Archives of Microbiology

, Volume 181, Issue 1, pp 8–16 | Cite as

Production of plantaricin NC8 by Lactobacillus plantarum NC8 is induced in the presence of different types of gram-positive bacteria

  • Antonio Maldonado
  • José Luis Ruiz-Barba
  • Rufino Jiménez-DíazEmail author
Original Paper


Lactobacillus plantarum NC8 was shown to produce plantaricin NC8 (PLNC8), a recently purified and genetically characterized inducible class IIb bacteriocin, only when it was co-cultured with other gram-positive bacteria. Among 82 strains belonging to the genera Bacillus, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Listeria, Pediococcus, Staphylococcus, and Streptococcus, 41 were shown to induce PLNC8 production in L. plantarum NC8. There was apparently no relationship between the sensitivity of the strains and their ability to induce the bacteriocin, indicating that the inducer and sensitive phenotypes may not be linked. In some instances, induction was promoted by both living and heat-killed cells of the inducing bacteria. However, no PLNC8-inducing activity was found in the respective cell-free, pure culture supernatants. Inducer strains also promoted the production of a PLNC8-autoinducing activity by L. plantarum NC8, which was found only in the cell-free co-culture supernatants showing inhibitory activity. This PLNC8-autoinducing activity was diffusible, heat resistant, and of a proteinaceous nature, and was different from the bacteriocin itself. Taken together, the results suggest that the presence of specific gram-positive bacteria acts as an environmental stimulus activating both PLNC8 production by L. plantarum NC8 and a PLNC8-autoinducing activity, which in turn triggers or maintains bacteriocin production in the absence of inducing cells.


Autoinduction Induction Lactobacillus plantarum Plantaricin NC8 



We thank L. Axelsson (MATFORSK, Norwegian Food Research Institute, Osloveien, Norway) for providing the L. plantarum NC8 strain, and Mrs. Belén Caballero Guerrero for her skillful assistance. This work was supported by the Spanish Government through MCYT project AGL2000-1611-CO3-01. A.M. was the recipient of a grant from MCYT, Spain.


