Skip to main content
Log in

Characterization of mesentericin ST99, a bacteriocin produced by Leuconostoc mesenteroides subsp. dextranicum ST99 isolated from boza

  • Original Paper
  • Published:
Journal of Industrial Microbiology and Biotechnology

Abstract

Lactic acid bacteria isolated from Boza, a cereal-fermented beverage from Belogratchik, Bulgaria, were screened for the production of bacteriocins. With the first screening, 13 of the 52 isolates inhibited the growth of Listeria innocua and Lactobacillus plantarum. The cell-free supernatant of one of these strains, classified as Leuconostoc mesenteroides subsp. dextranicum ST99, inhibited the growth of Bacillus subtilis, Enterococcus faecalis, several Lactobacillus spp., Lactococcus lactis subsp. cremoris, Listeria innocua, Listeria monocytogenes, Pediococcus pentosaceus, Staphylococcus aureus and Streptococcus thermophilus. Clostridium spp., Carnobacterium spp., L. mesenteroides and Gram-negative bacteria were not inhibited. Maximum antimicrobial activity, i.e. 6,400 arbitrary units (AU)/ml, was recorded in MRS broth after 24 h at 30°C. Incubation in the presence of protease IV and pronase E resulted in loss of antimicrobial activity, confirming that growth inhibition was caused by a bacteriocin, designated here as mesentericin ST99. No loss in activity was recorded after treatment with α-amylase, SDS, Tween 20, Tween 80, urea, Triton X-100, N-laurylsarcosin, EDTA and phenylmethylsulfonylfluoride. Mesentericin ST99 remained active after 30 min at 121°C and after 2 h of incubation at pH 2 to 12. Metabolically active cells of L. innocua treated with mesentericin ST99 did not undergo lysis. Mesentericin ST99 did not adhere to the cell surface of strain ST99. Precipitation with ammonium sulfate (70% saturation), followed by Sep-Pack C18 chromatography and reverse-phase HPLC on a C18 Nucleosil column yielded one antimicrobial peptide.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3a, b

Similar content being viewed by others

References

  1. Atrih A, Rekhif N, Milliere JB, Lefebvre G (1993) Detection and characterization of a bacteriocin produced by Lactobacillus plantarum C19. Can J Microbiol 39:1173–1179

    CAS  Google Scholar 

  2. Blom H, Katla T, Holck A, Sletten K, Axelsson L, Holo H (1999) Characterization, production and purification of leucocin H, a two-peptide bacteriocin from Leuconostoc MF215B. Curr Microbiol 39:43–48

    CAS  PubMed  Google Scholar 

  3. Budde BB, Horubaek T, Jacobsen T, Barkholt V, Koch AG (2003) Leuconostoc carnosum 4010 has the potential for use as a protective culture for vacuum-packed meats: culture isolation, bacteriocin identification, and meat application experiments. Int J Food Microbiol 83:171–184

    Article  PubMed  Google Scholar 

  4. Daba H, Lacroix C, Huang J, Simard RE (1993) Influence of growth conditions on production and activity of mesenterocin 5 by a strain of Leuconostoc mesenteroides. Appl Microbiol Biotechnol 39:166–173

    CAS  Google Scholar 

  5. Degnan AJ, Yousef AE, Luchansky JB (1992) Use of Pediococcus acidilactici to control Listeria monocytogenes in temperature-abused vacuum-packaged Wieners. J Food Protect 55:98–103

    Google Scholar 

  6. Delves-Broughton J (1990) Nisin and its uses of the DGHM preservative. Food Technol 44:100–117

    CAS  Google Scholar 

  7. De Man JC, Rogosa M, Sharpe E (1960) A medium for the cultivation of lactobacilli. J Appl Bacteriol 23:130–135

    Google Scholar 

  8. Elliker PR, Anderson AW, Hannesson G (1956) An agar culture medium for lactic acid streptococci and lactobacilli. J Dairy Sci 39:1611

    Google Scholar 

  9. Felix JV, Papathanasopoulos MA, Smith AA, Von Holy A, Hastings JW (1994) Characterization of leucocin B-Ta11a: a bacteriocin from Leuconostoc carnosum Ta11a isolated from meat. Curr Microbiol 29:207–212

