Skip to main content
SpringerLink
Log in

Characterization of the bacteriocin produced by Enterococcus italicus ONU547 isolated from Thai fermented cabbage

  • Original Article
  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Seventy-eight isolates of lactic acid bacteria from Ukraine and Thailand were screened for bacteriocinogenic activity against indicator strain Lactobacillus sakei subsp. sakei JCM 1157. One isolate showed an antagonistic activity of cell-free supernatant eliminated after the treatment with Proteinase K. Based on 16S rRNA gene sequence, this isolate was identified as Enterococcus italicus. Bacteriocin produced by this strain showed antimicrobial activity against L. sakei subsp. sakei JCM 1157, Brochothrix thermosphacta DSMZ 20171, and Listeria ivanovii subsp. ivanovii DSMZ 20750 in agar well diffusion assay. This bacteriocin was cationic and hydrophobic. The partially purified bacteriocin was thermostable, while heating of cell-free supernatant increased its activity more than twofold. Molecular mass of the partially purified bacteriocin as determined by SDS-PAGE differed from enterocin A and B previously known for E. italicus. Concentrated bacteriocin decreased the level of biofilm formation in L. sakei subsp. sakei JCM 1157 and Pseudomonas aeruginosa PAO1 in 52.5 and 48.0%, respectively (p < 0.05). We suggest that the studied bacteriocin could be a perspective antibiofilm agent in food conservation and medicine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Aguilar C, Klotz B (2010) Effect of the temperature on the antagonistic activity of lactic acid bacteria against Escherichia coli and Listeria monocytogenes. J Food Saf 30:996–1015. https://doi.org/10.1111/j.1745-4565.2010.00257.x

    Article  CAS  Google Scholar 

  • Al-Mathkhury HJ, Ali AS, Ghafil JA (2011) Antagonistic effect of bacteriocin against urinary catheter associated Pseudomonas aeruginosa biofilm. N Am J Med Sci 3:367–370. https://doi.org/10.4297/najms.2011.3367

    Article  PubMed  PubMed Central  Google Scholar 

  • Aymerich T, Holo H, Håvarstein LS, Hugas M, Garriga M, Nes IF (1996) Biochemical and genetic characterization of enterocin A from Enterococcus faecium, a new antilisterial bacteriocin in the pediocin family of bacteriocins. Appl Environ Microbiol 62:1676–1682

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bajpai VK, Han J-H, Rather IA, Park C, Lim J, Paek WK, Lu JS, Yoon J-I, Park Y-H (2016) Characterization and antibacterial potential of lactic acid bacterium Pediococcus pentosaceous 4I1 isolated from freshwater fish Zacco koreanus. Front Microbiol 7:1–15. https://doi.org/10.3389/fmicb.2016.02037

    Article  Google Scholar 

  • Belgacem ZB, Rehaiem A, Bernárdez PF, Manai M, Castro PL (2012) Interactive effects of pH and temperature on the bacteriocin stability by response surface analysis. Microbiologiia 81:195–200. https://doi.org/10.1134/S002626171201002X

    Article  CAS  Google Scholar 

  • Casaus P, Nilsen T, Cintas LM, Nes IF, Hernández PE, Holo H (1997) Enterocin B, a new bacteriocin from Enterococcus faecium T136 which can act synergistically with enterocin A. Microbiology 143:2287–2294. https://doi.org/10.1099/00221287-143-7-2287

    Article  CAS  PubMed  Google Scholar 

  • Christensen GD, Simpson WA, Younger JJ, Baddour LM, Barrett FF, Melton DM, Beachey EH (1985) Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 22:996–1006

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cirkovik I, Bozic DD, Draganic V, Lozo J, Beric T, Kojic M, Arsic B, Garalejic E, Djukic S, Stankovic S (2016) Licheniocin 50.2 and bacteriocins from Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 inhibit biofilms of coagulase negative staphylococci and Listeria monocytogenes clinical isolates. PLoS One 11:1–12. https://doi.org/10.1371/journal.pone.0167995

