Partial purification, characterization and mode of action of bacteriocins produced by three strains of Pediococcus sp.

  • Bishnu Charan Dey
  • Neekunj Rai
  • Saikat Das
  • Sukhendu Mandal
  • Vivekananda MandalEmail author
Original Article


The main objective of the study is to assess a comparative antibacterial potential of three new bacteriocins produced by Pediococcus sp. through partial characterization and mode of action against some food spoilage bacteria. The bacteriocins from three different Pediococcus sp. viz. Pediococcus sp. LAB 33 (HQ185406), Pediococcus sp. LAB 41 (HQ185407), and Pediococcus sp. LAB 51 (HQ184064) were partially purified by adsorption–desorption method and tested for autoclave heat, pH, detergent and enzymes stability. A comparative analysis by Tricin-SDS PAGE with MALDI-TOF MS was done to estimate their molecular weight. The mode of action studies were done by cell viability and lactate dehydrogenase assay against two food associated pathogens, viz. Listeria monocytogenes and Pseudomonas aeruginosa using standard protocols. The bacteriocins produced by the strains were resistant to autoclave heat, detergent, wide range of pH and were active against different food borne pathogens at a minimum dose of ~ 100 AU/ml. The mode of action studies showed bactericidal action with lysis of the targeted cells. Therefore, the selective low dose efficacy, heat and detergent stability of the bacteriocins produced by the three strains could be considered as potent bacteriocins for use as food preservatives.


Bacteriocins Pediococcus sp. Listeria monocytogenes Pseudomonas aeruginosa Tricin-SDS PAGE MALDI-TOF MS 



