Abstract

Soy continues to be one of the top sources of vegetable protein. Structurally modified soy proteins and processed products are used as part of functional foods. Enzymatic hydrolysates of food proteins have different degrees of hydrolysis and functional profiles, hence the constant search for the optimal hydrolysis parameters. The present research objective was to design a two-stage enzymatic conversion process of soy protein using mathematical methods, as well as to evaluate the antioxidant properties of the hydrolysate in laboratory conditions.
Soy protein isolate was tested to define the maximal value of the hydrolysis degree. It underwent a series of two-factor experiments in the presence of pepsin and trypsin. The study focused on the hydrolysis time and the enzyme-substrate ratio. The results were optimized using the response surface methodology in MathCad 15. The total antioxidant activity of the hydrolysate during hydrolysis was determined on a Tsvet-Yauza-01-AA chromatograph using the amperometric method.
For the pepsin test, the processing time was 7 h and the enzyme-to-substrate ratio was 1:22. For the trypsin test, the time was 7 h and the ratio was 1:30. The mathematical modeling revealed the following optimal parameters. The first stage involved hydrolysis with pepsin for 5 h at an enzyme-to-substrate ratio of 1:20; the second stage involved hydrolysis with trypsin for 3 h at an enzyme-to-substrate ratio of 1:19. The resulting hydrolysate demonstrated 88% hydrolysis. The highest summary antioxidant activity was registered after 5 h of hydrolysis and amounted to about 250 mg/100 mL.
The resulting enzymatic hydrolysate of soy protein can be used as a food component or an antioxidant feed additive. The obtained peptides can immobilize essential microelements, e.g., Zn, I, and Se, as well as produce polyvalent complexes. Further studies will be aimed at the residual antigenicity of the hydrolysate and other functional indicators.

Keywords

Soy, protein, pepsin, trypsin, hydrolysate, degree of hydrolys is, peptides, antioxidant activity

Contribution

All authors participated in the work in a substantial way and are prepared to take public responsibility for the work and manuscript contents. S.D. Zhamsaranova designed the research and analysis. S.N. Lebedeva, B.A. Bazhenova, D.V. Sokolov, and B.A. Bolkhonov collected the data, analyzed the results, and wrote the manuscript.

CONFLICTS OF INTEREST

The authors declare that there is no conflict of interests regarding the publication of this article.

FUNDING

The research was supported by the grant of the Russian Science Foundation (RSF) No. 23-26-00058.

