foodsystFood systemsПищевые системы2618-97712618-7272Федеральный научный центр пищевых систем им. В.М. Горбатова РАН10.21323/2618-9771-2026-9-1-32-43foodsyst-998Research ArticleСтатьиAmino acid composition and biological value of whey protein hydrolysates fractionated by cascade-selective membrane filtrationАминокислотный состав и биологическая ценность гидролизатов сывороточных белков, фракционированных каскадно-селективной мембранной фильтрациейhttps://orcid.org/0000-0002-3227-8133КручининА. Г.KruchininA. G.

Кручинин Александр Геннадьевич — доктор технических наук, старший научный сотрудник, экспериментальная клиника — лаборатория биологически активных веществ животного происхождения

109316, Москва, ул. Талалихина, 26

Aleksandr G. Kruchinin, Doctor of Technical Sciences, Senior Researcher, Experimental Clinic-Laboratory of Biologically Active Substances of Animal Origin

26, Talalikhina str., 109316, Moscow

 

a.kruchinin@fncps.ruhttps://orcid.org/0000-0003-4544-4433СпиринаМ. Е.SpirinaM. E.

Спирина Мария Евгеньевна — инженер-исследователь, экспериментальная клиника  — лаборатория биологически активных веществ животного происхождения

109316, Москва, ул. Талалихина, 26

Maria E. Spirina, Research Engineer, Experimental Clinic-Laboratory of Biologically Active Substances of Animal Origin

26, Talalikhina str., 109316, Moscow

m.spirina@fncps.ruФедеральный научный центр пищевых систем им. В. М. ГорбатоваРоссияV. M. Gorbatov Federal Research Center for Food SystemsRussian Federation202617042026913243Copyright © Kruchinin A.G., Spirina M.E., 20262026Кручинин А.Г., Спирина М.Е.Kruchinin A.G., Spirina M.E.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.fsjour.com/jour/article/view/998

This article discusses the application of industrial cascade-selective membrane filtration techniques for the purification of enzymatic hydrolysates of whey proteins and the fractionation of bioactive peptides. The main focus is on a comprehensive assessment of the impact of this technological process on the amino acid composition, biological value, and in vitro digestion kinetics of the obtained peptide fractions in comparison with intact proteins. Native and heat-denatured whey protein concentrates were hydrolyzed using a two-enzyme system (Alcalase and Formea CTL) and fractionated by ultrafiltration, nanofiltration, and reverse osmosis. The key results showed that the developed cascade-selective ultra-nanodiafiltration process effectively removes highmolecular-weight protein compounds, minerals, and free amino acids. The hydrolysates retain a peptide fraction with a molecular weight of 0.4–5 kDa, which exhibits maximal antioxidant (1280 and 979 µM TE/g protein equivalent) and ACE-inhibitory (IC50 =0.10 and 0.19 mg/mL) activities. The purified hydrolysates were characterized by a balanced amino acid profile (EAA/NEAA ratio of 0.77–0.78), a high content of BCAA (19.07–19.20 g/100 g protein), and an increased coefficient of utility (0.71–0.72) compared to the initial whey protein concentrates. Digestibility assessment using the INFOGEST 2.0 model confirmed high bioavailability, as well as faster and more complete digestion of the fractionated hydrolysates compared to intact proteins. The obtained results open prospects for the industrial scaling of the technology for producing whey protein hydrolysates and their use as functional ingredients. Such ingredients can be applied in the development of functional and specialized food products, including protein formulas for sports nutrition, nutritional support for metabolic syndrome and age-related sarcopenia, as well as for the correction of chronic oxidative stress and the prevention of hypertension.

В данной статье обсуждается вопрос использования промышленных методов каскадно-селективной мембранной биоактивные фильтрации для очистки ферментативных гидролизатов сывороточных белков и фракционирования биоактивных пептидов. Основное внимание уделено комплексной оценке влияния данного технологического процесса на аминокислотный состав, биологическую ценность и кинетику переваривания in vitro полученных пептидных фракций в сравнении с интактными белками. Нативный и термоденатурированный концентрат сывороточных белков гидролизовали биферментной композицией (Alcalase и Formea CTL) и фракционировали методами ультра-, нанофильтрации и обратного осмоса. Основные результаты показали, что разработанный процесс каскадно-селективной ультра-нанодиафильтрации позволяет эффективно удалять высокомолекулярные белковые соединения, минеральные вещества и свободные аминокислоты. При этом в составе гидролизатов сохраняется фракция пептидов с молекулярной массой 0,4–5 кДа, обладающая максимальными антиоксидантной (1280 и 979 мкМ ТЭ/г БЭ) и АПФ-ингибирующей (IC50 = 0,10 и 0,19 мг/мл) активностями. Очищенные гидролизаты характеризовались сбалансированным аминокислотным профилем (отношение НАК/ЗАК 0,77–0,78), высоким содержанием BCAA (19,07–19,20 г/100 г белка) и повышенным коэффициентом утилитарности (0,71–0,72) по сравнению с исходными концентратами сывороточных белков. Оценка перевариваемости на модели INFOGEST 2.0 подтвердила высокую биодоступность, более быстрое и полное переваривание фракционированных гидролизатов по сравнению интактными белками. Полученные результаты открывают перспективы промышленного масштабирования технологии производства гидролизатов сывороточных белков и их использования в качестве функциональных ингредиентов. Такие ингредиенты могут применяться при создании продуктов функционального и специализированного питания, включая протеиновые формулы для спортивного питания, нутритивной поддержки при метаболическом синдроме и возрастной саркопении, а также для коррекции хронического оксидативного стресса и профилактики гипертонии.

