Minsk, Belarus
Minsk, Belarus
Stavropol, Russian Federation
Enzymatic hydrolysis of whey proteins is aimed to obtain products with low allergenic potential and high nutritional value. Whey peptides are protein fraction that possesses a variety of physicochemical, immunochemical and bioactive properties (antioxidant, antibacterial, immunomodulatory effects). Controlled parameters of enzymatic hydrolysates are the degree of hydrolysis of protein substrates, peptide composition, residual antigenicity, antioxidant capacity, etc. The purpose of this work is to characterize peptide profile, antigenicity and free radical scavenging properties of experimental hydrolysate sample, toxicological and hygienic assessment of its impact on a test object (infusoria Tetrahymena pyriformis). The research of properties of whey proteins and their enzymatic hydrolysates was conducted using classical and modern methodological approaches: SDS electrophoresis in polyacrylamide gel, HPLC, mass-spectrometry, competitive ELISA, TEAC (Trolox Equivalent Capacity Assay) technique. Peptide composition, antigenic and antioxidant properties of obtained enzymatic hydrolysate were determined, toxicity of raw and digested whey proteins were examined. We established that the investigated hydrolysate sample falls under the category of partial hydrolysates for functional products according to its physicochemical, immunochemical, free radical scavenging and organoleptic properties. According to the results of toxicological and hygienic assessment using T. pyriformis model, whey protein concentrate and its hydrolysates are non-toxic and do not possess cumulative properties. Thus, we obtained partial enzymatic hydrolysate of whey proteins from milk, which can be used as physiologically active component in the development of new specialized food.
Allergenic milk proteins, protein hydrolysates, peptide composition, residual antigenicity, antioxidant properties, toxicological and hygienic assessment, infusoria Tetrahymena pyriformis
1. Hurley W.L. Milk protein. Vol. 1. Rijeka, Croatia: InTech, 2012. 352 p.
2. Wal J.M. Bovine milk allergenicity. Ann. Allergy Asthma Immunol., 2004, vol. 93, pp. 2-11.
3. Raikos V. and Dassios T. Health-promoting properties of bioactive peptides derived from milk proteins in infant food: a review. Dairy Sci. & Technol., 2014, vol. 94, pp. 91-101.
4. Mahmoud M.I. The physicochemical and functional properties of protein hydrolysates in nutritional products. Food Technol., 1994, vol. 48, no. 10, pp. 89-95.
5. Maldonado J., Gil A., Narbona E. and Molina J.A. Special formulas in infant nutrition: a review. Early Hum. Dev., 1998, vol. 53, no. 1, pp. 23-32.
6. Zulueta A., Maurizi A., Frígola A., Esteve M.J., Coli, R. and Burini G. Antioxidant capacity of cow milk, whey and milk deproteinized. Int. Dairy J., 2009, vol. 19, no. 6-7, pp. 380-385.
7. Hernández-Ledesma B., Dávalos A., Bartolomé B. and Amigo L. Preparation of antioxidant enzymatic hydrolysates from alpha- lactalbumin and beta-lactoglobulin. Identification of active peptides by HPLC-MS/MS. J. Agric. Food Chem., 2005, vol. 53, no. 3, pp. 588-593.
8. De Castro R.J.S. and Sato H.H. Comparison and synergistic effects of intact proteins and their hydrolysates on the functional properties and antioxidant activities in a simultaneous process of enzymatic hydrolysis. Food and Bioproducts Proc., 2014, vol. 92, no. 1, pp. 80-88.
9. Hernández-Ledesma B., Quirós A., Amigo L. and Recio I. Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin. Int. Dairy J., 2007, vol. 17, no. 1, pp. 42-49.
10. Sinks G. and Schultz T. Correlation of Tetrahymena periformis and Pimephales toxicity: evaluation of 100 additional compounds. Environ. Toxicol. Chem., 2001, vol. 20, no. 4, pp. 917-921.
11. Zhu H., Tropsha A., Fourches D., Varnek A., Papa E., Gramatica P., Oberg T., Dao P., Cherkasov A. and Tetko I.V. Combinatorial QSAR modeling of chemical toxicants tested against Tetrahymena periformis. J. Chem. Inf. Model., 2008, vol. 48, no. 4, pp. 766-784.
12. Kurchenko V.P., Kapustin M.A., Butkevich T.V., Rizevskiy S.V., Halavach T.N. and Strogiy V.N. Integrated technology of whey processing. Molochnaya Industriya, 2013, no. 2, pp. 38-41. (In Russian).
13. Osterman L.A. Research methods of proteins and nucleic acids: Electrophoresis and ultracentrifugation (practice note). Moscow: Nauka Publ., 1981. 288 p. (In Russian).
14. Kim S.B., Seo I.S., Khan A., Ki K.S., Lee W.S. and Lee H.J. Enzymatic hydrolysis of whey is heated: iron-binding ability of peptides and antigenic protein fractions. J. Dairy Sci., 2007, vol. 90, pp. 4033-4042.
15. Kim S.B., Ki K.S., Khan A., Lee W.S., Lee H.J. and Ahn B.S. Peptic and tryptic hydrolysis of native and heated whey protein to reduce its antigenicity. J. Dairy Sci., 2007, vol. 90, pp. 4043-4050.
16. Kruglik V.I., Zorin S.N., Gmoshinskiy I.V., Ponomarev D.V., Nikitina N.E., Abramova A.A., Volkova I.N. and Revyakina N.V. Sposob polucheniya fermentativnogo gidrolizata syvorotochnykh belkov so sredney stepen´yu gidroliza [Obtaining method of whey protein enzymatic hydrolysate with moderate degree of hydrolysis]. Patent RF, no. 2375910, 2009.
17. Kiselev P.A., Oreshko N.A., Bovdey N.A., Le Min Kha, Fam Kuok Long, Baranovskiy A.V., Khripach N.B., Khlebnikova T.S. and Lakhvich F.A. Obtaining and characterization of antioxidant activity of Iris Domestica extracts and their individual components. Proceedings of the Belarusian State University, 2012, vol. 7, pp. 228-237. (In Russian).
18. Kutsenko S.A. Toxicology basics: method. pub. St. Petersburg: Foliant Publ., 2004. 720 p. (In Russian).
19. Podunova L.G., Menshikova, T.A. and Dvoskin Ya.G. Alternative research methods (express methods) for toxicological-hygienic assessment of materials, products and environmental objects: method. pub. Moscow, 1999. 108 p. (In Russian).
20. Prozorovsky V.B. Least square method for probit analysis of mortality curves. Farmakologiya i toxikologiya, 1962, no. 1, pp. 41-63. (In Russian).
21. O´Keeffe M.B. and FitzGerald, J.R. Antioxidant effects of enzymatic hydrolysates of whey protein concentrate on cultured human endothelial cells. Int. Dairy J., 2014, vol. 36, no. 2, pp. 128-135.
22. Halavach T.M., Kurchenko V.P. and Albulov A.I. Enzymatic hydrolysis of milk proteins as a basis of specialized food products biotechnology. Ural Scientific Bulletin, 2014, vol. 104, no. 25, pp. 69-79.
