Abstract

Food allergy is a matter of global concern, proteins being a popular allergen worldwide. Enzymatic protein hydrolysates serve as hypoallergenic components in functional foods. This article describes enzymatic hydrolysates of egg and soy proteins as potential antigens, sensitizers, and antioxidants.
The research featured soy protein isolate and egg albumin. The protein hydrolysates were obtained in a two-step process with pepsin and trypsin to measure hydrolysis, total antioxidant activity, and residual antigenicity. The sensitizing ability of native proteins and their hydrolysates was studied using a delayed-type hypersensitivity (DTH) skin test of paw swelling in mice.
The egg albumin and soy protein were hydrolyzed by sequentially adding the proteases, i.e., pepsin and trypsin. The degree of protein hydrolysis was calculated against total nitrogen and its contents in non-hydrolyzed raw materials and hydrolysate. It was 82.6% for the egg hydrolysate and 88.3% for the soy hydrolysate, the total antioxidant activity being 114.3 and 91.4 mg/100 mL, respectively. The residual antigenicity of the hydrolysates determined by ELISA in a sandwich version was 1.55×10^–4 and 3.30×10^–4 RU, respectively. The native egg and soy proteins demonstrated good allergenic properties, and the DTH response index increased fivefold. The hydrolysates reduced the DTH response indices by an average of 3.5 for egg hydrolysate and 2.6 times for soy hydrolysate.
The resulting enzymatic hydrolysates of egg albumin and soy protein demonstrated a high degree of hydrolysis and antioxidant activity. They had low residual antigenicity (10^–4) and low sensitizing ability. The resulting enzymatic hydrolysates could be used as hypoallergenic components and antioxidants in new functional foods.

Keywords

Egg albumin, soy protein, hydrolysates, residual antigenic activity, sensitizing activity, antioxidant activity

FUNDING

The research was supported by the Russian Science Foundation (RSF), grant no. 23-26-00058, https://rscf.ru/project/23-26-00058