  1. Bhunia AK, Johnson MC, Ray B, Kalchayanand N (1991) Mode of action of pediocin AcH from Pediococcus acidilactici H on sensitive bacterial strains. J Appl Bacteriol 70:25–33CrossRefGoogle Scholar
  2. Cai Y, Ng LK, Farber JM (1997) Isolation and characterization of nisin-producing Lactococcus lactis subsp. lactis from bean-sprouts. J Appl Microbiol 83:499–507CrossRefGoogle Scholar
  3. Chikindas ML, García-Garcerá MJ, Driessen AJ et al (1993) Pediocin PA1, a bacteriocin from Pediococcus acidilactici PAC l.0, forms hydrophilic pores in the cytoplasmic membrane of target cells. Appl Environ Microbiol 59(11):3577–3584Google Scholar
  4. Chikindas ML, Weeks R, Drider D et al (2018) Functions and emerging applications of bacteriocins. Curr Opin Biotechnol 49:23–28. CrossRefGoogle Scholar
  5. Daw MA, Falkiner FR (1996) Bacteriocins: nature, function and structure. Micron 27:467–479. CrossRefGoogle Scholar
  6. De Vuyst L, Leroy F (2007) Bacteriocins from lactic acid bacteria: production, purification, and food applications. J Mol Microbiol Biotechnol 13:194–199CrossRefGoogle Scholar
  7. De Vuyst L, Vandamme EJ (1994) Antimicrobial potential of lactic acid bacteria. In: Bacteriocins of lactic acid bacteria. Springer, pp 91–142Google Scholar
  8. 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. Microbiol Read Engl 146(Pt 9):2155–2160. CrossRefGoogle Scholar
  9. Elegado FB, Kim WJ, Kwon DY (1997) Rapid purification, partial characterization, and antimicrobial spectrum of the bacteriocin, Pediocin AcM, from Pediococcus acidilactici M. Int J Food Microbiol 37:1–11CrossRefGoogle Scholar
  10. García-Cayuela T, Requena T, Martínez-Cuesta MC, Peláez C (2017) Rapid detection of Lactococcus lactis isolates producing the lantibiotics nisin, lacticin 481 and lacticin 3147 using MALDI-TOF MS. J Microbiol Methods 139:138–142. CrossRefGoogle Scholar
  11. Gough R, Gómez-Sala B, O’Connor PM et al (2017) A simple method for the purification of Nisin. Probiotics Antimicrob Proteins 9:363–369. CrossRefGoogle Scholar
  12. Guerra NP, Pastrana L (2003) Influence of pH drop on both nisin and pediocin production by Lactococcus lactis and Pediococcus acidilactici. Lett Appl Microbiol 37:51–55CrossRefGoogle Scholar
  13. Heng NCK, Tagg JR (2006) What’s in a name? Class distinction for bacteriocins. Nat Rev Microbiol 4:160. CrossRefGoogle Scholar
  14. Ikram-ul-Haq null, Mukhtar H (2006) Biosynthesis of protease from Lactobacillus paracasei: kinetic analysis of fermentation parameters. Indian J Biochem Biophys 43:377–381Google Scholar
  15. Kalchayanand N, Hanlin MB, Ray B (1992) Sublethal injury makes Gram-negative and resistant Gram-positive bacteria sensitive to the bacteriocins, pediocin AcH and nisin. Lett Appl Microbiol 15:239–243CrossRefGoogle Scholar
  16. Keren T, Yarmus M, Halevy G, Shapira R (2004) Immunodetection of the bacteriocin lacticin RM: analysis of the influence of temperature and Tween 80 on its expression and activity. Appl Environ Microbiol 70:2098–2104CrossRefGoogle Scholar
  17. Klaenhammer TR (1988) Bacteriocins of lactic acid bacteria. Biochimie 70:337–349CrossRefGoogle Scholar
  18. Kleerebezem M, Quadri LEN, Kuipers OP, de Vos WM (1997) Quorum sensing by peptide pheromones and two-component signal-transduction systems in Gram-positive bacteria. Mol Microbiol 24:895–904. CrossRefGoogle Scholar
  19. Kruger NJ (2009) Detection of polypeptides on immunoblots using enzyme- conjugated or radiolabeled secondary ligands. In: Walker JM (ed) Protein protocols handbook. Humana Press, New Jersey, pp 405–414Google Scholar
  20. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  21. Mandal V, Sen SK, Mandal NC (2008) Optimized culture conditions for bacteriocin production by Pediococcus acidilactici LAB 5 and its characterization. Indian J Biochem Biophys 45:106–110Google Scholar
  22. Mandal V, Sen SK, Mandal NC (2010) Assessment of antibacterial activities of Pediocin produced by Pediococcus acidilactici LAB 5. J Food Saf. Google Scholar
  23. Mandal V, Sen SK, Mandal NC (2011) Isolation and characterization of Pediocin NV 5 producing Pediococcus acidilactici LAB 5 from vacuum-packed fermented meat product. Indian J Microbiol 51:22–29. CrossRefGoogle Scholar
  24. Phumisantiphong U, Siripanichgon K, Reamtong O, Diraphat P (2017) A novel bacteriocin from Enterococcus faecalis 478 exhibits a potent activity against vancomycin-resistant enterococci. PLoS ONE 12:e0186415. CrossRefGoogle Scholar
  25. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. Google Scholar
  26. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  27. Schägger H, von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166:368–379CrossRefGoogle Scholar
  28. Silva CCG, Silva SPM, Ribeiro SC (2018) Application of bacteriocins and protective cultures in dairy food preservation. Front Microbiol 9:594. CrossRefGoogle Scholar
  29. Stockland AE, San Clemente CL (1968) Lactate dehydrogenase activity in certain strains of Staphylococcus aureus. J Bacteriol 95:74–80Google Scholar
  30. Suda S, Field D, Barron N (2017) Antimicrobial peptide production and purification. Methods Mol Biol (Clifton, NJ) 1485:401–410. CrossRefGoogle Scholar
  31. Tulini FL, De Martinis ECP (2010) Improved adsorption-desorption extraction applied to the partial characterization of the antilisterial bacteriocin produced by Carnobacterium maltaromaticum C2. Braz J Microbiol Publ Braz Soc Microbiol 41:493–496. CrossRefGoogle Scholar
  32. van Niel EWJ, Hahn-Hägerdal B (1999) Nutrient requirements of lactococci in defined growth media. Appl Microbiol Biotechnol 52:617–627. CrossRefGoogle Scholar
  33. Vera Pingitore E, Todorov SD, Sesma F, de Melo Gombossy, Franco BD (2012) Application of bacteriocinogenic Enterococcus mundtii CRL35 and Enterococcus faecium ST88Ch in the control of Listeria monocytogenes in fresh Minas cheese. Food Microbiol 32:38–47. CrossRefGoogle Scholar
  34. Yang R, Johnson MC, Ray B (1992) Novel method to extract large amounts of bacteriocins from lactic acid bacteria. Appl Environ Microbiol 58:3355–3359Google Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2019

Authors and Affiliations

  1. 1.Plant and Microbial Physiology and Biochemistry Laboratory, Department of BotanyUniversity of Gour BangaMaldaIndia
  2. 2.Department of BotanyDarjeeling Government CollegeDarjeelingIndia
  3. 3.Industrial Section Indian MuseumBotanical Survey of IndiaKolkataIndia
  4. 4.Department of MicrobiologyUniversity of CalcuttaKolkataIndia

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