  1. Ahn C, Stiles ME (1990) Plasmid-associated bacteriocin production by a strain of Carnobacterium piscicola from meat. Appl Environ Microbiol 56:2503–2510PubMedGoogle Scholar
  2. Aukrust T, Blom H (1992) Transformation of Lactobacillus strains used in meat and vegetable fermentations. Food Res Int 25:253–261Google Scholar
  3. Axelsson L, Holck A (1995) The genes involved in production of and immunity to sakacin A, a bacteriocin from Lactobacillus sake Lb706. J Bacteriol 177:2125–2137PubMedGoogle Scholar
  4. Barefoot SF, Grinstead GA (1993) Bacteriocins for dairy propionibacteria and inducible bacteriocins of lactic acid bacteria. In: Hoover DG, Steenson LR (eds) Bacteriocins of lactic acid bacteria. Academic, New York, pp 219–231Google Scholar
  5. Barefoot SF, Chen YR, Hughes TA, Bodine AB, Shearer MY, Hughes MD (1994a) Identification and purification of a protein that induces production of the Lactobacillus acidophilus bacteriocin lactacin B. Appl Environ Microbiol 60:3522–3528PubMedGoogle Scholar
  6. Barefoot SF, Nettles CG, Chen YR (1994b) Lactacin B, a bacteriocin produced by Lactobacillus acidophilus. In: de Vuyst L, Vandamme EJ (eds) Bacteriocins of lactic acid bacteria: microbiology, genetics and applications. Blackie, London, pp 353–376Google Scholar
  7. Biswas S R, Ray P, Johnson MC, Ray B (1991) Influence of growth conditions on the production of a bacteriocin, pediocin AcH, by Pediococcus acidilacti AcH. Appl Environ Microbiol 57:1265–1267Google Scholar
  8. Brurberg M B, Nes IF, Eijsink VGH (1997) Pheromone-induced production of antimicrobial peptides in Lactobacillus. Mol Microbiol 26:347–360PubMedGoogle Scholar
  9. De Vuyst, L (1994) Bacteriocins and bacteriocin-like substances from Lactobacillus. In: de Vuyst L, Vandamme EJ (eds) Bacteriocins of lactic acid bacteria: microbiology, genetics and applications. Blackie, London, pp 319–329Google Scholar
  10. De Vuyst L, Vandamme EJ (1994) Antimicrobial potential of lactic acid bacteria. In: de Vuyst L, Vandamme EJ (eds) Bacteriocins of lactic acid bacteria: microbiology, genetics and applications. Blackie Academic and Professional, London, United Kingdom, pp 91–142Google Scholar
  11. De Vuyst L, Callewaert R, Crabbe K (1996) Primary metabolite kinetics of bacteriocin biosynthesis by Lactobacillus amylovorus and evidence for stimulation of bacteriocin production under unfavourable growth conditions. Microbiology 142:817–827Google Scholar
  12. Diep DB, Håvarstein LS, Nissen-Meyer J, Nes IF (1994) The gene encoding plantaricin A, a bacteriocin from Lactobacillus plantarum C11, is located on the same transcription unit as an agr-like regulatory system. Appl Environ Microbiol 60:160–166PubMedGoogle Scholar
  13. Diep DB, Håvarstein LS, Nes IF (1995) A bacteriocin-like peptide induces bacteriocin synthesis in Lactobacillus plantarum C11. Mol Microbiol 18:631–639PubMedGoogle Scholar
  14. Diep DB, Håvarstein LS, Nes IF (1996) Characterization of the locus responsible for the bacteriocin production in Lactobacillus plantarum C11. J Bacteriol 178:4472–4483PubMedGoogle Scholar
  15. Diep DB, Axelsson L, Grefsli C, Nes IF (2000) The synthesis of the bacteriocin sakacin A is a temperature-sensitive process regulated by a pheromone peptide through a three-component regulatory system. Microbiology 146:2155–2160PubMedGoogle Scholar
  16. Dykes GA (1995) Bacteriocins: ecological and evolutionary significance. Tree 10:186–189CrossRefGoogle Scholar
  17. Dykes GA, Hastings JW (1997) Selection and fitness in bacteriocin-producing bacteria. Proc R Soc Lon B 264:683–687CrossRefGoogle Scholar
  18. Eijsink VGH, Brurberg MB, Middelhoven PH, Nes IF (1996) Induction of bacteriocin production in Lactobacillus sake by a secreted peptide. J Bacteriol 178:2232–2237PubMedGoogle Scholar
  19. Floriano B, Ruiz-Barba JL, Jiménez-Díaz R (1998) Purification and genetic characterization of enterocin I from Enterococcus faecium 6T1a, a novel antilisterial plasmid-encoded bacteriocin which does not belong to the pediocin family of bacteriocins. Appl Environ Microbiol 64:4883–4890PubMedGoogle Scholar
  20. Flynn S, van Sinderen D, Thornton GM, Holo H, Nes IF, Collins JK (2002) Characterization of the genetic locus responsible for the production of ABP-118, a novel bacteriocin produced by the probiotic bacterium Lactobacillus salivarius subsp. salivarius UCC118. Microbiology 148:973-984PubMedGoogle Scholar
  21. Franz CMAP, van Belkum MJ, Worobo RW, Vederas J, Stiles ME (2000) Characterization of the genetic locus responsible for production and immunity of carnobacteriocin A: the immunity gene confers cross-protection to enterocin B. Microbiology 146:621–631PubMedGoogle Scholar
  22. Geis A, Singh J, Teuber M (1983) Potential of lactic streptococci to produce bacteriocin. Appl Environ Microbiol 45:205–211Google Scholar
  23. Gutowski-Ecked Z, Klein C, Siegers K, Bohm K, Hammelmann M, Entian KD (1994) Growth phase-dependent regulation and membrane localization of Spa B, a protein involved in biosynthesis of the lantibiotic subtilin. Appl Environ Microbiol 60:1–11PubMedGoogle Scholar
  24. Jack R, Tagg JR, Ray B (1995) Bacteriocins of Gram-positive bacteria. Microbiol Rev 59:171–200PubMedGoogle Scholar
  25. Jiménez-Díaz R, Rios-Sánchez RM, Desmazeaud M, Ruiz-Barba JL, Piard JC (1993) Plantaricin S and T, two new bacteriocins produced by Lactobacillus plantarum LPCO10 isolated from a green olive fermentation. Appl Environ Microbiol 59:1416–1424Google Scholar