гидролизбиоактивные пептидыультрафильтрациянанодиафильтрациясвободные аминокислотыминеральные веществаперевариваемостьhydrolysisbioactive peptidesultrafiltrationnanodiafiltrationfree amino acidsmineralsdigestibilityИсследования проводились в рамках государственного задания ФГБНУ «ФНЦ пищевых систем им. В. М. Горбатова» РАН FGUS-2024.0001.The article was published as part of the research topic No. FGUS‑2024–0001 of the state assignment of the V. M. Gorbatov Federal Research Center for Food Systems of RAS.ReferencesMeganaharshini, M., Sudhakar, V., Dhivya Bharathi, N., Deepak, S. (2023). Review on recent trends in the application of protein concentrates and isolates — A food industry perspective. Food and Humanity, 1, 308–325. https://doi.org/10.1016/j.foohum.2023.05.022Meganaharshini, M., Sudhakar, V., Dhivya Bharathi, N., Deepak, S. (2023). Review on recent trends in the application of protein concentrates and isolates — A food industry perspective. Food and Humanity, 1, 308–325. https://doi.org/10.1016/j.foohum.2023.05.022Lagrange, V., Whitsett, D., Burris, C. (2015). Global market for dairy proteins. Journal of Food Science, 80(S1), A16-A22. https://doi.org/10.1111/1750-3841.12801Lagrange, V., Whitsett, D., Burris, C. (2015). Global market for dairy proteins. Journal of Food Science, 80(S1), A16-A22. https://doi.org/10.1111/1750-3841.12801Smithers, G. W. (2015). Whey-ing up the options — Yesterday, today and tomorrow. International Dairy Journal, 48, 2–14. https://doi.org/10.1016/j.idairyj.2015.01.011Smithers, G. W. (2015). Whey-ing up the options — Yesterday, today and tomorrow. International Dairy Journal, 48, 2–14. https://doi.org/10.1016/j.idairyj.2015.01.011Dullius, A., Goettert, M. I., de Souza, C. F. V. (2018). Whey protein hydrolysates as a source of bioactive peptides for functional foods — Biotechnological facilitation of industrial scale-up. Journal of Functional Foods, 42, 58–74. https://doi.org/10.1016/j.jff.2017.12.063Dullius, A., Goettert, M. I., de Souza, C. F. V. (2018). Whey protein hydrolysates as a source of bioactive peptides for functional foods — Biotechnological facilitation of industrial scale-up. Journal of Functional Foods, 42, 58–74. https://doi.org/10.1016/j.jff.2017.12.063Pan, D., Zhang, Z., Liu, N., Ashaolu, T. J. (2025). Whey protein nutrition in sports: Action mechanisms and gaps in research. Current Nutrition Reports, 14(1), Article 84. https://doi.org/10.1007/s13668-025-00673-wPan, D., Zhang, Z., Liu, N., Ashaolu, T. J. (2025). Whey protein nutrition in sports: Action mechanisms and gaps in research. Current Nutrition Reports, 14(1), Article 84. https://doi.org/10.1007/s13668-025-00673-wGonzález-Weller, D., Paz-Montelongo, S., Bethencourt-Barbuzano, E., Niebla-Canelo, D., Alejandro-Vega, S., Gutiérrez, Á. J. et al. (2023). Proteins and minerals in whey protein supplements. Foods, 12(11), Article 2238. https://doi.org/10.3390/foods12112238González-Weller, D., Paz-Montelongo, S., Bethencourt-Barbuzano, E., Niebla-Canelo, D., Alejandro-Vega, S., Gutiérrez, Á. J. et al. (2023). Proteins and minerals in whey protein supplements. Foods, 12(11), Article 2238. https://doi.org/10.3390/foods12112238Lam, F.C., Bukhsh, A., Rehman, H., Waqas, M. K., Shahid, N., Khaliel, A. M. et al. (2019). Efficacy and safety of whey protein supplements on vital sign and physical performance among athletes: A network meta-analysis. Frontiers in Pharmacology, 10, Article 317. https://doi.org/10.3389/fphar.2019.00317Lam, F.C., Bukhsh, A., Rehman, H., Waqas, M. K., Shahid, N., Khaliel, A. M. et al. (2019). Efficacy and safety of whey protein supplements on vital sign and physical performance among athletes: A network meta-analysis. Frontiers in Pharmacology, 10, Article 317. https://doi.org/10.3389/fphar.2019.00317Martinez Galan, B. S., Giolo De Carvalho, F., Carvalho, S. C., Cunha Brandao, C. F., Morhy Terrazas, S. I., Abud, G. F. et al. (2021). Casein and whey protein in the breast milk ratio: Could it promote protein metabolism enhancement in physically active adults? Nutrients, 13(7), Article 2153. https://doi.org/10.3390/nu13072153Martinez Galan, B. S., Giolo De Carvalho, F., Carvalho, S. C., Cunha Brandao, C. F., Morhy Terrazas, S. I., Abud, G. F. et al. (2021). Casein and whey protein in the breast milk ratio: Could it promote protein metabolism enhancement in physically active adults? Nutrients, 13(7), Article 2153. https://doi.org/10.3390/nu13072153Zhao, C., Chen, N., Ashaolu, T. J. (2022). Whey proteins and peptides in health-promoting functions–A review. International Dairy Journal, 126, Article 105269. https://doi.org/10.1016/j.idairyj.2021.105269Zhao, C., Chen, N., Ashaolu, T. J. (2022). Whey proteins and peptides in health-promoting functions–A review. International Dairy Journal, 126, Article 105269. https://doi.org/10.1016/j.idairyj.2021.105269Camargo, L.R., Doneda, D., Oliveira, V. R. (2020). Whey protein ingestion in elderly diet and the association with physical, performance and clinical outcomes. Experimental Gerontology, 137, Article 110936. https://doi.org/10.1016/j.exger.2020.110936Camargo, L.R., Doneda, D., Oliveira, V. R. (2020). Whey protein ingestion in elderly diet and the association with physical, performance and clinical outcomes. Experimental Gerontology, 137, Article 110936. https://doi.org/10.1016/j.exger.2020.110936Polberger, S., Räihä, N. C., Juvonen, P., Moro, G. E., Minoli, I., Warm, A. (1999). Individualized protein fortification of human milk for preterm infants: Comparison of ultrafiltrated human milk protein and a bovine whey fortifier. Journal of Pediatric Gastroenterology and Nutrition, 29(3), 332–338. https://doi.org/10.1002/j.1536-4801.1999.tb02424.xPolberger, S., Räihä, N. C., Juvonen, P., Moro, G. E., Minoli, I., Warm, A. (1999). Individualized protein fortification of human milk for preterm infants: Comparison of ultrafiltrated human milk protein and a bovine whey fortifier. Journal of Pediatric Gastroenterology and Nutrition, 29(3), 332–338. https://doi.org/10.1002/j.1536-4801.1999.tb02424.xde Figueiredo Furtado, G., da Silva Carvalho, A. G., Hubinger, M. D. (2021). Model