    CAS  PubMed  Google Scholar 

  10. Garver KI, Muriana M (1993) Detection and characterization of bacteriocin producing lactic acid bacteria from retail food products. Int J Food Microbiol 19:241–258

    Article  CAS  PubMed  Google Scholar 

  11. Geisen R, Becker B, Holzapfel WH (1993) Bacteriocin production of Leuconostoc carnosum LA54A at different combinations of pH and temperature. J Ind Microbiol 12:337–340

    Google Scholar 

  12. Gotcheva V, Pandiella SS, Angelov A, Roshkova ZG, Webb C (2000) Microflora identification of the Bulgarian cereal-based fermented beverage boza. Process Biochem 36:127–130

    Article  CAS  Google Scholar 

  13. Hancioglu O, Karapinar M (1997) Microflora of Boza, a traditional fermented Turkish beverage. Int J Food Microbiol 35:271–274

    Article  CAS  PubMed  Google Scholar 

  14. Harding CD, Shaw BG (1990) Antimicrobial activity of Leuconostoc gelidium against related species and Listeria monocytogenes. J Appl Bacteriol 69:648–654

    CAS  PubMed  Google Scholar 

  15. Hasting JW, Sailer M, Johnson K, Roy KL, Vederas JC, Stiles ME (1991) Characterization of leucocin AUAL 187 and cloning of the bacteriocin gene from Leuconostoc gelidium. J Bacteriol 173:7491–7500

    PubMed  Google Scholar 

  16. Hechard Y, Derijard B, Letellier F, Cenatiempo Y (1992) Characterization and purification of mesenterocin Y105, an anti-Listeria bacteriocin from Leuconostoc mesenteroides. J Gen Microbiol 138:2725–2731

    CAS  PubMed  Google Scholar 

  17. Kabadjova P, Gotcheva I, Ivanova I, Dousset X (2000) Investigation of bacteriocin activity of lactic acid bacteria isolated from boza. Biotechnol Biotechnol Eq 14:56–59

    CAS  Google Scholar 

  18. Keppler K, Geisen R, Holzapfel WH (1994) An α-amylase sensitive bacteriocin of Leuconostoc carnosum. Food Microbiol 11:39–45

    Article  CAS  Google Scholar 

  19. Klaenhammer TR (1988) Bacteriocins of lactic acid bacteria. Biochimie 70:337–349

    Article  CAS  PubMed  Google Scholar 

  20. Klaenhammer TR (1993) Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol Rev 12:39–86

    Article  CAS  PubMed  Google Scholar 

  21. Lewus CB, Kaiser A, Montville TJ (1991) Inhibition of food-borne bacterial pathogens by bacteriocins from lactic acid bacteria isolated from meat. Appl Environ Microbiol 57:1683–1688

    CAS  PubMed  Google Scholar 

  22. Lewus CB, Sun S, Montville TJ (1992) Production of an α-amylase bacteriocin by an atypical Leuconostoc paramesenteroides strain. Appl Environ Microbiol 58:143–149

    CAS  Google Scholar 

  23. Mataragas M, Metaxopoulos J, Galiotou M, Drosinos EH (2003) Influence of pH and temperature on growth and bacteriocin production by Leuconostoc mesenteroides L124 and Lactobacillus curvatus L442. Meat Sci 64:265–271

    Article  CAS  Google Scholar 

  24. Mathieu F, Sudirman Suwandhi I, Rekhif N, Milliere JB, Lefebvre G (1992) Mesenterocin 52, a bacteriocin produced by Leuconostoc mesenteroides ssp. mesenteroides FR52. J Appl Bacteriol 74:372–349

    Google Scholar 

  25. Müller T (1990) Comparison of methods for differentiation between homofermentative and heterofermentative lactic acid bacteria. Zentralbl Microbiol 145:363–366

    Google Scholar 

  26. Nettles CG, Barefoot SF (1993) Biochemical and genetic characteristics of bacteriocins of food-associated lactic acid bacteria. J Food Prot 56:338–356

    CAS  Google Scholar 

  27. Nielsen JW, Dickson JS, Crouse JD (1990) Use of a bacteriocin produced by Pediococcus acidilactici to inhibit Listeria monocytogenes associated with fresh meat. Appl Environ Microbiol 56:2142–2145

