Skip to main content
SpringerLink
Log in

Fermented milks from Enterococcus faecalis TH563 and Lactobacillus delbrueckii subsp. bulgaricus LA2 manifest different degrees of ACE-inhibitory and immunomodulatory activities

Enterococcus faecalis TH563 Lactobacillus delbrueckii subsp. bulgaricus LA2 ACE

Les laits fermentés par Enterococcus faecalis TH563 et Lactobacillus delbrueckii bulgaricus LA2 montrent différents degrés d’activités anti-ACE et immunomodulatrice

  • Note
  • Published:
Dairy Science & Technology

Abstract

Milk proteins are precursors of biologically active components that are released by enzymatic proteolysis. Among the biological activities recognised in milk components, the angiotensin-I converting enzyme (ACE)-inhibitory and immunomodulatory activities are of great interest. In the present work the ACE-inhibitory and immunomodulatory activities were analysed in milks fermented by two bacterial strains isolated from Italian dairy products, Enterococcus faecalis TH563 or Lactobacillus delbrueckii subsp. bulgaricus (L. delb. bulgaricus) LA2. ACE-inhibitory activity was determined by an already established enzymatic method and immunomodulatory activity by the proliferation of bovine peripheral blood lymphocytes (BPBL) taken from nine cows. BPBL were incubated for 48 h with increasing concentrations of peptide fractions (< 5000 g·mol−1) extracted from the two fermented milks. Concanavalin A (conA), a known activator of lymphocyte proliferation, was used as a positive control. Fermentation products from E. faecalis TH563 showed a significantly (P < 0.05) greater ACE-inhibitory activity than that obtained by L. delb. bulgaricus LA2 (69.43 ± 3.12% vs. 60.86 ± 1.01%). The immunomodulatory activity showed a large interanimal variability. Peptide fractions from milk fermented by L. delb. bulgaricus LA2 significantly inhibited BPBL proliferation at concentrations of 5, 25 and 50 μg·mL−1 in the presence of conA (P < 0.01). E. faecalis TH563 did not significantly modify BPBL proliferation at any peptide concentration used. In conclusion, L. delb. bulgaricus LA2-fermented milk showed ACE-inhibitory and immunomodulatory activities, while E. faecalis TH563-fermented milk had high ACE-inhibitory activity, suggesting a possible use of these strains for determining bioactive properties in dairy products.

Abstract

-I (ACE) Enterococcus faecalis TH563 Lactobacillus delbrueckii subsp. bulgaricus LA2 ACE ACE (BPBL) A (conA) 48 h E. faecalis TH563 L. delb. bulgaricus LA2 ACE (69.43 ± 3.12%; P < 0.05) conA L. delb. bulgaricus LA2 (5-25-50 μg·mL−1) BPBL (P < 0.01), E. faecalis TH563 L. delb. bulgaricus LA2 ACE E. faecalis TH563 ACE

Résumé

Les protéines du lait sont des précurseurs de composés à activité biologique, qui sont libérés par protéolyse enzymatique. L’inhibition de l’enzyme convertissant l’angiotensine-I (ACE) et l’activité immunomodulatrice sont des activités d’intérêt parmi les activités biologiques reconnues des composés du lait. Dans cette étude, les activités anti-ACE et immunomodulatrices ont été analysées dans du lait fermenté par deux souches bactériennes isolées de produits laitiers italiens, Enterococcus faecalis TH563 ou Lactobacillus delbrueckii ssp. bulgaricus LA2. L’activité anti-ACE était déterminée par une méthode enzymatique pré-établie, l’activité immunomodulatrice par la prolifération de lymphocytes de sang périphérique de bovin (BPBL), prélevés à partir de neuf vaches. Les BPBL étaient incubées pendant 48 h en présence de concentrations croissantes de fractions peptidiques (< 5000 g·mol−1) extraites des deux laits fermentés. La concanavalin A (conA), un activateur connu de la prolifération des lymphocytes, était utilisée comme témoin positif. Le produit fermenté par E. faecalis TH563 montrait une activité anti-ACE significativement (P < 0,05) plus élevée que celle obtenue avec L. delb. bulgaricus LA2 (69,43 ± 3,12 %, vs. 60,86 ± 1,01 %). L’activité immunomodulatrice montrait une forte variabilité inter-animal. Les fractions peptidiques issues du lait fermenté par L. delb. bulgaricus LA2 inhibaient significativement (P < 0,01) la prolifération des BPBL aux concentrations 5, 25, et 50 μ·gmL−1 en présence de conA. E. faecalis TH563 ne modifiait pas significativement la prolifération des BPBL quelle que soit la concentration en peptides mise en œuvre. En conclusion, L. delb. bulgaricus LA2 produisait un lait fermenté avec des activités anti-ACE et immunomodulatrices, alors que E. faecalis TH563 produisait un lait fermenté à forte activité anti-ACE, suggérant une utilisation possible de ces souches pour apporter des propriétés bioactives dans les produits laitiers.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Andrighetto C., Knijff E., Lombardi A., Torriani S., Vancanneyt M., Kersters K., Swings J., Dellaglio F., Phenotypic and genetic diversity of enterococci isolated from Italian cheeses, J. Dairy Res. 68 (2001) 303–316.