infant formulas: Influence of types of whey proteins and oil composition on emulsion and powder properties. Journal of Food Engineering, 292, Article 110256. https://doi.org/10.1016/j.jfoodeng.2020.110256de Figueiredo Furtado, G., da Silva Carvalho, A. G., Hubinger, M. D. (2021). Model infant formulas: Influence of types of whey proteins and oil composition on emulsion and powder properties. Journal of Food Engineering, 292, Article 110256. https://doi.org/10.1016/j.jfoodeng.2020.110256Chen, Y., Callanan, M., Shanahan, C., Tobin, J., Gamon, L. F., Davies, M. J. et al. (2021). The use of membrane filtration to increase native whey proteins in infant formula. Dairy, 2(4), 515–529. https://doi.org/10.3390/dairy2040041Chen, Y., Callanan, M., Shanahan, C., Tobin, J., Gamon, L. F., Davies, M. J. et al. (2021). The use of membrane filtration to increase native whey proteins in infant formula. Dairy, 2(4), 515–529. https://doi.org/10.3390/dairy2040041Chitti, W., Insin, P., Prueksaritanond, N. (2025). Effect of whey protein supplementation on postoperative outcomes after gynecological cancer surgery: A randomized controlled trial. World Journal of Oncology, 16(1), 70–82. https://doi.org/10.14740/wjon1990Chitti, W., Insin, P., Prueksaritanond, N. (2025). Effect of whey protein supplementation on postoperative outcomes after gynecological cancer surgery: A randomized controlled trial. World Journal of Oncology, 16(1), 70–82. https://doi.org/10.14740/wjon1990Khalooeifard, R., Oraee-Yazdani, S., Keikhaee, M., Shariatpanahi, Z. V. (2022). Protein supplement and enhanced recovery after posterior spine fusion surgery: A randomized, double-b lind, placebo-controlled trial. Clinical Spine Surgery, 35(3), E356-E362. https://doi.org/10.1097/BSD.0000000000001222Khalooeifard, R., Oraee-Yazdani, S., Keikhaee, M., Shariatpanahi, Z. V. (2022). Protein supplement and enhanced recovery after posterior spine fusion surgery: A randomized, double-b lind, placebo-controlled trial. Clinical Spine Surgery, 35(3), E356-E362. https://doi.org/10.1097/BSD.0000000000001222Niitsu, M., Ichinose, D., Hirooka, T., Mitsutomi, K., Morimoto, Y., Sarukawa, J. et al. (2016). Effects of combination of whey protein intake and rehabilitation on muscle strength and daily movements in patients with hip fracture in the early postoperative period. Clinical Nutrition, 35(4), 943–949. https://doi.org/10.1016/j.clnu.2015.07.006Niitsu, M., Ichinose, D., Hirooka, T., Mitsutomi, K., Morimoto, Y., Sarukawa, J. et al. (2016). Effects of combination of whey protein intake and rehabilitation on muscle strength and daily movements in patients with hip fracture in the early postoperative period. Clinical Nutrition, 35(4), 943–949. https://doi.org/10.1016/j.clnu.2015.07.006Kleekayai, T., O’Neill, A., Clarke, S., Holmes, N., O’Sullivan, B., FitzGerald, R. J. (2022). Contribution of hydrolysis and drying conditions to whey protein hydrolysate characteristics and in vitro antioxidative properties. Antioxidants, 11(2), Article 399. https://doi.org/10.3390/antiox11020399Kleekayai, T., O’Neill, A., Clarke, S., Holmes, N., O’Sullivan, B., FitzGerald, R. J. (2022). Contribution of hydrolysis and drying conditions to whey protein hydrolysate characteristics and in vitro antioxidative properties. Antioxidants, 11(2), Article 399. https://doi.org/10.3390/antiox11020399Pang, L., Huang, Z., Li, R., Su, Y., Shi, J., Yang, X. et al. (2025). Reducing the allergenicity of whey proteins while improving their functional properties and bioactivity using combined enzymes. Food Chemistry, 481, Article 144052. https://doi.org/10.1016/j.foodchem.2025.144052Pang, L., Huang, Z., Li, R., Su, Y., Shi, J., Yang, X. et al. (2025). Reducing the allergenicity of whey proteins while improving their functional properties and bioactivity using combined enzymes. Food Chemistry, 481, Article 144052. https://doi.org/10.1016/j.foodchem.2025.144052Kruchinin, A., Barkovskaya, I., Illarionova, E., Bolshakova, E., Turovskaya, S., Galstyan, A. (2025). Effect of enzymatic degradation of proteins on technological properties of whey powdered products. International Journal of Dairy Technology, 78(2), Article e70005. https://doi.org/10.1111/1471-0307.70005Kruchinin, A., Barkovskaya, I., Illarionova, E., Bolshakova, E., Turovskaya, S., Galstyan, A. (2025). Effect of enzymatic degradation of proteins on technological properties of whey powdered products. International Journal of Dairy Technology, 78(2), Article e70005. https://doi.org/10.1111/1471-0307.70005Cui, Q., Li, Y., Li, T., Yu, J., Shen, G., Sun, X. et al. (2024). Characterization of peptide profiles and the hypoallergenic and high antioxidant activity of whey protein hydrolysate prepared using different hydrolysis modes. Foods, 13(18), Article 2978. https://doi.org/10.3390/foods13182978Cui, Q., Li, Y., Li, T., Yu, J., Shen, G., Sun, X. et al. (2024). Characterization of peptide profiles and the hypoallergenic and high antioxidant activity of whey protein hydrolysate prepared using different hydrolysis modes. Foods, 13(18), Article 2978. https://doi.org/10.3390/foods13182978Espindola, J. S., Taccola, M. F., da Silva, V. S. N., Dos Santos, L. D., Rossini, B. C., Mendonca, B. C. et al. (2023). Digestion-resistant whey peptides promote antioxidant effect on Caco-2 cells. Food Research International, 173, Article 113291. https://doi.org/10.1016/j.foodres.2023.113291Espindola, J. S., Taccola, M. F., da Silva, V. S. N., Dos Santos, L. D., Rossini, B. C., Mendonca, B. C. et al. (2023). Digestion-resistant whey peptides promote antioxidant effect on Caco-2 cells. Food Research International, 173, Article 113291. https://doi.org/10.1016/j.foodres.2023.113291Cui, L., Yang, G., Lu, S., Zeng, X., He, J., Guo, Y. et al. (2022). Antioxidant peptides derived from hydrolyzed milk proteins by Lactobacillus strains: A BIOPEPUWM database-b ased analysis. Food Research International, 156, Article 111339. https://doi.org/10.1016/j.foodres.2022.111339Cui, L., Yang, G., Lu, S., Zeng, X., He, J., Guo, Y. et al. (2022). Antioxidant peptides derived from hydrolyzed milk proteins