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Differentiated assessment of Lactococcus enzymatic activity for the creation of starter cultures of direct application

https://doi.org/10.21323/2618-9771-2025-8-4-566-575

Abstract

Representatives of the genus Lactococcus are one of the key starter microorganisms in the dairy industry. They play a leading role in lactic acid fermentation in the production of fermented dairy products and cheeses. The article presents the results of a differentiated assessment of the enzymatic activity of nine strains of Lactococcus lactis subsp. lactis in milk at a cultivation temperature of (30 ± 1) °C, as well as during cooling and storage at a temperature of (6 ± 1) °C for 21 days. All studied parameters of acid formation (an increase in titratable acidity during fermentation and storage) demonstrated statistically significant differences between the strains (ANOVA, P < 0.05). The most pronounced variability of the strains was observed in their ability to acidify milk during storage: a decrease in the actual acidity over 21 days ranged from 0.19 to 0.49 pH units. This may be due to their genetic differences in enzymatic activity and resistance to low temperatures, which requires further study. The highest activity at (30 ± 1) °C was observed in four strains (663–12, 792–7, 618–5 and 549–1), which increased the titratable acidity of milk by 69–70 °T. Three of these strains (792–7, 618–5, and 549–1) slowly accumulated acid during storage, increasing the acidity by 4.7–7.3 °T over 21 days. Strains 637–4 and 429–6 showed a low fermentation rate at optimal temperature (an increase in acidity was 56.7 and 54.3 °T, respectively), but a high one during storage (an increase in acidity was 18.3 and 16.7 °T, respectively). Cluster analysis by the Ward method and the Euclidean distance metric performed in the R4.3.1 software environment allowed us to divide the studied L. lactis cultures into three groups according to the nature of the enzymatic profile at the temperature of active fermentation and under the storage conditions. The isolated clusters represent various metabolic profiles that determine the potential use of strains in technological processes. The results obtained indicate that for a comprehensive assessment of the acid-forming activity of cultures when selecting them for using in the composition of starter cultures of direct application for fermented dairy products and cheeses, it is necessary to study both the rate of acid formation and the post-enzymatic potential.

About the Authors

A. Yu. Duganova
V. M. Gorbatov Federal Scientific Center for Food Systems of the Russian Academy of Sciences
Russian Federation

Anna Yu. Duganova, Junior Researcher, Department of Microbiology

55, Liteiny prospekt, 191014, Saint Petersburg



N. P. Sorokina
V. M. Gorbatov Federal Scientific Center for Food Systems of the Russian Academy of Sciences
Russian Federation

Ninel P. Sorokina, Candidate of Technical Sciences, Leading Researcher, Department of Microbiology

55, Liteiny prospekt, 191014, Saint Petersburg



D. S. Mamykin
V. M. Gorbatov Federal Scientific Center for Food Systems of the Russian Academy of Sciences
Russian Federation

Denis S. Mamykin, Junior Researcher, Department of Microbiology

55, Liteiny prospekt, 191014, Saint Petersburg



A. A. Semenova
V. M. Gorbatov Federal Scientific Center for Food Systems of the Russian Academy of Sciences
Russian Federation

Anastasia A. Semenova, Doctor of Technical Sciences, Deputy Director for Scientific Work

55, Liteiny prospekt, 191014, Saint Petersburg



G. N. Rogov
V. M. Gorbatov Federal Scientific Center for Food Systems of the Russian Academy of Sciences
Russian Federation

Grigory N. Rogov, Candidate of Technical Sciences, Director of the Brunch

55, Liteiny prospekt, 191014, Saint Petersburg



A. A. Belenko
V. M. Gorbatov Federal Scientific Center for Food Systems of the Russian Academy of Sciences
Russian Federation

Andrey A. Belenko, Candidate of Biological Sciences, Head of the Laboratory of Genetic Research

55, Liteiny prospekt, 191014, Saint Petersburg



References

1. Hutkins, R. W. (2018). Microbiology and Technology of Fermented Foods. WileyBlackwell, 2018. https://doi.org/10.1002/9780470277515

2. Altieri, C., Ciuffreda, E., Di Maggio, B., Sinigaglia, M. (2017). Lactic acid bacteria as starter cultures. Chapter in a book: Starter Cultures in Food Production. Weinheim, Germany: John Wiley & Sons., 2017.

3. Fusieger, A., Martins, M. C. F., de Freitas, R., Nero, L. A., de Carvalho, A. F. (2020). Technological properties of Lactococcus lactis subsp. lactis bv. diacetylactis obtained from dairy and non-dairy niches. Brazilian Journal of Microbiology, 51(1), 313–321. https://doi.org/10.1007/s42770-019-00182-3

4. Kamartdinova, D. R., Kitaevskaya, S. V., Reshetnik, O. A., Turin, M. Yu. (2025). Development of a lactic acid bacteria consortium with antioxidant properties. Processes and Food Production Equipment, 2, 15–23. (In Russian). https://doi.org/10.17586/2310-1164-2025-18-2-14-23

