The Effect of Non-enzymatic Glycosylation on the Physicochemical Properties and Immunoreactivity of Chicken Egg Ovomucoid

DOI: 10.18805/ijar.B-1232    | Article Id: B-1232 | Page : 1286-1292
Citation :- The Effect of Non-enzymatic Glycosylation on the Physicochemical Properties and Immunoreactivity of Chicken Egg Ovomucoid.Indian Journal of Animal Research.2021.(55):1286-1292
Lyayla K. Bupebayeva, Miciñski Bartosz, Miciñski Jan, Milewski Stanisław micinsk@uwm.edu.pl
Address : Department of Sheep and Goat Breeding, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-719 Olsztyn, Poland. 
Submitted Date : 8-12-2019
Accepted Date : 9-06-2020


Background: Research studies conducted with the involvement of various molecular techniques have demonstrated that ovomucoid, a protein found in chicken egg whites, is also a major food allergen.  Many studies have shown that about 40% of patients with allergy against egg proteins were sensitive to ovomucoid. Egg whites are popularly used in the world to enhance the nutritional value of food products (as additives in pastry, meats, sauces, salads and creams) and the production and use of eggs in the world is increasing. Their complete elimination from the diet is highly challenging. Ovomucoid comprises 186 amino acids and has antibacterial properties, and inhibits the activity of microbial enzymes. In this experiment, chicken egg ovomucoid was isolated and polyclonal antibodies were obtained by immunizing a rabbit with the ovomucoid solution to analyze the effect of non-enzymatic glycosylation on the physicochemical properties and immunoreactivity of the studied ovomucoid.
Methods: The study analyzed ovomucoid – a protein isolated from the whites of the analyzed eggs. Ovomucoid was isolated by the method proposed by Roy et al. (2003). The next procedure was rabbit immunization. Polyclonal antibodies were obtained by subcutaneously immunizing a male rabbit with a solution of chicken egg glycated or native (a 100 ìg protein dose) in 1 ml PBS with pH 7.2. All measurements were performed in triplicate. The Fisher’s test was conducted to analyze variations in immunoreactivity and changes in glycation degree.
Result: Research has shown that it is needed to research the toxicity of proteins, in this case ovomucoid. Glycation may be recommended as a way to reduce or inhibit immunoreactivity. In case of allergies, such action is necessary and purposeful and it is essential to choose such reaction conditions which do not cause products harmful to human’s health.


