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Fast protein liquid chromatography purification of hydrophobic fraction of bovine milk proteose-peptone and characterization by bidimensional electrophoresis

  • Jean-Michel Girardet (a1), Abderrahmane Mati (a1), Tibogo Sanogo (a1), Luc Etienne (a1) and Guy Linden (a1)...

Summary

Bovine milk Hydrophobic fraction of proteose-peptone was prepared by hydrophobic interaction fast protein liquid chromatography. This method has several advantages such as high rapidity, simple good reproducibility and less denaturation. The proteose-peptone was eluted from a TSK-Phenyl-5PW column with a 1 M-0 M ionic strength gradient of NaH2PO4, pH 6·8, using a 6 ml/min flow rate for 56 min. The quantity of protein injected was 62·5 mg; however, it could be increased up to 100 mg. The elution order was β-CN-4P < BSA (1·6% of total N) < β-CN-5P < β-CN-1P. The hydrophobic fraction was obtained in pure water at the end of the gradient (17·3% of total N). A proteose-peptone cartograph was achieved by bidimensional electrophoresis. This hydrophobic fraction represented three principal zones of Mr 30000–28000, 19000 and 11000, which were respectively composed of 13, 4 and 2 principal spots distributed between 4·9 and 6·1 isoelectric points (IP). These spots corresponded to glycoproteins. ·7, 5·0 and 5·1 IP which migrated to Mr 18000 while β-CN-1P was identified as Mr 9000 in two spots of 5·1 and 5·3 IP.

