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Characterization of multiple antilisterial peptides produced by sakacin P-producing Lactobacillus sakei subsp. sakei 2a

Archives of Microbiology volume 200, pages 635–644 (2018)Cite this article

Abstract

Antimicrobial compounds produced by lactic acid bacteria can be explored as natural food biopreservatives. In a previous report, the main antimicrobial compounds produced by the Brazilian meat isolate Lactobacillus sakei subsp. sakei 2a, i.e., bacteriocin sakacin P and two ribosomal peptides (P2 and P3) active against Listeria monocytogenes, were described. In this study, we report the spectrum of activity, molecular mass, structural identity and mechanism of action of additional six antilisterial peptides produced by Lb. sakei 2a, detected in a 24 h-culture in MRS broth submitted to acid treatment (pH 1.5) and proper fractionation and purification steps for obtention of free and cell-bound proteins. The six peptides presented similarity to different ribosomal proteins of Lb. sakei subsp sakei 23K and the molecular masses varied from 4.6 to 11.0 kDa. All peptides were capable to increase the efflux of ATP and decrease the membrane potential in Listeria monocytogenes. The activity of a pool of the obtained antilisterial compounds [enriched active fraction (EAF)] against Listeria monocytogenes in a food model (meat gravy) during refrigerated storage (4 °C) for 10 days was also tested and results indicated that the populations of L. monocytogenes in the food model containing the acid extract remained lower than those at time 0-day, evidencing that the acid extract of a culture of Lb. sakei 2a is a good technological alternative for the control of growth of L. monocytogenes in foods.

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References

  • Abee T (1995) Pore-forming bacteriocins of gram-positive bacteria and self-protection mechanisms of producer organisms. FEMS Microbiol Lett 129:1–10

    CAS  Article  PubMed  Google Scholar 

  • Alves VF, Lavrador MAS, De Martinis ECPP. (2003) Bacteriocin exposure and food ingredients influence on growth and virulence of Listeria monocytogenes in a model meat gravy system. J Food Saf 23:201–217

    CAS  Article  Google Scholar 

  • Barbosa MS, Todorov SD, Ivanova I, Chobert JM, Haertl T, Franco BDGM. (2015) Improving safety of salami by application of bacteriocins produced by an autochthonous Lactobacillus curvatus isolate. Food Microbiol 46:254–262

    Article  Google Scholar 

  • Bradford MM (1976) Rapid and sensitive method for quantification of microgram quantities of proteins utilizing principle of protein–dye binding. Anal Biochem 72:248–254

    CAS  Article  PubMed  Google Scholar 

  • Buchanan RL, Gorris L, Hayman MM, Jackson TC, Whiting RC (2017) A review of Listeria monocytogenes: an update on outbreaks, virulence, dose–response, ecology, and risk assessments. Food Control 75:1–13

    Article  Google Scholar 

  • Carvalho KG, Bambirra FHS, Kruger MF, Barbosa MS, Oliveira JS, Santos AMC, Nicoli JR, Bemquerer MP, Miranda A, Salvucci EJ, Sesma FJM, Franco BDGM. (2010) Antimicrobial compounds produced by Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian meat product. J Ind Microbiol Biotechnol 37:381–390

    CAS  Article  PubMed  Google Scholar 

  • Castellano P, Mora l, Escudero E, Vignolo G, Aznar R, Toldra F (2016) Antilisterial peptides from Spanish dry-cured hams: purification and identification. Food Microbiol 59:133–141

    CAS  Article  PubMed  Google Scholar 

  • Chaillou S, Champomier-Verges M, Cornet M, Crutz Lecoq AM, Dudez AM, Martin V, Beaufils S, Darbon-Rongere E, Bossy R, Loux V, Zagorec M (2005) The genome sequence of the meat-borne lactic acid bacterium Lactobacillus sakei 23K. Nat Biotechnol 23:1527–1533

    CAS  Article  PubMed  Google Scholar 

  • Chen Y, Montville TJ (1995) Efflux of ions and ATP depletion induced by pediocin PA-1 are concomitant with cell death in Listeria monocytogenes Scott A. J Appl Bacteriol 79:684–690

