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Sequential Separation of Lysozyme and Ovalbumin from Chicken Egg White

  • Abeyrathne, Nalaka Sandun (Department of Agricultural Biotechnology, Major in Biomodulation, Seoul National University) ;
  • Lee, Hyun Yong (Department of Agricultural Biotechnology, Major in Biomodulation, Seoul National University) ;
  • Ahn, Dong Uk (Department of Agricultural Biotechnology, Major in Biomodulation, Seoul National University)
  • 투고 : 2013.05.22
  • 심사 : 2013.08.12
  • 발행 : 2013.08.31

초록

Lysozyme was trapped from $2{\times}$ diluted egg white using Amberlite FPC 3500 ion exchange resin (1 g/10mL of egg white). The lysozyme bound to the resin was recovered using 0.1 N glycine-NaOH buffers, pH 9.0, containing 0.5 M NaCl. After separating lysozyme, the pH of the egg white solution was adjusted to 4.75 and centrifuged to remove interfering proteins. The supernatant was collected, added with 2.5% citric acid and 5.0% ammonium sulfate combination to precipitate egg white proteins, except for ovalbumin. After centrifugation, both supernatant (S1) and precipitant were collected. The precipitant was dissolved with 4 volumes of distilled water, and then 2.0% ammonium sulfate and 1.5% citric acid combinations added, stirred overnight in a cold room, and centrifuged. The resulting supernatant (S2) was pooled with the first supernatant (S1), desalted using an ultrafiltration unit, heat-treated at $70^{\circ}C$ for 15 min, and then centrifuged. The supernatant was collected as an ovalbumin fraction and lyophilized. The separated proteins were confirmed using Western blotting. The yield of lysozyme and ovalbumin was > 88.9% and > 97.7%, respectively, and the purity of lysozyme and ovalbumin was > 97% and 87%, respectively. The results indicated that the protocol was simple, and separated lysozyme and ovalbumin effectively.

키워드

참고문헌

  1. Abeyrathne, E. D. N. S., Lee, H. Y, Ham, J. S., and Ahn, D. U. (2013) Separation of ovotransferrinfrom chicken egg white without using organic solvents. Poultry Sci. 92, 1091-1097. https://doi.org/10.3382/ps.2012-02654
  2. Alderton, G., Ward, W. H., and Fevold, H. L. (1945) Ioslation of lysozyme from egg white. J. Biol. Chem. 157, 43-58.
  3. Awade, A. C. and Efstahiou, T. (1999). Comparison of three liquid chromatography methods for egg-white protein analysis. J. Chromatogr. B, 723, 69-74. https://doi.org/10.1016/S0378-4347(98)00538-6
  4. Chang, H. M., Yang, C. C., and Chang, Y. C. (2000) Rapid separation of lysozyme from chicken egg white by reductants and thermal treatment. J. Agric. Food Chem. 48, 161-164. https://doi.org/10.1021/jf9902797
  5. Chick, H. and Martin, C. J. (1913). The Precipitation of egg albumin by ammonium sulfate.A contribution to the theory of the "Salting Out" of proteins. Biochem. J. 7, 380-398.
  6. Datta, D., Bhattacharjee, S., Nath, A., Das, R., Bhattacharjee, C., and Datta, S. (2009). Separation of ovalbumin from chicken egg white using two-stage ultrafiltration technique. Separ. Purif. Technol. 66, 353-361. https://doi.org/10.1016/j.seppur.2008.12.016
  7. Desert, C., Dubiard, C. G., Nau, F., Jan, G., Val, F., and Mallard, J. (2001) Comparison of different electrophoretic separations of hen egg white proteins. J. Agric. Food Chem. 49, 4553-4561. https://doi.org/10.1021/jf001423n
  8. Hopkins, F. G. (1900) On the separation of a pure albumin from egg white. J. Physiol. 25, 306-330. https://doi.org/10.1113/jphysiol.1900.sp000799
  9. Huopalahti, R., Fandino, R. L., Anton, M., and Schade, R. (2007) Bioactive egg compounds. Springer, NY. pp. 3-66.
  10. Manso, M. A., Miguel, M., Even, J., Heranandez, R., Aleixandre, A., and Fandino, R. L. (2008) Effect of the long-term intake of egg white hydrolysates on the oxidative status and blood lipid profile of spontaneously hypertensive rats. Food Chem. 109, 361-367. https://doi.org/10.1016/j.foodchem.2007.12.049
  11. Miguel, M. and Aleixandre, A. (2006) Antihypertensive peptides derived from egg proteins. J. Nutr. 136, 1457-1460.
  12. Miguel, M., Alonso, M. J., Salaices, M., Aleixandre, A., and Fandino, R. L. (2007) Antihypertensive, ACE-inhibitory and vasodilator properties of egg white hydrolysates: Effect of a simulated intestinal digestion. Food Chem. 104, 163-168. https://doi.org/10.1016/j.foodchem.2006.11.016
  13. Mine, Y., Ma, F., and Lauriau, S. (2004) Antimicrobial peptides released by enzymatic hydrolysis of hen egg white lysozyme. J. Agric. Food Chem. 52, 1088-1094. https://doi.org/10.1021/jf0345752
  14. Price, N. C. and Nairn, J. (2009) Exploring proteins: A student's guide to experimental skills and methods. Oxford University Press, pp. 259-278.
  15. Radziejewska, R. C., Leoenierowski, G., and Kijowski, J. (2008) Properties and application of egg white lysozyme and its modified preparations-A Review. Polish J. Food Nutr. Sci. 58, 5-10.
  16. Safarik, I., Sabatkova, Z., Oldrich, T., and Safarikov, M. (2007) Magnetic cation exchange isolation of lysozyme from native hen egg white. Food Technol. Biotechnol. 45, 355-359.
  17. Stadelman, W. J. and Cotterill, O. J. (2001) Egg science and technology. 4th ed. Avi. Publ. Co., Westport, CT.
  18. Strang, R. H. (1984) Purification of egg white lysozyme by ion exchange chromatography. Biochem. Edu. 12, 57-59. https://doi.org/10.1016/0307-4412(84)90003-7
  19. Tankrathok, A., Daduang, S., Patramanon, R., Arakai, T., and Thammasirirak, S. (2009) Purification process for the preparation and characterization of hen egg white ovalbumin, lysozyme, ovotransferrin and ovomucoid. Prep. Biochem. Biotechnol. 39, 380-399. https://doi.org/10.1080/10826060903209646
  20. Wan, Y., Lu, J., and Cui, Z. (2006) Separation of lysozyme from chicken egg white using ultrafiltration. Separ. Purif. Technol. 48, 133-142. https://doi.org/10.1016/j.seppur.2005.07.003
  21. Ward, T. M., Edwards, R. A., and Tanner, R. D. (2007) Separating a mixture of egg yolk and egg white using foam fractionation. Appl. Biochem. Biotechnol. 136, 927-934.
  22. Xie, H., Huff, G. R., Huff, W. E., Balog, J. M., Holt, P., and Rath, N. C. (2002) Identification of ovotransferrin as an acute phase protein in chickens. Poultry Sci. 81, 112-120. https://doi.org/10.1093/ps/81.1.112

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