A novel enterocin T1 with anti-Pseudomonas activity produced by Enterococcus faecium T1 from Chinese Tibet cheese

  • Hui Liu
  • Lanwei ZhangEmail author
  • Huaxi YiEmail author
  • Xue Han
  • Wei Gao
  • Chunliang Chi
  • Wei Song
  • Haiying Li
  • Chunguang Liu
Original Paper


An enterocin-producing Enterococcus faecium T1 was isolated from Chinese Tibet cheese. The enterocin was purified by SP-Sepharose and reversed phase HPLC. It was identified as unique from other reported bacteriocins based on molecular weight (4629 Da) and amino acid compositions; therefore it was subsequently named enterocin T1. Enterocin T1 was stable at 80–100 °C and over a wide pH range, pH 3.0–10.0. Protease sensitivity was observed to trypsin, pepsin, papain, proteinase K, and pronase E. Importantly, enterocin T1 was observed to inhibit the growth of numerous Gram-negative and Gram-positive bacteria including Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas fluorescens, Escherichia coli, Salmonella typhimurium, Shigella flexneri, Shigella sonnei, Staphylococcus aureus, Listeria monocytogenes. Take together, these results suggest that enterocin T1 is a novel bacteriocin with the potential to be used as a bio-preservative to control Pseudomonas spp. in food.


Enterocin Pseudomonas Charaterization Biopreservation 



This work was financially supported by the Research Fund for the Doctoral Program of Higher Education of China (No. 20112302110051), National Science Foundation, China (Nos. 31271906/C2002204, 31571850/C200502).

Compliance with ethical standards

Conflict of interest

No conflict of interest is declared.