    Article  CAS  Google Scholar 

  2. Andrighetto C., Marcazzan G., Cariolato D., Storti A., Cattelan A., Lombardi A., Isolation and characterization of microrganisms from traditional Triveneto cheeses, Sci. Tecn. Latt-Cas. 57 (2006) 309–318.

    CAS  Google Scholar 

  3. Cariolato D., Andrighetto C., Lombardi A., Occurrence of virulence factors and antibiotic resistance in Enterococcus faecalis and Enterococcus faecium collected from dairy and human samples in North of Italy, Food Control 19 (2008) 886–892.

    Article  CAS  Google Scholar 

  4. Cushman D.W., Cheung H.S., Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung, Biochem. Pharmacol. 20 (1971) 1637–1648.

    Article  CAS  Google Scholar 

  5. De Simone C., Bianchi Salvadori B., Negri R., Ferrazzi M., Baldinelli M., Vesely R., The adjuvant effect of yogurt on production of gamma-interferon by con A-stimulated human peripheral blood lymphocytes, Nutr. Rep. Int. 33 (1986) 419–433.

    Google Scholar 

  6. Donkor O.N., Henriksson A., Vasiljevic T., Shah N.P., Proteolytic activity of dairy lactic acid bacteria and probiotics as determinant of growth and in vitro angiotensin-converting enzyme inhibitory activity in fermented milk, Lait 87 (2007) 21–38.

    Article  CAS  Google Scholar 

  7. Ewaschuk J.B., Walker J.W., Diaz H., Madsen K.L., Bioproduction of conjugated linoleic acid by probiotic bacteria occurs in vitro and in vivo in mice, J. Nutr. 136 (2006) 1483–1487.

    CAS  Google Scholar 

  8. Franz C.M., Holzapfel W.H., Stiles M.E., Enterococci at the crossroads of food safety?, Int. J. Food Microbiol. 47 (1999) 1–24.

    Article  CAS  Google Scholar 

  9. Fujiwara S., Kadooka Y., Hirita T., Nakazato H., Screening for mitogenic activity of food microorganisms and their skim milk culture supernatants, J. Jpn. Soc. Nutr. Food Sci. 43 (1990) 203–208.

    Article  Google Scholar 

  10. Gobbetti M., Ferranti P., Smacchi E., Goffredi F., Addeo F., Production of angiotensin-I-converting-enzyme-inhibitory peptides in fermented milks started by Lactobacillus delbrueckii subsp. bulgaricus SS1 and Lactococcus lactis subsp. cremoris FT4, Appl. Environ. Microbiol. 66 (2000) 3898–3904.

    Article  CAS  Google Scholar 

  11. Hayes M., Ross R.P., Fitzgerald R.J., Stanton C., Putting microbes into work: dairy fermentation, cell factories and bioactive peptides. Part I: overview, Biotechnol. J. 2 (2007) 435–439.

    Article  CAS  Google Scholar 

  12. Hernandez-Ledesma B., Amigo L., Ramos M., Recio I., Angiotensin converting enzyme inhibitory activity in commercial fermented products. Formation of peptides under simulated gastrointestinal digestion, J. Agric. Food Chem. 52 (2004) 1504–1510.

    Article  CAS  Google Scholar 

  13. Hull M.E., Studies on milk proteins. Colorimetric determination of the partial hydrolysis of the proteins in milk, J. Dairy Sci. 30 (1947) 881–884.

    Article  CAS  Google Scholar 

  14. IDF, Dairy Starter Cultures of Lactic Acid Bacteria (LAB) — Standard of Identity, Standard 149A, International Dairy Federation, Brussels, Belgium, 1997.

    Google Scholar 

  15. Layne E., Spectrophotometric and turbidimetric methods for measuring proteins, in: Colowick S.P., Kaplan N.O. (Eds.), Methods in Enzymology, Academic Press Inc., New York, USA, 1957, pp. 447–455.

    Chapter  Google Scholar 

  16. Maeno M., Yamamoto N., Takano T., Identification of an antihypertensive peptide from casein hydrolysate produced by a proteinase from Lactobacillus helveticus CP790, J. Dairy Sci. 79 (1996) 1316–1321.