REFERENCES

1. Sampath V, Abrams EM, Adlou B, Akdis C, Akdis M, Brough H, et al. Food Allergy Across the Globe. Journal of Allergy and Clinical Immunology. 2021;148(6):1347–1364. https://doi.org/10.1016/j.jaci.2021.10.018
2. Revyakina VA. The Problem of Food Allergies at the Present Stage. Problems of Nutrition. 2020;89(4):186–192. (In Russ.). https://doi.org/10.24411/0042-8833-2020-10052; https://www.elibrary.ru/IAVQHX
3. Chittoor RI, Saraswathi HTB. A Review of Naturally Occuring Food Allergens and Their Impact on Health. Biosciences Biotechnology Research Asia. 2022;19(1):13–35. https://doi.org/10.13005/bbra/2965
4. Dolganyuk V, Sukhikh S, Kalashnikova O, Ivanova S, Kashirskikh E, Prosekov A, et al. Food Proteins: Potential Resources. Sustainability. 2023;15(7):5863. https://doi.org/10.3390/su15075863
5. Fedotova MM, Fedorova OS, Konovalova UV, Kamaltynova EM, Nagaeva TA Ogorodova LM. Hen's Egg Allergy: An Update. Bulletin of Siberian Medicine. 2018;17(2):156–166. https://doi.org/10.20538/1682-0363-2018-2-156-166; https://www.elibrary.ru/UTINVZ
6. Monaci L, Pilolli R, de Angelis E, Crespo JF, Novak N, Cabanillas B. Food Allergens: Classification, Molecular Properties, Characterization, and Detection in Food Sources. Advances in Food and Nutrition Research. 2020;93:113–146. https://doi.org/10.1016/bs.afnr.2020.03.001
7. Gromov DA, Borisova AV, Bakharev VV. Food Allergens and Methods for Producing Hypoallergenic Foods. Food Processing: Techniques and Technology. 2021;51(2):232–247. (In Russ.). https://doi.org/10.21603/2074-9414-2021-2-232-247; https://elibrary.ru/MHOLPD
8. Zorin SN, Sidorova YuS, Mazo VK. Enzymatic Hydrolysates of Whey Protein and Chicken Egg Ppotein: Production, Physical-Chemical and Immunochemical Characteristics. Problems of Nutrition. 2020;89(1):64–68. (In Russ.). https://doi.org/10.24411/0042-8833-2020-10007; https://elibrary.ru/EYXSNG
9. Zorin SN. Enzymatic Hydrolysates of Food Proteins for Specialized Foods for Therapeutic and Prophylactic Nutrition. Problems of Nutrition. 2019;88(3):23–31. (In Russ.). https://doi.org/10.24411/0042-8833-2019-10026; https://elibrary.ru/JMLHXW
10. Clare Mills EN, Sancho AI, Rigby NM, Jenkins JA, Mackie AR. Impact of Food Processing on the Structural and Allergenic Properties of Food Allergens. Molecular Nutrition and Food Research. 2009;53(8):963–969. https://doi.org/10.1002/mnfr.200800236
11. Mahler V, Goodman RE. Definition and Design of Hypoallergenic Foods. In: Kleine-Tebbe J, Jakob T, editors. Molecular Allergy Diagnostics. Innovation for a Better Patient Management. Cham: Springer; 2017. pp. 487–511. https://doi.org/10.1007/978-3-319-42499-6_27
12. Verhoeckx KCM, Vissers YM, Baumert JL, Faludi R, Feys M, Flanagan S, et al. Food Processing and Allergenicity. Food and Chemical Toxicology. 2015;80:223–240. https://doi.org/10.1016/j.fct.2015.03.005
13. Prikhodko DV, Krasnoshtanova AA. Using casein and gluten protein fractions to obtain functional ingredients. Foods and Raw Materials. 2023;11(2):223–231. https://doi.org/10.21603/2308-4057-2023-2-569
14. Rahaman T, Vasiljevic T, Ramchandran L. Effect of Processing on Conformational Changes of Food Proteins Related to Allergenicity. Trends in Food Science and Technology. 2016;49:24–34. https://doi.org/10.1016/j.tifs.2016.01.001
15. Andjelkovic U. Food Allergy and Food Allergens. In: Cifuentes A, editor. Comprehensive Foodomics. Elsevier; 2021. pp. 154–174. https://doi.org/10.1016/B978-0-08-100596-5.22844-8
16. Semenova ES, Simonenko ES, Simonenko SV, Zorin SN, Petrov NA, Mazo VK. Study of the Process of Hydrolysis of Milk Proteins Using Enzyme Preparations of Domestic and Production. Food Systems. 2023;6(2):224–232. (In Russ.). https://doi.org/10.21323/2618-9771-2023-6-2-224-232; https://elibrary.ru/VQTKGN
17. Sviridenko YuYa, Myagkonosov DS, Abramov DV, Ovchinnikova EG. Theoretical and Practical Aspects of Development Technology of Manufacturing Protein Hydrolyzates for Special Nutrition Use. Part 1. Technology of Production and Technical Characteristics of Hydrolysates. Food Industry. 2017;(5):48–51. (In Russ.). https://elibrary.ru/YPIXGD
