foodsystFood systemsПищевые системы2618-97712618-7272Федеральный научный центр пищевых систем им. В.М. Горбатова РАН10.21323/2618-9771-2023-6-1-36-45foodsyst-227Research ArticleСтатьиEnzymatic proteolysis during the conversion of milk into cheeseФерментативный протеолиз при преобразовании молока в сырhttps://orcid.org/0000-0002-2375-3959ЛепилкинаО. В.LepilkinaO. V.

Лепилкина Ольга Валентиновна  — доктор технических наук, главный научный сотрудник, отдел физической химии

152613, Ярославская обл., Углич, Красноармейский бульвар, 19

Tel.: +7–910–965–51–61

Olga. V. Lepilkina, Doctor of Technical Sciences, Leading Scientific Worker, Department of Physical Chemistry

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

Tel.: +7–910–965–51–61

ov.lepilkina@fncps.ruhttps://orcid.org/0000-0003-4364-0342ГригорьеваА. И.GrigorievaA. I.

Григорьева Анастасия Игоревна — младший научный сотрудник, отдел физической химии

152613, Ярославская обл., Углич, Красноармейский бульвар, 19

Tel.: +7–910–965–51–61

Anastasija I. Grigorieva, Junior Researcher, Department of Physical Chemistry

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

Tel.: +7–910–965–51–61

a.grigoriyeva@fncps.ruВсероссийский научно-исследовательский институт маслоделия и сыроделияРоссияAll-Russian Scientific Research Institute of Butter- and CheesemakingRussian Federation202307042023613645Copyright © Lepilkina O.V., Grigorieva A.I., 20232023Лепилкина О.В., Григорьева А.И.Lepilkina O.V., Grigorieva A.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.fsjour.com/jour/article/view/227

The transformation of milk into cheese occurs under the influence of many physicochemical, biochemical and microbiological processes, among which proteolysis plays a very important role. Proteolysis belongs to the most complex type of irreversible post-translational modification of proteins. Enzymatic proteolysis catalysts at different stages of cheese production are native milk enzymes, exo- and endopeptidases of starter and non-starter microorganisms, and milk-clotting enzymes. The article presents a brief overview of modern ideas about the properties, mechanism of action and specificity of the main representatives of enzymes that hydrolyze milk proteins at the stages of preparing milk for coagulation, during rennet coagulation and subsequent maturation of cheeses. These include the plasmin system of milk, enzymes of psychrotrophic bacteria and lactic acid microorganisms that enter milk both accidentally (non-starter microflora) and planned in the form of starter cultures from specially selected strains. Milk-clotting enzymes, having fulfilled their main function — milk coagulation — partially pass into cheese and, along with enzymes of starter microorganisms and plasmin, participate in proteolytic processes during cheese ripening. It is generally accepted that proteolysis in ripening cheeses is the most significant biochemical process that affects the formation of taste, aroma and texture along with lipolysis and glycolysis. The combination of proteolysis products (peptides, amino acids, amines, etc.) is individual for different types of cheese and varies depending on the technological parameters of production, including the duration of maturation. Proteolysis in cheeses has been studied by many scientists in various aspects. This review supplements the known information with new information, without claiming to be comprehensive.

Преобразование молока в сыр происходит под влиянием множества физико-химических, биохимических и микробиологических процессов, среди которых протеолизу отводится очень важная роль. Протеолиз относится к наиболее сложному типу необратимой посттрансляционной модификации белков. Катализаторами ферментативного протеолиза на разных стадиях производства сыра являются нативные ферменты молока, экзо- и эндопептидазы заквасочных и незаквасочных микроорганизмов, молокосвертывающие ферменты. В статье представлен краткий обзор современных представлений о свойствах, механизме действия и специфичности основных представителей ферментов, гидролизующих молочные белки на стадиях подготовки молока к свертыванию, во время сычужного свертывания и последующего созревания сыров. К ним относятся плазминовая система молока, ферменты психротрофных бактерий и молочнокислых микроорганизмов, попадающих в молоко как случайно (незаквасочная микрофлора), так и планируемо в виде заквасок из специально подобранных штаммов. Молокосвертывающие ферменты, выполнив свою основную функцию — свертывание молока,— частично переходят в  сыр и  наряду с  ферментами заквасочных микроорганизмов и плазмином участвуют в протеолитических процессах при созревании сыра. Общепризнано, что протеолиз в созревающих сырах является наиболее значимым биохимическим процессом, влияющим на формирование вкуса, аромата и консистенции наряду с липолизом и гликолизом. Сочетание продуктов протеолиза (пептидов, аминокислот, аминов и др.) индивидуально для разных видов сыров и меняется в зависимости от технологических параметров изготовления, в том числе от продолжительности созревания. Протеолиз в сырах исследовался многими учеными в различных аспектах. Этот обзор дополняет известные сведения новой информацией, не претендуя на всеобъемлемость.