Chicken ovomucoid Glycation Immunoreactivity Inactivation Maillard reaction


  1. Aabin, B., Poulsen, L.K., Ebbehøj, K., Nørgaard, A., Frøkiœr, H., Bindslev-Jensen, C., Barkholt, V. (1996). Identification of IgE-binding egg white proteins: comparison of results obtained by different methods. International Archives Allergy and Immunology. 109: 50-57. https://doi.org/10.1159/000237231. 
  2. Ashraf, J.M., Ahmad, S., Choi, I., Ahmad, N., Farhan, M., Godovikova, T., Shahab, U. (2015). Recent advances in detection of AGEs: immunochemical, bioanalytical and biochemical approaches. International Union of Biochemistry and Molecular Biology (IUBMB) Life. 67: 897-913. Doi.org/10.1002/iub.1450.
  3. Bielikowicz, K., Kostyra, H., Kostyra, E., Teodorowicz, M., Rigby, N., Wojtacha, P. (2012). The influence of non-enzymatic glycosylation on physicochemical and biological properties of pea globulin 7S. Food Research International. 48: 831-838. https://doi.org/10.1016/j.foodres.2012.06.028.
  4. Cazacu-Davidescu, L.M., Kostyra, H., Marciniak-Darmochwal, K., Kostyra, E. (2005). Influence of non-enzymatic glycosylation (glycation) of pea (Pisum sativum) albumins on their enzymatic hydrolysis. Journal of the Science of Food and Agriculture. 85: 948-954. doi: 10.1002/jsfa.2047.
  5. Chaudhari D.J., Tingree A.S. (2015). Forecasting eggs production in India. Indian Journal of Animal Research. 49(3): 367-372. DOI: 10.5958/0976-0555-2015.00143.0.
  6. Chiou, S.H., Wu, S.H. (1999). Evaluation of commonly used electrophoretic methods for the analysis of proteins and peptides and their application to biotechnology. Analytica Chimica Acta. 383: 47-60. Doi: 10.1016/S0003-2670 (98)00487-5.
  7. Czarnecki, T., Targoñski, Z. (2002). Allergens and food allergies. Ýywnoúã: nauka- technologia-jakoúã. 1: 19-33.
  8. Dall’Antonia, F., Pavkov-Keller, T., Zangger, K., Keller, W. (2014). Structure of allergens and structure based epitope predictions. Methods. 66: 3-21. Doi: 10.1016/j.ymeth. 2013.07.024.
  9. Davis, K.E., Prasad, C., Vijayagopal, P., Juma, S., Imrhan, V. (2016). Advanced glycation end products, inflammation and chronic metabolic diseases: links in a chain? Critical Reviews in Food Science and Nutrition. 56: 989-998. Doi: 10.1080/10408398.2012.744738.
  10. Falade, K.O., Adeyanju, K.I., Uzo-Peters, P.I. (2003). Foam-mat drying of cowpea (Vigna unguiculata) using glyceryl monostearate and egg albumin as foaming agents. European Food Research and Technology. 217(6). pp. 486-491. https://doi.org/10.1007/s00217-003-0775-3. 
  11. Frederico, E., Deutsch, H.F. (1949). Studies on ovomucoid. The Journal of Biological Chemistry. 181: 499-510.
  12. Go³¹b, K., Warwas, M. (2005). Chicken egg proteins – biochemical properties and applications. Advances in Clinical and Experimental Medicine. 14(5): 1001-1010.
  13. Gupta, R.K., Gupta, K., Sharma, A., Das, M., Ansari, I.A., Dwivedi, P.D. (2016). Maillard reaction in food allergy: pros and cons. Critical Reviewes in Food Science and Nutrition. 58(2): 208-226. Doi: 10.1080/10408398.2016.1152949.
  14. Jain, E., Bairoch, A., Duvaud, S., Phan, I., Redaschi, N., Suzek, B.E., Martin, M.J., McGarvey, P., Gasteiger, E. (2009). Infrastructure for the life sciences: design and implementation of the UniProt website. BMC Bioinformatics. 8(10): 136. Doi: 10.1186/1471-2105-10-136.
  15. Jiménez-Saiz, R., Belloque, J., Molina, E., López-Fandiño, R. (2011). Human immunoglobulin E (IgE) binding to heated and glycated ovalbumin and ovomucoid before and after in vitro digestion. Journal of Agricultural and Food Chemistry. 59(18): 10044-10051. Doi: 10.1021/jf2014638.
  16. Kato, Y., Matsuda, T. (1997). Glycation of proteinous inhibitors: loss in trypsin inhibitory activity by the blocking of arginine and lysine residues at their reactive sites. Journal of Agricultural and Food Chemistry. 45: 3826-3831. https://doi.org/10.1021/jf970310+.
  17. Konisi, Y., Kurisaki, J., Kaminogawa, S., Yamauchi, K. (1985). Determination of antigenicity by radioimmunoassay and of trypsin inhibitory activities in heat or enzyme denatured ovomucoid. Journal of Food Science. 50: 1422-1426. Doi. 10.1111/j.1365-2621.1985.tb10491.x. 