5. Wang, Y., Wu, J., Lv, M., Shao, Z., Hungwe, M., Wang, J. et al. (2021). Metabolism characteristics of lactic acid bacteria and the expanding applications in food industry. Frontiers in Bioengineering and Biotechnology, 9, Article 612285. https://doi.org/10.3389/fbioe.2021.612285

6. Tamime, A. Y., Robinson, R. K. (2007). Tamime and Robinson’s Yoghurt: Science and Technology. Woodhead Publishing, 2007. https://doi.org/10.1533/9781845692612

7. De Melo Pereira, G. V., De Carvalho Neto, D. P., De O. Junqueira, A. C., Karp, S. G., Letti, L. A. J., Magalhães Júnior, A.I. et al. (2019). A review of selection criteria for starter culture development in the food fermentation industry. Food Reviews International, 36(2), 135–167. https://doi.org/10.1080/87559129.2019.1630636

8. Shukhalova, О. М. (2021). Main criteria for selection of microorganisms in the composition of bacterial starter for ripening cheeses. Food Systems, 4(3S), 315–320. (In Russian) https://doi.org/10.21323/2618-9771-2021-4-3S-315-320

9. Kelleher, P., Mahony, J., Bottacini, F., Lugli, G.A., Ventura, M., van Sinderen, D. (2019). The Lactococcus lactis Pan-Plasmidome. Frontiers in Microbiology, 10, Article 707. https://doi.org/10.3389/fmicb.2019.00707

10. White, K., Eraclio, G., McDonnell, B., Bottacini, F., Lugli, G. A., Ventura, M. et al. (2024). A multifaceted investigation of lactococcal strain diversity in undefined mesophilic starter cultures. Applied and Environmental Microbiology, 90(3), Article e0215223. https://doi.org/10.1128/aem.02152-23

11. Panebianco, F., Giarratana, F., Caridi, A., Sidari, R., De Bruno, A., Giuffrida, A. (2021) Lactic acid bacteria isolated from traditional Italian dairy products: Activity against Listeria monocytogenes and modelling of microbial competition in soft cheese. LWT, 137(17), Article 110446. https://doi.org/10.1016/j.lwt.2020.110446

12. De Vos, P., Garrity, G. M., Jones, D., Krieg, N. R., Ludwig, W., Rainey, F. A. et al. (2009). Bergey’s manual® of systematic bacteriology. Firmicutes. Springer New York, NY, 2009.

13. Iskandar, C. F., Cailliez-Grimal, C., Borges, F., Revol-Junelles, A.-M. (2019) Review of lactose and galactose metabolism in lactic acid bacteria dedicated to expert genomic annotation. Trends in Food Science and Technology, 88, 121–132. https://doi.org/10.1016/j.tifs.2019.03.020

14. Pohanka, M. (2020). D-Lactic acid as a metabolite: Toxicology, diagnosis, and detection. BioMed Research International, 2020, Article 3419034. https://doi.org/10.1155/2020/3419034

15. Ma´slak, E., Złoch, M., Arendowski, A., Sugajski, M., Janczura, I., Rudnicka, J. et al. (2022). Isolation and identification of Lactococcus lactis and Weissella cibaria strains from fermented beetroot and an investigation of their properties as potential starter cultures and probiotics. Foods, 11(15), Article 2257. https://doi.org/10.3390/foods11152257

16. Bintsis, T. (2018). Lactic acid bacteria as starter cultures: An update in their metabolism and genetics. AIMS Microbiology, 4(4), 665–684. https://doi.org/10.3934/microbiol.2018.4.665

17. Li, T. T., Tian, W. L., Gu, C. T. (2019). Elevation of Lactococcus lactis subsp. cremoris to the Species Level as Lactococcus cremoris sp. nov. and Transfer of Lactococcus lactis subsp. tructae to Lactococcus cremoris as Lactococcus cremoris subsp. tructae comb. nov. International Journal of Systematic and Evolutionary Microbiology, 71(3), Article 004727. https://doi.org/10.1099/ijsem.0.004727

18. Manno, M. T., Zuljan, F., Alarcón, S., Esteban, L., Blancato, V., Espariz, M. et al. (2018). Genetic and phenotypic features defining industrial relevant Lactococcus lactis, L. cremoris and L. lactis biovar. diacetylactis strains. Journal Biotechnology, 282, 25–31. https://doi.org/10.1016/j.jbiotec.2018.06.345

19. Kim, W. S., Ren, J., Dunn, N. W. (1999). Differentiation of Lactococcus lactis subspecies lactis and subspecies cremoris strains by their adaptive response to stresses. FEMS Microbiology Letters, 171(1), 57–65. https://doi.org/0.1111/j.1574-6968.1999.tb13412.x

20. Zeidan, A. A., Poulsen, V. K., Janzen, T. Buldo, P., Derkx, P. M. F., Øregaard, G. et al. (2017). Polysaccharide production by lactic acid bacteria: From genes to industrial applications. FEMS Microbiology Reviews, 41, 168–200. https://doi.org/10.1093/femsre/fux017

