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Probiotics and Antimicrobial Proteins

, Volume 5, Issue 3, pp 206–215 | Cite as

Anti-listerial Bactericidal Activity of Lactobacillus plantarum DM5 Isolated from Fermented Beverage Marcha

  • Deeplina Das
  • Arun Goyal
Article

Abstract

The strain Lactobacillus plantarum DM5 was isolated from fermented beverage Marcha of Sikkim and explored for its antagonistic activity against food-borne pathogens. The cell-free supernatant of L. plantarum DM5 showed antibacterial activity of 6,400 AU/mL in MRS medium (pH 6.0) against the indicator strain Staphylococcus aureus. MRS medium supplemented with 15 g/L of maltose at 37 °C under static condition yielded highest antimicrobial activity (6,400 AU/mL) with 3 % increase in specific activity when compared to 20 g/L glucose. The antimicrobial compound was heat stable (60 min at 100 °C) and was active over a wide pH range. It showed bactericidal effect on S. aureus and Listeria monocytogenes by causing 96 and 98 % of cell lysis, respectively. The cell morphology of the treated S. aureus and L. monocytogenes was completely deformed as revealed by scanning electron microscopy, suggesting the high potential of L. plantarum DM5 as natural preservatives in food industry. The antimicrobial compound was purified by 80 % ammonium sulphate precipitation and showed antimicrobial activity of 12,800 AU/mL with 19-fold purification and a molecular mass of 15.2 kDa, indicating the proteinaceous nature of the compound.

Keywords

Lactobacillus plantarum Antagonistic activity Pathogens Marcha 

Notes

Acknowledgments

The research work was supported by a project grant from Department of Biotechnology (DBT), New Delhi, India, to A. Goyal. The fellowship of D. Das was supported by Council of Industrial and Scientific Research (CSIR), New Delhi, India. The authors are thankful to the Indian Institute of Technology, Guwahati, India, for providing the research facilities.

Conflict of interest

Arun Goyal has received the project grant for this research from Department of Biotechnology (DBT), New Delhi, India; Deeplina Das has received fellowship from by Council of Industrial and Scientific Research (CSIR), New Delhi, India; the authors certify that there is no other conflict of interest with any other financial organization regarding this research.

Ethical standard

This article does not contain any studies with human or animal subjects. The indicator strains used in this study were procured from Microbial Type Culture Collection (MTCC), Institute of Microbial Technology, Chandigarh, India.

