Hostname: page-component-cd9895bd7-jn8rn Total loading time: 0 Render date: 2024-12-18T07:53:02.888Z Has data issue: false hasContentIssue false

Isolation of bovine milk fat globule membrane material from cream without prior removal of caseins and whey proteins

Published online by Cambridge University Press:  01 June 2009

Avis V. McPherson
Affiliation:
Otto Madsen Dairy Research Laboratory, Department of Primary Industries, Hamilton, Brisbane 4007Australia
Mary C. Dash
Affiliation:
Otto Madsen Dairy Research Laboratory, Department of Primary Industries, Hamilton, Brisbane 4007Australia
Barry J. Kitchen
Affiliation:
Otto Madsen Dairy Research Laboratory, Department of Primary Industries, Hamilton, Brisbane 4007Australia

Summary

Bovine milk fat globule membrane (MFGM) material was isolated from cream by a new technique which did not involve removal of caseins and whey proteins before destabilization of the fat globules. These components were removed by centrifugation of the membrane material extract through a concentrated sucrose solution (52·5% (w/v) sucrose in 10 mM-Tris HCl buffer, pH 7·5). Membranes collected at the sample–sucrose interface, while whey proteins remained in the supernatant and caseins migrated into the concentrated sucrose solution. The yield of membrane material using this procedure was 25% less than that from conventional methods. This reduced yield was due mainly to lower levels of lipid material, in particular triglyceride. Electrophoretic analysis showed that the polypeptide composition of ‘unwashed’ membrane material was similar to that of conventionally prepared MFGM. This method is particularly suitable for the isolation of membrane material from milks and dairy products in which the fat globule stability is reduced or unknown.

Type
Original Articles
Copyright
Copyright © Proprietors of Journal of Dairy Research 1984