by Lactobacillus strains: A BIOPEPUWM database-b ased analysis. Food Research International, 156, Article 111339. https://doi.org/10.1016/j.foodres.2022.111339Jiang, S., Mo, F., Li, W., Yang, S., Li, C., Jiang, L. (2024). Deep learning-driven optimization of antihypertensive properties from whey protein hydrolysates: A multienzyme approach. Journal of Agricultural and Food Chemistry, 73(2), 1373–1388. https://doi.org/10.101610.1021/acs.jafc.4c10830Jiang, S., Mo, F., Li, W., Yang, S., Li, C., Jiang, L. (2024). Deep learning-driven optimization of antihypertensive properties from whey protein hydrolysates: A multienzyme approach. Journal of Agricultural and Food Chemistry, 73(2), 1373–1388. https://doi.org/10.101610.1021/acs.jafc.4c10830Dou, P., Li, X., Zou, X., Wang, K., Yao, L., Sun, Z. et al. (2024). Antihypertensive effects of whey protein hydrolysate involve reshaping the gut microbiome in spontaneously hypertension rats. Food Science and Human Wellness, 13(4), 1974–1986. https://doi.org/10.26599/FSHW.2022.9250164Dou, P., Li, X., Zou, X., Wang, K., Yao, L., Sun, Z. et al. (2024). Antihypertensive effects of whey protein hydrolysate involve reshaping the gut microbiome in spontaneously hypertension rats. Food Science and Human Wellness, 13(4), 1974–1986. https://doi.org/10.26599/FSHW.2022.9250164Mansinhbhai, C. H., Sakure, A., Liu, Z., Maurya, R., Das, S., Basaiawmoit, B. et al. (2023). Anti-inflammatory, ACE inhibitory, antioxidative activities and release of novel antihypertensive and antioxidative peptides from whey protein hydrolysate with molecular interactions. Journal of the American Nutrition Association, 42(4), 371–385. https://doi.org/10.1080/07315724.2022.2052201Mansinhbhai, C. H., Sakure, A., Liu, Z., Maurya, R., Das, S., Basaiawmoit, B. et al. (2023). Anti-inflammatory, ACE inhibitory, antioxidative activities and release of novel antihypertensive and antioxidative peptides from whey protein hydrolysate with molecular interactions. Journal of the American Nutrition Association, 42(4), 371–385. https://doi.org/10.1080/07315724.2022.2052201Tovar-Jiménez, X., Téllez-J urado, A., Gómez-Aldapa, C. A., Mercado-Flores, Y., Arana-Cuenca, A. (2021). Antioxidant and antihypertensive activity of bovine whey protein concentrate enzymatic hydrolysates. Biotecnia, 23(1), 161–169. https://doi.org/10.18633/biotecnia.v23i1.1321Tovar-Jiménez, X., Téllez-J urado, A., Gómez-Aldapa, C. A., Mercado-Flores, Y., Arana-Cuenca, A. (2021). Antioxidant and antihypertensive activity of bovine whey protein concentrate enzymatic hydrolysates. Biotecnia, 23(1), 161–169. https://doi.org/10.18633/biotecnia.v23i1.1321Alvarez-Chávez, S. I., Reyes-Ramírez, I., González-Olivares, L. G., Pérez-Flores, J. G., Contreras-López, E., Olvera-Rosales, L. B. et al. (2025). Hydrolysates from a whey protein concentrate are a promising functional ingredient for diabetes control via DPP-IV inhibition. Biology and Life Sciences Forum, 40(1), Article 10. https://doi.org/10.3390/blsf2024040010Alvarez-Chávez, S. I., Reyes-Ramírez, I., González-Olivares, L. G., Pérez-Flores, J. G., Contreras-López, E., Olvera-Rosales, L. B. et al. (2025). Hydrolysates from a whey protein concentrate are a promising functional ingredient for diabetes control via DPP-IV inhibition. Biology and Life Sciences Forum, 40(1), Article 10. https://doi.org/10.3390/blsf2024040010Bunsroem, K., Prinyawiwatkul, W., Thaiudom, S. (2022). The influence of whey protein heating parameters on their susceptibility to digestive enzymes and the antidiabetic activity of hydrolysates. Foods, 11(6), Article 829. https://doi.org/10.3390/foods11060829Bunsroem, K., Prinyawiwatkul, W., Thaiudom, S. (2022). The influence of whey protein heating parameters on their susceptibility to digestive enzymes and the antidiabetic activity of hydrolysates. Foods, 11(6), Article 829. https://doi.org/10.3390/foods11060829Vadher, K. R., Sakure, A. A., Mankad, P. M., Rawat, A., Bishnoi, M., Kondepudi, K. K. et al. (2025). A comparative study on antidiabetic and anti-inflammatory activities of fermented whey and soy protein isolates and the release of biofunctional peptides: An in vitro and in silico studies. Journal of the Science of Food and Agriculture, 105(7), 3826–3842. https://doi.org/10.1002/jsfa.14154Vadher, K. R., Sakure, A. A., Mankad, P. M., Rawat, A., Bishnoi, M., Kondepudi, K. K. et al. (2025). A comparative study on antidiabetic and anti-inflammatory activities of fermented whey and soy protein isolates and the release of biofunctional peptides: An in vitro and in silico studies. Journal of the Science of Food and Agriculture, 105(7), 3826–3842. https://doi.org/10.1002/jsfa.14154Li, Y., Fan, Y., Liu, J., Meng, Z., Huang, A., Xu, F. et al. (2023). Identification, characterization and in vitro activity of hypoglycemic peptides in whey hydrolysates from rubing cheese by-product. Food Research International, 164, Article 112382. https://doi.org/10.1016/j.foodres.2022.112382Li, Y., Fan, Y., Liu, J., Meng, Z., Huang, A., Xu, F. et al. (2023). Identification, characterization and in vitro activity of hypoglycemic peptides in whey hydrolysates from rubing cheese by-product. Food Research International, 164, Article 112382. https://doi.org/10.1016/j.foodres.2022.112382Singh, A., Duche, R. T., Wandhare, A. G., Sian, J. K., Singh, B. P., Sihag, M. K. et al. (2023). Milk-derived antimicrobial peptides: Overview, applications, and future perspectives. Probiotics and Antimicrobial Proteins, 15(1), 44–62. https://doi.org/10.1007/s12602-022-10004-ySingh, A., Duche, R. T., Wandhare, A. G., Sian, J. K., Singh, B. P., Sihag, M. K. et al. (2023). Milk-derived antimicrobial peptides: Overview, applications, and future perspectives. Probiotics and Antimicrobial Proteins, 15(1), 44–62. https://doi.org/10.1007/s12602-022-10004-yInnocente, N., Calligaris, S., Di Filippo, G., Melchior, S., Marino, M., Nicoli, M. C. (2023). Process design for the production of peptides from whey protein isolate with targeted antimicrobial functionality. International Journal of Food Science and Technology, 58(5), 