21. Kelleher, P., Mahony, J., Bottacini, F., Lugli, G. A., Ventura, M., van Sinderen, D. (2019). The Lactococcus lactis Pan-Plasmidome. Frontiers in Microbiology, 10, Article 707. https://doi.org/10.3389/fmicb.2019.00707

22. Sviridenko, G. M., Shukhalova, O. M. (2019). Lactic acid lactococci as a main acid forming component. Dairy Industry, 4, 30–33. (In Russian) https://doi.org/10.31515/1019-8946-2019-4-30-33

23. Zapaśnik, A., Sokołowska, B., Bryła, M. (2022). Role of lactic acid bacteria in food preservation and safety. Foods, 11(9), Article 1283. https://doi.org/10.3390/foods11091283

24. Guan, Y., Cui, Y., Qu, X., Li, B., Zhang, L. (2024). Post-acidification of fermented milk and its molecular regulatory mechanism. International Journal of Food Microbiology, 426, Article 110920. https://doi.org/10.1016/j.ijfoodmicro.2024.110920

25. Gudkov, A. V. (2004). Cheese making: Technological, biological and physico-chemical aspects. Moscow: Dely print, 2004. (In Russian)

26. McSweeney, P. L. N., Cotter, P. D., Everett, D. W. (2017). Cheese: Chemistry, Physics and Microbiology. Academic Press, 2017.

27. Kok, J., van Gijtenbeek, L. A., de Jong, A. van der Meulen, S. B., Solopova, A., Kuipers, O. P. (2017). The evolution of gene regulation research in Lactococcus lactis. FEMS Microbiology Reviews, 41(Supp_1), S220-S243. https://doi.org/10.1093/femsre/fux028

28. Sviridenko, G. M., Shukhalova, O. M. (2020). The effect of temperature on the development and metabolism of the main acid-forming starter microflora. Dairy Industry, 7, 49–51. (In Russian) https://doi.org/10.31515/1019-8946-2020-07-49-51

29. Poudel, R., Thunell, R. K., Oberg, C. J., Lefevre, M., Oberg, T. S., McMahon, G. J. (2022). Comparison of growth and survival of single strains of Lactococcus lactis and Lactococcus cremoris during Cheddar cheese manufacture. Journal of Dairy Science, 105(3), 2069–2081. https://doi.org/10.3168/jds.2021-20958

30. Deshwal, G. K., Tiwari, S., Kumar, A., Raman, R. K., Kadyan, S. (2021). Review on factors affecting and control of post-acidification in yoghurt and related products. Trends in Food Science and Technology, 109, 499–512. https://doi.org/10.1016/j.tifs.2021.01.057

31. Prosekov, A. Yu., Ostroumov, L. A. (2016). Innovation management biotechnology of starter cultures. Food Processing: Techniques and Technology, 43(4), 64–69 (In Russian)

32. Mahony, J., Bottacini, F., van Sinderen, D. (2023). Towards the diversification of lactococcal starter and non-starter species in mesophilic dairy culture systems. Microbial Biotechnology, 16(9), 1745–1754. https://doi.org/10.1111/1751-7915.14320

33. Dorofeev, R. V., Kuznetsova, T. N., Ott, E. F., Funk, I. A. (2023). Isolation of lactococci promising for dairy industry. Polzunovskiy Vestnik, 4, 24–28. (In Russian). https://doi.org/10.25712/ASTU.2072-8921.2023.04.003

34. Cavanagh, D., Casey, A., Altermann, E., Cotter, P. D., Fitzgerald, G. F., McAuliffe, O. (2015). Evaluation of Lactococcus lactis isolates from non-dairy sources with potential dairy applications reveals extensive phenotype-genotype disparity and implications for a revised species. Applied Environmental Microbiology, 81(12), 3961–3972. https://doi.org/10.1128/AEM.04092-14

35. Semenova, V. A., Mitrova V. A., Kishilova, S. A., Rozhkova, I. V., Petrov, A. N. (2025). Comparative assessment of properties of industrially important Lactococcus strains. Food Industry, 8, 82–87. (In Russian). https://doi.org/10.52653/PPI.2025.8.8.022

36. Trubitsyna, Ju. M., Ott, E. F., Dorofeev, R.V., Shevchenko, K. E., Gryankina, T. V. (2024). Biotechnological properties of lactococci isolated from natural sources. Polzunovskiy Vestnik, 3, 29–35. (In Russian) https://doi.org/10/25712/ASTU.2072–8921.2024.03.004

37. Li, W., Ren, M., Duo, L., Li, J., Wang, S., Sun, Y. et al. (2020). Fermentation Characteristics of Lactococcus lactis subsp. lactis isolated from naturally fermented dairy products and screening of potential starter isolates. Frontiers in Microbiology, 11, Article 1794. https://doi.org/10.3389/fmicb.2020.01794


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For citations:


Duganova A.Yu., Sorokina N.P., Mamykin D.S., Semenova A.A., Rogov G.N., Belenko A.A. Differentiated assessment of Lactococcus enzymatic activity for the creation of starter cultures of direct application. Food systems. 2025;8(4):566-575. (In Russ.) https://doi.org/10.21323/2618-9771-2025-8-4-566-575

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