  26. Jiménez-Díaz R, Ruiz-Barba JL, Cathcart DP, Holo H, Nes IF, Sletten KH, Warner PJ (1995) Purification and partial amino acid sequence of plantaricin S, a bacteriocin produced by Lactobacillus plantarum LPCO10, the activity of which depends on the complementary action of two peptides. Appl Environ Microbiol 61:4459–4463PubMedGoogle Scholar
  27. Kaiser AL, Montville TJ (1993) The influence of pH and growth rate on production of the bacteriocin, bavaricin MN, in batch and continuous fermentations. J Appl Bacteriol 75:536–540Google Scholar
  28. Klaenhammer TR (1993) Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol Rev 12:39–86PubMedGoogle Scholar
  29. Kleerebezem M, Quadri LEN (2001) Peptide pheromone-dependent regulation of antimicrobial peptide production in Gram-positive bacteria: a case of multicellular behavior. Peptides 22:1579–1596CrossRefPubMedGoogle Scholar
  30. Kleerebezem M, Quadri LEN, Kuipers O.P, de Vos WM (1997) Quorum sensing by peptide pheromones and two-component signal-transduction systems in gram-positive bacteria. Mol Microbiol 24:895-904PubMedGoogle Scholar
  31. Kleerebezem M, Kuipers OP, de Vos WM, Stiles ME, Quadri LEN (2001) A two-component signal transduction cascade in Carnobacterium piscicola LV17B: two signaling peptides and one sensor-transmitter. Peptides 22:1597–1601CrossRefPubMedGoogle Scholar
  32. Klein C, Kaletta C, Entian KD (1993) Biosynthesis of the lantibiotic subtilin is regulated by a histidine kinase/response regulator system. Appl Environ Microbiol 59:296–303PubMedGoogle Scholar
  33. Kuipers OP, Beerthuyzen MB, de Ruyter PGGA, Luesink EJ, de Vos WM (1995) Autoregulation of nisin biosynthesis in Lactococcus lactis by signal transduction. J Biol Chem 270:27299–27304PubMedGoogle Scholar
  34. Kuipers OP, de Ruyter PGGA, Kleerebezem M, de Vos WM (1998) Quorum sensing-controlled gene expression in lactic acid bacteria. J Biotechnol 64:15–21Google Scholar
  35. Leal-Sánchez MV, Jiménez-Díaz R, Maldonado-Barragán A, Garrido-Fernández A, Ruiz-Barba JL (2002) Optimization of bacteriocin production by batch fermentation of Lactobacillus plantarum LPCO10. Appl Environ Microbiol 68:4465-4471CrossRefPubMedGoogle Scholar
  36. Maldonado A, Ruiz-Barba JL, Jiménez-Díaz R (2003) Purification and genetic characterization of plantaricin NC8, a novel coculture-inducible two-peptide bacteriocin from Lactobacillus plantarum NC8. Appl Environ Microbiol 69:383–389CrossRefPubMedGoogle Scholar
  37. Moll GN, van den Akker E, Hauge HH, Nissen-Meyer J, Nes IF, Konings WN, Driessen AJM (1999) Complementary and overlapping selectivity of the two-peptide bacteriocins plantaricin EF and JK. J Bacteriol 181:4848–4852PubMedGoogle Scholar
  38. Mørtvedt-Abildgaard CI, Nissen-Meyer J, Jelle B, Grenov B, Skaugen M, Nes IF (1995) Production and pH-dependent bactericidal activity of lactococin S, a lantibiotic from Lactobacillus sake L45. Appl Environ Microbiol 61:175–179Google Scholar
  39. Nes IF, Eijsink VGH (1999) Regulation of group II peptide bacteriocin synthesis by quorum sensing mechanisms. In: Dunny GM, Winans SC (eds) Cell-cell signaling in bacteria. American Society for Microbiology, Washington, DC, pp 175–192Google Scholar
  40. Nes IF, Diep DB, Håvarstein LS, Brurberg MB, Eijsink VGH, Holo H (1996) Biosynthesis of bacteriocins in lactic acid bacteria. Antonie van Leeuwenhoek 70:113–128PubMedGoogle Scholar
  41. Nilsen T, Nes IF, Holo H (1997) An exported inducer peptide regulates bacteriocin production in Enterococcus faecium CTC492. J Bacteriol 180:1848–1854Google Scholar
  42. Quadri LEN, Kleerebezem M, Kuipers OP, de Vos WM, Roy KL, Vederas JC, Stiles ME (1997) Characterization of a locus from Carnobacterium piscicola LV17B involved in bacteriocin production and inmunity: evidence for global inducer-mediated transcriptional regulation. J Bacteriol 179:6163–6171PubMedGoogle Scholar
  43. Riley MA (1998) Molecular mechanisms of bacteriocin evolution. Annu Rev Genet 32:255–278CrossRefPubMedGoogle Scholar
  44. Ruiz-Barba JL, Cathcart DP, Warner PJ, Jiménez-Díaz R (1994) Use of Lactobacillus plantarum LPCO10, a bacteriocin producer, as a starter culture in spanish-style green olive fermentations. Appl Environ Microbiol 60:2059–2064Google Scholar
  45. Saucier L, Poon A, Stiles ME (1995) Induction of bacteriocin in Carnobacterium piscicola LV17. J Appl Bacteriol 78:684–690Google Scholar
  46. Saucier L, Paradkar AS, Frost LS, Jensen SE, Stiles ME (1997) Transcriptional analysis and regulation of carnobacteriocin production in Carnobacterium piscicola LV17. Gene 188:271–277CrossRefPubMedGoogle Scholar
  47. Sip A, Grajek W, Boyaval P (1998) Enhancement of bacteriocin production by Carnobacterium divergens AS7 in the presence of a bacteriocin-sensitive strain Carnobacterium piscicola. Int Food Microbiol 42:63–69CrossRefGoogle Scholar
  48. Tagg JR, Dajani AS, Wannamaker LW (1976) Bacteriocins of gram-positive bacteria. Bacteriol Rev 40:722–756PubMedGoogle Scholar
  49. Tichaczeck PS, Nissen-Meyer J, Nes IF, Vogel RF, Hammes WP (1992) Characterization of the bacteriocins curvacin A from Lactobacillus curvatus LTH1174 and sakacin P from L. sake LTH673. Syst Appl Microbiol 15:460–468Google Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Antonio Maldonado
    • 1
  • José Luis Ruiz-Barba
    • 1
  • Rufino Jiménez-Díaz
    • 1
    Email author
  1. 1.Departamento de Biotecnología de Alimentos, Instituto de la GrasaConsejo Superior de Investigaciones CientíficasSevilleSpain

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