  1. Achemchem F, Martínez-Bueno M, Abrini J, Valdivia E, Maqueda M (2005) Enterococcus faecium F58, a bacteriocinogenic strain naturally occurring in Jben, a soft, farmhouse goat’s cheese made in Morocco. J Appl Microbiol 99:141–150CrossRefGoogle Scholar
  2. Alvarado C, Garcia-Almendarez B, Martin S, Regalado C (2005) Anti-Listeria monocytogenes bacteriocin-like inhibitory substances from Enterococcus faecium UQ31 isolated from artisan Mexican-style cheese. Curr Microbiol 51:110–115CrossRefGoogle Scholar
  3. 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 62:1676–1682Google Scholar
  4. Batdorj B et al (2006) Purification and characterization of two bacteriocins produced by lactic acid bacteria isolated from Mongolian airag. J Appl Microbiol 101:837–848CrossRefGoogle Scholar
  5. Chatterjee C, Paul M, Xie LL, van der Donk WA (2005) Biosynthesis and mode of action of lantibiotics. Chem Rev 105:633–683CrossRefGoogle Scholar
  6. Cintas LM, Casaus P, Håvarstein LS, Hernandez PE, Nes IF (1997) Biochemical and genetic characterization of enterocin P, a novel sec-dependent bacteriocin from Enterococcus faecium P13 with a broad antimicrobial spectrum. Appl Environ Microbiol 63:4321–4330Google Scholar
  7. Cintas LM, Casaus P, Herranz C, Håvarstein LS, Holo H, Hernández PE, Nes IF (2000) Biochemical and genetic evidence that Enterococcus faecium L50 produces enterocins L50A and L50B, thesec-dependent enterocin P, and a novel bacteriocin secreted without an N-terminal extension termed enterocin Q. J Bacteriol 182:6806–6814CrossRefGoogle Scholar
  8. Cotter PD, Hill C, Ross RP (2005) Bacteriocins: developing innate immunity for food. Nat Rev Microbiol 3:777–788CrossRefGoogle Scholar
  9. Craven H, Macauley B (1992) Microorganisms in pasteurised milk after refrigerated storage. 1. Identification of types. Aust J Dairy Technol 47:38–45Google Scholar
  10. Drider D, Rebuffat S (2011) Prokaryotic antimicrobial peptides: from genes to applications. Springer Science & Business Media, BerlinCrossRefGoogle Scholar
  11. Drider D, Fimland G, Héchard Y, McMullen LM, Prévost H (2006) The continuing story of class IIa bacteriocins. Microbiol Mol Biol Rev 70:564–582CrossRefGoogle Scholar
  12. Eneroth Å, Ahrné S, Molin G (2000) Contamination of milk with Gram-negative spoilage bacteria during filling of retail containers. Int J Food Microbiol 57:99–106CrossRefGoogle Scholar
  13. Ennahar S, Deschamps N (2000) Anti-Listeria effect of enterocin A, produced by cheese-isolated Enterococcus faecium EFM01, relative to other bacteriocins from lactic acid bacteria. J Appl Microbiol 88:449–457CrossRefGoogle Scholar
  14. Farias ME, De Ruiz H, Aida A, Sesma F (1994) Bacteriocin production by lactic acid bacteria isolated from regional cheeses: inhibition of foodborne pathogens. J Food Prot® 57:1013–1015Google Scholar
  15. Foulquié Moreno M, Sarantinopoulos P, Tsakalidou E, De Vuyst L (2006) The role and application of enterococci in food and health. Int J Food Microbiol 106:1–24CrossRefGoogle Scholar
  16. Franz C, Schillinger U, Holzapfel W (1996) Production and characterization of enterocin 900, a bacteriocin produced by Enterococcus faecium BFE 900 from black olives. Int J Food Microbiol 29:255–270CrossRefGoogle Scholar
  17. Franz CM, Van Belkum MJ, Holzapfel WH, Abriouel H, Gálvez A (2007) Diversity of enterococcal bacteriocins and their grouping in a new classification scheme. FEMS Microbiol Rev 31:293–310CrossRefGoogle Scholar
  18. Gálvez A, Abriouel H, López RL, Omar NB (2007) Bacteriocin-based strategies for food biopreservation. Int J Food Microbiol 120:51–70CrossRefGoogle Scholar
  19. Garcia M, Sanz B, Garcia-Collia P, Ordonez J (1989) Activity and thermostability of the extracellular lipases and proteinases from pseudomonads isolated from raw milk. Milchwissenschaft 44:547–550Google Scholar
  20. Ghrairi T, Frere J, Berjeaud J, Manai M (2008) Purification and characterisation of bacteriocins produced by Enterococcus faecium from Tunisian rigouta cheese. Food Control 19:162–169CrossRefGoogle Scholar
  21. Heng NC, Tagg JR (2006) What’s in a name?. Class distinction for bacteriocins, Nat Rev Microbiol 4Google Scholar
  22. Herranz C et al (1999) Biochemical and genetic evidence of enterocin P production by two Enterococcus faecium-like strains isolated from fermented sausages. Curr Microbiol 39:282–290CrossRefGoogle Scholar
  23. Hickey RM, Twomey DP, Ross RP, Hill C (2003) Production of enterolysin A by a raw milk enterococcal isolate exhibiting multiple virulence factors. Microbiol 149:655–664CrossRefGoogle Scholar
  24. Jang S, Lee J, Jung U, Choi HS, Suh HJ (2014) Identification of an anti-listerial domain from Pediococcus pentosaceus T1 derived from Kimchi, a traditional fermented vegetable. Food Control 43:42–48CrossRefGoogle Scholar
  25. Kang J, Lee M (2005) Characterization of a bacteriocin produced by Enterococcus faecium GM-1 isolated from an infant. J Appl Microbiol 98:1169–1176CrossRefGoogle Scholar
  26. Kaur G et al (2011) Nisin and class IIa bacteriocin resistance among Listeria and other foodborne pathogens and spoilage bacteria. Microb Drug Resist 17:197–205CrossRefGoogle Scholar
  27. Khalil R, Elbahloul Y, Djadouni F, Omar S (2009) Isolation and partial characterization of a bacteriocin produced by a newly isolated Bacillus megaterium 19 strain. Pak J Nutr 8:242–250CrossRefGoogle Scholar