    Article  CAS  Google Scholar 

  17. Miguel M., Muguerza B., Sanchez E., Delgado M.A., Recio I., Ramos M., Aleixandre M.A., Changes in arterial blood pressure in hypertensive rats caused by long-term intake of milk fermented by Enterococcus faecalis CECT 5728, Br. J. Nutr. 94 (2005) 36–43.

    Article  CAS  Google Scholar 

  18. Moller N.P., Scholz-Ahrens K.E., Roos N., Schrezenmeir J., Bioactive peptides and proteins from foods: indication for health effects, Eur. J. Nutr. 47 (2008) 171–182.

    Article  CAS  Google Scholar 

  19. Muguerza B., Ramos M., Sanchez E., Manso M.A., Miguel M., Aleixandre A., Delgado M.A., Recio I., Antihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk, Int. Dairy J. 16 (2006) 61–69.

    Article  CAS  Google Scholar 

  20. Nakamura Y., Yamamoto N., Sakai K., Okubo A., Yamazaki S., Takano T., Purification and characterization of angiotensin I-converting enzyme-inhibitors from sour milk, J. Dairy Sci. 78 (1995) 777–783.

    Article  CAS  Google Scholar 

  21. Nakamura Y., Yamamoto N., Sakai K., Takano T., Antihypertensive effect of sour milk and peptides isolated from it that are inhibitors to angiotensin I-converting enzyme, J. Dairy Sci. 78 (1995) 1253–1257.

    Article  CAS  Google Scholar 

  22. Narva M., Halleen J., Vaananen K., Korpela R., Effects of Lactobacillus helveticus fermented milk on bone cells in vitro, Life Sci. 75 (2004) 1727–1734.

    Article  CAS  Google Scholar 

  23. Nielsen M.S., Martinussen T., Flambart B., Sorensen K.I., Otte J., Peptide profiles and angiotensin-I-converting enzyme inhibitory activity of fermented milk products: effect of bacterial strain, fermentation pH, and storage time, Int. Dairy J. 19 (2009) 155–165.

    Article  CAS  Google Scholar 

  24. Parra M.D., Martinez de Morentin B.E., Cobo J.M., Mateos A., Martinez J.A., Daily ingestion of fermented milk containing Lactobacillus casei DN114001 improves innatedefense capacity in healthy middle-aged people, J. Physiol. Biochem. 60 (2004) 85–91.

    Article  CAS  Google Scholar 

  25. Quiros A., Ramos M., Muguerza B., Delgado M.A., Miguel M., Aleixandre A., Recio I., Identification of novel antihypertensive peptides in milk fermented with Enterococcus faecalis, Int. Dairy J. 17 (2007) 33–41.

    Article  CAS  Google Scholar 

  26. Rachid M., Matar C., Duarte J., Perdigon G., Effect of milk fermented with a Lactobacillus helveticus R389(+) proteolytic strain on the immune system and on the growth of 471 breast cancer cells in mice, FEMS Immunol. Med. Microbiol. 47 (2006) 242–253.

    Article  CAS  Google Scholar 

  27. Sarantinopoulos P., Andrighetto C., Georgalaki M.D., Rea M.C., Lombardi A., Cogan T.M., Kalantzopoulos G., Tsakalidou E., Biochemical properties of enterococci relevant to their technological performance, Int. Dairy J. 11 (2001) 621–647.

    Article  CAS  Google Scholar 

  28. Templer S.P., Rohner P., Baumgartner A., Relation of Enterococcus faecalis and Enterococcus faecium isolates from foods and clinical specimens, J. Food Prot. 71 (2008) 2100–2104.

    CAS  Google Scholar 

  29. Wu J.P., Aluko R.E., Nakai S., Structural requirements of angiotensin I-converting enzyme inhibitory peptides: quantitative structure-activity relationship study of di- and tripeptides, J. Agric. Food Chem. 54 (2006) 732–738.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Experimental Veterinary Science, University of Padua, Legnaro, Padua, Italy

    Daniela Regazzo, Laura Da Dalt, Alessandro Negro & Gianfranco Gabai

  2. Istituto per la Qualità e le Tecnologie Agroalimentari, Veneto Agricoltura, Thiene, Vicenza, Italy

    Angiolella Lombardi & Christian Andrighetto

Authors
  1. Daniela Regazzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laura Da Dalt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Angiolella Lombardi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christian Andrighetto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alessandro Negro
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gianfranco Gabai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daniela Regazzo.

About this article

Cite this article

Regazzo, D., Da Dalt, L., Lombardi, A. et al. Fermented milks from Enterococcus faecalis TH563 and Lactobacillus delbrueckii subsp. bulgaricus LA2 manifest different degrees of ACE-inhibitory and immunomodulatory activities. Dairy Sci. Technol. 90, 469–476 (2010). https://doi.org/10.1051/dst/2010009

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1051/dst/2010009

Search

Navigation