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Potential of enterocin from Enterococcus durans MF5 in controlling Listeria species

Published online by Cambridge University Press:  13 March 2025

Nathália Aparecida Andrade de Souza ,
Luana de Carvalho ,
Matheus Henrique Nogueira ,
Márcia Cristina Furlaneto  and
Luciana Furlaneto Maia Open the ORCID record for Luciana Furlaneto Maia [Opens in a new window]
Nathália Aparecida Andrade de Souza
Affiliation:
Department of Microbiology, Center of Biological Sciences, Paraná State University of Londrina, Rodovia Celso Garcia Cid, Pr 445 Km 380, C.P. 6001, 86051990 Londrina, Paraná, Brazil
Luana de Carvalho
Affiliation:
Department of Food Technology, Technological Federal University of Paraná, Londrina, Pioneiros Avenue 3131, Jardim Morumbi, 86036-370 Londrina, Paraná, Brazil
Matheus Henrique Nogueira
Affiliation:
Department of Food Technology, Technological Federal University of Paraná, Londrina, Pioneiros Avenue 3131, Jardim Morumbi, 86036-370 Londrina, Paraná, Brazil
Márcia Cristina Furlaneto
Affiliation:
Department of Microbiology, Center of Biological Sciences, Paraná State University of Londrina, Rodovia Celso Garcia Cid, Pr 445 Km 380, C.P. 6001, 86051990 Londrina, Paraná, Brazil
Luciana Furlaneto Maia*
Affiliation:
Department of Food Technology, Technological Federal University of Paraná, Londrina, Pioneiros Avenue 3131, Jardim Morumbi, 86036-370 Londrina, Paraná, Brazil
*
Corresponding author: Luciana Furlaneto Maia; Email: lucianamaia@utfpr.edu.br
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Abstract

This research paper presents the characterization of an enterocin-producing Enterococcus durans MF5 isolate and the determination of the in vitro antilisterial activity of enterocin produced by this isolate, named Ent-MF5. PCR-based screening for bacteriocin biosynthetic genes revealed that E. durans MF5 harbors multiple enterocin-encoding genes (ent A, B, P and X), classified as class II bacteriocins and enterocin-P of Enterococcus faecium (sharing up to 99% similarity at the genetic level). E. durans MF5 is sensitive to eight clinically important antibiotics and does not possess cytolysin activator -cylA, gelatinase -gelE and hyaluronidase -hylA virulence genes. The antilisterial activity of Ent-MF5 was abolished by trypsin, α-chymotrypsin, protease and proteinase-K. Ent-MF5 showed thermal and pH stability. In addition, the activity of Ent-MF5 was unaffected in the presence of various surfactants (1% SDS, Triton X-100, Tween 20, and Tween 80). Ent-MF5 exhibited antimicrobial activity against Listeria monocytogenes, Listeria innocua, Listeria ivanovii and Listeria seeligeri at concentrations as low as 0.13 μg/ml. Ent-MF5 had a bactericidal effect against L. monocytogenes with a significant reduction in surviving cells at concentrations equal to or greater than 0.13 μg/ml. A 75–100% reduction in L. monocytogenes growth and bactericidal effect determined by CFU counts was observed following treatment with Ent-MF5 at 4.47 μg/ml at time points starting at 2 and 4 h, respectively. Ent-MF5 action is associated with Listeria cell membrane damage, as observed by flow cytometry and fluorescence microscopy. Thus, the effective antilisterial activity and stability of Ent-MF5 presents promising perspectives for application as biopreservatives in the food industry.

Keywords

Bacteriocinbiopreservativesfood safetyfoodborne pathogensListeria

Information

Type
Research Article
Information
Journal of Dairy Research , Volume 91 , Issue 4 , November 2024 , pp. 516 - 524
Copyright
Copyright © The Author(s), 2025. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

