Estimation of the kinetic parameters of whey proteins proteolysis in the UF-concentrate of cheese whey
https://doi.org/10.20914/2310-1202-2020-4-107-112
Abstract
The purpose of the research is to substantiate the choice of enzyme preparations Promod 523MDP, Promod 439L, Flavorpro 766MDP, and Flavorpro 750MDP (Biocatalysts Limited, UK) and to determine the effective time of whey proteins hydrolysis in an ultrafiltration concentrate (UF-concentrate) of cheese whey for reducing their allergenicity based on the analysis of kinetic constants of the proteolysis reaction. Experimental studies were carried out with samples of cheese whey UF-concentrate with a total protein mass fraction at least 3.0% obtained with the use of industrial ultrafiltration unit MMS Swissflow UF with ceramic membranes under the conditions of the PSC Dairy Plant “Voronezhskii”. They were preliminarily subjected to enzymatic hydrolysis for 8 hours at a constant temperature, based on the dosage data, the optimum pH and the temperature of the used enzymes, recommended by the manufacturer. The specificity constant Vmax/Km was used to estimate the effectiveness of the enzyme preparations, which characterizes the constants of all stages of the hydrolysis reaction. The highest proteolysis rate has a mixture of Promod 439L and Flavorpro 766MDP in the ratio of 1.5 and 3.0%, respectively, of the total protein content in the substrate. Microscopy results showed an increasing in the solubility of nitrogen-containing components after hydrolysis due to a decreasing in hydrophobic areas on the surface of peptides. The resulting hydrolysate can be applied in the technology of a wide range of dairy products to reduce their residual antigenicity by partially replacing skim milk in the formulation.
About the Authors
E. I. Melnikova
Voronezh State University of Engineering Technologies
Russian Federation
Dr. Sci. (Engin.), professor, animal-derived food technology department, Revolution Av., 19 Voronezh, 394036, Russia
Russian Federation
Dr. Sci. (Engin.), professor, animal-derived food technology department, Revolution Av., 19 Voronezh, 394036, Russia
E. V. Bogdanova
Voronezh State University of Engineering Technologies
Cand. Sci. (Engin.), associate professor, animal-derived food technology department, Revolution Av., 19 Voronezh, 394036, Russia
Cand. Sci. (Engin.), associate professor, animal-derived food technology department, Revolution Av., 19 Voronezh, 394036, Russia
References
1. Contesini F.J., de Melo R.R., Sato H.H. An overview of Bacillus proteases: from production to application. Critical Reviews in Biotechnology. 2018. vol. 38. no. 3. pp. 321–334. doi:10.1080/07388551.2017.1354354.
2. Kostenko K., Bratsikhin A., Borisenko A., Salmanova D. et al. Computer modeling of whey protein ?-lactoglobulin behavior in the activated liquid systems. Journal of Hygienic Engineering and Design. 2017. vol. 20. pp. 70–74.
3. Vorob’ev M.M. Proteolysis of ?-lactoglobulin by trypsin: simulation by two-step model and experimental verification by intrinsic tryptophan. Fluorescence Symmetry. 2019. vol. 11. pp. 153. doi:10.3390/sym11020153.
4. Tolkacheva A.A., Cherenkov D.A., Korneeva O.S., Ponomarev P.G. Industrial enzymes – overview of the market for enzyme preparations and prospects for its development. Proceedings of VSUET. 2017. vol. 79. no. 4 (74). pp. 197–203. doi: 10.20914/2310–1202–2017–4–197–203. (in Russian).
5. Ryabtseva S.A., Khramtsov A.G., Budkevich R.O., Anisimov G.S. et. al. Physiological effects, mechanisms of action and application of lactulose. Voprosy Pitaniya. 2020. vol. 89. no. 2. pp. 5–20. doi: 10.24411/0042–8833–2020–10012. (in Russian).
6. Vorob’ev M.M., Raob N.M., Kochetkova K.A. Kinetic modeling of demasking and hydrolysis of peptide bonds during proteolysis of ?-lactoglobulin by trypsin. Doklady Akademii Nauk. 2016. vol. 471. no. 4. pp. 487–491.
7. Ponomareva N.V., Mel'nikova E.I., Bogdanova E.V. Bioconversion of milk proteins in order to reduce residual antigenicity. Biotechnology. 2015. vol. 31. no. 1. pp. 70–74.
8. Ponomareva N.V. Melnikova E.I. Composition and properties of ?-lactoglobulin hydrolysate with reduced residual antigenicity. Dairy Industry. 2015. no. 8. pp. 46–47. (in Russian).
9. Souza P.M., Aliakbarian B., Ferreira Filho E.X., Magalh?es P.O. et al. Kinetic and thermodynamic studies of a novel acid protease from Aspergillus foetidus. International Journal of Biological Macromolecules. 2015. vol. 81. no. 11. pp. 17–21.
10. Choi B., Rempala G.A., Kim J.K. Beyond the Michaelis-Menten equation: accurate and efcient estimation of enzyme kinetic parameters. Scientific Reports. 2017. vol. 17018. doi:10.1038/s41598–017–17072 z.
11. Schulte P.M. The effects of temperature on aerobic metabolism: towards a mechanistic understanding of the responses of ectotherms to a changing environment. Journal of Experimental Biology. 2015. vol. 218. pp. 1856–1866. doi: 10.1242/jeb.118851.
12. Vinnov A., Bal'-Prilipko L. Selection of enzymes for hydrolysis of industrial protein substrates. Food Industry of the Agroindustrial Complex. 2013. no. 3 (23). pp. 9–13. (in Russian).
13. Zainol N., Ismail S.N. Evaluation of enzyme kinetic parameters to produce methanol using Michaelis-Menten equation. Bulletin of Chemical Reaction Engineering & Catalysis. 2019. vol. 14. no. 2. pp. 436–442. doi: 10.9767/bcrec.14.2.3317.436–442.
14. Acquah C., Stefano E.D., Udenigwe C.C. Role of hydrophobicity in food peptide functionality and bioactivity. Journal of Food Bioactives. 2018. vol. 4. pp. 88–98. doi: 10.31665/JFB.2018.4164.
15. MacDonald A., Singh R.H., Rocha J.C., van Spronsen F.J. Optimising amino acid absorption: essential to improve nitrogen balance and metabolic control in phenylketonuria. Nutrition Research Reviews. 2019. vol. 32 (1). pp. 70–78. doi: 10.1017/S0954422418000173.
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For citations:
Melnikova E.I., Bogdanova E.V. Estimation of the kinetic parameters of whey proteins proteolysis in the UF-concentrate of cheese whey. Proceedings of the Voronezh State University of Engineering Technologies. 2020;82(4):107-112. (In Russ.) https://doi.org/10.20914/2310-1202-2020-4-107-112
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