молокосырпротеолизплазминферменты молочнокислых бактерийсычужный ферментmilkcheeseproteolysisplasminlactic acid bacteria enzymesrennetСтатья подготовлена в рамках выполнения исследований по государственному заданию № FNEN‑2019–0010 Федерального научного центра пищевых систем им. В. М. Горбатова Российской академии наукThe article was published as part of the research topic No. FNEN‑2019–0010 of the state assignment of the V. M. Gorbatov Federal Research Center for Food Systems of RAS.ReferencesThompson, А., Boland, M., Singh, Н. (2009). Milk proteins: from expression to food. Academic Press, 2009. https://doi.org/10.1016/B978–0–12–374039–7.X0001–3Thompson, А., Boland, M., Singh, Н. (2009). Milk proteins: from expression to food. Academic Press, 2009. https://doi.org/10.1016/B978–0–12–374039–7.X0001–3D’Ambrosio, C., Arena, S., Salzano, A. M., Renzone, G., Ledda, L., Scaloni, A. (2008). A proteomic characterization of water buffalo milk fractions describing PTM of major species and the identification of minor components involved in nutrient delivery and defense against pathogens. Proteomics, 8(17), 3657–3666. https://doi.org/10.1002/pmic.200701148D’Ambrosio, C., Arena, S., Salzano, A. M., Renzone, G., Ledda, L., Scaloni, A. (2008). A proteomic characterization of water buffalo milk fractions describing PTM of major species and the identification of minor components involved in nutrient delivery and defense against pathogens. Proteomics, 8(17), 3657–3666. https://doi.org/10.1002/pmic.200701148Rout, Р.К., Verma, М. (2021). Post translational modifications of milk proteins in geographically diverse goat breeds. Scientific Reports, 11, Article 5619. https://doi.org/10.1038/s41598–021–85094–9Rout, Р.К., Verma, М. (2021). Post translational modifications of milk proteins in geographically diverse goat breeds. Scientific Reports, 11, Article 5619. https://doi.org/10.1038/s41598–021–85094–9Baptista, D.P., Gigante, M.L. (2021). Bioactive peptides in ripened cheeses: release during technological processes and resistance to the gastrointestinal tract. Journal of the Science of Food and Agriculture, 101(10), 4010–4017. https://doi.org/10.1002/jsfa.11143Baptista, D.P., Gigante, M.L. (2021). Bioactive peptides in ripened cheeses: release during technological processes and resistance to the gastrointestinal tract. Journal of the Science of Food and Agriculture, 101(10), 4010–4017. https://doi.org/10.1002/jsfa.11143Goulding, D.A., Fox, P.F., O’Mahony, J.A. (2020). Milk proteins: An overview. Chapter in a book: Milk Proteins: From Expression to Food. Amsterdam: Elsevier, 2020. https://doi.org/10.1016/b978–0–12–815251–5.00002–5Goulding, D.A., Fox, P.F., O’Mahony, J.A. (2020). Milk proteins: An overview. Chapter in a book: Milk Proteins: From Expression to Food. Amsterdam: Elsevier, 2020. https://doi.org/10.1016/b978–0–12–815251–5.00002–5Fox, P.F., Uniacke-Lowe, Т., McSweeney, P.L.H., O’Mahony, J.A. (2015). Milk proteins. Chapter in a book: Dairy Chemistry and Biochemistry. Springer International Publishing Switzerland, 2015. https://doi.org/10.1007/978–3–319–14892–2Fox, P.F., Uniacke-Lowe, Т., McSweeney, P.L.H., O’Mahony, J.A. (2015). Milk proteins. Chapter in a book: Dairy Chemistry and Biochemistry. Springer International Publishing Switzerland, 2015. https://doi.org/10.1007/978–3–319–14892–2Горбатова, К.К., Гунькова, П.И. (2010). Биохимия молока и молочных продуктов. СПб.: ГИОРД, 2010.Gorbatova, K.K., Gunkova, P.I. (2010). Biochemistry of milk and dairy products. Saint-Petersburg: GIORD, 2010. (In Russian)Holland, J.W. (2008). Post-translational modifications of caseins. Chapter in a book: Milk Proteins: From Expression to Food. Amsterdam: Elsevier, 2008. https://doi.org/10.1016/B978–0–12–374039–7.00004–0Holland, J.W. (2008). Post-translational modifications of caseins. Chapter in a book: Milk Proteins: From Expression to Food. Amsterdam: Elsevier, 2008. https://doi.org/10.1016/B978–0–12–374039–7.00004–0O’Mahony, J. A., Fox, P. F. (2012). Milk Proteins: Introduction and Historical Aspects. Chapter