    Article  CAS  Google Scholar 

  • Cocolin L, Rantsiou K, Iacumin L, Urso R, Cantoni C, Comi G (2004) Study of the ecology of fresh sausages and characterization of populations of lactic acid bacteria by molecular methods. Appl Environ Microbiol 70:1883–1894. https://doi.org/10.1128/AEM.70.4.1883-1894.2004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dezwaan DC, Mequio MJ, Littell JS, Allen JP, Rossbach S, Pybus V (2007) Purification and characterization of enterocin 62-6, a two-peptide bacteriocin produced by a vaginal strain Enterococcus faecium: potential significance in bacterial vaginosis. Microb Ecol Health Dis 19:241–250. https://doi.org/10.1080/08910600701538240

    Article  CAS  PubMed  Google Scholar 

  • Dias FS, Ramos CL, Schwan RF (2013) Characterization of spoilage bacteria in pork sausage by PCR-DGGE analysis. Food Sci Technol 33:468–474. https://doi.org/10.1590/S0101-20612013005000079

    Article  Google Scholar 

  • Djadouni F, Kihal M (2012) Antimicrobial activity of lactic acid bacteria and the spectrum of their biopeptides against spoiling germs in foods. Braz Arch Biol Technol 55:435–443

    Article  CAS  Google Scholar 

  • Eid R, El Jakee J, Rashidy A, Asfour H, Omara S, Kandil MM, Mahmood Z, Hahne J, Seida AA (2016) Potential antimicrobial activities of probiotic Lactobacillus strains isolated from raw milk. J Probiotics Health 4:1–8. https://doi.org/10.4172/2329-8901.1000138

    Article  CAS  Google Scholar 

  • El-Deeb N, Sharaf MM, El-Adawi H (2015) Antibacterial and plasmid curing activity of lactic acid bacteria against multidrug resistant bacteria strains. Int J Pharmacol 11:114–121. https://doi.org/10.3923/ijp.2015.114.121

    Article  CAS  Google Scholar 

  • Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791

    Article  PubMed  Google Scholar 

  • Fortina MG, Ricci G, Mora D, Manachini PL (2004) Molecular analysis of artisanal Italian cheeses reveals Enterococcus italicus sp. nov. Int J Syst Evol Microbiol 54:1717–1721. https://doi.org/10.1099/ijs.0.63190-0

    Article  CAS  PubMed  Google Scholar 

  • Franciosi E, Carafa I, Nardin T, Schiavon S, Poznanski E, Cavazza A, Larcher R, Tuohy KM (2015) Biodiversity and γ-aminobutyric acid production by lactic acid bacteria isolated from traditional alpine raw cow’s milk cheeses. BioMed Res Int 2015:1–11. https://doi.org/10.1155/2015/625740

    Article  CAS  Google Scholar 

  • Gaaloul N, Braiek OB, Berjeaud JM, Arthur T, Cavera VL, Chikindas ML, Hani K, Ghrairi T (2014) Evaluation of antimicrobial activity and safety aspect of Enterococcus italicus GGN 10 strain isolated from Tunisian bovine raw milk. J Food Saf 34:1–12. https://doi.org/10.1111/jfs.12126

    Article  CAS  Google Scholar 

  • Guillet C, Join-Lambert O, Le Monnier A, Leclercq A, Mechaї F, Mamzer-Bruneel MF, Bielecka MK, Scortti M, Disson O, Berche P, others (2010) Human listeriosis caused by Listeria ivanovii. Emerg Infect Dis 16:136–138. https://doi.org/10.3201/eid1601.091155

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guttiérrez J, Criado R, Martín M, Herranz C, Cintas LM, Hernández PE (2005) Production of enterocin P, an antilisterial pediocin-like bacteriocin from Enterococcus faecium P13, in Pichia pastoris. Antimicrob Agents Chemother 49:3004–3008. https://doi.org/10.1128/AAC.49.7.3004-3008.2005

    Article  CAS  Google Scholar 

  • H-Kittikun A, Biscola V, El-Ghaish S, Jaffrès E, Dousset X, Pillot G, Haertlé T, Chobert J-M, Hwanhlem N (2015) Bacteriocin-producing Enterococcus faecalis KT2W2G isolated from mangrove forests in southern Thailand: purification, characterization and safety evaluation. Food Control 54:126–134. https://doi.org/10.1016/j.foodcont.2014.12.037

