COMPARATIVE ANALYSIS AND ANTIGENIC PROPERTIES EVALUATION OF WHEY PROTEINS HYDROLYSATES WITH SERINE PROTEASES TRYPSIN AND ALCALASE
Abstract
Degree of proteolysis and antigenic properties of whey proteins hydrolysates with application of serine proteases (alcalase and trypsin) and their complex were investigated. At application of both proteases by SDS–PAGE and high performance liquid chromatography the most complete proteolysis of β–lg and α–la to peptides was demonstrated, while deleting of high molecular BSA fraction was achieved by filtration of hydrolysates. By double–immunodiffusion methods for products received with both serine proteases antigenic properties were detected; by ELISA reduction of ability to bind antibodies on 60% was reported. Filtration of hydrolysate provided obtaining of peptide fraction with Mr ≤10 kDa that wasn’t able to form immune complexes and had antigenic potential <10%.
About the Authors
T. HalavachRussian Federation
N. Zhabanos
Russian Federation
V. Kurchenko V. Kurchenko
Russian Federation
References
1. Smithers, G.F. Advances in dairy foods processing and engineering: symposium / G.F. Smithers [et al.] // Journal of Dairy Science. – 1996. –Vol. 79. – P. 1454–1459.
2. Cavagni, G. Allergy to cow’s milk proteins in childhood: the author’s personal experience and new diagnostic and therapeutic proposals / G. Cavagni [et al.] // Pediatr. Med. Chir. – 1994. – Vol.16. – № 5. –P. 413–419.
3. Foegeding, E. A. Advances in modifying and understanding whey protein functionality / E. A.Foegeding [et al.] // Trends in Food Science and Technology. – 2002. – Vol. 13. – P. 151–159.
4. Wal, J.M. Bovine milk allergenicity / J.M. Wal // Annals of Allergy, Asthma, and Immunology. – 2004. – Vol. 93. – P. 2–11.
5. Nakamura, T. Production of low antigenic whey protein hydrolysates by enzymatic hydrolysis and denaturation with high pressure / T. Nakamura, H. Sado, Y. Syukunobe // Milchwissenschaft. – 1993. – Vol. 48. – № 3. –P. 141–144.
6. Mullally, M.M. Angiotensin-iconverting enzyme inhibitory activities of gastric and pancreatic proteinase digest of whey proteins / M.M. Mullally, H. Meisel, R.J. Fitzgerald // International Dairy Journal. – 1997. – Vol. 7. –P. 299–303.
7. Korhonen, H. Impact of processing on bioactive proteins and peptides / H. Korhonen [et al.] // Trends in Food Science and Technology. – 1998. –Vol. 9. – P. 307–319.
8. Bertrand-Harb, C. Thermal modifications of structure and co-denaturation of a-lactalbumin and b-lactoglobulin induce changes of solubility and susceptibility to proteases / C. Bertrand-Harb [et al.] // Nahrung. – 2002. –Vol. 46. – P. 283–289.
9. Protein measurement with the Folin phenol reagent / O.H. Lowry [et al.] // J. Biol. Chem. – 1951. – P. 265–275.
10. Остерман, Л.А. Методы исследования белков и нуклеиновых кислот: Электрофорез и ультрацентрифугирование: практ. пособие / Л.А. Остерман. – М.: Наука, 1981. – 288 c.
11. Kim, S.B. Peptic and tryptic hydrolysis of native and heated whey protein to reduce its antigenicity / S.B. Kim [et al.] // J. Dairy. Sci. – 2007. – Vol. 90. – P. 4043–4050.
12. Иммунологические методы / под ред. Х. Фримеля. – М.: Мир, 1979. –518 c.
13. Gjesing, B. Allergen-specific IgE antibodies against antigenic components in cow milk and milk substitutes / B. Gjesing [et al.] // Allergy. – 1986. –Vol. 41. – P. 51–56.
14. Haddad, Z.H. IgE antibodies to peptic and peptic-tryptic digest of β-lactoglobulin: significance in food hypersensitivity Z.H. Haddad, V. Kalra, S. Verma // Ann. Allergy. – 1979. – Vol. 42. – P. 368–371.
15. Wong, D.W.S. Structures and functionalities of milk proteins / D.W.S. Wong; W.M. Camirand; A.E. Paviath // Crit. Rev. Food Sci. Nutr. – 1996. – Vol. 36. – P. 807–844.
16. Papiz, M. Z. The structure of β-lactoglobulin and its similarity to plasma retinol-binding protein / M.Z. Papiz // Nature. – 1986. – Vol. 324. – P. 383–385.
17. Brew, K. α–lactalbumin / K. Brew, J.A. Grobler // P. Fox, Editor, Advances in Dairy Chemistry-1. – Elsevier. – Amsterdam, 1992. – P. 191–229.
18. Peters, T.Jr. Serum albumin / T.Jr. Peters // Adv. Protein Chem. – 1985. –Vol. 3. – P. 161–245.
19. Doucet, D. Gel formation of peptides produced by extensive enzymatic hydrolysis of β-lactoglobulin / D. Doucet, E.A. Foegeding // Biomacromolecules. – 2005. – Vol. 6. – P. 1140–1148.
20. Диксон, М. Ферменты / М. Диксон, Э. Уэбб; пер. с англ. – М., 1982. –C. 370–376.
21. Besler, M. Stability of food allergens and allergenicity of processed foods / M. Besler, H. Steinhart, A. Paschke // J. Chromatogr. Biomed. Sci. Appl. –2001. – Vol. 756. – P. 207–228.
22. Restani, P. Evaluation by SDS-PAGE and immunoblotting of residual antigenicity in hydrolysed protein formulas / P. Restani [et al.] // Clinical and Experimental Allergy: Journal of the British Society for Allergy and Clinical Immunology. – Vol. 25. – P. 651–658.
Review
For citations:
Halavach T., Zhabanos N., V. Kurchenko V. COMPARATIVE ANALYSIS AND ANTIGENIC PROPERTIES EVALUATION OF WHEY PROTEINS HYDROLYSATES WITH SERINE PROTEASES TRYPSIN AND ALCALASE. Topical Issues of Processing of Meat and Milk Raw Materials. 2009;(4):102-113. (In Russ.)