in a book: Advanced Dairy Chemistry. Springer, Boston, 2012. https://doi.org/10.1007/978–1–4614–4714–6_2O’Mahony, J. A., Fox, P. F. (2012). Milk Proteins: Introduction and Historical Aspects. Chapter in a book: Advanced Dairy Chemistry. Springer, Boston, 2012. https://doi.org/10.1007/978–1–4614–4714–6_2Guerin, J., Burgain, J., Gomand, F., Scher, J., Gaiani, C. (2017). Milk fat globule membrane glycoproteins: Valuable ingredients for lactic acid bacteria encapsulation? Critical Reviews in Food Science and Nutrition, 59(4), 639–651. https://doi.org/10.1080/10408398.2017.1386158Guerin, J., Burgain, J., Gomand, F., Scher, J., Gaiani, C. (2017). Milk fat globule membrane glycoproteins: Valuable ingredients for lactic acid bacteria encapsulation? Critical Reviews in Food Science and Nutrition, 59(4), 639–651. https://doi.org/10.1080/10408398.2017.1386158Lopez, C., Cauty, C., Guyomarc’h, F. (2018). Unravelling the complexity of milk fat globules to tailor bioinspired emulsions providing health benefits: the key role played by the biological membrane. European Journal of Lipid Science and Technology, 121(10), Article 1800201. https://doi.org/10.1002/ejlt.201800201Lopez, C., Cauty, C., Guyomarc’h, F. (2018). Unravelling the complexity of milk fat globules to tailor bioinspired emulsions providing health benefits: the key role played by the biological membrane. European Journal of Lipid Science and Technology, 121(10), Article 1800201. https://doi.org/10.1002/ejlt.201800201Ельчанинов, В.В. (2019). Белки мембраны молочной жировой глобулы. 1. Генез и структура жировой глобулы молока, номенклатура белков мембраны. Молочная промышленность, 7, 24–27.Elchaninov, V.V. (2019). The proteins of milk fat globule membrane. 1. Genesis and structure of the milk fat globule, nomenclature of proteins of milk fat globule membrane. Dairy Industry, 7, 24–27. (In Russian)Rogers, L.D., Overall, C.M. (2013). Proteolytic post-translational modification of Pproteins: Proteomic tools and methodology. Molecular and Cellular Proteomics, 12(12), 3532–3542. https://doi.org/10.1074/mcp.M113.031310Rogers, L. D., Overall, C. M. (2013). Proteolytic post-translational modification of P\proteins: Proteomic tools and methodology. Molecular and Cellular Proteomics, 12(12), 3532–3542. https://doi.org/10.1074/mcp.M113.031310Sousa, M.J., Ardö, Y., McSweeney, P.L.H. (2001). Advances in the study of proteolysis during cheese ripening. International Dairy Journal, 11(4–7), 327–345. https://doi.org/10.1016/S0958–6946(01)00062–0Sousa, M.J., Ardö, Y., McSweeney, P.L.H. (2001). Advances in the study of proteolysis during cheese ripening. International Dairy Journal, 11(4–7), 327–345. https://doi.org/10.1016/S0958–6946(01)00062–0Ward, O.P. (2011). 3.49 — Proteases. Comprehensive Biotechnology, 571–582. PMCID: PMC7152071. https://doi:10.1016/b978–0–08–088504–9.00222–1Ward, O.P. (2011). 3.49 — Proteases. Comprehensive Biotechnology, 571–582. PMCID: PMC7152071. https://doi:10.1016/b978–0–08–088504–9.00222–1Ardö, Y. (2021). Enzymes in cheese ripening. Chapter in a book: Agents of Change. Enzymes in Milk and Dairy Products. Switzerland: Springer, Cham., 2021. https://doi.org/10.1007/978–3–030–55482–8_15Ardö, Y. (2021). Enzymes in cheese ripening. Chapter in a book: Agents of Change. Enzymes in Milk and Dairy Products. Switzerland: Springer, Cham., 2021. https://doi.org/10.1007/978–3–030–55482–8_15Webb, E.C. (1992). Enzyme nomenclature 1992. Recommendations of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology on the Nomenclature and Classification of Enzymes. San Diego: Academic Press, 1992.Webb, E.C. (1992). Enzyme nomenclature 1992. Recommendations of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology on the Nomenclature and Classification of Enzymes. San Diego: Academic Press, 1992.France, T.C., O’Mahony, J.A., Kelly, A.L. (2021). The plasmin system in milk and dairy products. Chapter in a book: Agents of Change. Food Engineering Series. Springer, Cham., 2021. https://doi.org/10.1007/978–3–030–5548–8_2France, T.C., O’Mahony, J.A., Kelly, A.L. (2021). The