Archives of Microbiology

, Volume 200, Issue 5, pp 771–782 | Cite as

Bacteriocinogenic properties of Escherichia coli P2C isolated from pig gastrointestinal tract: purification and characterization of microcin V

  • Mohammed Tahar Boubezari
  • Tayeb Idoui
  • Riadh Hammami
  • Benoît Fernandez
  • Ahmed Gomaa
  • Ismail Fliss
Original Paper


The aim of this study was to isolate and investigate the bacteriocinogenic and probiotic potential of new Gram-negative isolates. Of 22 bacterial isolates from pig intestine and chicken crops, ten isolates had demonstrated a good activity, and the most potent five strains were identified as four E. coli and one as Proteus sp. No virulence factors were detected for E. coli strains isolated from pig intestine. The semi-purified microcins proved to be resistant to temperature and pH variation, but sensitive to proteolytic enzymes. Of particular interest, strain E. coli P2C was the most potent, free of virulence genes and sensitive to tested antibiotics. Purification procedure revealed the presence of a single pure peak having a molecular mass of 8733.94 Da and matching microcin V (MccV). The sequence obtained by LC–MS/MS confirmed the presence of MccV. Purified MccV showed a good activity against pathogenic coliforms, especially E. coli O1K1H7 involved in avian colibacillosis. The present study provides evidence that E. coli strains isolated from pig intestine produce microcin-like substances. E. coli P2C is a safe MccV producer that could be a good candidate for its application as novel probiotic strain to protect livestock and enhance growth performance.


Bacteriocin Probiotic HPLC Microcin V Antimicrobial activity 



This project was financially supported by a research grant from the Natural Sciences and Engineering Research Council of Canada (NSERC). Mohammed Tahar Boubezari was supported by Ph.D. scholar fellowship from the Ministry of Higher Education and Scientific Research, Democratic Republic of Algeria.