2505–2517. https://doi.org/10.1111/ijfs.16393Innocente, N., Calligaris, S., Di Filippo, G., Melchior, S., Marino, M., Nicoli, M. C. (2023). Process design for the production of peptides from whey protein isolate with targeted antimicrobial functionality. International Journal of Food Science and Technology, 58(5), 2505–2517. https://doi.org/10.1111/ijfs.16393Dalaka, E., Stefos, G. C., Politis, I., Theodorou, G. (2025). Immunomodulatory properties of sweet whey-derived peptides in THP-1 macrophages. Molecules, 30(6), Article 1261. https://doi.org/10.3390/molecules30061261Dalaka, E., Stefos, G. C., Politis, I., Theodorou, G. (2025). Immunomodulatory properties of sweet whey-derived peptides in THP-1 macrophages. Molecules, 30(6), Article 1261. https://doi.org/10.3390/molecules30061261Kiewiet, M. B., Dekkers, R., Ulfman, L. H., Groeneveld, A., de Vos, P., Faas, M. M. (2018). Immunomodulating protein aggregates in soy and whey hydrolysates and their resistance to digestion in an in vitro infant gastrointestinal model: New insights in the mechanism of immunomodulatory hydrolysates. Food Function, 9(1), 604–613. https://doi.org/10.1039/C7FO01537BKiewiet, M. B., Dekkers, R., Ulfman, L. H., Groeneveld, A., de Vos, P., Faas, M. M. (2018). Immunomodulating protein aggregates in soy and whey hydrolysates and their resistance to digestion in an in vitro infant gastrointestinal model: New insights in the mechanism of immunomodulatory hydrolysates. Food Function, 9(1), 604–613. https://doi.org/10.1039/C7FO01537BReyes-Díaz, A., González-Córdova, A. F., Hernández-Mendoza, A., Reyes-Díaz, R., Vallejo-Cordoba, B. (2018). Immunomodulation by hydrolysates and peptides derived from milk proteins. International Journal of Dairy Technology, 71(1), 1–9. https://doi.org/10.1111/1471-0307.12421Reyes-Díaz, A., González-Córdova, A. F., Hernández-Mendoza, A., Reyes-Díaz, R., Vallejo-Cordoba, B. (2018). Immunomodulation by hydrolysates and peptides derived from milk proteins. International Journal of Dairy Technology, 71(1), 1–9. https://doi.org/10.1111/1471-0307.12421Mora, L., Toldrá, F. (2023). Advanced enzymatic hydrolysis of food proteins for the production of bioactive peptides. Current Opinion in Food Science, 49, Article 100973. https://doi.org/10.1016/j.cofs.2022.100973Mora, L., Toldrá, F. (2023). Advanced enzymatic hydrolysis of food proteins for the production of bioactive peptides. Current Opinion in Food Science, 49, Article 100973. https://doi.org/10.1016/j.cofs.2022.100973Martínez-Araiza, G., Castaño-Tostado, E., Amaya-Llano, S. L., Regalado-González, C., Martínez-Vera, C., Ozimek, L. (2012). Modeling of enzymatic hydrolysis of whey proteins. Food and Bioprocess Technology, 5(6), 2596–2601. https://doi.org/10.1007/s11947-011-0624-5Martínez-Araiza, G., Castaño-Tostado, E., Amaya-Llano, S. L., Regalado-González, C., Martínez-Vera, C., Ozimek, L. (2012). Modeling of enzymatic hydrolysis of whey proteins. Food and Bioprocess Technology, 5(6), 2596–2601. https://doi.org/10.1007/s11947-011-0624-5Morifuji, M., Ishizaka, M., Baba, S., Fukuda, K., Matsumoto, H., Koga, J. et al. (2010). Comparison of different sources and degrees of hydrolysis of dietary protein: Effect on plasma amino acids, dipeptides, and insulin responses in human subjects. Journal of Agricultural and Food Chemistry, 58(15), 8788–8797. https://doi.org/10.1021/jf101912nMorifuji, M., Ishizaka, M., Baba, S., Fukuda, K., Matsumoto, H., Koga, J. et al. (2010). Comparison of different sources and degrees of hydrolysis of dietary protein: Effect on plasma amino acids, dipeptides, and insulin responses in human subjects. Journal of Agricultural and Food Chemistry, 58(15), 8788–8797. https://doi.org/10.1021/jf101912nBoza, J. J., Moennoz, D., Vuichoud, J., Jarret, A. R., Gaudard-de-Weck, D., Ballevre, O. (2000). Protein hydrolysate vs free amino acid-based diets on the nutritional recovery of the starved rat. European Journal of Nutrition, 39(6), 237–243. https://doi.org/10.1007/s003940070001Boza, J. J., Moennoz, D., Vuichoud, J., Jarret, A. R., Gaudard-de-Weck, D., Ballevre, O. (2000). Protein hydrolysate vs free amino acid-based diets on the nutritional recovery of the starved rat. European Journal of Nutrition, 39(6), 237–243. https://doi.org/10.1007/s003940070001Douwes, J., Thorne, P., Pearce, N., Heederik, D. (2003). Bioaerosol health effects and exposure assessment: Progress and prospects. Annals of Occupational Hygiene, 47(3), 187–200. https://doi.org/10.1093/annhyg/meg032Douwes, J., Thorne, P., Pearce, N., Heederik, D. (2003). Bioaerosol health effects and exposure assessment: Progress and prospects. Annals of Occupational Hygiene, 47(3), 187–200. https://doi.org/10.1093/annhyg/meg032Yike, I. (2011). Fungal proteases and their pathophysiological effects. Mycopathologia, 171(5), 299–323. https://doi.org/10.1007/s11046-010-9386-2Yike, I. (2011). Fungal proteases and their pathophysiological effects. Mycopathologia, 171(5), 299–323. https://doi.org/10.1007/s11046-010-9386-2Henriques, M., Gomes, D., Pereira, C. (2017). Liquid whey protein concentrates produced by ultrafiltration as primary raw materials for thermal dairy gels. Food Technology and Biotechnology, 55(4), Article 454. https://doi.org/10.17113/ftb.55.04.17.5248Henriques, M., Gomes, D., Pereira, C. (2017). Liquid whey protein concentrates produced by ultrafiltration as primary raw materials for thermal dairy gels. Food Technology and Biotechnology, 55(4), Article 454. https://doi.org/10.17113/ftb.55.04.17.5248Курченко, В. П., Симоненко, Е. С., Сушинская, Н. В., Головач, Т. Н., Петров, А. Н., Симоненко, С. В. (2021). Идентификация кобыльего молока и его смеси с коровьим молоком методом ВЭЖХ-анализа. Техника и технология пищевых производств, 51(2), 402–412 https://doi.org/10.21603/2074-9414-2021-2-402-412Kurchenko, V. P., Simonenko, E. S., Sushynskaya, N. V., Halavach, T. N., Petrov, A. N., Simonenko, S. V. (2021). HPLC identification of mare’s milk and its mix with cow’s milk. Food Processing: Techniques and Technology, 51(2), 402–412. (In Russian)] https://doi.org/10.21603/2074-9414-2021-2-402-412Spellman, D., McEvoy, E., O’cuinn, G., FitzGerald, R. J. (2003). Proteinase and exopeptidase hydrolysis of whey protein: Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis. International Dairy Journal, 13(6), 447–453. https://doi.org/10.1016/S0958-6946(03)00053-0Spellman, D., McEvoy, E., O’cuinn, G., FitzGerald, R. J. (2003). Proteinase and exopeptidase hydrolysis of whey protein: Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis. International Dairy Journal, 13(6), 447–453. https://doi.org/10.1016/S0958-6946(03)00053-0Acid, T. (1979). Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. Journal of Agricultural and Food Chemistry, 27(6), 1257–1262. https://doi.org/10.1021/jf60226a042Acid, T. (1979). Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. Journal of Agricultural and Food Chemistry, 27(6), 1257–1262. https://doi.org/10.1021/jf60226a042Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9–10), 1231–1237. https://doi.org/10.1016/S0891-5849(98)00315-3Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9–10), 1231–1237. https://doi.org/10.1016/S0891-5849(98)00315-3Hernández-Ledesma, B., Martín-Álvarez, P. J., Pueyo, E. (2003). Assessment of the spectrophotometric method for determination of angiotensin-convertingenzyme activity: Influence of the inhibition type. Journal of Agricultural and Food Chemistry, 51(15), 4175–4179. https://doi.org/10.1021/jf034148oHernández-Ledesma, B., Martín-Álvarez, P. J., Pueyo, E. (2003). Assessment of the spectrophotometric method for determination of angiotensin-convertingenzyme activity: Influence of the inhibition type. Journal of Agricultural and Food Chemistry, 51(15), 4175–4179. https://doi.org/10.1021/jf034148oAraujo, M. C., Melo, R. L., Cesari, M. H., Juliano, M. A., Juliano, L., Carmona, A. K. (2000). Peptidase specificity characterization of C-and N-terminal catalytic sites of angiotensin I-converting enzyme. Biochemistry, 39(29), 8519–8525. https://doi.org/10.1021/bi9928905Araujo, M. C., Melo, R. L., Cesari, M. H., Juliano, M. A., Juliano, L., Carmona, A. K. (2000). Peptidase specificity characterization of C-and N-terminal catalytic sites of angiotensin I-converting enzyme. Biochemistry, 39(29), 8519–8525. https://doi.org/10.1021/bi9928905Vafin, R. R., Radaeva, I. A., Kruchinin, A. G., Illarionova, E. E., Bigaeva, A. V., Turovskaya, S. N. et al. (2021). κ-casein polymorphism effect on technological properties of dried milk. Foods and Raw Materials, 9(1), 95–105. http://doi.org/10.21603/2308-4057-2021-1-95-105Vafin, R. R., Radaeva, I. A., Kruchinin, A. G., Illarionova, E. E., Bigaeva, A. V., Turovskaya, S. N. et al. (2021). κ-casein polymorphism effect on technological properties of dried milk. Foods and Raw Materials, 9(1), 95–105. http://doi.org/10.21603/2308-4057-2021-1-95-105Brodkorb, A., Egger, L., Alminger, M., Alvito, P., Assunção, R., Ballance, S. et al. (2019). INFOGEST static in vitro simulation of gastrointestinal food digestion. Nature Protocols, 14(4), 991–1014. https://doi.org/10.1038/s41596-018-0119-1Brodkorb, A., Egger, L., Alminger, M., Alvito, P., Assunção, R., Ballance, S. et al. (2019). INFOGEST static in vitro simulation of gastrointestinal food digestion. Nature Protocols, 14(4), 991–1014. https://doi.org/10.1038/s41596-018-0119-1Чернуха, И. М., Василевская, Е. Р., Арюзина, М. А., Кусай, А., Полищук, Е. К. (2023). Комплексная модель in vitro и ex vivo для изучения трансформации белков в процессе пищеварения. Все о мясе, 4, 40–45. https://doi.org/10.21323/2071-2499-2023-4-10-10Chernukha, I. M., Vasilevskaya, E. R., Aryuzina, M. A., Qusay, A., Polishchuk, E. K. (2023). Complex in vitro and ex vivo model to study protein transformation during digestion. Vsyo o Myase, 4, 40–45. (In Russian)] https://doi.org/10.21323/2071-2499-2023-4-10-10Neelima, Sharma, R., Rajput, Y. S., Mann, B. (2013). Chemical and functional properties of glycomacropeptide (GMP) and its role in the detection of cheese whey adulteration in milk: A review. Dairy Science Technology, 93(1), 21–43. https://doi.org/10.1007/s13594-012-0095-0Neelima, Sharma, R., Rajput, Y. S., Mann, B. (2013). Chemical and functional properties of glycomacropeptide (GMP) and its role in the detection of cheese whey adulteration in milk: A review. Dairy Science Technology, 93(1), 21–43. https://doi.org/10.1007/s13594-012-0095-0Feeney, S., Joshi, L., Hickey, R. M. (2018). Biological roles and production technologies associated with bovine glycomacropeptide. Chapter in a book: Novel Proteins for Food, Pharmaceuticals and Agriculture: Sources, Applications and Advances. Wiley-Blackwell, 2018. https://doi.org/10.1002/9781119385332.ch1Feeney, S., Joshi, L., Hickey, R. M. (2018). Biological roles and production technologies associated with bovine glycomacropeptide. Chapter in a book: Novel Proteins for Food, Pharmaceuticals and Agriculture: Sources, Applications and Advances. Wiley-Blackwell, 2018. https://doi.org/10.1002/9781119385332.ch1Kolar, C. W., Brunner, J. R. (1970). Proteose-peptone fraction of bovine milk: Lacteal serum components 5 and 8-casein-associated glycoproteins. Journal of Dairy Science, 53(8), 997–1008. https://doi.org/10.3168/jds.S0022-0302(70)86335-4Kolar, C. W., Brunner, J. R. (1970). Proteose-peptone fraction of bovine milk: Lacteal serum components 5 and 8-casein-associated glycoproteins. Journal of Dairy Science, 53(8), 997–1008. https://doi.org/10.3168/jds.S0022-0302(70)86335-4Karamoko, G., Anihouvi, P., Blecker, C. (2013). The development of knowledge regarding the characteristics of the proteose peptone fraction of milk: Techno-functional and biological properties. A review. Biotechnology, Agronomy, and Society and Environment, 17(2), 373–382.Karamoko, G., Anihouvi, P., Blecker, C. (2013). The development of knowledge regarding the characteristics of the proteose peptone fraction of milk: Techno-functional and biological properties. A review. Biotechnology, Agronomy, and Society and Environment, 17(2), 373–382.Alavi, F., Ciftci, O. N. (2023). Purification and fractionation of bioactive peptides through membrane filtration: A critical and application review. Trends in Food Science Technology, 131, 