plasmin system in milk and dairy products. Chapter in a book: Agents of Change. Food Engineering Series. Springer, Cham., 2021. https://doi.org/10.1007/978–3–030–5548–8_2Nielsen, S.S. (2002). Plasmin system and microbial proteases in milk: Characteristics, roles, and relationship. Journal of Agricultural and Food Chemistry, 50(22), 6628–6634. https://doi.org/10.1021/jf0201881Nielsen, S.S. (2002). Plasmin system and microbial proteases in milk: Characteristics, roles, and relationship. Journal of Agricultural and Food Chemistry, 50(22), 6628–6634. https://doi.org/10.1021/jf0201881Crudden, A., Kelly, A.L. (2003). Studies of plasmin activity in whey. International Dairy Journal, 13(12), 987–993. https://doi.org/10.1016/s0958–6946(03)00140–7Crudden, A., Kelly, A.L. (2003). Studies of plasmin activity in whey. International Dairy Journal, 13(12), 987–993. https://doi.org/10.1016/s0958–6946(03)00140–7Srinivasan, M., Lucey, J.A. (2002). Effects of added plasmin on the formation and rheological properties of rennet-induced skim milk gels. Journal of Dairy Science, 85(5), 1070–1078. https://doi.org/10.3168/jds.s0022–0302(02)74167–2Srinivasan, M., Lucey, J.A. (2002). Effects of added plasmin on the formation and rheological properties of rennet-induced skim milk gels. Journal of Dairy Science, 85(5), 1070–1078. https://doi.org/10.3168/jds.s0022–0302(02)74167–2Borda, D., Indrawati, Smout, C., Van Loey, A., Hendrickx, M. (2004). High pressure thermal inactivation kinetics of a plasmin system. Journal of Dairy Sciеnse, 87(8), 2351–2358. https://doi.org/10.3168/jds.s0022–0302(04)73357–3Borda, D., Indrawati, Smout, C., Van Loey, A., Hendrickx, M. (2004). High pressure thermal inactivation kinetics of a plasmin system. Journal of Dairy Sciеnse, 87(8), 2351–2358. https://doi.org/10.3168/jds.s0022–0302(04)73357–3Budkevich, R.O., Eremina, A.I., Evdokimov, I.A., Fedortsov, N.M., Martak, A.A., Budkevich, E.V. (2018). The physical properties of the casein in solution: effect of ultra-high pressure. Food Systems, 1(3), 4–12. https://doi.org/10.21323/2618–9771–2018–1–3–4–12Budkevich, R.O., Eremina, A.I., Evdokimov, I.A., Fedortsov, N.M., Martak, A.A., Budkevich, E.V. (2018). The physical properties of the casein in solution: effect of ultra-high pressure. Food Systems, 1(3), 4–12. https://doi.org/10.21323/2618–9771–2018–1–3–4–12Villalobos, J.C., Sigler, A.I.G., Oliete, B., Sánchez, R.A., Jiménez, L., Sánchez, N.N. et al. (2015). Relationship of somatic cell count and composition and coagulation properties of ewe’s milk. Mljekarstvo, 65(2), 138–143. https://doi.org/10.15567/mljekarstvo.2015.0208Villalobos, J.C., Sigler, A.I.G., Oliete, B., Sánchez, R.A., Jiménez, L., Sánchez, N.N. et al. (2015). Relationship of somatic cell count and composition and coagulation properties of ewe’s milk. Mljekarstvo, 65(2), 138–143. https://doi.org/10.15567/mljekarstvo.2015.0208Villalobos, J.С., Garzón, A.I., Martínez Marín, A.L., Arias, R., Ciocia, F., McSweeney, P.L.H. (2018). Plasmin activity in Manchega ewe milk: The effect of lactation, parity and health of the udder, and its influence on milk composition and rennet coagulation. Small Ruminant Research, 158, 57–61. https://doi.org/10.1016/j.smallrumres.2017.10.005Villalobos, J.С., Garzón, A.I., Martínez Marín, A.L., Arias, R., Ciocia, F., McSweeney, P.L.H. (2018). Plasmin activity in Manchega ewe milk: The effect of lactation, parity and health of the udder, and its influence on milk composition and rennet coagulation. Small Ruminant Research, 158, 57–61. https://doi.org/10.1016/j.smallrumres.2017.10.005Sánchez, A.F., Muñoz, J.P., Villalobos, J.C., Sánchez, R.A., Garzón, A., de Pedro, E.A.S. (2021). Coagulation process in Manchega sheep milk from Spain: A path analysis approach. Journal of Dairy Science, 104(7), 7544–7554. https://doi.org/10.3168/jds.2020–19187Sánchez, A.F., Muñoz, J.P., Villalobos, J.C., Sánchez, R.A., Garzón, A., de Pedro, E.A.S. (2021). Coagulation process in Manchega sheep milk from Spain: A path analysis approach. Journal