  18. Kovacs-Nolan, J., Zhang, J.W., Hayakawa, S., Mine, Y. (2000). Immunochemical and structural analysis of pepsin-digested egg white ovomucoid. Journal of Agricultural and Food Chemistry. 48: 6261-6266. Doi.10.1021/jf000358e.
  19. Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685. https://www.nature.com/articles/227680a0. 
  20. Ma, X., Gao, J., Chen, H. (2013). Combined effect of glycation and sodium carbonate–bicarbonate buffer concentration on IgG binding, IgE binding and conformation of ovalbumin. Journal of the Science of Food and Agriculture. 93(13): 3209-3215. Doi: 10.1002/jsfa.6157.
  21. Maillard, L.C. (1912). Action des acides amines sur les sucres; formation des mela-noidines par voie methodique. Comptes Rendus de l’Académie des Sciences. 154: 66-68.
  22. Mari, A., Rasi, C., Palazzo, P., Scala, E. (2009). Allergen databases: current status and perspectives. Current Allergy and Asthma Reports. 9: 376-383. Doi: 10.1007/s11882-009-0055-9.
  23. Martins, S.I.F.S., Jongen, W.M.F., van Boekel, M.A.J.S. (2001). A review of Maillard reaction in food and implications to kinetic modeling. Trends in Food Science and Technology. 11: 364-373. https://ucanr.edu/datastoreFiles/608-648. pdf.
  24. Mills, E.N.C., Breiteneder, H. (2005). Food allergy and its relevance to industrial food proteins. Biotechnology Advances. 23: 409-414. Doi: 10.1016/j.biotechadv.2005.05.006.
  25. Mine, Y., Yang, M. (2008). Recent advances in the understanding of egg allergens: basic, industrial and clinical perspectives. Journal of Agricultural and Food Chemistry. 56: 4874-4900. Doi: 10.1021/jf8001153. 
  26. Nielsen, P.M., Petersen, D., Dambmann, C. (2001). Improved method for determining food protein degree of hydrolysis. Journal of Food Science. 66: 642-646. https://doi.org/10.1111/j.1365-2621.2001.tb04614.x.
  27. Oliver, C.O., Melton, L.D., Stanley, R.A. (2006). Creating Proteins with Novel Functionality via the Maillard Reaction: A Review. Critical Reviews in Food Science and Nutrition. 46(4): 337-350. Doi: 10.1080/10408690590957250.
  28. Roy, I., Rao, M.V.S., Gupta, M.N. (2003). An integrated process of purification of lysozyme, ovalbumin and ovomucoid from hen egg white. Applied Biochemistry and Biotechnology. 111: 55-63. http://people.uleth.ca/~steyqj/Bchm3300Lab/ Paper_1.pdf.
  29. Seidler, N.W., Yeargans, G.S. (2002). Effects of thermal denaturation on protein glycation. Life Science. 70: 1789-1799.
  30. Staliñska, K. (2001). Receptors for advanced glycosylation end products. Advances in Cell Biology. 28(s16): 35-42.
  31. Thorpe, S.R., Baynes, J.W. (2003). Maillard reaction products in tissue proteins: new products and new perspectives. Amino Acids. 25(3-4): 275-281. Doi: 10.1007/s00726-003-0017-9.
  32. Tong, J.C., Lim, S.J., Muh, H.C., Chew, F.T., Tammi, M.T. (2009). Allergen atlas: A comprehensive knowledge center and analysis resource for allergen information. Bioinformatics. 25: 979-980.
  33. UniProt Consortium. (2015). UniProt: a hub for protein information. Nucleic Acids Research. 43(Database issue): D204-D212. Doi: 10.1093/nar/qku989.
  34. Vita, R., Overton, J.A., Greenbaum, J.A., Ponomarenko, J., Clark, J.D., Cantrell, J.R., Wheeler, D.K., Gabbard, J.L., Hix, D., Sette, A., Peters, B. (2015). The immune epitope database (IEDB) 3.0. Nucleic Acids Research. 43 (Database issue): D405-D412. Doi: 10.1093/nar/gku938.
  35. Vita, R., Zarêbski, L., Greenbaum, J.A., Emami, H., Hoof, I., Salimi, N., Damle, R., Sette, A., Peters, B. (2010).The Immune Epitope Database 2.0. Nucleic Acids Research. 38 (Database issue): D854-D862. Doi: 10.1093/nar/gkp1004.
  36. Wal, J.M. (2003). Thermal processing and allergenicity of foods. Allergy. 58(8): 727-729. Doi: 10.1034/j.1398-9995.2003. 00225.x.
  37. Whittow, G.C. (Ed). (2000). Sturkie’s Avian Physiology 5th Edition. Academic Press, New York. pp. 569-596.
  38. Worobiej, E., Wo³osiak, R., Chwalisz, M. (2006). The properties of egg white protein preparations in an oxidation process. Ýywnoúã: nauka-technologia - jakoúã. 4(49): 136-144.
  39. Zhang, J.W., Mine, Y. (1999). Characterization of residues in human IgE and IgG binding site by chemical modification of ovomucoid third domain. Biochemical and Biophysical Research Communications. 261: 610-613.

Global Footprints