    CAS  PubMed  Google Scholar 

  28. Papathanasopoulos MA, Krier F, Revol Junelles AM, Lefebvre G, Caer JP, Von Holy A, Hastings JW (1997) Multiple bacteriocin production by Leuconostoc mesenteroides TA33a and other Leuconostoc/Weissella strains. Curr Microbiol 35:331–335

    Article  CAS  PubMed  Google Scholar 

  29. Papathanasopoulos MA, Dykes GA, Revol Junelles AM, Delfour A, Von Holy A, Hastings JW (1998) Sequence and structural relationships of leucocins A-, B- and C-TA33a from Leuconostoc mesenteroides TA33a. Microbiology 144:1343–1348

    CAS  PubMed  Google Scholar 

  30. Parente E, Moles M, Ricciardi A (1996) Leucocin F10, a bacteriocin from Leuconostoc carnosum. Int J Food Microbiol 33:231–243

    Article  CAS  PubMed  Google Scholar 

  31. Requena T, Pelàez C (1995) Actividad antimicrobiana de bacterias lacticas. Produccion de bacteriocinas. Rev Esp Cienc Tecnol Aliment 35:19–44

    Google Scholar 

  32. Revol Junelles AM, Mathis R, Krier F, Fleury Y, Delfour A, Lefebvre G (1996) Leuconostoc mesenteroides subsp. mesenteroides FR52 synthesizes two distinct bacteriocins. Lett Appl Microbiol 23:120–124

    CAS  PubMed  Google Scholar 

  33. Schillinger U, Lücke FK (1989) Antibacterial activity of Lactobacillus sake isolated from meat. J Appl Bacteriol 70:473–478

    Google Scholar 

  34. Sneath PHA, Mair NS, Sharpe ME, Holt JG (1986) In: Sneath PHA, Hold JG (eds) Bergey’s manual of systematic bacteriology. Williams and Wilkins, Baltimore, pp 1071–1075

    Google Scholar 

  35. Stiles ME, Hastings JW (1991) Bacteriocin production by lactic acid bacteria: potential for use in meat preservation. Trends Food Sci Technol 2:247–251

    Article  CAS  Google Scholar 

  36. Tagg JR, McGiven AR (1971) Assay system for bacteriocins. Appl Microbiol 21:943

    CAS  PubMed  Google Scholar 

  37. Tagg JR, Dajani AS, Wannamaker LW (1976) Bacteriocins of Gram-positive bacteria. Bacteriol Rev 40:722–756

    CAS  PubMed  Google Scholar 

  38. Van Laack RL, Schillinger U, Holzapfel WH (1992) Characterization and partial purification of a bacteriocin produced by Leuconostoc carnosum LA44A. Int J Food Microbiol 16:183–195

    Article  PubMed  Google Scholar 

  39. Van Reenen CA, Dicks LMT, Chikindas ML (1998) Isolation, purification and partial characterization of plantaricin 423, a bacteriocin produced by Lactobacillus plantarum. J Appl Microbiol 84:1131–1137

    Article  PubMed  Google Scholar 

  40. Yang R, Johnson M, Ray B (1992) A novel method to extract large amounts of bacteriocins from lactic acid bacteria. Appl Environ Microbiol 58:3355–3359

    CAS  PubMed  Google Scholar 

  41. Zorba M, Hancioglu O, Genc M, Karapinar M, Ova G (2003) The use of starter culture in the fermentation of boza, a traditional Turkish beverage. Process Biochem 38:1405–1411

    Article  CAS  Google Scholar 

Download references

Acknowledgement

This research was funded by the French Embassy (Service Culturel) in Sofia, Bulgaria.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Leon M. T. Dicks.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Todorov, S.D., Dicks, L.M.T. Characterization of mesentericin ST99, a bacteriocin produced by Leuconostoc mesenteroides subsp. dextranicum ST99 isolated from boza. J IND MICROBIOL BIOTECHNOL 31, 323–329 (2004). https://doi.org/10.1007/s10295-004-0153-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-004-0153-6

Keywords

Navigation