of Dairy Science, 104(7), 7544– 7554. https://doi.org/10.3168/jds.2020–19187Somers, J.M., Guinee, T. P., Kelly, A.L. (2002). The effect of plasmin activity and cold storage of cheese milk on the composition, ripening and functionality of mozzarella-type cheese. International Journal of Dairy Technology, 55(1), 5–11. https://doi.org/10.1046/j.1471–0307.2002.00030.xSomers, J.M., Guinee, T. P., Kelly, A.L. (2002). The effect of plasmin activity and cold storage of cheese milk on the composition, ripening and functionality of mozzarella-type cheese. International Journal of Dairy Technology, 55(1), 5–11. https://doi.org/10.1046/j.1471–0307.2002.00030.xМироненко, И.М. (2021). Функции ионного кальция и нативных протеаз молока в процессе сычужного свертывания. Сыроделие и маслоделие, 1, 25–28. https://doi.org/10.31515/2073–4018–2021–25–28Mironenko, I.M. (2021). Functions of ionic calcium and native milk proteases in the process of rennet clotting. Cheesemaking and Buttermaking, 1, 25–28. https://doi.org/10.31515/2073–4018–2021–25–28 (In Russian)Мироненко, И.М. (2019). Вероятные участники процесса сычужного свертывания молока. Сыроделие и маслоделие, 4, 20–23.Mironenko, I.M. (2019). Probable participants in the process of rennet coagulation of milk. Cheesemaking and Buttermaking, 4, 20–23. (In Russian)Ardö, Y., McSweeney, P.L.H, Magboul, A.A., Upadhyay, V.K., Fox, P.F. (2017). Biochemistry of cheese ripening: Proteolysis. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press. https://doi.org/10.1016/B978–0–12–417012–4.00018_1Ardö, Y., McSweeney, P.L.H, Magboul, A.A., Upadhyay, V.K., Fox, P.F. (2017). Biochemistry of cheese ripening: Proteolysis. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press. https://doi.org/10.1016/B978–0–12–417012–4.00018_1Ozer, В.В., Akdemir-Evrendilek, G. (2014). Microbiology of Raw Milk. Chapter in a book: Dairy Microbiology and Biochemistry. Boca-Raton: CRC Press, 2017.Ozer, В.В., Akdemir-Evrendilek, G. (2014). Microbiology of Raw Milk. Chapter in a book: Dairy Microbiology and Biochemistry. Boca-Raton: CRC Press, 2017.Glück, С., Stressler, Т., Fischer, L. (2021). Heat-stable microbial peptidases associated with the microbiota of raw milk. Chapter in a book: Agents of Change. Enzymes in Milk and Dairy Products. Switzerland: Springer, Cham., 2021. https://doi.org/10.1007/978–3–030–55482–8_11Glück, С., Stressler, Т., Fischer, L. (2021). Heat-stable microbial peptidases associated with the microbiota of raw milk. Chapter in a book: Agents of Change. Enzymes in Milk and Dairy Products. Switzerland: Springer, Cham., 2021. https://doi.org/10.1007/978–3–030–55482–8_11Pukančíková, L., Lipničanová, S., Kačániová, M., Chmelová, D., Ondrejovič, M. (2016). Natural microflora of raw cow milk and their enzymatic spoilage potential. Nova Biotechnologica et Chimica, 15(2), 142–155. https://doi.org/10.1515/nbec‑2016–0015Pukančíková, L., Lipničanová, S., Kačániová, M., Chmelová, D., Ondrejovič, M. (2016). Natural microflora of raw cow milk and their enzymatic spoilage potential. Nova Biotechnologica et Chimica, 15(2), 142– 155. https://doi.org/10.1515/nbec‑2016–0015Baur, C., Krewinkel, M., Kranz, B., von Neubeck, M., Wenning, M., Scherer, S. et al. (2015). Quantification of the proteolytic and lipolytic activity of microorganisms isolated from raw milk. International Dairy Journal, 49, 23–29. https://doi.org/10.1016/j.idairyj.2015.04.005Baur, C., Krewinkel, M., Kranz, B., von Neubeck, M., Wenning, M., Scherer, S. et al. (2015). Quantification of the proteolytic and lipolytic activity of microorganisms isolated from raw milk. International Dairy Journal, 49, 23–29. https://doi.org/10.1016/j.idairyj.2015.04.005Datta, N, Deeth, H.C. (2003). Diagnosing the cause of proteolysis in UHT milk. LWT — Food Science and Technology, 36(2), 173–182. https://doi.org/10.1016/S0023–6438(02)00214–1Datta, N, Deeth, H.C. (2003). Diagnosing the cause of proteolysis in UHT milk. LWT — Food Science and Technology, 36(2), 173–182. https://doi.org/10.1016/S0023–6438(02)00214–1Kelly, A.L., Larsen, L.B. (2021). Agents