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Fluorescence-Based Comparative Evaluation of Bactericidal Potency and Food Application Potential of Anti-listerial Bacteriocin Produced by Lactic Acid Bacteria Isolated from Indigenous Samples

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Abstract

The aim of the present study was to ascertain the potency of anti-listerial bacteriocin produced by lactic acid bacteria (LAB) isolated from indigenous samples of dahi, dried fish, and salt-fermented cucumber. A total of 231 LAB isolates were obtained from the samples, of which 51 isolates displayed anti-listerial activity. The anti-listerial LAB were identified by PCR as Lactobacillus sp., Pediococcus sp., Enterococcus sp., and Lactococcus sp. PCR also enabled the detection of Class IIa bacteriocin-encoding genes such as enterocin A, pediocin, and plantaricin A in some of the LAB isolates. The culture filtrate from anti-listerial LAB isolates demonstrated bacteriocin-like inhibitory substance (BLIS) against common Gram-positive pathogenic bacteria such as Staphylococcus aureus, Enterococcus faecalis, and Bacillus cereus, and partial characterization of BLIS confirmed the production of bacteriocin by the LAB isolates. Sensitive fluorescence-based assays employing specific probes indicated the comparative potencies of the bacteriocin and clearly revealed the membrane-targeted anti-listerial activity of the purified bacteriocin produced by selected LAB isolates. The food application potential of plantaricin A produced by a native isolate Lactobacillus plantarum CRA52 was evidenced as the bacteriocin suppressed the growth of Listeria monocytogenes Scott A inoculated in paneer samples that were stored at 8 °C for 5 days.

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References

  1. Albano H, Pinho C, Leite D, Barbosa J, Silva J, Carneiro L, Magalhaes R, Hogg T, Teixeira P (2009) Evaluation of a bacteriocin-producing strain of Pediococcus acidilactici as a biopreservative for ‘‘Alheira”, a fermented meat sausage. Food Control 20:764–770

    Article  CAS  Google Scholar 

  2. Albano H, Oliveira M, Aroso R, Cubero N, Hogg T, Teixeira P (2007) Antilisterial activity of lactic acid bacteria isolated from ‘‘Alheiras’’ (traditional Portuguese fermented sausages): In situ assays. Meat Sci 76:796–800

    Article  CAS  Google Scholar 

  3. Allende A, Martinez B, Selma V, Gila MI, Suarez JE, Rodriguez A (2007) Growth and bacteriocin production by lactic acid bacteria in vegetable broth and their effectiveness at reducing Listeria monocytogenes in vitro and in fresh-cut lettuce. Food Microbiol 24:759–766

    Article  CAS  Google Scholar 

  4. Atrih A, Rekhif N, Moir AJG, Lebrihi A, Lefebvre G (2001) Mode of action, purification and amino acid sequence of plantaricin C19, an anti-listeria bacteriocin produced by Lactobacillus plantarum C19. Int J Food Microbiol 68:93–104

    Article  CAS  Google Scholar 

  5. Belgacem BZ, Ferchichi M, Prévost H, Dousset X, Manai M (2008) Screening for anti-listerial bacteriocin-producing lactic acid bacteria from ‘‘Gueddid’’ a traditionally Tunisian fermented meat. Meat Sci 78:513–521

    Article  Google Scholar 

  6. Bhunia AK, Johnson MC, Ray B (1987) Direct detection of an antimicrobial peptide of Pediococcus acidilactici in sodium dodecyl sulphate-polyacrylamide gel electrophoresis. J Ind Microbiol 2:319–322

    Article  CAS  Google Scholar 

  7. Budde BB, Rasch M (2001) A comparative study on the use of flow cytometry and colony forming units for assessment of the antibacterial effect of bacteriocins. Int J Food Microbiol 63:65–72

    Article  CAS  Google Scholar 

  8. Castellano P, Belfiore C, Fadda S, Vignolo G (2008) A review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina. Meat Sci 79:483–499