REFERENCES

1. Fahed G, Aoun L, Zerdan MB, Allam S, Zerdan MB, Bouferraa Y, et al. Metabolic syndrome: Updates on pathophysiology and management in 2021. International Journal of Molecular Sciences. 2022;23(2). https://doi.org/10.3390/ijms23020786
2. Castro-Barquero S, Ruiz-León AM, Sierra-Pérez M, Estruch R, Casas R. Dietary strategies for metabolic syndrome: A comprehensive review. Nutrients. 2020;12(10). https://doi.org/10.3390/nu12102983
3. Vorobyeva VM, Vorobyeva IS, Kochetkova AA, Mazo VK, Zorin SN, Sharafetdinov KhKh. Specialised hypocholesteremic foods: Ingredients, technology, effects. Foods and Raw Materials. 2020;8(1):20–29. https://doi.org/10.21603/2308-4057-2020-1-20-29
4. Sadovoy VV, Shchedrina TV, Trubina IA, Morgunova AV, Franko EP. Cooked sausage enriched with essential nutrients for the gastrointestinal diet. Foods and Raw Materials. 2021;9(2):345–353. https://doi.org/10.21603/2308-4057-2021-2-345-353.
5. Sui X, Zhang T, Jiang L. Soy protein: Molecular structure revisited and recent advances in processing technologies. Annual Review of Food Science and Technology. 2021;12:119–147. https://doi.org/10.1146/annurev-food-062220-104405
6. Zhang T, Dou W, Zhang X, Zhao Y, Zhang Y, Jiang L, et al. The development history and recent updates on soy protein-based meat alternatives. Trends in Food Science and Technology. 2021;109:702–710. https://doi.org/10.1016/j.tifs.2021.01.060
7. Qin P, Wang T, Luo Y. A review on plant-based proteins from soybean: Health benefits and soy product development. Journal of Agriculture and Food Research. 2022;7. https://doi.org/10.1016/j.jafr.2021.100265
8. Wu Y-HS, Chen Y-C. Trends and applications of food protein-origin hydrolysates and bioactive peptides. Journal of Food and Drug Analysis. 2022;30(2):172–184. https://doi.org/10.38212/2224-6614.3408
9. Ewert J, Eisele T, Stressler T. Enzymatic production and analysis of antioxidative protein hydrolysates. European Food Research and Technology. 2022;248:2167–2184. https://doi.org/10.1007/s00217-022-04022-x
10. Agarkova EYu, Kruchinin AG. Enzymatic conversion as a method of producing biologically active peptides. Vestnik of MSTU. Scientific Journal of Murmansk State Technical University. 2018;21(3):412–419. https://doi.org/10.21443/1560-9278-2018-21-3-412-419
11. Cruz-Casas DE, Aguilar CN, Ascacio-Valdés JA, Rodríguez-Herrera R, Chávez-González ML, Flores-Gallegos AC. Enzymatic hydrolysis and microbial fermentation: The most favorable biotechnological methods for the release of bioactive peptides. Food Chemistry: Molecular Sciences. 2021;3. https://doi.org/10.1016/j.fochms.2021.100047
12. 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.).
13. 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
14. Sidorova YuS, Mazo VK, Kochetkova AA. Experimental evaluation of hypolipidemic properties of soy and rice proteins and their enzyme hydrolysates. Problems of Nutrition. 2018;87(2):77–84. (In Russ.). https://doi.org/10.24411/0042-8833-2018-10021
15. Grishin DV, Podobed OV, Gladilina YuA, Pokrovskaya MV, Aleksandrova SS, Pokrovsky VS, et al. Bioactive proteins and peptides: Current state and new trends of practical application in the food industry and feed production. Problems of Nutrition. 2017;86(3):19–31. (In Russ.).
16. Caponio GR, Wang DQ-H, Di Ciaula A, De Angelis M, Portincasa P. Regulation of cholesterol metabolism by bioactive components of soy proteins: Novel translational evidence. International Journal of Molecular Sciences. 2020;22(1). https://doi.org/10.3390/ijms22010227
17. Ashaolu TJ. Applications of soy protein hydrolysates in the emerging functional foods: A review. International Journal of Food Science and Technology. 2020;55(2):421–428. https://doi.org/10.1111/ijfs.14380
18. Nasri M. Protein hydrolysates and biopeptides: Production, biological activities, and applications in foods and health benefits. A Review. Advances in Food and Nutrition Research. 2017;81:109–159. https://doi.org/10.1016/bs.afnr.2016.10.003
19. Morgan PT, Breen L. The role of protein hydrolysates for expertise-induced skeletal muscle recovery and adaptation: A current perspective. Nutrition and Metabolism. 2021;18(44). https://doi.org/10.1186/s12986-021-00574-z