of change: Enzymes in milk and dairy products. Springer Cham, 2021. https://doi.org/10.1007/978–3–030–55482–8Kelly, A.L., Larsen, L.B. (2021). Agents of change: Enzymes in milk and dairy products. Springer Cham, 2021. https://doi.org/10.1007/978–3–030–55482–8Settanni, L., Moschetti, G. (2010). Non-starter lactic acid bacteria used to improve cheese quality and provide health benefits. Food Microbiology, 27(6), 691–697. https://doi.org/10.1016/j.fm.2010.05.023Settanni, L., Moschetti, G. (2010). Non-starter lactic acid bacteria used to improve cheese quality and provide health benefits. Food Microbiology, 27(6), 691–697. https://doi.org/10.1016/j.fm.2010.05.023Pogačić, T., Mancini, A., Santarelli, M., Bottari, B., Lazzi, C., Neviani, E. et al. (2013). Diversity and dynamic of lactic acid bacteria strains during aging of a long ripened hard cheese produced from raw milk and undefined natural starter. Food Microbiology, 36(2), 207–215. https://doi.org/10.1016/j.fm.2013.05.009Pogačić, T., Mancini, A., Santarelli, M., Bottari, B., Lazzi, C., Neviani, E. et al. (2013). Diversity and dynamic of lactic acid bacteria strains during aging of a long ripened hard cheese produced from raw milk and undefined natural starter. Food Microbiology, 36(2), 207–215. https://doi.org/10.1016/j.fm.2013.05.009Wouters, J. T.M., Ayad, E.H.E., Hugenholtz, J., Smit, G. (2002) Microbes from raw milk for fermented dairy products. International Dairy Journal, 12(2–3), 91–109. https://doi.org/10.1016/s0958–6946(01)00151–0Wouters, J. T.M., Ayad, E.H.E., Hugenholtz, J., Smit, G. (2002) Microbes from raw milk for fermented dairy products. International Dairy Journal, 12(2–3), 91–109. https://doi.org/10.1016/s0958–6946(01)00151–0Bluma, A., Ciprovica, I. (2015). Diversity of lactic acid bacteria in raw milk. Research for Rural Development, 1, 157–161.Bluma, A., Ciprovica, I. (2015). Diversity of lactic acid bacteria in raw milk. Research for Rural Development, 1, 157–161.Hernandez-Valdes, J.A.; van Gestel, J.; Kuipers, O.P. (2020). A riboswitch gives rise to multi-generational phenotypic heterogeneity in an auxotrophic bacterium. Nature Communications, 11(1), e00133. https://doi:10.1038/s41467–020–15017–1Hernandez-Valdes, J.A.; van Gestel, J.; Kuipers, O.P. (2020). A riboswitch gives rise to multi-generational phenotypic heterogeneity in an auxotrophic bacterium. Nature Communications, 11(1), e00133. https://doi:10.1038/s41467–020–15017–1Tagliazucchi, D., Martini, S., Solieri, L. (2019). Bioprospecting for bioactive peptide production by lactic acid bacteria isolated from fermented dairy food. Fermentation, 5(96), Article 96.. https://doi.org/10.3390/fermentation5040096Tagliazucchi, D., Martini, S., Solieri, L. (2019). Bioprospecting for bioactive peptide production by lactic acid bacteria isolated from fermented dairy food. Fermentation, 5(96), Article 96.. https://doi.org/10.3390/fermentation5040096Parente, E., Cogan, T.M., Powell, I.B. (2017). Starter cultures: General aspects. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press, 2017. https://doi.org/10.1016/B978–0–12–417012–4.00008_1Parente, E., Cogan, T.M., Powell, I.B. (2017). Starter cultures: General aspects. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press, 2017. https://doi.org/10.1016/B978–0–12–417012–4.00008_1Китаевская, С.В., Пономарев, В.Я., Решетник, О.А. (2022). Оценка протеолитической активности новых штаммов лактобацилл с криорезистентными свойствами. Известия вузов. Прикладная химия и биотехнология, 12(1), 76–86. https://doi.org/10.21285/2227–2925–2022–12–1–76–86Kitaevskaya, S.V., Ponomarev, V.Y., Reshetnik, O.A. (2022). Evaluation of the proteolytic activity of new cryoresistant lactobacillus strains. Proceedings of Universities. Applied Chemistry and Biotechnology, 12(1), 76–86. https://doi.org/10.21285/2227–2925–2022–12–1–76–86 (In Russian)Kieliszek, M, Pobiega, K, Piwowarek, K, Kot, AM. (2021). Characteristics of the proteolytic enzymes produced by lactic acid bacteria. Molecules, 26(7), Article 