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References

Abanoz, HS and Kunduhoglu, B (2018) Antimicrobial activity of a bacteriocin produced by Enterococcus faecalis KT11 against some pathogens and antibiotic-resistant bacteria. Korean Journal for Food Science of Animal Resources 38, 1064–1079.CrossRefGoogle ScholarPubMed
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle ScholarPubMed
CDC (2022) Centers for Disease Control and Prevention. Listeria outbreaks. Available at https://www.cdc.gov/listeria/outbreaks/index.htmlGoogle Scholar
Chen, R, Skeens, J, Orsi, RH, Wiedmann, M and Guariglia-Oropeza, V (2020) Pre-growth conditions and strain diversity affect nisin treatment efficacy against Listeria monocytogenes on cold-smoked salmon. International Journal of Food Microbiology 333, 108793.CrossRefGoogle ScholarPubMed
Chen, R, Skeens, JW and Wiedmann, M (2022) The efficacy of nisin against Listeria monocytogenes on cold-smoked salmon at natural contamination levels is concentration-dependent and varies by serotype. Frontiers in Microbiology 3, 930400.CrossRefGoogle Scholar
CLSI (2020) Performance Standards for Antimicrobial Susceptibility Testing. 30th ed. CLSI Supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute.Google Scholar
Comerlato, CB, Zhang, X, Walker, K, Mayne, J, Figeys, D and Brandelli, A (2023) The influence of protein secretomes of Enterococcus durans on ex vivo human gut microbiome. Probiotics and Antimicrobial Proteins 16, 1954–1965.CrossRefGoogle ScholarPubMed
Creti, R, Imperi, M, Bertuccini, L, Fabretti, F, Orefici, G, Di Rosa, R and Baldassarri, L (2004) Survey for virulence determinants among Enterococcus faecalis isolated from different sources. Journal of Medical Microbiology 53, 13–20.CrossRefGoogle ScholarPubMed
Daba, G, Mostafa, FA, Saleh, SAA, Elkhateeb, WA, Awad, G, Nomiyama, T, Zendo, T and El-Dein, AN (2022) Purification, amino acid sequence, and characterization of bacteriocin GA15, a novel class IIa bacteriocin secreted by Lactiplantibacillus plantarum GCNRC_GA15. International Journal of Biological Macromolecules 213, 651–662.CrossRefGoogle ScholarPubMed
Darbandi, A, Asadi, A, Mahdizade, AM, Ohadi, E, Talebi, M, Halaj, ZM, Darb, EA, Ghanavati, R and Kakanj, M (2022) Bacteriocins: properties and potential use as antimicrobials. Journal of Clinical Laboratory Analysis 36, e24093.CrossRefGoogle ScholarPubMed
Du, Z, Su, H, Wang, W, Ye, L, Hong Wei, H, Peng, Z, Anishchenko, I, Baker, D and Yang, J (2021) The trRosetta server for fast and accurate protein structure prediction. Nature Protocols 16, 5634–5651.CrossRefGoogle ScholarPubMed
Duze, ST, Marimani, M and Patel, M (2021) Tolerance of Listeria monocytogenes to biocides used in food processing environments. Food Microbiology 97, 103758.CrossRefGoogle ScholarPubMed
Furlaneto-Maia, L, Ramalho, R, Rocha, KR and Furlaneto, MC (2020) Antimicrobial activity of enterocins against Listeria sp. and other food spoilage bacteria. Biotechnology Letters 42, 797–806.CrossRefGoogle ScholarPubMed
Gómez, NC, Abriouel, H, Grande, MAJ, Pulido, RP and Gálvez, A (2012) Effect of enterocin AS-48 in combination with biocides on planktonic and sessile Listeria monocytogenes. Food Microbiology 30, 51–58.CrossRefGoogle ScholarPubMed
Gonelimali, FD, Lin, J, Miao, W, Xuan, J, Charles, F, Chen, M and Hatab, SR (2018) Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms. Frontiers in Microbiology 9, 1639.CrossRefGoogle ScholarPubMed
Hanchi, H, Mottawea, W, Sebei, K and Hammami, R (2018) The genus Enterococcus: between probiotic potential and safety concerns – an update. Frontiers in Microbiology 9, 1791.CrossRefGoogle ScholarPubMed
Ibarguren, C, Guitián, MV, Lenz, RM, Cecilia, SM and Audisio, MC (2022) Response of sensitive and resistant Listeria monocytogenes strains against bacteriocins produced by different Enterococcus spp. strains. International Journal of Food Microbiology 382, 109928.CrossRefGoogle Scholar