    Article  CAS  Google Scholar 

  9. Cleveland J, Montville TJ, Nes IF, Chikindas ML (2001) Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 71:1–20

    Article  CAS  Google Scholar 

  10. Coolbear T, Crow V, Harnett J, Harvey S, Holland R, Martley F (2008) Developments in cheese microbiology in New Zealand—use of starter and non-starter lactic acid bacteria and their enzymes in determining flavour. Int Dairy J 18:705–713

    Article  CAS  Google Scholar 

  11. Cotter PD, Hill C, Ross RP (2005) Bacteriocins: developing innate immunity for food. Nat Rev Microbiol 3:777–788

    Article  CAS  Google Scholar 

  12. Deasy BM, Rea MC, Fitzgerald GF, Cogan TM, Beresford TP (2000) A rapid PCR based method to distinguish between Lactococcus and Enterococcus. Syst Appl Microbiol 23:510–522

    Article  CAS  Google Scholar 

  13. Drider D, Fimland G, Hechard Y, Mcmullen LM, Prévost H (2006) The continuing story of Class IIa bacteriocins. Microbiol Mol Biol Rev 70:564–582

    Article  CAS  Google Scholar 

  14. Halami PM, Ramesh A, Chandrashekar A (2000) Megaplasmid encoding novel sugar utilizing phenotypes, pediocin production and immunity in Pediococcus acidilactici C20. Food Microbiol 17:475–483

    Article  CAS  Google Scholar 

  15. Halami PM, Ramesh A, Chandrashekar A (2005) Fermenting cucumber, a potential source for the isolation of pediocin like bacteriocin producers. World J Microbiol Biotechnol 21:1351–1358

    Article  CAS  Google Scholar 

  16. Hoefel D, Groobya WL, Monisa PT, Andrews S, Saint CP (2003) A comparative study of carboxyfluorescein diacetate and carboxyfluorescein diacetate succinimidyl ester as indicators of bacterial activity. J Microbiol Methods 52:379–388

    Article  CAS  Google Scholar 

  17. Ibrahim HR, Sugimoto Y, Aoki T (2000) Ovotransferrin antimicrobial peptide (OTAP-92) kills bacteria through a membrane damage mechanism. BBA 1523:196–205

    CAS  Google Scholar 

  18. Jagannath A, Ramesh A, Ramesh MN, Chandrashekar A, Varadaraj MC (2001) Predictive model for the behavior of Listeria monocytogenes Scott A in Shrikhand, prepared with a biopreservative pediocin K7. Food Microbiol 18:335–343

    Article  CAS  Google Scholar 

  19. Jones RJ, Hussein HM, Zagorec M, Brightwell G, Tagg JR (2008) Isolation of lactic acid bacteria with inhibitory activity against pathogens and spoilage organisms associated with fresh meat. Food Microbiol 25:228–234

    Article  CAS  Google Scholar 

  20. Knoll C, Divol B, du Toit M (2008) Genetic screening of lactic acid bacteria of oenological origin for bacteriocin-encoding genes. Food Microbiol 25:983–991

    Article  CAS  Google Scholar 

  21. Kristiansen PE, Fimland G, Mantzilas D, Nissen-Meyer J (2005) Structure and mode of action of the membrane-permeabilizing antimicrobial peptide pheromone plantaricin A. J Biol Chem 280:22945–22950

    Article  CAS  Google Scholar 

  22. Le Marc Y, Hucket V, Bourfeois CM, Guyonnet JP, Mafart P (2002) Modelling the growth kinetics of Listeria as a function of temperature, pH and organic acid concentration. Int J Food Microbiol 73:219–237

    Article  Google Scholar 

  23. Lee YK, Ho PS, Low CS, Arvilommi H, Salminen S (2004) Permanent colonization by Lactobacillus casei is hindered by the low rate of cell division in mouse gut. Appl Environ Microbiol 70:670–674