1858. https://doi.org/10.3390/molecules26071858Kieliszek, M, Pobiega, K, Piwowarek, K, Kot, AM. (2021). Characteristics of the proteolytic enzymes produced by lactic acid bacteria. Molecules, 26(7), Article 1858. https://doi.org/10.3390/molecules26071858Korhonen, H., Pihlanto, A. (2003). Food — derived bioactive peptidesopportunities for designing future foods. Current Pharmaceutical Design, 9(16), 1297–1308. https://doi.org/10.2174/1381612033454892Korhonen, H., Pihlanto, A. (2003). Food — derived bioactive peptidesopportunities for designing future foods. Current Pharmaceutical Design, 9(16), 1297–1308. https://doi.org/10.2174/1381612033454892Головач, Т.Н., Жабанос, Н.К., Фурик, Н.Н., Курченко, В.П., Ризевский, С.В. (2013). Субстратная специфичность и уровень ферментной активности при расщеплении белковых фракций молока пробиотическими микроорганизмами. Актуальные вопросы переработки мясного и молочного сырья, 8, 130–142.Halavach, T.N., Zhabanos, N.K., Furik, N.N., Kurchenko, V.P., Rizevsky, S.V. (2013). Substrate specificity and enzymatic activity level in the cleavage of milk protein fractions with probiotic microorganisms. Topical Issues of Processing of Meat and Milk Raw Materials, 8, 130–142. (In Russian)Одегов, Н. И. Гришкова, А.В., Белов, А.Н. (2019). К вопросу направленного регулирования протеолитических процессов в сырах. Сыроделие и маслоделие, 5, 24–26. https://doi.org/10.31515/2073–4018–2019–5–24–26Odegov, N.I., Grishkova, A.V., Belov, A.N. (2019). To the question of directed regulation of proteolytic processes in cheeses. Cheesemaking and Buttermaking, 5, 24–26. https://doi.org/10.31515/2073–4018–2019–5–24–26 (In Russian)Мягконосов, Д.С., Мордвинова, В.А., Абрамов, Д.В., Делицкая, И.Н. (2014). Особенности протеолиза у сыров различных видовых групп. Сыроделие и маслоделие, 2, 24–27.Myagkonosov, D.S., Mordvinova, V.A., Abramov, D.V., Delitskaya, I.N. (2014). Special features of proteolysis in different groups of cheese types. Cheesemaking and Buttermaking, 2, 24–27. (In Russian)Franco, I., Prieto, B., Urdiales, R., Fresno, J.М., Carballo, J. (2001). Study of the biochemical changes during ripening of Ahumado de Áliva cheese: a Spanish traditional variety. Food Chemistry, 74(4), 463–469. https://doi:10.1016/s0308–8146(01)00164–9Franco, I., Prieto, B., Urdiales, R., Fresno, J.М., Carballo, J. (2001). Study of the biochemical changes during ripening of Ahumado de Áliva cheese: a Spanish traditional variety. Food Chemistry, 74(4), 463–469. https://doi:10.1016/s0308–8146(01)00164–9Rampilli, M., Larsen, R., Harboe, M. (2005). Natural heterogeneity of chymosin and pepsin in extracts of bovine stomachs. International Dairy Journal, 15(11), 1130–1137. https://doi.org/10.1016/j.idairyj.2004.10.003Rampilli, M., Larsen, R., Harboe, M. (2005). Natural heterogeneity of chymosin and pepsin in extracts of bovine stomachs. International Dairy Journal, 15(11), 1130–1137. https://doi.org/10.1016/j.idairyj.2004.10.003Horne, D.S., Lucey, J.A. (2017). Rennet-induced coagulation of milk. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press, 2017. https://doi.org/10.1016/b978–0–12–417012–4.00005–3Horne, D.S., Lucey, J.A. (2017). Rennet-induced coagulation of milk. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press, 2017. https://doi.org/10.1016/b978–0–12–417012–4.00005–3Britten, M., Giroux, H.J. (2021). Rennet coagulation of heated milk: A review. International Dairy Journal, 124, Article 105179. https://doi.org/10.1016/j.idairyj.2021.105179Britten, M., Giroux, H.J. (2021). Rennet coagulation of heated milk: A review. International Dairy Journal, 124, Article 105179. https://doi.org/10.1016/j.idairyj.2021.105179Fox, P.F., Guinee, T.P., Cogan, T.M., McSweeney, P.L.H. (2016). Enzymatic coagulation of milk. Chapter in a book: Fundamentals of Cheese Science, New York: Springer, 2016. https://doi.org/10.1007/978–1–4899–7681–9_7Fox, P.F., Guinee, T.P., Cogan, T.M., McSweeney, P.L.H. (2016). Enzymatic coagulation of milk. Chapter in a book: Fundamentals of Cheese Science, New York: Springer, 2016. https://doi.org/10.1007/978–1–4899–7681–9_7Jaros, D., Rohm, H. (2017). Rennets: Applied aspects. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press, 2017. https://doi.org/10.1016/B978–0–12–417012–4.00003_1Jaros, D., Rohm, H. (2017). Rennets: Applied aspects. Chapter in a book: Cheese. Chemistry, Physics and Microbiology. Academic Press, 2017. https://doi.org/10.1016/B978–0–12–417012–4.00003_1Кригер, А.В., Белов, А.Н. (2010). Влияние ферментных композиций на протеолиз в сырах. Сыроделие и маслоделие, 3, 38–40.Kriger, A.V., Belov, A.N. (2010). Effect of enzyme compositions on cheese proteolysis. Cheesemaking and Buttermaking, 3, 38–40. (In Russian)Ye, А., Cui, J., Singh, H. (2011). Proteolysis of milk fat globule membrane proteins during in vitro gastric digestion of milk. Journal of Dairy Science, 94(6), 2762–2770. https://doi.org/10.3168/jds.2010–4099Ye, А., Cui, J., Singh, H. (2011). Proteolysis of milk fat globule membrane proteins during in vitro gastric digestion of milk. Journal of Dairy Science, 94(6), 2762–2770. https://doi.org/10.3168/jds.2010–4099Абрамов, Д.В., Мягконосов, Д.С., Делицкая, И.Н., Мордвинова, В.А., Муничева, Т.Э., Овчинникова, Е.Г. (2019). Перспективы применения комплексных МФП для производства созревающих сычужных сыров. Сыроделие и маслоделие, 1, 24–26. https://doi.org/10.31515/2073–4018–2019–1–24–26Abramov, D.V., Myagkonosov, D.S., Delitskaya, I.N., Mordvinova, V.A., Municheva, T.E., Ovchinnikova E. G. (2019). Perspectives of using complex milk-clotting enzyme preparations for ripening rennet cheeses production. Cheesemaking and Buttermaking, 1, 24–26. https://doi.org/10.31515/2073–4018–2019–1–24–26 (In Russian)Мягконосов, Д.С., Абрамов, Д.В., Овчинникова, Е.Г., Муничева, Т.Э. (2019). Перспективы использования микробных заменителей химозина в сыроделии. Сыроделие и маслоделие, 4, 14–17.Myagkonosov, D.S., Abramov, D.V., Ovchinnikova, E.G., Municheva, T.E. (2019). Prospects of using microbial substitutes of chymosin in cheesemaking. Cheesemaking and Buttermaking, 4, 14–17. (In Russian)Amira, A. B., Besbes, S., Attia, H., Blecker, C. (2017). Milk-clotting properties of plant rennets and their enzymatic, rheological, and sensory role in cheese making: A review. International Journal of Food Properties, 20(sup1), S76–S93. https://doi.org/10.1080/10942912.2017.1289959Amira, A. B., Besbes, S., Attia, H., Blecker, C. (2017). Milk-clotting properties of plant rennets and their enzymatic, rheological, and sensory role in cheese making: A review. International Journal of Food Properties, 20(sup1), S76–S93. https://doi.org/10.1080/10942912.2017.1289959Мягконосов, Д.С., Абрамов, Д.В., Делицкая, И.Н., Овчинникова, Е.Г. (2022). Протеолитическая активность молокосвертывающих ферментов разного происхождения. Пищевые системы, 5(1), 47–54. https://doi.org/10.21323/2618–9771–2022–5–1–47–54Myagkonosov, D. S., Abramov, D. V., Delitskaya, I. N., Ovchinnikova, E. G. (2022). Proteolytic activity of milk-clotting enzymes of different origin. Food Systems, 5(1), 47–54. https://doi.org/10.21323/2618–9771–2022–5–1–47–54 (In Russian)Khattab, A. R., Guirguis, H. A., Tawfik, S. M., Farag, M. A. (2019). Cheese ripening: A review on modern technologies towards flavor enhancement, process acceleration and improved quality assessment. Trends in Food Science and Technology, 88, 343–360. https://doi.org/10.1016/j.tifs.2019.03.009Khattab, A. R., Guirguis, H. A., Tawfik, S. M., Farag, M. A. (2019). Cheese ripening: A review on modern technologies towards flavor enhancement, process acceleration and improved quality assessment. Trends in Food Science and Technology, 88, 343–360. https://doi.org/10.1016/j.tifs.2019.03.009Fox, P.F. (1989). Proteolysis during cheese manufacture and ripening. Journal of Dairy Science, 72(6), 1379–1400. https://doi.org/10.3168/jds.S0022–0302(89)79246–8Fox, P.F. (1989). Proteolysis during cheese manufacture and ripening. Journal of Dairy Science, 72(6), 1379–1400. https://doi.org/10.3168/jds.S0022–0302(89)79246–8

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