    Article  CAS  Google Scholar 

  • Hor YY, Liong MT (2014) Use of extracellular extracts of lactic acid bacteria and bifidobacteria for the inhibition of dermatological pathogen Staphylococcus aureus. Dermatologica Sinica 32:141–147. https://doi.org/10.1016/j.dsi.2014.03.001

    Article  Google Scholar 

  • Hwanhlem N, Biscola V, El-Ghaish S, Jaffrès E, Dousset X, Haertlé T, H-Kittikun A, Chobert JM (2013) Bacteriocin-producing lactic acid bacteria isolated from mangrove forests in southern Thailand as potential bio-control agents: purification and characterization of bacteriocin produced by Lactococcus lactis subsp. lactis KT2W2L. Probiotics Antimicro Prot 5:264–278. https://doi.org/10.1007/s12602-013-9150-2

    Article  CAS  Google Scholar 

  • Hwanhlem N, Chobert J-M, H-Kittikun A (2014) Bacteriocin-producing lactic acid bacteria isolated from mangrove forests in southern Thailand as potential bio-control agents in food: isolation, screening and optimization. Food Control 41:202–211. https://doi.org/10.1016/j.foodcont.2014.01.021

    Article  CAS  Google Scholar 

  • Hwanhlem N, Jaffrès E, Dousset X, Pillot G, Choiset Y, Haertlé T, H-Kittikun A, Chobert J-M (2015) Application of a nisin Z-producing Lactococcus lactis subsp. lactis KT2W2L isolated from brackish water for biopreservation in cooked, peeled and ionized tropical shrimps during storage at 8 °C under modified atmosphere packaging. Eur Food Res Technol 240:1259–1269. https://doi.org/10.1007/s00217-015-2428-8

    Article  CAS  Google Scholar 

  • Kalschne DL, Womer R, Mattana A, Sarmento MP, Colla LM, Colla E (2015) Characterization of the spoilage lactic acid bacteria in «sliced vacuum-packed ham». Braz J Microbiol 64:173–181. https://doi.org/10.1590/S1517-838246120130019

    Article  Google Scholar 

  • Kimura M (1980) A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120

    Article  CAS  PubMed  Google Scholar 

  • Klewicka E, Libudzisz Z (2004) Antagonistic activity of Lactobacillus acidophilus bacteria towards selected food-contaminating bacteria. Pol J food Nutr Sci 13/54(2):169–174

    Google Scholar 

  • Lengkey HAW, Balia RL, Togoe I, Taşbac BA, Ludong M (2009) Isolation and identification of lactic acid bacteria from raw poultry meat. Biotechnol Anim Husb 25:1071–1077

    Google Scholar 

  • Lim H-J, Kim S-Y, Lee W-K (2004) Isolation of cholesterol-lowering lactic acid bacteria from human intestine for probiotic use. J Vet Sci 5(4):391–395

    Article  PubMed  Google Scholar 

  • Line JE, Svetoch EA, Eruslanov BV, Perelygin VV, Mitsevich EV, Mitsevich IP, Levchuk VP, Svetoch OE, Seal BS, Siragusa GR and others (2008) Isolation and purification of enterocin E-760 with broad antimicrobial activity against gram-positive and gram-negative bacteria. Antimicrob Agents Chemother 52:1094–1100. https://doi.org/10.1128/AAC.01569-06

  • Mahdavi M, Jalali M, Kermanshahi RK (2007) The effect of nisin on biofilm formating foodborne bacteria using microtiter plate method. Res Pharm Sci 2:113–118

    Google Scholar 

  • Malini M, Savitha J (2012) Heat stable bacteriocin from Lactobacillus paracasei subsp. tolerans isolated from locally available cheese: an in vitro study. E3 J Biotechnol Pharm Res 3:28–41

    Google Scholar 

  • Merlich AG, Zhunko ID, Limanska NV, Ivanytsia VO (2017) Antagonistic activity of metabolic products of bacteria Lactobacillus plantarum and Enterococcus italicus with joint action against phytopathogenic bacteria. Microbiologiia i Biotechnologiia 3:45–54 (in Ukrainian. https://doi.org/10.18524/2307-4663.2017.3(39).110876