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Addeo, F., Mauriello, R. & Di Luccia, A. 1988 A gel electrophoretic study of caprine casein. Journal of Dairy Research 55 413421
Anderson, M. 1981 Inhibition of lipolysis in bovine milk by proteose-peptone. Journal of Dairy Research 48 247252
Andrews, A. T. 1978 The composition, structure and origin of proteose-peptone component 5 of bovine milk. European Journal of Biochemistry 90 5965
Andrews, A. T. 1979 The formation and structure of some proteose-peptone components. Journal of Dairy Research 46 215218
Andrews, A. T. 1981 Detection and measurement by the electrophoretic pattern of peptides produced by caseinolysis. In Methods of Enzymatic Analysis, 3rd ed, vol. 5, pp. 277285 (Ed. Bergmeyer, H. U.) New York: Academic Press
Andrews, A. T. & Alichanidis, E. 1983 Proteolysis of caseins and the proteose-peptone fraction of bovine milk. Journal of Dairy Research 50 275290
Cartier, P. & Chilliard, Y. 1986 Effects of different skim milk fractions on activity of cow milk purified lipoprotein lipase. Journal of Dairy Science 69 951955
Chaplin, L. C. 1986 Hydrophobic interaction fast protein liquid chromatography of milk proteins. Journal of Chromatography 363 329335
Edge, A. S. B. & Spiro, R. G. 1987 Selective deglycosylation of the heparan sulfate proteoglycan of bovine glomerular basement membrane and identification of the core protein. Journal of Biological Chemistry 262 68936898
Eigel, W. N. 1981 Identification of proteose-peptone component 5 as a plasmin-derived fragment of bovine β-casein. International Journal of Biochemistry 13 10811086
Eigel, W. N., Butler, J. E., Ernstrom, C. A., Farrell, H. M., Harwalkar, V. R., Jenness, R. & Whitney, R. McL. 1984 Nomenclature of proteins of cow's milk: fifth revision. Journal of Dairy Science 67 15991631
Eigel, W. N. & Keenan, T. W. 1979 Identification of proteose-peptone component 8-slow as a plasmin-derived fragment of bovine β-casein. International Journal of Biochemistry 10 529535
Frank, R. N. & Rodbard, D. 1975 Precision of sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the molecular weight estimation of a membrane glycoprotein: studies on bovine rhodopsin. Archives of Biochemistry and Biophysics 171 113
Girardet, J. M., Pâquet, D. & Linden, G. 1989 Effects of Chromatographic parameters on the fractionation of whey proteins by anion exchange FPLC. Milchwissenschaft 44 692696
Gu, J., Matsuda, T., Nakamura, R., Ishiguro, H., Ohkudo, I., Sasaki, M. & Takahashi, N. 1989 Chemical deglycosylation of hen ovomucoid: protective effect of carbohydrate moiety on tryptic hydrolysis and heat denaturation. Journal of Biochemistry 106 6670
Holt, D. L. & Zeece, M. G. 1988 Two-dimensional electrophoresis of bovine milk proteins. Journal of Dairy Science 71 20442050
International Dairy Federation 1962 Determination of the total nitrogen content of milk by the Kjeldahl method. (FIL-IDF Standard 20)
Kanno, C. 1989 a Purification and separation of multiple forms of lactophorin from bovine milk whey and their immunological and electrophoretic properties. Journal of Dairy Science 72 883891
Kanno, C. 1989 b Characterization of multiple forms of lactophorin isolated from bovine milk whey. Journal of Dairy Science 72 17321739
Kapitany, R. A. & Zebrowski, E. J. 1973 A high resolution PAS stain for polyacrylamide gel electrophoresis. Analytical Biochemistry 56 361369
Karman, A. H. & Van Boekel, M. A. J. S. 1986 Evaluation of the Kjeldahl factor for conversion of the nitrogen content of milk and milk products to protein content. Netherlands Milk and Dairy Journal 40 315336
Kasper, G. A. & Brunner, J. R. 1978 Electrophoretic characterization of the proteose-peptone fraction of cow's milk. Journal of Dairy Science 61 (Suppl. 1) 112–112
Kester, J. J. & Brunner, J. R. 1982 Milk fat-globule membrane as possible origin of proteose-peptone glycoproteins. Journal of Dairy Science 65 22412252
Laemmli, U. K. & Favre, M. 1973 Maturation of the head of bacteriophage T4. I. DNA packaging events. Journal of Molecular Biology 80 575599
Nejjar, Y., Pâquet, D., Godbillon, G. & Le Deaut, J. Y. 1986 Immunological relationship between the Hydrophobic fraction of proteose-peptone and the milk fat globule membrane of bovine milk. International Journal of Biochemistry 18 893900
Ng-Kwai-Hang, K. F., Hayes, J. F., Moxley, J. E. & Monardes, H. G. 1984 Association of genetic variants of casein and milk serum proteins with milk, fat, and protein production by dairy cattle. Journal of Dairy Science 67 835840
Pâquet, D. 1989 [Review: the proteose-peptone fraction of milk.] Lait 69 121
Pâquet, D. & Alais, C. 1982 [Purification and some molecular characteristics of component 3 of proteose-peptones.] Lait 62 338349
Pâquet, D., Nejjar, Y. & Linden, G. 1988 Study of a hydrophobic protein fraction isolated from milk proteose-peptone. Journal of Dairy Science 71 14641471
Poduslo, J. F. 1981 Glycoprotein molecular-weight estimation using sodium dodecyl sulfate-pore gradient electrophoresis: comparison of Tris-glycine and Tris-borate-EDTA buffer systems. Analytical Biochemistry 114 131139
Sanogo, T., Pâquet, D., Aubert, F. & Linden, G. 1989 Purification of αs1-casein by fast protein liquid chromatography. Journal of Dairy Science 72 22422246
Sanogo, T., Pâquet, D., Aubert, F. & Linden, G. 1990 Proteolysis of αs1-casein by papain in a complex environment. Influence of ionic strength on the reaction products. Journal of Food Science 55 796800
Shimizu, M., Yamauchi, K. & Saito, M. 1989 Emulsifying properties of the proteose-peptone fraction obtained from milk. Milchwissenschaft 44 497500
Shimomura, K. & Bremel, R. D. 1988 Characterization of bovine placental lactogen as a glycoprotein with N-linked and O-linked carbohydrate side chains. Molecular Endocrinology 2 845853
Trieu-Cuot, P. & Gripon, J.-C. 1981 Electrofocusing and two-dimensional electrophoresis of bovine caseins. Journal of Dairy Research 48 303310
Trieu-Cuot, P. & Gripon, J.-C. 1982 A study of proteolysis during Camembert cheese ripening using isoelectric focusing and two-dimensional electrophoresis. Journal of Dairy Research 49 501510
Welsch, U., Buchheim, W., Schumacher, U., Schinko, I. & Patton, S. 1988 Structural, histochemical and biochemical observations on horse milk-fat-globule membranes and casein micelles. Histochemistry 88 357365
Yoshida, S. 1989 Preparation of lactoferrin by hydrophobic interaction chromatography from milk acid whey. Journal of Dairy Science 72 14461450

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