    CAS  Article  Google Scholar 

  • De Martinis ECP, Franco BDGM. (1998) Inhibition of Listeria monocytogenes in a pork product by a Lactobacillus sake strain. Int J Food Microbiol 42:119–126

    Article  PubMed  Google Scholar 

  • Drider D, Fimland G, Héchard Y, McMullen LM, Prévost H (2006) The continuing story of class IIa bacteriocins. Microbiol Mol Biol Rev 70:564–582

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Farias LM, Totola AH, Miranda CMS, Carvalho MAR, Damasceno CAV (1994) Extraction, partial purification and characterization of a bacteriocin (fragilicin) produced by a strain of Bacteroides fragilis isolated from Callithrix penicillata. Res Microbiol 145:9–16

    CAS  Article  PubMed  Google Scholar 

  • Gaaloul N, Braiek OB, Hani K, Volski A, Chikindas ML, Ghrairi T (2014) Isolation and characterization of large spectrum and multiple bacteriocin-producing Enterococcus faecium strain from raw bovine milk. J Appl Microbiol 118:343–355

    Article  PubMed  Google Scholar 

  • Guihard G, Benedettig SH, Besnard M, Letelliern L (1993) Phosphate efflux through the channels formed by colicins and phage T5 in Escherichia coli cells is responsible for the fall in cytoplasmic ATP. J Biol Chem 268:17775–17780

    CAS  PubMed  Google Scholar 

  • Herranz C, Cintas LM, Hernandez PE, Moll GN, Driessen AJ (2001a) Enterocin P causes potassium ion efflux from Enterococcus faecium T136 cells. Antimicrob Agents Chemother 45:901–904

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Herranz C, Chen Y, Chung HJ, Cintas LM, Hernandez PE, Montville TJ, Chikindas ML (2001b) Enterocin P selectively dissipates the membrane potential of Enterococcus faecium T136. Appl Environ Microbiol 67:1689–1692

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Masuda Y, Perez RH, Zendo T, Sonomoto K (2015) Nutrition-adaptative control of multiple bacteriocin production by Weissella hellenica QU 13. J Appl Microbiol 120:70–79

    Article  Google Scholar 

  • Miao J, Liu G, Ke C, Fan W, Li C, Chen Y, Dixon W, Song M, Cao Y, Xiao H (2016) Inhibitory effects of a novel antimicrobial peptide from kefir against Escherichia coli. Food Control 65:63–72

    CAS  Article  Google Scholar 

  • Mirkovic N, Polovic N, Vukotic G, Jovcic B, Miljkovic M, Rdaulovic Z, Diep DB, Kojic M (2016) Lactococcus lactis LMG2081 produces two bacteriocins, a nonlantibiotic and a novel lantibiotic. Appl Environ Microbiol 82:2555–2562

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • O’Connor PM, Ross RP, Hill C, Cotter PD (2015) Antimicrobial antagonists against food pathogens: a bacteriocin perspective. Curr Opin Food Sci 2:51–57

    Article  Google Scholar 

  • Rosa CM, Franco BDGM., Montville TJ, Chikindas M (2002) Purification and mechanistic action of a bacteriocin produced by a Brazilian sausage isolate, Lactobacillus sakei 2a. J Food Saf 22:39–54

    CAS  Article  Google Scholar 

  • Sawa N, Koga S, Okamura K, Ishibashi N, Zendo T, Sonomoto K (2013) Identification and characterization of novel multiple bacteriocins produced by Lactobacillus sakei D98. J Appl Microbiol 115:61–69

    CAS  Article  PubMed  Google Scholar 

  • Snyder AB, Worobo RW (2014) Chemical and genetic characterization of bacteriocins: antimicrobial peptides for food safety. J Sci Food Agric 94:28–44