  1. Andersson DI, Hughes D, Kubicek-Sutherland JZ (2016) Mechanisms and consequences of bacterial resistance to antimicrobial peptides. Drug Resist Updates 26:43–57. CrossRefGoogle Scholar
  2. Andrews JM (2001) Determination of minimum inhibitory concentrations. J Antimicrob Chemother 48:5–16CrossRefPubMedGoogle Scholar
  3. Baquero F, Moreno F (1984) The microcins. FEMS Microbiol Lett 23:117–124. CrossRefGoogle Scholar
  4. Chalasani AG, Dhanarajan G, Nema S, Sen R, Roy U (2014) An antimicrobial metabolite from Bacillus sp.: significant activity against pathogenic bacteria including multidrug-resistant clinical strains. Front Microbiol 6:1335–1335. CrossRefGoogle Scholar
  5. Chalón MC, Acuña L, Morero RD, Minahk CJ, Bellomio A (2012) Membrane-active bacteriocins to control Salmonella in foods: are they the definite hurdle? Food Res Int 45:735–744. CrossRefGoogle Scholar
  6. Chandran A, Mazumder A (2013) Prevalence of diarrhea-associated virulence genes and genetic diversity in Escherichia coli isolates from fecal material of various animal hosts. Appl Environ Microbiol 79:7371–7380. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Chehade H, Braun V (1988) Iron-regulated synthesis and uptake of colicin V. FEMS Microbiol Lett 52:177–181. CrossRefGoogle Scholar
  8. Cherif A, Rezgui W, Raddadi N, Daffonchio D, Boudabous A (2008) Characterization and partial purification of entomocin 110, a newly identified bacteriocin from Bacillus thuringiensis subsp. Entomocidus HD110. Microbiol Res 163:684–692. CrossRefPubMedGoogle Scholar
  9. The Clinical and Laboratory Standards Institute (CLSI) (2012) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Approved Standard—Ninth Edition. M07-A9 32Google Scholar
  10. Cotter PD, Ross RP, Hill C (2013) Bacteriocins—a viable alternative to antibiotics? Nat Rev Microbiol 11:95–105. CrossRefPubMedGoogle Scholar
  11. Davies J, Davies D (2010) Origins and evolution of antibiotic resistance. Microbiol Mol Biol Rev 74:417–433. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Deslouches B, Steckbeck JD, Craigo JK, Doi Y, Burns JL, Montelaro RC (2015) Engineered cationic antimicrobial peptides to overcome multidrug resistance by ESKAPE pathogens. Antimicrob Agents Chemother 59:1329–1333. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Dho-Moulin M, Fairbrother JM (1999) Avian pathogenic Escherichia coli (APEC). Vet Res 30:299–316PubMedGoogle Scholar
  14. Dobson A, Cotter PD, Ross RP, Hill C (2012) Bacteriocin production: a probiotic trait? Appl Environ Microbiol 78:1–6. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Drider D, Rebuffat S (2011) Prokaryotic antimicrobial peptides: from genes to applications. Springer Science Business Media.
  16. Duquesne S, Destoumieux-Garzón D, Peduzzi J, Rebuffat S (2007) Microcins, gene-encoded antibacterial peptides from enterobacteria. Nat Prod Rep 24:708–734. CrossRefPubMedGoogle Scholar
  17. Elayaraja S, Annamalai N, Mayavu P, Balasubramanian T (2014) Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum. Asian Pac J Trop Biomed 4:S305–S311. CrossRefPubMedPubMedCentralGoogle Scholar
  18. EUCAST (2016) Breakpoint tables for interpretation of MICs and zone diameters. Version 6.0.
  19. Fath M, Zhang L, Rush J, Kolter R (1994) Purification and characterization of colicin V from Escherichia coli culture supernatants. Biochemistry 33:6911. CrossRefPubMedGoogle Scholar
  20. Gaaloul N, Braiek O, Hani K, Volski A, Chikindas M, Ghrairi T (2015) Isolation and characterization of large spectrum and multiple bacteriocin-producing Enterococcus faecium strain from raw bovine milk. J Appl Microbiol 118:343–355. CrossRefPubMedGoogle Scholar
  21. Gao Q, Jia X, Wang X, Xiong L, Gao S, Liu X (2015) The avian pathogenic Escherichia coli O2 strain E058 carrying the defined aerobactin-defective iucD or iucDiutA mutation is less virulent in the chicken. Infect Genet Evol 30:267–277. CrossRefPubMedGoogle Scholar
  22. Ge J, Sun Y, Xin X, Wang Y, Ping W (2016) Purification and partial characterization of a novel bacteriocin synthesized by Lactobacillus paracasei HD1-7 isolated from Chinese sauerkraut juice. Sci Rep. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Goh HF, Philip K (2015) Purification and characterization of bacteriocin produced by Weissella confusa A3 of dairy origin. PloS one 10:e0140434. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Hammami R, Zouhir A, Hamida JB, Neffati M, Vergoten G, Naghmouchi K, Fliss I (2009) Antimicrobial properties of aqueous extracts from three medicinal plants growing wild in arid regions of Tunisia. Pharm Biol 47:452–457. CrossRefGoogle Scholar