References

  1. 1.
    Klein G, Pack A, Bonaparte C, Reuter G (1998) Taxonomy and physiology of probiotic lactic acid bacteria. Int J Food Microbiol 41:103–125CrossRefGoogle Scholar
  2. 2.
    Yoon KY, Woodams EE, Hang YD (2006) Production of probiotic cabbage juice by lactic acid bacteria. Bioresour Technol 97:1427–1430CrossRefGoogle Scholar
  3. 3.
    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 24:343–362CrossRefGoogle Scholar
  4. 4.
    Galvez A, Abriouel H, Lopez RL, Omar NB (2007) Bacteriocin-based strategies for food biopreservation. Int J Food Microbiol 120:51–70CrossRefGoogle Scholar
  5. 5.
    Klaenhammer TR (1993) Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol Rev 12:39–86Google Scholar
  6. 6.
    Delves-Broughton J (1990) Nisin and its uses as a food preservative. Food Technol 44:100–117Google Scholar
  7. 7.
    Ahern M, Verschueren S, van-Sinderen D (2003) Isolation and characterisation of a novel bacteriocin produced by Bacillus thuringiensis strain B439. FEMS Microbiol Lett 220:127–131CrossRefGoogle Scholar
  8. 8.
    Nithya V, Halami PM (2012) Antibacterial peptides, probiotic properties and biopreservative efficacy of native Bacillus species isolated from different food sources. Probiotics Antimicrobial Prot 4:279–290CrossRefGoogle Scholar
  9. 9.
    Carolissen-Mackay V, Arendse G, Hastings JW (1997) Purification of bacteriocins of lactic acid bacteria: problems and pointers. Int J Food Microbiol 34:1–16CrossRefGoogle Scholar
  10. 10.
    Todorov SD, Dicks LMT (2009) Effect of modified MRS medium on production and purification of antimicrobial peptide ST4SA produced by Enterococcus mundtii. Food Microbiol 15:65–73Google Scholar
  11. 11.
    Nissen L, Chingwaru W, Sgorbati B, Biavati B, Cencic A (2009) Gut health promoting activity of new putative probiotic protective Lactobacillus spp. strains: a functional study in the small intestinal cell model. Int J Food Microbiol 135:288–294CrossRefGoogle Scholar
  12. 12.
    Todorov SD, Dicks LMT (2005) Effect of growth medium on bacteriocin production by Lactobacillus plantarum ST194BZ, a Strain isolated from Boza. Food Technol Biotechnol 43:165–173Google Scholar
  13. 13.
    Messi P, Bondi M, Sabia C, Battini R, Manicardi G (2001) Detection and preliminary characterization of a bacteriocin (plantaricin 35d) produced by a Lactobacillus plantarum strain. Int J Food Microbiol 64:193–198CrossRefGoogle Scholar
  14. 14.
    Ross PP, Morgan S, Hill C (2002) Preservation and fermentation: past, present and future. Int J Food Microbiol 79:3–16CrossRefGoogle Scholar
  15. 15.
    Marth EH (1998) Extended shelf life refrigerated foods: microbiological quality and safety. Food Technol 52:57–62Google Scholar
  16. 16.
    Das D, Goyal A (2010) Characterization and screening of antimicrobial activity of lactic acid bacterium isolated from a traditional beverage Marcha of Sikkim. J Pharm Chem 4:136–139Google Scholar
  17. 17.
    Tamang JP, Dewan S, Tamang B, Rai A, Schillinger U, Holzapfel WH (2007) Lactic acid bacteria in Hamei and Marcha of North East India. Indian J Microbiol 47:119–125CrossRefGoogle Scholar
  18. 18.
    Tsuyoshi N, Fudou R, Yamanaka S, Kozaki M, Tamang N, Thapa S, Tamang JP (2005) Identification of yeast strains isolated from Marcha in Sikkim, a microbial starter for amylolytic fermentation. Int J Food Microbiol 99:135–146CrossRefGoogle Scholar
  19. 19.
    De Man JC, Ragosa M, Sharpe E (1960) A medium for the cultivation of lactobacilli. J Appl Bact 23:130–135CrossRefGoogle Scholar
  20. 20.
    Todorov SD, Dicks LMT (2009) Bacteriocin production by Pediococcus pentosaceus isolated from marula (Scerocarya birrea). Int J Food Microbiol 132:117–126CrossRefGoogle Scholar
  21. 21.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  22. 22.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  23. 23.
    Bhunia AK, Johnson MC, Ray B (1987) Direct detection of an antimicrobial peptide of Pediococcus acidilactici in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Ind Microbiol 2:319–322CrossRefGoogle Scholar
  24. 24.
    Enan G, Essawy AA, Uyttendaele M, Debevere J (1996) Antibacterial activity of Lactobacillus plantarum UG1 isolated from dry sausage: characterization, production and bactericidal action of plantaricin UG1. Int J Microbiol 30:189–215CrossRefGoogle Scholar
  25. 25.
    Gonzalez B, Glaasker E, Kunji ERS, Driessen AJM, Suarez JE, Konings WN (1996) Bactericidal mode of action of plantaricin C. Appl Environ Microbiol 62:2701–2709Google Scholar
  26. 26.
    Todorov S, Onno B, Sorokine O, Chobert JM, Ivanova I, Dousset X (1999) Detection and characterization of a novel antibacterial substance produced by Lactobacillus plantarum ST31 isolated from sourdough. Int J Food Microbiol 48:167–177CrossRefGoogle Scholar
  27. 27.
    Rekhif N, Atrih A, Michel M, Lefebvre G (1995) Activity of plantaricin SA6, a bacteriocin produced by Lactobacillus plantarum SA6 isolated from fermented sausages. J Appl Bacteriol 78:349–358CrossRefGoogle Scholar
  28. 28.
    Mook P, O’Brien SJ, Gillespie IA (2011) Concurrent conditions and human listeriosis, England, 1999–2009. Emerg Infect Dis 17:38–43CrossRefGoogle Scholar
  29. 29.
    Loir YL, Baron F, Gautier M (2003) Staphylococcus aureus and food poisoning. Genet Mol Res 2:63–76Google Scholar
  30. 30.
    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 84:1131–1137CrossRefGoogle Scholar
  31. 31.
    Mandal V, Sen SK, Mandal NC (2008) Optimized culture conditions for bacteriocin production by Pediococcus acidilactici LAB5 and its characterization. Indian J Biochem Biophys 45:106–110Google Scholar
  32. 32.
    Piva A, Headon DR (1994) Pediocin A, a bacteriocin produced by Pediococcus pentosaceus FBB61. Microbiology 140:697–702CrossRefGoogle Scholar
  33. 33.
    Wu CW, Yin LJ, Jiang ST (2004) Purification and characterization of bacteriocin from Pediococcus pentosaceus ACCEL. J Agric Food Chem 52:1146–1151CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Department of BiotechnologyIndian Institute of Technology GuwahatiGuwahatiIndia

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