118–128. https://doi.org/10.1016/j.tifs.2022.11.024Alavi, F., Ciftci, O. N. (2023). Purification and fractionation of bioactive peptides through membrane filtration: A critical and application review. Trends in Food Science Technology, 131, 118–128. https://doi.org/10.1016/j.tifs.2022.11.024Yang, G., Xing, W., Xu, N. (2003). Concentration polarization in spiral-wound nanofiltration membrane elements. Desalination, 154(1), 89–99. https://doi.org/10.1016/S0011-9164(03)00210-8Yang, G., Xing, W., Xu, N. (2003). Concentration polarization in spiral-wound nanofiltration membrane elements. Desalination, 154(1), 89–99. https://doi.org/10.1016/S0011-9164(03)00210-8Suwal, S., Roblet, C., Doyen, A., Amiot, J., Beaulieu, L., Legault, J. et al. (2014). Electrodialytic separation of peptides from snow crab by-product hydrolysate: Effect of cell configuration on peptide selectivity and local electric field. Separation and Purification Technology, 127, 29–38. https://doi.org/10.1016/j.seppur.2014.02.018Suwal, S., Roblet, C., Doyen, A., Amiot, J., Beaulieu, L., Legault, J. et al. (2014). Electrodialytic separation of peptides from snow crab by-product hydrolysate: Effect of cell configuration on peptide selectivity and local electric field. Separation and Purification Technology, 127, 29–38. https://doi.org/10.1016/j.seppur.2014.02.018Poulin, J. -F., Amiot, J., Bazinet, L. (2006). Simultaneous separation of acid and basic bioactive peptides by electrodialysis with ultrafiltration membrane. Journal of Biotechnology, 123(3), 314–328. https://doi.org/10.1016/j.jbiotec.2005.11.016Poulin, J. -F., Amiot, J., Bazinet, L. (2006). Simultaneous separation of acid and basic bioactive peptides by electrodialysis with ultrafiltration membrane. Journal of Biotechnology, 123(3), 314–328. https://doi.org/10.1016/j.jbiotec.2005.11.016Pramanik, B. K., Thangavadivel, K., Shu, L., Jegatheesan, V. (2016). A critical review of membrane crystallization for the purification of water and recovery of minerals. Reviews in Environmental Science and Biotechnology, 15(3), 411–439. https://doi.org/10.1007/s11157-016-9403-0Pramanik, B. K., Thangavadivel, K., Shu, L., Jegatheesan, V. (2016). A critical review of membrane crystallization for the purification of water and recovery of minerals. Reviews in Environmental Science and Biotechnology, 15(3), 411–439. https://doi.org/10.1007/s11157-016-9403-0Soral-Smietana, M., Zduñczyk, Z., Wronkowska, M., Juskiewicz, J., Zander, L. (2013). Mineral composition and bioavailability of calcium and phosphorus from acid whey concentrated by various membrane processes. Journal of Elementology, 18(1), 115–125. https://doi.org/10.5601/JELEM.2013.18.1.10Soral-Smietana, M., Zduñczyk, Z., Wronkowska, M., Juskiewicz, J., Zander, L. (2013). Mineral composition and bioavailability of calcium and phosphorus from acid whey concentrated by various membrane processes. Journal of Elementology, 18(1), 115–125. https://doi.org/10.5601/JELEM.2013.18.1.10Price, D., Jackson, K. G., Lovegrove, J. A., Givens, D. I. (2022). The effects of whey proteins, their peptides and amino acids on vascular function. Nutrition Bulletin, 47(1), 9–26. https://doi.org/10.1111/nbu.12543Price, D., Jackson, K. G., Lovegrove, J. A., Givens, D. I. (2022). The effects of whey proteins, their peptides and amino acids on vascular function. Nutrition Bulletin, 47(1), 9–26. https://doi.org/10.1111/nbu.12543Simonson, M., Boirie, Y., Guillet, C. (2020). Protein, amino acids and obesity treatment. Reviews in Endocrine and Metabolic Disorders, 21(3), 341–353. https://doi.org/10.1007/s11154-020-09574-5Simonson, M., Boirie, Y., Guillet, C. (2020). Protein, amino acids and obesity treatment. Reviews in Endocrine and Metabolic Disorders, 21(3), 341–353. https://doi.org/10.1007/s11154-020-09574-5Marshall, K. (2004). Therapeutic applications of whey protein. Alternative Medicine Review, 9(2), 136–156.Marshall, K. (2004). Therapeutic applications of whey protein. Alternative Medicine Review, 9(2), 136–156.Melnik, V., Ryabinina, E., Rodionova, K., Nalivayko, L., Кhimych, M. (2023). Chemical composition and amino acid profile of goose meat (Ukrainian large gray and large white breeds) in semi-intensive system of growing. Journal of Microbiology, Biotechnology and Food Sciences, 12(6), Article e9828. https://doi.org/10.55251/jmbfs.9828Melnik, V., Ryabinina, E., Rodionova, K., Nalivayko, L., Кhimych, M. (2023). Chemical composition and amino acid profile of goose meat (Ukrainian large gray and large white breeds) in semi-intensive system of growing. Journal of Microbiology, Biotechnology and Food Sciences, 12(6), Article e9828. https://doi.org/10.55251/jmbfs.9828Li, C., Nielsen, S. B., Engholm-K eller, K., Lund, M. N. (2022). Oxidation of whey proteins during thermal treatment characterized by a site-specific LC–MS/MSbased proteomic approach. Journal of Agricultural and Food Chemistry, 70(14), 4391–4406. https://doi.org/10.1021/acs.jafc.1c07946Li, C., Nielsen, S. B., Engholm-K eller, K., Lund, M. N. (2022). Oxidation of whey proteins during thermal treatment characterized by a site-specific LC–MS/MSbased proteomic approach. Journal of Agricultural and Food Chemistry, 70(14), 4391–4406. https://doi.org/10.1021/acs.jafc.1c07946Halabi, A., Croguennec, T., Bouhallab, S., Dupont, D., Deglaire, A. (2020). Modification of protein structures by altering the whey protein profile and heat treatment affects in vitro static digestion of model infant milk formulas. Food Function, 11(8), 6933–6945. https://doi.org/10.1039/D0FO01362EHalabi, A., Croguennec, T., Bouhallab, S., Dupont, D., Deglaire, A. (2020). Modification of protein structures by altering the whey protein profile and heat treatment affects in vitro static digestion of model infant milk formulas. Food Function, 11(8), 6933–6945. https://doi.org/10.1039/D0FO01362EdeWit, J. N., Klarenbeek, G. (1984). Effects of various heat treatments on structure and solubility of whey proteins. Journal of Dairy Science, 67(11), 2701–2710. https://doi.org/10.3168/jds.S0022-0302(84)81628-8deWit, J. N., Klarenbeek, G. (1984). Effects of various heat treatments on