    Article  CAS  Google Scholar 

  24. Leroy F, De Vuyst L (2004) Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci Technol 15:67–78

    Article  CAS  Google Scholar 

  25. Montville TJ, Chen Y (1998) Mechanistic action of pediocin and nisin: recent progress and unresolved questions. Appl Microbiol Biotechnol 50:511–519

    Article  CAS  Google Scholar 

  26. Naghmouchi K, Kheadra E, Lacroix C, Fliss I (2007) Class I/Class IIa bacteriocin cross-resistance phenomenon in Listeria monocytogenes. Food Microbiol 24:718–727

    Article  CAS  Google Scholar 

  27. Pucci MJ, Vedamuthu ER, Kunka BS, Vandenbergh PA (1988) Inhibition of Listeria monocytogenes by using bacteriocin PA-1 produced by Pediococcus acidilactici PAC 1.0. Appl Environ Microbiol 54:2349–2353

    CAS  Google Scholar 

  28. Schagger 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 166:368–379

    Article  CAS  Google Scholar 

  29. Settani L, Corsetti A (2008) Application of bacteriocin in vegetable food biopreservation. Int J Food Microbiol 121:123–138

    Article  Google Scholar 

  30. Singh AK, Ramesh A (2008) Succession of dominant and antagonistic lactic acid bacteria in fermented cucumber: insights from a PCR-based approach. Food Microbiol 25:278–287

    Article  CAS  Google Scholar 

  31. Singh AK, Ramesh A (2009) Evaluation of a facile method of template DNA preparation for PCR-based detection and typing of lactic acid bacteria. Food Microbiol 26:504–513

    Article  CAS  Google Scholar 

  32. Thapa N, Pal N, Tamang JP (2006) Phenotypic identification and technological properties of lactic acid bacteria isolated from traditionally processed fish products of the Eastern Himalayas. Int J Food Microbiol 107:33–38

    Article  CAS  Google Scholar 

  33. Todorov SD, Dicks LMT (2006) Screening for bacteriocin-producing lactic acid bacteria from boza, a traditional cereal beverage from Bulgaria. Comparison of the bacteriocins. Process Biochem 41:11–19

    Article  CAS  Google Scholar 

  34. Tome E, Teixeira P, Gibbs PA (2006) Anti-listerial inhibitory lactic acid bacteria isolated from commercial cold smoked salmon. Food Microbiol 23:399–405

    Article  CAS  Google Scholar 

  35. Virto R, Manas P, Alvarez I, Condon S, Raso J (2005) Membrane damage and microbial inactivation by chlorine in the absence and presence of a chlorine-demanding substrate. Appl Environ Microbiol 71:5022–5028

    Article  CAS  Google Scholar 

  36. Vooturi SK, Cheung CM, Rybak MJ, Firestine SM (2009) Design, synthesis, and structure-activity relationships of benzophenone-based tetraamides as novel antibacterial agents. J Med Chem 52:5020–5031

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

Download references

Acknowledgments

We thank the Council of Scientific and Industrial Research (CSIR), New Delhi, Government of India for a research grant [No. 38(1251)/10/EMR-II]. We thank the National Facility of Automated DNA Sequencing, Department of Biochemistry, Delhi University, South campus for their support in nucleic acid sequencing.

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  1. Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, 781 039, Assam, India

    Atul Kumar Singh, Sandipan Mukherjee, Manab Deb Adhikari & Aiyagari Ramesh

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  1. Atul Kumar Singh
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  2. Sandipan Mukherjee
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  4. Aiyagari Ramesh
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Correspondence to Aiyagari Ramesh.

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Singh, A.K., Mukherjee, S., Adhikari, M.D. et al. Fluorescence-Based Comparative Evaluation of Bactericidal Potency and Food Application Potential of Anti-listerial Bacteriocin Produced by Lactic Acid Bacteria Isolated from Indigenous Samples. Probiotics & Antimicro. Prot. 4, 122–132 (2012). https://doi.org/10.1007/s12602-012-9100-4

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