMP by proteinase treatment. A sample of GMP was hydrolyzed with papain, and the obtained hydrolysate was chromatographed on a column of diethylaminoethyl-Sephacel to obtain a glycopeptide fraction (GPF). This product accounted for average 48.1% dry weight of GMP or 81.1% total recovered sialic acid from GMP. The content of sialic acid (expressed as % dry weight) was 1.7 times higher in GPF (22.6) than in unhydrolyzed GMP (13.4). Major differences in amino acid composition between GPF and GMP were found in the contents (mol%) of: lysine (<1 and 4.5, respectively), serine (20.3 and 10.3, approximately twice higher in GPF), asparagine/aspartic acid and isoleucine. The contents of the last two amino acids were approximately twice lower in GPF. On gel filtration chromatography with Sephacryl S-100, GMP was eluted as a single peak with elution volume similar to that of dimeric β-lactoglobulin (36.6 kDa) whereas GPF was eluted in two peaks both with elution volumes greater than that of α-lactalbumin (14.2 kDa). These peak fractions containing high (fraction I) and low (fraction II) molecular size glycopeptides gave different sialic acid to peptide ratio, which was 1.7 times higher in fraction I than in fraction II. Results of size exclusion HPLC on Superdex-75 were consistent with those of gel filtlation chromatography. On cellulose acetate electrophoresis, the mobility of GPF relative to that of GMP as 1.0 was found to average 1.2, suggesting a higher negative charge density in GPF than in GMP. It was concluded that papain digestion of GMP is an efficient method to produce glycopeptide with high sialic acid content.

Keywords

Caseinomacropeptidepapain digestionsialic acid rich glycopeptidesialic acidκ-casein glycomacropeptide
Type
Research Article
Information
Journal of Dairy Research , Volume 87 , Issue 3 , August 2020 , pp. 364 - 367
Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

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References

Brody, EP (2000) Biological activities of bovine glycomacropeptide. British Journal of Nutrition 84(suppl. 1), S39–S46.CrossRefGoogle ScholarPubMed
Cheng, X, Gao, D-X, Song, J-J, Ren, F-Z and Mao, X-Y (2015) Casein glycomacropeptide hydrolysate exerts cytoprotection against H2O2-induced oxidative stress in RAW 264.7 macrophages via ROS-dependent heme oxygenase-1 expression. RSC Advances 5, 4511–4523.CrossRefGoogle Scholar
Dziuba, J and Minkiewicz, P (1996) Influence of glycosylation on micelle-stabilizing ability and biological properties of C-terminal fragments of cow's κ-casein. International Dairy Journal 6, 1017–1044.CrossRefGoogle Scholar
Eigel, WN, Butler, JE, Ernstrom, CA, Farrel, HM Jr, Harwalker, VR, Jeness, R and Whitney, RMCL (1984) Nomenclature of proteins of cow's milk. Fifth revision. Journal of Dairy Science 67, 1599–1631.CrossRefGoogle Scholar
Idota, T, Kawakami, H and Nakajima, I (1994) Growth-promoting effects of N-acetylneuraminic acid-containing substances on bifidobacteria. Bioscience, Biotechnology, and Biochemistry 58, 1720–1722.CrossRefGoogle Scholar
Iijima, R, Takahashi, H, Namme, R, Ikegami, S and Yamazaki, M (2004) Novel biological function of sialic acid (N-acetylneuraminic acid) as a hydrogen peroxide scavenger. FEBS Letters 561, 163–166.CrossRefGoogle Scholar
Kawasaki, Y, Isoda, H, Tanimoto, M, Dosako, S, Idota, T and Ahiko, K (1992) Inhibition by Lactoferrin and κ-casein glycomacropeptide of binding of Cholera Toxin to its receptor. Bioscience, Biotechnology, and Biochemistry 56, 195–198.CrossRefGoogle ScholarPubMed
Kawasaki, Y, Isoda, H, Shinmoto, H, Tanimoto, M, Dosako, S, Idota, T and Nakajima, I (1993) Inhibition by κ-casein glycomacropeptide and lactoferrin of influenza virus hemagglutination. Bioscience, Biotechnology, and Biochemistry 57, 1214–1215.CrossRefGoogle ScholarPubMed
Nakano, T and Betti, M (2020) Isolation of κ-casein glycomacropeptide from bovine whey fraction using food grade anion exchange resin and chitin as an adsorbent. Journal of Dairy Research 87, 127–133.CrossRefGoogle ScholarPubMed
Nakano, T and Ozimek, L (1999) Bovine milk glycosaminoglycans. Milchwissenschaft 54, 373–377.Google Scholar
Nakano, T and Ozimek, L (2015) Selective removal of phenylalanine impurities from commercial κ-casein glycomacropeptide by anion exchange chromatography. Journal of Food Processing and Technology 7, 537.Google Scholar
Nakano, K, Nakano, T, Ahn, DU and Sim, JS (1994) Sialic acid contents in chicken eggs and tissues. Canadian Journal of Animal Science 74, 601–606.CrossRefGoogle Scholar
Nakano, T, Ikawa, N and Ozimek, L (2007) Detection of sialylated phosphorylated κ-casein glycomacropeptide electrophoresed on polyacrylamide gels and cellulose acetate strips by the thiobarbituric acid and malachite green dye reactions. Journal of Agricultural and Food Chemistry 55, 2714–2726.CrossRefGoogle ScholarPubMed
Tian, Q, Wang, T-T, Tang, X, Han, M-Z, Leng, X-J and Mao, X-Y (2015) Developing a potential prebiotic of yogurt: growth of Bifidobacterium and yogurt cultures with addition of glycomacropeptide hydrolysate. International Journal of Food Science and Technology 50, 120–127.CrossRefGoogle Scholar
Van Veldhoven, PP and Mannaerts, GP (1987) Inorganic and organic phosphate measurements in the nanomolar range. Analytical Biochemistry 161, 45–48.CrossRefGoogle ScholarPubMed
Wang, B and Brand-Miller, J (2003) The role and potential of sialic acid in human nutrition. European Journal of Clinical Nutrition 57, 1351–1369.Google ScholarPubMed
Wang, B, Yu, B, Karim, M, Hu, H, Sun, Y, McGreeby, P, Petocz, P, Held, S and Brand-Miller, J (2007) Dietary sialic acid supplementation improves learning and memory in piglets. American Journal of Clinical Nutrition 85, 561–569.CrossRefGoogle ScholarPubMed
Warren, L (1959) The thiobarbituric acid assay of sialic acids. Journal of Biological Chemistry 234, 1971–1975.Google ScholarPubMed
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