Evaluation of the purification process of phosvitin extracted from chicken egg yolk using liquid chromatography
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Abstract
Phosvitin from chicken egg yolk, known as a phosphoglycoprotein, owns a very strong metal chelating property due to its polyanionic character. This study aimed to evaluate the factors affecting the purification process and suitable conditions to increase phosvitin’s purity. Phosvitin was separated from yolk granules by using 10% of NaCl solution in 0.05 M NaOH solution and heat treatment which removes lipoprotein from the extracted solution. The highest phosphorus content (58.14 mg) and phosphorus recovery rate (32.4%) were obtained at thermal treatment of 30℃ for 30 minutes. In addition, phosvitin was purified using anion-exchange chromatography (AEC) and gel-filtration chromatography (GFC). The fraction 1 (F1) obtained from AEC using UNO-Sphere Q at pH 8 had the recovery rate of phosvitin approximately 72.73%. Furthermore, fraction F1 was separated on GFC to obtain two main sub-fractions (F1 and F2). Sub-fraction F1 from gel filtration was composed mostly of β-phosvitin with a high recovery rate (81.93%) while F2 was dominant with α-phosvitin in a lower phosvitin recovery rate (16.89%). These findings will provide useful information for further researches on other properties of phosvitin so that it can be applied widely in human needs.
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References
Abe, Y., Itoh, T., & Adachi, S. (1982). Fractionation and characterization of hen’s egg yolk phosvitin. Journal of Food Science, 47(6), 1903-1907.
Anton, M., Castellani, O., & Guérin-Dubiard, C. (2007). Phosvitin: Bioactive Egg Compounds. Springer Press. Europe, pp. 17-24.
Bertrand, C. P. T. (2012). Development of antioxidant peptide fractions from egg yolk protein using enzymatic hydrolysis and ultrafiltration membranes. Science of Food and Nutrition Department of Agriculture and Food Faculty, Laval university.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of Mirogram quantities of protein utilizing the principle of protein - dye binding. Analytical Biochemistry, 72(1-2), 248-254.
Burgess, R. R. (2009). Protein precipitation techniques. In: Methods in Enzymology: Guide to protein purificaion. Academic Press. USA, pp. 331-342.
Burley, R. W., & Cook, W. H. (1961). Isolation and composition of avian egg yolk granules and their constituents α- and β-lipovitellins. Canadian Journal of Biochemistry and Physiology, 39(8), 1295-1307.
Castellani, O., Martinet, V., David-Briand, E., Guerin-Dubiard, C., & Anton, M. (2003). Egg yolk phosvitin: preparation of metal-free purified protein by fast protein liquid chromatography using aqueous solvents. Journal of Chromatography B, 791(1-2), 273-284.
Causeret, D., Matringe, E., & Lorient, D. (1991). Ionic strength and pH effects on composion and microstructure of yolk granules. Journal of Food Science, 56(6), 1532-1536.
Ishikawa, S., Yano, Y., Arihara, K., & Itoh, M. (2004). Egg yolk phosvitin inhibits hydroxyl radical formation from the Fenton reaction. Bioscience, Biotechnology, and Biochemistry, 68(6), 1324-1331.
Jung, S., Nam, K. C., Ahn, D. U., Kim, H. J., & Jo, C. (2013). Effect of phosvitin on lipid and protein oxidation in ground beef treated with high hydrostatic pressure. Meat Science, 95(1), 8-13.
Khan, S. M. A., Nakamura, S., Ogawa, M., Akita, E., Azakami, H., & Kato, A. (2000). Bactericidal action of egg yolk phosvitin against Escherichia coli under thermal stress. Journal of Agricultural and Food Chemistry, 48(5), 1503-1506.
Ko, K. Y., Nam, K., Jo, C., Lee, E. J., & Ahn, D. U. (2011). A simple and efficient method for preparing partially purified phosvitin from egg yolk using ethanol and salts. Poultry Science, 90(5), 1096-1104.
Lee, H. Y., Abeyrathne, E. D. N. S, Inwook, C., Suh, J. W., & Ahn, D. U. (2014). Sequential separation of immunoglobulin Y and phosvitin from chicken egg yolk without using organic solvents. Poultry Science, 93(10), 2668-2677.
Lei, B., & Wu, J. (2012). Purification of egg yolk phosvitin by anion exchange chromatography. Journal of Chromatography A, 1223, 41-46.
Mecham, D. K., & Olcott, H. S. (1949). Phosvitin, the principal phosphoprotein of egg yolk. Journal of American Chemistry Society, 71, 3670-3679.
Pulliainen, T. K., & Wallin, H. C. (1994). Determination of total phosphorus in foods by colorimetric measurement of phosphorus as molybdenum blue after dry-ashing: NMKL interlaboratory study. Journal of AOAC International, 77, 1557.
Tsutsui, T., & Obara, T. (1982). Hydrophobic components in delipidated granule of egg-yolk. Agricultural and Biological Chemistry, 46(10), 2587-2589.
Zhang, X., Ning, Q., Feng, G., & Meihu, M. (2011). Simply and effectively preparing high-purity phosvitin using polyethylene glycol and anion-exchange chromatography. Journal of Separation Science, 34(22), 3295-3301.