    CAS  Article  PubMed  Google Scholar 

  • Suzuki M, Yamamoto T, Kawai Y, Inoue N, Yamazaki K (2005) Mode of action of piscicocin CS526 produced by Carnobacterium piscicola CS526. J Appl Microbiol 98:1146–1151

    CAS  Article  PubMed  Google Scholar 

  • Todorov SD, Vaz-Velho M, Franco BDGM., Holzapfel WH (2013) Partial characterization of bacteriocins produced by three strains of Lactobacillus sakei, isolated from salpicao, a fermented meat product from North-West of Portugal. Food Control 30:111–121

    CAS  Article  Google Scholar 

  • Urso R, Rantsiou K, Cantoni C, Comi G, Cocolin L (2006) Technological characterization of a bacteriocin-producing Lactobacillus sakei and its use in fermented sausages production. Int J Food Microbiol 110:232–239

    CAS  Article  PubMed  Google Scholar 

  • Ustyugova EA, Timofeeva AV, Stoyanova LG, Netrusov AI, Katrukha GS (2012) Characteristics and identification of bacteriocins produced by Lactococcus lactis subsp lactis 194-K. Appl Biochem Microbiol 48:557–563

    CAS  Article  Google Scholar 

  • Xiao J, Niu L (2015) Antilisterial peptides released by enzymatic hydrolysis from grass carp proteins and activity on controlling Listeria monocytogenes inoculated in surimi noodle. J Food Sci 80:M2564–M2569

    Article  Google Scholar 

  • 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  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The study was supported by FAPESP (Processes 2004/08041-9, 2005/60619-8 and 2013/07914-8) and FAPEMIG (EDT 24,000) Grants.

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Affiliations

  1. Food Research Center, Departamento de Alimentos e Nutrição Experimental, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, SP, Brazil

    Bernadette D. G. M. Franco

  2. Planta Piloto de Procesos Industriales Microbiológicos (PROIMI), CONICET, Av. Belgrano y Pje. Caseros, San Miguel de Tucumán, 4000, Tucumán, Argentina

    Kátia G. Carvalho

  3. Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil

    Felipe H. S. Bambirra & Jacques R. Nicoli

  4. Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil

    Jamil S. Oliveira & Alexandre M. C. Santos

  5. EMBRAPA Recursos Genéticos e Biotecnologia, Parque Estação Biológica, Brasília, DF, Brazil

    Marcelo P. Bemquerer

  6. Departamento de Ciências Fisiológicas, Universidade Federal do Espírito Santo, Vitória, ES, Brazil

    Alexandre M. C. Santos

  7. Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brazil

    Antonio Miranda

Authors
  1. Kátia G. Carvalho
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  2. Felipe H. S. Bambirra
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  3. Jacques R. Nicoli
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  4. Jamil S. Oliveira
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  5. Alexandre M. C. Santos
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  6. Marcelo P. Bemquerer
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  7. Antonio Miranda
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  8. Bernadette D. G. M. Franco
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Corresponding author

Correspondence to Kátia G. Carvalho.

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No conflict of interest declared.

Additional information

Chemical compounds studied in this article: sodium chloride (PubChem CID: 5234), phosphoric acid (PubChem CID: 1004), trifluoroacetic acid (PubChem CID: 45039676), acetonitrile (PubChem CID: 6342), hepes potassium Salt (PubChem CID: 23702134), glucose (PubChem CID: 82400), nisin (PubChem CID: 16130280), valinomycin (PubChem CID: 5649), potassium chloride (PubChem CID: 4873).

Communicated by Erko Stackebrandt.

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Carvalho, K.G., Bambirra, F.H.S., Nicoli, J.R. et al. Characterization of multiple antilisterial peptides produced by sakacin P-producing Lactobacillus sakei subsp. sakei 2a. Arch Microbiol 200, 635–644 (2018). https://doi.org/10.1007/s00203-018-1477-3

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  • DOI: https://doi.org/10.1007/s00203-018-1477-3

Keywords

  • Lactobacillus sakei subsp. sakei 2a
  • Listeria monocytogenes
  • Multiple antimicrobial peptides
  • Meat gravy
  • Biopreservation