    Article  Google Scholar 

  • Okuda K, Zendo T, Sugimoto S, Iwase T, Tajima A, Yamada S, Sonomoto K, Mizunoe Y (2013) Effects of bacteriocins on methicillin-resistant Staphylococcus aureus biofilm. Antimicrob Agents Chemother 57:5572–5579. https://doi.org/10.1128/AAC.00888-13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Perez RH, Zendo T, Sonomoto K (2014) Novel bacteriocins from lactic acid bacteria (LAB): various structures and applications. Microb Cell Factories 13:1–8. https://doi.org/10.1186/1475-2859-13-S1-S3

    Article  Google Scholar 

  • Pholsen S, Khota W, Pang H, Higgs D, Cai Y (2016) Characterization and application of lactic acid bacteria for tropical silage preparation. Anim Sci J 87:1202–1211. https://doi.org/10.1111/asj.12534

    Article  CAS  PubMed  Google Scholar 

  • Saad MA, Abdelsamei HM, Ibrahim EMA, Abdou AM, Sohaimy SAE (2015) Effect of pH, heat treatments and proteinase K enzyme on the activity of Lactobacillus acidophilus bacteriocin. Benha Vet Med J 28:210–215

    Article  Google Scholar 

  • Sadekuzzaman M, Yang S, Mizan MFR, Ha SD (2015) Current and recent advanced strategies for combating biofilms. Compr Rev Food Sci Food Saf 14:491–509. https://doi.org/10.1111/1541-4337.12144

    Article  Google Scholar 

  • Saeed S, Ahmad S, Rasool SA (2004) Antimicrobial spectrum, production and mode of action of staphylococcin 188 produced by Staphylococcus aureus 188. Pak J Pharm Sci 17:1–8

    CAS  PubMed  Google Scholar 

  • Saelim K, Kaewsuwan S, Tani A, Maneerat S (2015) Physical, biochemical and genetic characterization of enterocin CE5-1 produced by Enterococcus faecium CE5-1 isolated from Thai indigenous chicken intestinal tract. Songklanakarin J Sci Technol 37:299–307

    CAS  Google Scholar 

  • Şahingil D, Işleroǧlu H, Yildirim Z, Akçelik M, Yildirim M (2011) Characterization of lactococcinBZ produced by Lactococcus lactis subsp. lactis BZ isolated from boza. Tukr J Biol 35:21–33. https://doi.org/10.3906/biy-0906-48

    Article  CAS  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454

    Article  CAS  PubMed  Google Scholar 

  • Salman M, Shahid M, Jamil A, Ur Rahman S (2014) Effect of Lactobacillus acidophilus bacteriocin on Bacillus cereus biofilms. J Pure Appl Microbio 8:3721–3728

    CAS  Google Scholar 

  • Samelis J, Kakouri A, Rementzis J (2000) Selective effect of the product type and the packaging conditions on the species of lactic acid bacteria dominating the spoilage microbial association of cooked meats at 4 °C. Food Microbiol 17:329–340. https://doi.org/10.1006/fmic.1999.0316

    Article  Google Scholar 

  • Satish Kumar R, Kanmani P, Yuvaraj N, Paari KA, Pattukumar V, Arul V (2011) Purification and characterization of enterocin MC13 produced by a potential aquaculture probiont Enterococcus faecium MC13 isolated from the gut of Mugil cephalus. Can J Microbiol 57:993–1001. https://doi.org/10.1139/W11-092

    Article  CAS  PubMed  Google Scholar 

  • Sawa N, Wilaipun P, Kinoshita S, Zendo T, Leelawatcharamas V, Nakayama J, Sonomotoa K (2012) Isolation and characterization of Enterocin W, a novel two-peptide lantibiotic produced by Enterococcus faecalis NKR-4-1. Appl Environ Microbiol 78:900–903. https://doi.org/10.1128/AEM.06497-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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 166:368–379

    Article  PubMed  Google Scholar 

  • Schillinger U, Lüke F-K (1989) Antibacterial activity of Lactobacillus sake isolated from meat. Appl Environ Microbiol 55(8):1901–1906

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sifour M, Ouled-Haddar H, Idoui T, Aissaoui S, Namous H (2014) Screening of some factors affecting bacteriocin production from Lactobacillus curvatus G6 using Plackett-Burman design res rev. Biosci 8:386–393

    Google Scholar 

  • Simões LC, Simões M, Vieira MJ (2011) The effects of metabolite molecules produced by drinking water-isolated bacteria on their single and multispecies biofilms. Biofouling 27:685–699. https://doi.org/10.1080/08927014.2011.597502