20. Kim M-S, Kim B, Park H, Ji Y, Holzapfel W, Kim D-Y, et al. Longterm fermented soybean paste improves metabolic parameters associated with non-alcoholic fatty liver disease and insulin resistance in high-fat diet-induced obese mice. Biochemical and Biophysical Research Communications. 2018;495(2):1744–1751. https://doi.org/10.1016/j.bbrc.2017.12.003
21. Tutelian VA, Khavinson VKh, Ryzhak GA, Linkova NS. Short peptides as components of nutrition: Molecular bases of gomeostasis regulation. Uspekhi Sovremennoi Biologii. 2014;134(3):227–235. (In Russ.).
22. Yu YM, Fukagawa NK. Protein and amino acid. In: Marriott BP, Birt DF, Stallings VA, Yates AA, editors. Present knowledge in nutrition. Volume 1: Basic nutrition and metabolism. Academic Press; 2020. pp. 15–35. https://doi.org/10.1016/B978-0-323-66162-1.00002-0
23. Hajam YA, Rani R, Ganie SY, Sheikh TA, Javaid D, Qadri SS, et al. Oxidative stress in human pathology and aging: Molecular mechanisms and perspectives. Cells. 2022;11(3). https://doi.org/10.3390/cells11030552
24. Lv R, Dong Y, Bao Z, Zhang S, Lin S, Sun N. Advances in the activity evaluation and cellular regulation pathways of food-derived antioxidant peptides. Trends in Food Science and Technology. 2022;122:171–186. https://doi.org/10.1016/j.tifs.2022.02.026
25. Wen C, Zhang J, Zhang H, Duan Y, Ma H. Plant protein-derived antioxidant peptides: Isolation, identification, mechanism of action and application in food systems: A review. Trends in Food Science and Technology. 2020;105:308–322. https://doi.org/10.1016/j.tifs.2020.09.019
26. Montesano D, Gallo M, Blasi F, Cossignani L. Biopeptides from vegetable proteins: New scientific evidences. Current Opinion in Food Science. 2020;31:31–37. https://doi.org/10.1016/j.cofs.2019.10.008
27. Aguilar-Toalá JE, Liceaga AM. Cellular antioxidant effect of bioactive peptides and molecular mechanisms underlying: Beyond chemical properties. International Journal of Food Science and Technology. 2020;56(5):2193–2204. https://doi.org/10.1111/ijfs.14855
28. Elam E, Feng J, Lv Y-M, Ni Z-J, Sun P, Thakur K, et al. Recent advances on bioactive food derived anti-diabetic hydrolysates and peptides from natural resources. Journal of Functional Foods. 2021;86. https://doi.org/10.1016/j.jff.2021.104674
29. Daroit DJ, Brandelli A. In vivo bioactivities of food protein-derived peptides – a current review. Current Opinion in Food Science. 2021;39:120–129. https://doi.org/10.1016/j.cofs.2021.01.002
30. Nwachukwu ID, Aluko RE. Structural and functional properties of food protein-derived antioxidant peptides. Journal of Food Biochemistry. 2019;43(1). https://doi.org/10.1111/jfbc.12761
31. Kim J-H, Jang H-J, Cho W-Y, Yeon S-J, Lee C-H. In vitro antioxidant actions of sulfur-containing amino acids. Arabian Journal of Chemistry. 2020;13(1):1678–1684. https://doi.org/10.1016/j.arabjc.2017.12.036
32. Liu J, Zhang D, Zhu Y, Wang Y, He S, Zhang T. Enhancing the in vitro Antioxidant Capacities via the interaction of amino acids. Emirates Journal of Food and Agriculture. 2018;30(3):224–231. https://doi.org/10.9755/ejfa.2018.v30.i3.1641
33. Zhamsaranova SD, Lebedeva SN, Bolkhonov BA, Sokolov DV. Enzymatic food protein conversion and assessment of antioxidant activity of peptides. Bulletin of ESSTUM. 2021;83(4):5–14. (In Russ.). https://doi.org/ 10.53980/24131997_2021_4_5
34. Babin AV, Rakipov DF. Organization and mathematical planning of the experiment. Ekaterinburg: Ural Federal University; 2014. 113 p. (In Russ.).
35. Dorovskikh VI, Milushev RK, Shulaev GM, Zharikov VS. Using a generic function the Harrington's desirability in the evaluation of quality of feed additives. Science in the Central Russia. 2020;45(3):79–85. (In Russ.). https://doi.org/10.35887/2305-2538-2020-3-79-85
36. Yashin AYa, Yashin YaI, Chernousova NI, Pakhomov VP. TsvetYauza-01AA: A new device for the determination of antioxidants in medicines, dietary supplements, foods, and drinks. Moscow: NPO “Khimavtomatika”: 2005. 100 p. (In Russ.).
37. Milentyeva IS, Davydenko NI, Rasshchepkin AN. Casein proteolysis in bioactive peptide production: Optimal operating parameters. Food Processing: Techniques and Technology. 2020;50(4):726–735. (In Russ.). https://doi.org/10.21603/2074-9414-2020-4-726-735