IFSAC – Interagency Food Safety Analytics Collaboration (2022) Foodborne illness source attribution estimates for 2020 for Salmonella, Escherichia coli O157, and Listeria monocytogenes using multi-year outbreak surveillance data, United States. Available at https://www.cdc.gov/foodsafety/ifsac/pdf/P19-2020-reportTriAgency-508.pdfGoogle Scholar
Johnson, MB and Criss, AK (2013) Fluorescence microscopy methods for determining the viability of bacteria in association with mammalian cells. Journal of Visualized Experiments 9, 50729.Google Scholar
Kang, BS, Seo, JG, Lee, GS, Kim, JH, Kim, SY, Han, YW, Kang, H, Kim, HO, Rhee, JH, Chung, MJ and Park, YM (2009) Antimicrobial activity of enterocins from Enterococcus faecalis SL-5 against Propionibacterium acnes, the causative agent in acne vulgaris and its therapeutic effect. The Journal of Microbiology 47, 101–109.CrossRefGoogle ScholarPubMed
Kaszoni-Rückerl, I, Mustedanagic, A, Muri-Klinger, S, Brugger, K, Wagner, KH, Wagner, M and Stessl, B (2020) Predominance of distinct Listeria innocua and Listeria monocytogenes in recurrent contamination events at dairy processing facilities. Microorganisms 8, 234.CrossRefGoogle ScholarPubMed
Masias, E, Dupuy, FG, da Silva Sanches, PR, Farizano, JV, Cilli, E, Bellomio, A and Minahk, C (2017) Impairment of the class IIa bacteriocin receptor function and membrane structural changes are associated to enterocin CRL35 high resistance in Listeria monocytogenes. Biochimica et Biophysica Acta 1861, 1770–1776.CrossRefGoogle ScholarPubMed
Mazaheri, T, Cervantes-Huamán, BRH, Bermúdez-Capdevila, M, Ripolles-Avila, C and Rodríguez-Jerez, JJ (2021) Listeria monocytogenes biofilms in the food industry: is the current hygiene program sufficient to combat the persistence of the pathogen? Microorganisms 9, 181.CrossRefGoogle Scholar
Melero, B, Stessl, B, Manso, B, Wagner, M, Esteban-Carbonero, OJ, Hernandez, M, Rovira, J and Rodriguez-Lazaro, D (2019) Listeria monocytogenes colonization in a newly established dairy processing facility. International Journal of Food Microbiology 289, 64–71.CrossRefGoogle Scholar
Minahk, CJ, Farías, ME, Sesma, F and Morero, RD (2000) Effect of enterocin CRL35 on Listeria monocytogenes cell membrane. FEMS Microbiology Letters 192, 79–83.CrossRefGoogle ScholarPubMed
Muthuvelu, KS, Ethiraj, B, Pramnik, S, Raj, NK, Venkataraman, S, Rajendran, DS, Bharathi, P, Palanisamy, E, Narayanan, AS, Vaidyanathan, VK and Muthusamy, S (2023) Biopreservative technologies of food: an alternative to chemical preservation and recent developments. Food Science and Biotechnology 32, 1337–1350.CrossRefGoogle ScholarPubMed
Nüesch-Inderbinen, M, Bloemberg, GV, Müller, A, Stevens, MJA, Cernela, N, Kollöffel, B and Stephan, R (2021) Listeriosis caused by persistence of Listeria monocytogenes serotype 4b sequence type 6 in cheese production environment. Emerging Infectious Diseases 27, 284–288.CrossRefGoogle ScholarPubMed
Orsi, RH and Wiedmann, M (2016) Characteristics and distribution of Listeria spp., including Listeria species newly described since 2009. Applied Microbiology and Biotechnology 100, 5273–5287.CrossRefGoogle ScholarPubMed
Osek, J, Lachtara, B and Wieczorek, K (2022) Listeria monocytogenes – how this pathogen survives in food-production environments? Frontiers in Microbiology 26, 866462.CrossRefGoogle Scholar
Perumal, V and Venkatesan, A (2017) Antimicrobial, cytotoxic effect and purification of bacteriocin from vancomycin susceptible Enterococcus faecalis and its safety evaluation for probiotization. Lebensmittel-Wissenschaft + Technologie 78, 303–310.CrossRefGoogle Scholar