structure and solubility of whey proteins. Journal of Dairy Science, 67(11), 2701–2710. https://doi.org/10.3168/jds.S0022-0302(84)81628-8Hensel, M., Steurer, R., Fichtl, J., Elger, C., Wedekind, F., Petzold, A. et al. (2011). Identification of potential sites for tryptophan oxidation in recombinant antibodies using tert-butylhydroperoxide and quantitative LC–MS. PLoS One, 6(3), Article e17708. https://doi.org/10.1371/journal.pone.0017708Hensel, M., Steurer, R., Fichtl, J., Elger, C., Wedekind, F., Petzold, A. et al. (2011). Identification of potential sites for tryptophan oxidation in recombinant antibodies using tert-butylhydroperoxide and quantitative LC–MS. PLoS One, 6(3), Article e17708. https://doi.org/10.1371/journal.pone.0017708Aledo, J. C., Cantón, F. R., Veredas, F. J. (2015). Sulphur atoms from methionines interacting with aromatic residues are less prone to oxidation. Scientific Reports, 5(1), Article 16955. https://doi.org/10.1038/srep16955Aledo, J. C., Cantón, F. R., Veredas, F. J. (2015). Sulphur atoms from methionines interacting with aromatic residues are less prone to oxidation. Scientific Reports, 5(1), Article 16955. https://doi.org/10.1038/srep16955Xiao, Q., Woo, M. W., Hu, J., Xiong, H., Zhao, Q. (2021). The role of heating time on the characteristics, functional properties and antioxidant activity of enzyme-hydrolyzed rice proteins-g lucose Maillard reaction products. Food Bioscience, 43, Article 101225. https://doi.org/10.1016/j.fbio.2021.101225Xiao, Q., Woo, M. W., Hu, J., Xiong, H., Zhao, Q. (2021). The role of heating time on the characteristics, functional properties and antioxidant activity of enzyme-hydrolyzed rice proteins-g lucose Maillard reaction products. Food Bioscience, 43, Article 101225. https://doi.org/10.1016/j.fbio.2021.101225Nagashima, T., Watanabe, A., Kise, H. (1992). Peptide synthesis by proteases in organic solvents: Medium effect on substrate specificity. Enzyme and Microbial Technology, 14(10), 842–847. https://doi.org/10.1016/0141-0229(92)90102-TNagashima, T., Watanabe, A., Kise, H. (1992). Peptide synthesis by proteases in organic solvents: Medium effect on substrate specificity. Enzyme and Microbial Technology, 14(10), 842–847. https://doi.org/10.1016/0141-0229(92)90102-TPeters, K., Pauli, D., Hache, H., Boteva, R. N., Genov, N. C., Fittkau, S. (1989). Subtilisin DY — kinetic characterization and comparison with related proteinases. Current Microbiology, 18(3), 171–177. https://doi.org/10.1007/BF01569566Peters, K., Pauli, D., Hache, H., Boteva, R. N., Genov, N. C., Fittkau, S. (1989). Subtilisin DY — kinetic characterization and comparison with related proteinases. Current Microbiology, 18(3), 171–177. https://doi.org/10.1007/BF01569566Pattabiraman, T. N., Lawson, W. B. (1972). Comparative studies of the specificities of α-chymotrypsin and subtilisin BPN′. Studies with flexible substrates. Biochemical Journal, 126(3), 645–657. https://doi.org/10.1042/bj1260645Pattabiraman, T. N., Lawson, W. B. (1972). Comparative studies of the specificities of α-chymotrypsin and subtilisin BPN′. Studies with flexible substrates. Biochemical Journal, 126(3), 645–657. https://doi.org/10.1042/bj1260645Yamawaki, Y., Yufu, T., Kato, T. (2021). The effect of a peptide substrate containing an unnatural branched amino acid on chymotrypsin activity. Processes, 9(2), Article 242. https://doi.org/10.3390/pr9020242Yamawaki, Y., Yufu, T., Kato, T. (2021). The effect of a peptide substrate containing an unnatural branched amino acid on chymotrypsin activity. Processes, 9(2), Article 242. https://doi.org/10.3390/pr9020242Shuai, X., Gao, L., Geng, Q., Li, T., He, X., Chen, J. et al. (2022). Effects of moderate enzymatic hydrolysis on structure and functional properties of pea protein. Foods, 11(15), Article 2368. https://doi.org/10.3390/foods11152368Shuai, X., Gao, L., Geng, Q., Li, T., He, X., Chen, J. et al. (2022). Effects of moderate enzymatic hydrolysis on structure and functional properties of pea protein. Foods, 11(15), Article 2368. https://doi.org/10.3390/foods11152368Food and Agriculture Organization of the United Nations (Rome). (2013). Dietary Protein Quality Evaluation in Human Nutrition: Report of an FAO Expert Consultation, 31 March-2 April, 2011, Auckland, New Zealand. Retrieved from https://www.fao.org/3/i3124e/i3124e.pdf. Accessed October 2, 2022Food and Agriculture Organization of the United Nations (Rome). (2013). Dietary Protein Quality Evaluation in Human Nutrition: Report of an FAO Expert Consultation, 31 March-2 April, 2011, Auckland, New Zealand. Retrieved from https://www.fao.org/3/i3124e/i3124e.pdf. Accessed October 2, 2022Melchior, S., Moretton, M., Alongi, M., Calligaris, S., Nicoli, M. C., Anese, M. (2023). Comparison of protein in vitro digestibility under adult and elderly conditions: The case study of wheat, pea, rice, and whey proteins. Food Research International, 163, Article 112147. https://doi.org/10.1016/j.foodres.2022.112147Melchior, S., Moretton, M., Alongi, M., Calligaris, S., Nicoli, M. C., Anese, M. (2023). Comparison of protein in vitro digestibility under adult and elderly conditions: The case study of wheat, pea, rice, and whey proteins. Food Research International, 163, Article 112147. https://doi.org/10.1016/j.foodres.2022.112147Broersen, K. (2020). Milk processing affects structure, bioavailability and immunogenicity of β-lactoglobulin. Foods, 9(7), Article 874. https://doi.org/10.3390/foods9070874Broersen, K. (2020). Milk processing affects structure, bioavailability and immunogenicity of β-lactoglobulin. Foods, 9(7), Article 874. https://doi.org/10.3390/foods9070874Peram, M. R., Loveday, S. M., Ye, A., Singh, H. (2013). In vitro gastric digestion of heat-induced aggregates of β-lactoglobulin. Journal of Dairy Science, 96(1), 63–74. https://doi.org/10.3168/jds.2012-5896Peram, M. R., Loveday, S. M., Ye, A., Singh, H. (2013). In vitro gastric digestion of heat-induced aggregates of β-lactoglobulin. Journal of Dairy Science, 96(1), 63–74. https://doi.org/10.3168/jds.2012-5896

The authors declare that there are no conflicts of interest present.