Applied Microbiology and Biotechnology

, Volume 65, Issue 4, pp 433–439 | Cite as

Isolation and partial characterization of bacteriocins from Pediococcus species

Applied Microbial and Cell Physiology


Lactic acid bacteria have received increased attention as a potential food preservative due to their strong antagonistic activity against many food-spoilage and pathogenic organisms. Three Pediococcus species, P. acidilactici NCIM 2292, P. pentosaceous. NCIM 2296 and P. cervisiae NCIM 2171, were evaluated for bacteriocin production. Inhibitory substance were produced during the late growth phase and maximum production occurred at 37 °C after 36–48 h of incubation. Bacteriocins partially purified from these species by cold-acetone precipitation at 0 °C and cell adsorption desorption techniques have a broad inhibitory spectrum against microorganisms, including gram-negative bacteria such as Escherichia coli and Pseudomonas. Proteolytic enzymes inactivated these peptides, but amylase and lipase did not show any effect. The bacteriocins were stable over a wide pH range (3–8) and apparently most active at pH 4.0–5.0. They were heat-stable (1 h at ~80 °C and autoclaving) at pH 5.0. No loss in activity was observed when stored under refrigeration (4–8 °C). Tris-Tricine SDS-PAGE revealed the molecular masses of these peptides to be between 3.5 and 5.0 kDa.


  1. Bhunia AK, Johnson MG (1992) A modified method to directly detect in SDS-PAGE the bacteriocin of Pediococcus acidilactici. Lett Appl Microbiol 41:84–89Google Scholar
  2. Bhunia AK, Johnson MC, Ray B (1988) Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici. J Appl Bacteriol 65:261–268PubMedGoogle Scholar
  3. Cintas LM, Rodriguez JM, Fernandez MF, Sletten K, Nes IF, Hernandez PE, Holo H (1995) Isolation and characterization of pediocin L50, a new bacteriocin from Pediococcus acidilactici with a broad inhibitory spectrum. Appl Environ Microbiol 61:2643–2648PubMedGoogle Scholar
  4. Cintas LM, Casaus P, Fernandez MF, Hernandez PE (1998) Comparative antimicrobial activity of enterocin L50, pediocin PA-1, nisin A and lactocin S against spoilage and food borne pathogenic bacteria. Food Microbiol 62:1764–1769Google Scholar
  5. Cleveland J, Montville TJ, Nes IF, Chikindas ML (2001) Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 71:1–20Google Scholar
  6. Daba H, Lacroix C, Huang J, Simrad RE, Lemieux L (1994) Simple method of purification and sequencing of a bacteriocin produced by Pediococcus acidilactici UL5. J Appl Bacteriol 77:682–688Google Scholar
  7. Eijsink VG, Skeie M, Middelhoven PH, Brurberg MB, Nes IF (1998) Comparative studies of Class IIa bacteriocins of lactic acid bacteria. Appl Environ Microbiol 64:3275–3281PubMedGoogle Scholar
  8. Gornall AG, Bardwill GJ, David MM (1949) Determination of serum proteins by means of biuret reaction. J Biol Chem 117:751–766Google Scholar
  9. Green G, Dicks LMT, Bruggeman G, Vandamme EJ, Chikindas ML (1997) Pediocion-PD-1, a bacteriocidal antimicrobial peptide from Pediococcus damnosus NCFB 1832. J Appl Microbiol 83:127–132PubMedGoogle Scholar
  10. Hanlin MB, Kalchayanand N, Ray P, Ray B (1993) Bacteriocins of lactic acid bacteria in combination have a greater antibacterial activity. J Food Prot 56:252–255Google Scholar
  11. Holzapfel WH, Geisen R, Schillinger U (1995) Biological preservation of foods with reference to protective cultures, bacteriocins and food grade enzymes. Int J Food Microbiol 24:343–362CrossRefPubMedGoogle Scholar
  12. Kim CH, Ji GE, Ahn C (2000) Purification and molecular characterization of a bacteriocin from Pediococcus sp. KCA 1303–10 isolated from fermented flatfish. Food Sci Biotechnol l9:270–276Google Scholar
  13. Mulet-Powell N, Lactoste-Armynot AM, Vinas M, Simeon De Buochberg M (1998) Interactions between pairs of bacteriocins from Lactic acid Bacteria. J Food Prot 61:1210–1212Google Scholar
  14. Ray B, Miller KW (2000) Pediocin natural food antimicrobial systems. CRC, Boca Raton, FloridaGoogle Scholar
  15. Rekhif N, Atrih A, Lefebvre G (1994) Characterization and partial purification of plantaricin LC74, a bacteriocin produced by Lactobacillus plantarum LC74. Biotechnol Lett 16:771–776Google Scholar
  16. Schagger H, Von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1–100 kDa. Anal Biochem 166:368–379PubMedGoogle Scholar
  17. Schved F, Lalazar A, Henis Y, Juven BJ (1993) Purification, partial characterization and plasmid-linkage of pediocin SJ-1, a bacteriocin produced by Pediococcus damnosus acidilactici. J Appl Bacteriol 74:67–77PubMedGoogle Scholar
  18. Scopes RK (1984) Protein purification: principles and practice. Springer, New YorkGoogle Scholar
  19. Skytta E, Haikara A, Tiina Mattila- Sandholm (1993) Production and characterization of antibacterial compounds produced by Pediococcus damnosus and Pediococcus pentosaceus. J Appl Bacteriol 74 :134 −142Google Scholar
  20. Tagg JR, McGiven AR (1971) Assay systems for bacteriocins. Applied Microbiol 21:125Google Scholar
  21. Yang R, Johnson MC, Ray B (1992) Novel method to extract large amounts of bacteriocins from lactic acid bacteria. Appl Environ Microbiol 58:3353–3359Google Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Biochemistry and Nutrition DisciplineDefence Food Research LaboratoryMysoreIndia

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