18. Sviridenko YuYa, Myagkonosov DS, Abramov DV, Ovchinnikova EG, Simonenko SV, Antipova TA. Technological Methods for Improving the Functional Properties of Hydrolysates for Specialized and Infant Nutrition. Food Industry. 2020;(10):12–17. (In Russ.). https://doi.org/10.24411/0235-2486-2020-10113; https://elibrary.ru/MXLHHE
19. Kostyleva EV, Sereda AS, Velikoretskaya IA, Kurbatova EI, Tsurikova NV. Proteases For Obtaining of Food Protein Hydrolysates from Proteinaceous By-Products. Problems of Nutrition. 2023;92(1):116–132. (In Russ.). https://doi.org/10.33029/0042-8833-2023-92-1-116-132; https://elibrary.ru/PUGROG
20. Pan M, Liu K, Yang J, Liu S, Wang S, Wang S. Advances on Food-Derived Peptidic Antioxidants – A Review. Antioxidants. 2020;9(9):799. https://doi.org/10.3390/antiox9090799
21. Dziuba M, Darewicz M. Food Proteins as Precursors of Bioactive Peptides – Classification into Families. Food Science and Technology International. 2007;13(6):393–404. https://doi.org/10.1177/1082013208085933
22. Sarmadi BH, Ismail A. Antioxidative Peptides from Food Proteins: A Review. Peptides. 2010;31(10):1949–1956. https://doi.org/10.1016/j.peptides.2010.06.020
23. Benedé S, Molina E. Chicken Egg Proteins and Derived Peptides with Antioxidant Properties. Foods. 2020;9(6):735. https://doi.org/10.3390/foods9060735
24. López-García G, Dublan-García O, Arizmendi-Cotero D, González-Olivan LM. Antioxidant and Antimicrobial Peptides Derived from Food Proteins. Molecules. 2022;27(4):1343. https://doi.org/10.3390/molecules27041343
25. Du Z, Li Y. Review and Perspective on Bioactive Peptides: A Roadmap for Research, Development, and Future Opportunities. Journal of Agriculture and Food Research. 2022;9:100353. https://doi.org/10.1016/j.jafr.2022.100353
26. Wong F-C, Xiao J, Wang S, Ee K-Y, Chai T-T. Advances on the Antioxidant Peptides from Edible Plant Sources. Trends in Food Science and Technology. 2020;99:44–57. https://doi.org/10.1016/j.tifs.2020.02.012
27. Zhamsaranova SD, Lebedeva SN, Bolkhonov BA, Sokolov DV. Enzymatic Food Protein Conversion and Assessment of Antioxidant Activity of Peptides. ESSUTM Bulletin. 2021;(4):5–14. (In Russ.). https://doi.org/10.53980/24131997_2021_4_5; https://elibrary.ru/TNSCKR
28. Bolkhonov BA, Sokolov DV, Zhamsaranova SD, Lebedeva SN, Qian Ch. Selection of Working Parameters for the Production of Egg Protein Peptides. ESSUTM Bulletin. 2022;(4):15–23. (In Russ.). https://doi.org/10.53980/24131997_2022_4_15; https://elibrary.ru/UICXSZ
29. Zhamsaranova SD, Lebedeva SN, Bolkhonov BA, Sokolov DV, Bazhenova BA. Hydrolysate of Ovalbumin: Production and Evaluation of the Functional Properties of Peptides. Theory and Practice of Meat Processing. 2023;8(1):34–42. https://doi.org/10.21323/2414-438X-2023-8-1-34-42; https://elibrary.ru/EQUMSB
30. Sokolov DV, Bolkhonov BA, Zhamsaranova SD, Lebedeva SN, Bazhenova BA. Enzymatic Hydrolysis of Soy Protein. Food Processing: Techniques and Technology. 2023;53(1):86–96. https://doi.org/10.21603/2074-9414-2023-1-2418; https://elibrary.ru/DHXSAX
31. Akbarian M, Khani A, Eghbalpour S, Uversky VN. Bioactive Peptides: Synthesis, Sources, Applications, and Proposed Mechanisms of Action. International Journal of Molecular Sciences. 2022;23(3):1445. https://doi.org/10.3390/ijms23031445
32. Manzoor M, Singh J, Gani A. Exploration of Bioactive Peptides from Various Origin as Promising Nutraceutical Treasures: In Vitro, In Silico and In Vivo Studies. Food Chemistry. 2021;373:131395. https://doi.org/10.1016/j.foodchem.2021.131395
33. Aider M. Potential Applications of Ficin in The Production of Traditional Cheeses and Protein Hydrolysates. JDS Communications. 2021;2(5):233–237. https://doi.org/10.3168/jdsc.2020-0073
34. Pang L, Liu M, Li X, Guo L, Man C, Yang X, et al. Effect of Enzymatic Hydrolysis Combined with Processing on Allergenicity of Food Allergens. Trends in Food Science and Technology. 2024;143:104248 https://doi.org/10.1016/j.tifs.2023.104248
35. Ahmed I, Chen H, Li J, Wang B, Li Z, Huang G. Enzymatic Crosslinking and Food Allergenicity: A Comprehensive Review. Comprehensive Reviews in Food Science and Food Safety. 2021;20(6):5856–5879. https://doi.org/10.1111/1541-4337.12855
36. El Mecherfi K-E, Todorov SD, de Albuquerque MAC, Denery-Papini S, Lupi R, Haertlé T, et al. Allergenicity of Fermented Foods: Emphasis on Seeds Protein-Based Products. Foods. 2020;9(6):792. https://doi.org/10.3390/foods9060792