  28. Khay EO, Idaomar M, Castro L, Bernárdez P, Senhaji N, Abrini J (2013) Antimicrobial activities of the bacteriocin-like substances produced by lactic acid bacteria isolated from Moroccan dromedary milk. Afr J Biotechnol 10:10447–10455Google Scholar
  29. Klaenhammer TR (1993) Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol Rev 12:39–85CrossRefGoogle Scholar
  30. Klein G, Rüben C, Upmann M (2013) Antimicrobial activity of essential oil components against potential food spoilage microorganisms. Curr Microbiol 67:200–208CrossRefGoogle Scholar
  31. Liu X et al (2011) Identification of an N-terminal formylated, two-peptide bacteriocin from Enterococcus faecalis 710C. J Agric Food Chem 59:5602–5608CrossRefGoogle Scholar
  32. Losteinkit C, Uchiyama K, Ochi S, Takaoka T, Nagahisa K, Shioya S (2001) Characterization of bacteriocin N15 produced by Enterococcus faecium N15 and cloning of the related genes. J Biosci Bioeng 91:390–395CrossRefGoogle Scholar
  33. Lü X, Yi L, Dang J, Dang Y, Liu B (2014) Purification of novel bacteriocin produced by Lactobacillus coryniformis MXJ 32 for inhibiting bacterial foodborne pathogens including antibiotic-resistant microorganisms. Food Control 46:264–271CrossRefGoogle Scholar
  34. Maqueda M, Sánchez-Hidalgo M, Fernández M, Montalbán-López M, Valdivia E, Martínez-Bueno M (2008) Genetic features of circular bacteriocins produced by Gram-positive bacteria. FEMS Microbiol Rev 32:2–22CrossRefGoogle Scholar
  35. Messaoudi S et al (2012) Purification and characterization of a new bacteriocin active against Campylobacter produced by Lactobacillus salivarius SMXD51. Food Microbiol 32:129–134CrossRefGoogle Scholar
  36. Moreno M et al (2002) Microbial analysis of Malaysian tempeh, and characterization of two bacteriocins produced by isolates of Enterococcus faecium. J Appl Microbiol 92:147–157CrossRefGoogle Scholar
  37. Nes IF, Diep DB, Håvarstein LS, Brurberg MB, Eijsink V, Holo H (1996) Biosynthesis of bacteriocins in lactic acid bacteria. Antonie Van Leeuwenhoek 70:113–128CrossRefGoogle Scholar
  38. Nes IF, Diep DB, Holo H (2007) Bacteriocin diversity in Streptococcus and Enterococcus. J Bacteriol 189:1189–1198CrossRefGoogle Scholar
  39. Nes IF, Diep DB, Ike Y (2014) Enterococcal bacteriocins and antimicrobial proteins that contribute to niche control. In: Gilmore MS, Clewell DB, Ike Y, Shankar N (eds) Enterococci: from commensals to leading causes of drug resistant infection. Massachusetts Eye and Ear Infirmary, BostonGoogle Scholar
  40. Nissen-Meyer J, Oppegård C, Rogne P, Haugen HS, Kristiansen PE (2010) Structure and mode-of-action of the two-peptide (Class-IIb) bacteriocins. Probiot Antimicrob Proteins 2:52–60CrossRefGoogle Scholar
  41. Pringsulaka O, Thongngam N, Suwannasai N, Atthakor W, Pothivejkul K, Rangsiruji A (2012) Partial characterisation of bacteriocins produced by lactic acid bacteria isolated from Thai fermented meat and fish products. Food Control 23:547–551CrossRefGoogle Scholar
  42. Saavedra L, Sesma F (2011) Purification techniques of bacteriocins from lactic acid bacteria and other Gram-positive bacteria. In: Prokaryotic antimicrobial peptides. Springer, Berlin, pp 99–113Google Scholar
  43. Samelis J, Roller S, Metaxopoulos J (1994) Sakacin B, a bacteriocin produced by Lactobacillus sake isolated from Greek dry fermented sausages. J Appl Bacteriol 76:475–486CrossRefGoogle Scholar
  44. Snyder AB, Worobo RW (2014) Chemical and genetic characterization of bacteriocins: antimicrobial peptides for food safety. J Sci Food Agric 94:28–44CrossRefGoogle Scholar
  45. Sperber WH, Doyle MP (2009) Compendium of the microbiological spoilage of food and beverages. Springer, BerlinCrossRefGoogle Scholar
  46. Svetoch EA et al (2008) Diverse antimicrobial killing by Enterococcus faecium E 50–52 bacteriocin. J Agric Food Chem 56:1942–1948CrossRefGoogle Scholar
  47. Svetoch EA, Eruslanov BV, Perelygin VV, Levchuk VP, Seal BS, Stern NJ (2010) Inducer bacteria, unique signal peptides, and low-nutrient media stimulate in vitro bacteriocin production by Lactobacillus spp. and Enterococcus spp. strains. J Agric Food Chem 58:6033–6038CrossRefGoogle Scholar
  48. Tiwari BK, Valdramidis VP, O’Donnell CP, Muthukumarappan K, Bourke P, Cullen P (2009) Application of natural antimicrobials for food preservation. J Agric Food Chem 57:5987–6000CrossRefGoogle Scholar
  49. Todokoro D, Tomita H, Inoue T, Ike Y (2006) Genetic analysis of bacteriocin 43 of vancomycin-resistant Enterococcus faecium. Appl Environ Microbiol 72:6955–6964CrossRefGoogle Scholar
  50. Todorov SD et al (2010) Characterisation of an antiviral pediocin-like bacteriocin produced by Enterococcus faecium. Food Microbiol 27:869–879CrossRefGoogle Scholar
  51. Vithanage NR, Yeager TR, Jadhav SR, Palombo EA, Datta N (2014) Comparison of identification systems for psychrotrophic bacteria isolated from raw bovine milk. Int J Food Microbiol 189:26–38CrossRefGoogle Scholar
  52. Yang R, Johnson MC, Ray B (1992) Novel method to extract large amounts of bacteriocins from lactic acid bacteria. Appl Environ Microbiol 58:3355–3359Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Hui Liu
    • 1
  • Lanwei Zhang
    • 1
    Email author
  • Huaxi Yi
    • 1
    Email author
  • Xue Han
    • 1
  • Wei Gao
    • 1
  • Chunliang Chi
    • 1
  • Wei Song
    • 1
  • Haiying Li
    • 2
  • Chunguang Liu
    • 2
  1. 1.School of Chemical Engineering and TechnologyHarbin Institute of TechnologyHarbinChina
  2. 2.College of Life SciencesHeilongjiang UniversityHarbinChina

Personalised recommendations