Qiao, X, Du, R, Wang, Y, Han, Y and Zhou, Z (2020) Purification, characterization and mode of action of enterocin, a novel bacteriocin produced by Enterococcus faecium TJUQ1. International Journal of Biological Macromolecules 144, 151–159.CrossRefGoogle ScholarPubMed
Quereda, JJ, Moron-Garcia, A, Palacios-Gorba, C, Dessaux, C, Garcia-Del Portillo, F, Pucciarelli, MG and Ortega, AD (2021) Pathogenicity and virulence of Listeria monocytogenes: a trip from environmental to medical microbiology. Virulence 12, 2509–2545.CrossRefGoogle ScholarPubMed
Ramalho, R, de Souza, NAA, Moreira, TFM, De Oliveira, A, Perini, HF, Furlaneto, MC, Leimann, FV and Furlaneto-Maia, L (2023) Antibacterial efficacy of Enterococcus microencapsulated bacteriocin on Listeria monocytogenes, Listeria innocua and Listeria ivanovii. Journal of Food Science and Technology 60, 262–271.CrossRefGoogle Scholar
Rocha, KR, Perini, HF, Souza, CM, Schueler, J, Tosoni, NF, Furlaneto, MC and Furlaneto-Maia, L (2019) Inhibitory effect of bacteriocins from enterococci on developing and preformed biofilms of Listeria monocytogenes, Listeria ivanovii and Listeria innocua. World Journal of Microbiology and Biotechnology 35, 96.CrossRefGoogle ScholarPubMed
Rossi, F, Giaccone, V, Colavita, G, Amadoro, C, Pomilio, F and Catellani, P (2022) Virulence characteristics and distribution of the pathogen Listeria ivanovii in the environment and in food. Microorganisms 10, 1679.CrossRefGoogle ScholarPubMed
Sameli, N, Skandamis, PN and Samelis, J (2021) Application of Enterococcus faecium KE82, an enterocin A-B-P-producing strain, as an adjunct culture enhances inactivation of Listeria monocytogenes during traditional protected designation of origin galotyri processing. Journal of Food Protection 84, 87–98.CrossRefGoogle Scholar
Sharma, P, Kaur, S, Chadha, BS, Kaur, R, Kaur, M and Kaur, S (2021) Anticancer and antimicrobial potential of enterocin 12a from Enterococcus faecium. BMC Microbiology 4, 39.CrossRefGoogle Scholar
Silva, CCG, Silva, SPM and Ribeiro, SC (2018) Application of bacteriocins and protective cultures in dairy food preservation. Frontiers in Microbiology 9, 594.CrossRefGoogle ScholarPubMed
Sioziou, E, Kakouri, A, Bosnea, L and Samelis, J (2024) Antilisterial activity of raw sheep milk from two native Epirus breeds: culture-dependent identification, bacteriocin gene detection and primary safety evaluation of the antagonistic LAB biota. Current Research in Microbial Sciences 10, 100209.CrossRefGoogle Scholar
Tosoni, NF, Perini, HF, Terra, MR, Katsuda, MS, Furlaneto, MC and Furlaneto-Maia, L (2019) Antimicrobial activity of enterocin obtained from Enterococcus durans on Shiga-like toxin-producing Escherichia coli. Ciência Rural 49, e20190297.CrossRefGoogle Scholar
Vasilchenko, AS and Valyshev, AV (2019) Pore-forming bacteriocins: structural–functional relationships. Archives of Microbiology 201, 147–154.CrossRefGoogle ScholarPubMed
Vijayendra, SV, Rajashree, K and Halami, PM (2010) Characterization of a heat stable anti-listerial bacteriocin produced by vancomycin sensitive Enterococcus faecium isolated from idli batter. Indian Journal of Microbiology 50, 243–246.CrossRefGoogle ScholarPubMed
Wu, M, Dong, Q, Ma, Y, Yang, S, Aslam, MZ, Liu, Y and Li, Z (2022a) Potential antimicrobial activities of probiotics and their derivatives against Listeria monocytogenes in food field: a review. Food Research International 160, 111733.CrossRefGoogle Scholar
Wu, Y, Pang, X, Wu, Y, Liu, X and Zhang, X (2022b) Enterocins: classification, synthesis, antibacterial mechanisms and food applications. Molecules 27, 2258.CrossRefGoogle Scholar
Yan, H, Aizhan, R, Lu, YY, Li, X, Wang, X, Yi, YL, Shan, YY, Liu, BF, Zhou, Y and Lü, X (2021) A novel bacteriocin BM1029: physicochemical characterization, antibacterial modes and application. Journal of Applied Microbiology 130, 755–768.CrossRefGoogle ScholarPubMed