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Allain, C. C., Poon, L. S., Chan, C. S. G., Richmond, W. & Fu, P. C. 1974 Enzymatic determination of total serum cholesterol. Clinical Chemistry 20 470475Google Scholar
Ames, B. N. & Dubin, D. T. 1960 The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. Journal of Biological Chemistry 235 769775Google Scholar
Anderson, M. & Brooker, B. E. 1975 Loss of material during the isolation of milk fat globule membrane. Journal of Dairy Science 58 14421448Google Scholar
Anderson, M., Cheeseman, G. C. & Knight, D. J. 1972 Location of proteins within the milk fat globule membrane. Journal of Dairy Research 39 409411Google Scholar
Bhavadasan, M. K. & Ganguli, N. C. 1976 Dependence of enzyme activities associated with milk fat globule membrane on the procedure used for membrane isolation. Indian Journal of Biochemistry and Biophysics 13 252254Google Scholar
Briley, M. S. & Eisenthal, R. 1974 Association of xanthine oxidase with the bovine milk-fat-globule membrane. Catalytic properties of the free and membrane-bound enzyme. Biochemical Journal 143 149157Google Scholar
Bruder, G., Heid, H., JaraschE,-D. E,-D., Keenan, T. W. & Mather, I. H. 1982 Characteristics of membrane-bound and soluble forms of xanthine oxidase from milk and endothelial cells of capillaries. Biochimica et Biophysica Acta 701 357369Google Scholar
Dulley, J. R. & Grieve, P. A. 1975 A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Analytical Biochemistry 64 136141Google Scholar
Folch, J., Lees, M. & Sloane Stanley, G. H. 1957 A simple method for the isolation and purification of total lipid from animal tissues. Journal of Biological Chemistry 226 497509Google Scholar
Giegel, J. L., Ham, A. B. & Clema, W. 1975 Manual and semi-automated procedures for measurement of triglycerides in serum. Clinical Chemistry 21 1575–1581Google Scholar
Glossmann, H. & Neville, D. M. 1971 Glycoproteins of cell surfaces. A comparative study of three different cell surfaces of the rat. Journal of Biological Chemistry 246 63396346Google Scholar
Herald, C. T. & Brunner, J. R. 1957 The fat-globule membrane of normal cow's milk. I. The isolation and characterization of two membrane-protein fractions. Journal of Dairy Science 40 948956CrossRefGoogle Scholar
Huang, C. M. & Keenan, T. W. 1972 Preparation and properties of 5′-nucleotidases from bovine milk fat globule membranes. Biochimica et Biophysica Acta 274 246257Google Scholar
Hwang, Q. -S., Ramachandran, K. S. & Whitney, R. McL. 1967 Presence of inhibitors and activators of xanthine oxidase in milk. Journal of Dairy Science 50 17231737Google Scholar
Keenan, T. W., Morré, D. J., Olson, D. E., Yunghans, W. N. & Patton, S. 1970 Biochemical and morphological comparison of plasma membrane and milk fat globule membrane from bovine mammary gland. Journal of Cell Biology 44 8093Google Scholar
Kitchen, B. J. 1974 A comparison of the properties of membranes isolated from bovine skim milk and cream. Biochimica et Biophysica Acta 356 257269Google Scholar
Kitchen, B. J. 1977 Fractionation and characterization of the membranes from bovine milk fat globules. Journal of Dairy Research 44 469482Google Scholar
Kobylka, D. & Carraway, K. L. 1972 Proteins and glycoproteins of the milk fat globule membrane. Biochimica et Biophysica Acta 288 282295Google Scholar
Laemmli, U. K. 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277 680685Google Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. 1951 Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193 265275Google Scholar
McPherson, A. V. & Kitchen, B. J. 1981 The proteins and lipids of the aqueous phase of butter. Australian Journal of Dairy Technology 36 1720Google Scholar
McPherson, A. V. & Kitchen, B. J. 1983 Reviews of the progress of Dairy Science: The bovine milk fat globule membrane – its formation, composition, structure and behaviour in milk and dairy products. Journal of Dairy Research 50 107133Google Scholar
Mather, I. H. & Keenan, T. W. 1975 Studies on the structure of milk fat globule membrane. Journal of Membrane Biology 21 6585Google Scholar
Mather, I. H., Tamplin, C. H. & Irving, M. G. 1980 Separation of the proteins of the bovine milk-fat-globule membrane by electrofocusing with retention of enzymatic and immunological activity. European Journal of Biochemistry 110 327336Google Scholar
Mather, I. H., Weber, K. & Keenan, T. W. 1977 Membranes of mammary gland. XII. Loosely associated proteins and compositional heterogeneity of bovine milk fat globule membrane. Journal of Dairy Science 60 394402Google Scholar
Spiro, R. G. 1966 Analysis of sugars found in glycoprotein. Methods in Enzymology 8 326Google Scholar
Standards Association of Australia. 1971 Microbiological methods for the dairy industry. Count techniques. AS 1095 Part I Section 2Google Scholar
Swope, F. C. & Brunner, J. R. 1968 The fat globule membrane of cow's milk: a reassessment of isolation procedures and mineral composition. Milchwissenschaft 23 470473Google Scholar
Swope, F. C. & Brunner, J. R. 1970 Characteristics of the fat globule membrane of cow's milk. Journal of Dairy Science 53 691699Google Scholar
Vasić, J. & DeMan, J. M. 1966 High melting glycerides and the milk fat globule membrane. 17th International Dairy Congress, Munich C 167172Google Scholar
Walstra, P. 1974 High-melting triglycerides in the fat globule membrane: an artefact? Netherlands Milk and Dairy Journal 28 39Google Scholar
Warren, L. 1959 The thiobarbituric acid assay of sialic acids. Journal of Biological Chemistry 234 19711975Google Scholar
Zikakis, J. P. & Wooters, S. C. 1980 Activity of xanthine oxidase in dairy products. Journal of Dairy Science 63 893904Google Scholar