  25. Hammami R, Zouhir A, Le Lay C, Hamida JB, Fliss I (2010) BACTIBASE second release: a database and tool platform for bacteriocin characterization. BMC Microbiol 10:22. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Hammami R, Fernandez B, Lacroix C, Fliss I (2013) Anti-infective properties of bacteriocins: an update. Cell Mol Life Sci 70(16):2947–2967. CrossRefPubMedGoogle Scholar
  27. Hanchi H, Hammami R, Kourda R, Hamida JB, Fliss I (2014) Bacteriocinogenic properties and in vitro probiotic potential of enterococci from Tunisian dairy products. Arch Microbiol 196:331–344. CrossRefPubMedGoogle Scholar
  28. Hanchi H, Hammami R, Gingras H, Kourda R, Bergeron MG, Ben Hamida J, Ouellette M, Fliss I (2017) Inhibition of MRSA and of Clostridium difficile by durancin 61A: synergy with bacteriocins and antibiotics. Future Microbiol 12:205–212. CrossRefPubMedGoogle Scholar
  29. Håvarstein LS, Holo H, Nes IF (1994) The leader peptide of colicin V shares consensus sequences with leader peptides that are common among peptide bacteriocins produced by gram-positive bacteria. Microbiology 140:2383–2389. CrossRefPubMedGoogle Scholar
  30. Jacobi CA, Malfertheiner P (2011) Escherichia coli Nissle 1917 (Mutaflor): new insights into an old probiotic bacterium. Dig Dis 29:600–607. CrossRefPubMedGoogle Scholar
  31. Johnson J, Moseley S, Roberts P, Stamm W (1988) Aerobactin and other virulence factor genes among strains of Escherichia coli causing urosepsis: association with patient characteristics. Infect Immun 56:405–412PubMedPubMedCentralGoogle Scholar
  32. Kostakioti M, Stathopoulos C (2004) Functional analysis of the Tsh autotransporter from an avian pathogenic Escherichia coli strain. Infect Immun 72:5548–5554. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Kumarasamy KK et al (2010) Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study. Lancet Infect Dis 10:597–602. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Liu G, Ren L, Song Z, Wang C, Sun B (2015) Purification and characteristics of bifidocin A, a novel bacteriocin produced by Bifidobacterium animals BB04 from centenarians’ intestine. Food Control 50:889–895. CrossRefGoogle Scholar
  35. Mainil J (2013) Escherichia coli virulence factors. Vet Immunol Immunopathol 152:2–12. CrossRefPubMedGoogle Scholar
  36. Morin N, Lanneluc I, Connil N, Cottenceau M, Pons AM, Sablé S (2011) Mechanism of bactericidal activity of microcin L in Escherichia coli and Salmonella enterica. Antimicrob Agents Chemother 55:997–1007. CrossRefPubMedGoogle Scholar
  37. Naghmouchi K, Belguesmia Y, Baah J, Teather R, Drider D (2011) Antibacterial activity of class I and IIa bacteriocins combined with polymyxin E against resistant variants of Listeria monocytogenes and Escherichia coli. Res Microbiol 162:99–107. CrossRefPubMedGoogle Scholar
  38. Pinou T, Riley MA (2001) Nucleotide polymorphism in microcin V plasmids. Plasmid 46:1–9. CrossRefPubMedGoogle Scholar
  39. Pons A-M, Lanneluc I, Cottenceau G, Sable S (2002) New developments in non-post translationally modified microcins. Biochimie 84:531–537. CrossRefPubMedGoogle Scholar
  40. Sablé S, Duarte M, Bravo D, Lanneluc I, Pons A, Cottenceau G, Moreno F (2003) Wild-type Escherichia coli producing microcins B17, D93, J25, and L; cloning of genes for microcin L production and immunity. Can J Microbiol 49:357–361. CrossRefPubMedGoogle Scholar
  41. Shanahan F (2010) Probiotics in perspective. Gastroenterology 139:1808–1812. CrossRefPubMedGoogle Scholar
  42. Verso LL, Lessard M, Talbot G, Fernandez B, Fliss I (2017) Isolation and selection of potential probiotic bacteria from the pig gastrointestinal tract. Probiotics Antimicrobial Proteins. CrossRefGoogle Scholar
  43. Waters VL, Crosa JH (1991) Colicin V virulence plasmids. Microbiol Rev 55:437–450PubMedPubMedCentralGoogle Scholar
  44. Zhang LH, Fath MJ, Mahanty H, Tai P, Kolter R (1995) Genetic analysis of the colicin V secretion pathway. Genetics 141:25–32PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Dairy Research Center (STELA) and Department of Food Sciences, Institute of Nutrition and Functional Foods (INAF)Université LavalQuébecCanada
  2. 2.Research Laboratory of Biotechnology, Environment, and HealthJijel UniversityJijelAlgeria
  3. 3.Faculty of Health Sciences, School of Nutrition SciencesUniversity of OttawaOttawaCanada

Personalised recommendations