    Article  CAS  PubMed  Google Scholar 

  • Song D-F, Zhu M-Y, Gu Q (2014) Purification and characterization of plantaricin ZJ5, a new bacteriocin produced by Lactobacillus plantarum ZJ5. PLoS one 9:1–8. https://doi.org/10.1371/journal.pone.0105549

    Article  CAS  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tiwari A, Singh NP, Sharma G, Dang S, Gupta S, Gabrani R (2015) Isolation and screening of lactic acid bacteria producing bacteriocin like inhibitory substance from soil. Int J Pharmacol Pharm Sci 2:32–36

    CAS  Google Scholar 

  • Trias R, Bañeras L, Montesinos E, Badosa E (2008) Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi. Int Microbiol 11:231–236. https://doi.org/10.2436/20.1501.01.66

    Article  CAS  PubMed  Google Scholar 

  • Vázquez-Boland JA, Kuhn M, Berche P, Chakraborty T, Domínguez-Bernal G, Goebel W, González-Zorn B, Wehland J, Kreft J (2001) Listeria pathogenesis and molecular virulence determinants. Clin Microbiol Rev 14:584–640. https://doi.org/10.1128/CMR.14.3.584-640.2001

    Article  PubMed  PubMed Central  Google Scholar 

  • Visser R, Holzapfel WH (1992) Lactic acid bacteria in the control of plant pathogens. In: Wood BJB (ed) The lactic acid bacteria, vol 1. Elsevier Applied Science, London and New York, pp 193–210

    Google Scholar 

  • Wang T, Liu M (2016) The effect of bacteriocins derived from lactic acid bacteria on growth and biofilm formation of clinical pathogenic strains. Int J Clin Exp Med 9:7343–7348

    CAS  Google Scholar 

  • Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Winkelströter LK, Gomes BC, Thomaz MRS, Souza VM, De Martinis ECP (2011) Lactobacillus sakei 1 and its bacteriocin influence adhesion of Listeria monocytogenes on stainless steel surface. Food Control 22:1404–1407. https://doi.org/10.1016/j.foodcont.2011.02.021

    Article  CAS  Google Scholar 

  • Zmantar T, Slama RB, Fahila K, Kouidhi B, Bakhrouf A, Chaieb K (2016) Modulation of drug resistance and biofilm formation of Staphylococcus aureus isolated from the oral cavity of Tunisian children. Braz J Infect Dis 21:27–34. https://doi.org/10.1016/j.bjid.2016.10.009

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

Andrii Merlich was supported by PhD scholarship from Campus France. We are grateful to Dr. Noraphat Hwanhlem for his kind advice during the work. We thank Hanitra Rabesona for her kind assistance.

Funding

The work was supported by Bilateral French-Ukrainian project «Dnipro» granted by the Ministry of Foreign and European Affairs of France and the Ministry of Education and Science of Ukraine.

Author information

Authors and Affiliations

  1. Microbiology, Virology and Biotechnology Chair, Biology Department, Odessa National I.I. Mechnikov University, 65082 Dvorianska str., 2, Odessa, Ukraine

    Andrii Merlich, Mykola Galkin, Nataliia Limanska, Nataliia Vasylieva & Volodymyr Ivanytsia

  2. UR 1268 INRA Biopolymères Interactions Assemblages, BP 71627, rue de la Géraudière, 44316, Nantes Cedex 3, France

    Yvan Choiset & Thomas Haertlé

  3. Department of Animal Nutrition and Feed Management, ul, Poznan University of Life Sciences, Wołyńska 33, 60-637, Poznan, Poland

    Thomas Haertlé

  4. Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran

    Thomas Haertlé

Authors
  1. Andrii Merlich
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mykola Galkin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yvan Choiset
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nataliia Limanska
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nataliia Vasylieva
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Volodymyr Ivanytsia
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Thomas Haertlé
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andrii Merlich.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Merlich, A., Galkin, M., Choiset, Y. et al. Characterization of the bacteriocin produced by Enterococcus italicus ONU547 isolated from Thai fermented cabbage. Folia Microbiol 64, 535–545 (2019). https://doi.org/10.1007/s12223-019-00677-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12223-019-00677-4

Search

Navigation