Purification and Characterization of a Novel Bacteriocin Against Vancomycin Resistant Enterococci Produced by Enterococcus hirae HM02-04
Abstract
Enterococcus hirae HM02-04 isolated from breast milk exerted growth inhibition against especially vancomycin resistance enterococci (VRE). The aims of this study were to purify and characterize the antimicrobial substance produced by the strain HM02-04. One active peptide was successfully purified by 3 steps of Amberlite XAD-16 adsorption-desorption, cation exchange chromatography, and reverse-phase HPLC. It had 2605.298 Da peptide by MALDI-TOF MS analysis. The peptide sequence determined by LC-MS/MS contained only 23 amino acid residues providing the molecular mass of 2312.67 Da by in silico analysis. However, its amino acid sequence obtained showed no homology to other bacteriocins including enterocins previously reported and proposed as a novel one named Hiracin HM02-04. This is the first report to discover E. hirae isolated from breast milk that was able to produce a bacteriocin against VRE. Hiracin HM02-04 was stable at a high temperature of 121°C for 15 min and at a wide pH range of 3–9. It was sensitive to actinase E, pepsin, proteinase K and trypsin. The Hiracin HM02-04 has the narrow inhibition spectrum and no inhibition against Listeria. This bacteriocin was also found to have a bactericidal mode of action with concomitant cell lysis against the strain VRE 426. The present research addresses Hiracin HM02-04 as a promising alternative to conventional antibiotics in the treatment of enterococcal infections.
Keywords
[1] A. T. Ulijasz, A. Grenader, and B. Weisblum, “A vancomycin-inducible lacZ reporter system in Bacillus subtilis: Induction by antibiotics that inhibit cell wall synthesis and by lysozyme,” Journal of Bacteriology, vol. 178, no. 21, pp. 6305–6309, Nov. 1996.
[2] W. R. Miller, B. E. Murray, L. B. Rice, and C. A. Arias, “Vancomycin-resistant enterococci: Therapeutic challenges in the 21st century,” Infectious Disease Clinics, vol. 30, no. 2, pp. 415–439, 2016.
[3] R. M. van Harten, R. J. Willems, N. I. Martin, and A. P. Hendrickx, “Multidrug-resistant enterococcal infections: New compounds, novel antimicrobial therapies?,” Trends in Microbiology, vol. 25, no. 6, pp. 467–479, Jun. 2017.
[4] V. Yarlagadda, P. Sarkar, G. B. Manjunath, and J. Haldar, “Lipophilic vancomycin aglycon dimer with high activity against vancomycin-resistant bacteria,” Bioorganic & Medicinal Chemistry Letters, vol. 25, no. 23, pp. 5477–5480, Dec. 2015.
[5] P. D. Cotter, C. Hill, and R. P. Ross, “Food microbiology: Bacteriocins: Developing innate immunity for food,” Nature Reviews Microbiology, vol. 3, no. 10, pp. 777–778, Oct. 2005.
[6] R. Aunpad and D. Pipatsatitpong, “Isolation and characterization of a bacteriocin with Anti- MRSA activity from Bacillus sp. strain WASM9- 25M,” Thai Journal of Pharmacology, vol. 36, no. 2, pp. 19–28, Dec. 2014.
[7] F. Lara-Villoslada, M. Olivares, S. Sierra, J. M. Rodriguez, J. Boza, and J. Xaus, “Beneficial effects of probiotic bacteria isolated from breast milk,” The British Journal of Nutrition, vol. 98, pp. 96–100, Oct. 2007.
[8] V. Savini, T. Bonfini, R. Marrollo, A. V. Argentieri, S. Riccioni, D. Astolfi, P. Fazii, D. D. Antonio, and G. Gherardi, “Enterococcus hirae: A zoonotic microorganism in human umbilical cord blood,” World Journal of Microbiology and Biotechnology, vol. 30, no. 4, pp. 1423–1426, Nov. 2013.
[9] N. Bourafa, L. Loucif, N. Boutefnouchet, and J. M. Rolain, “Enterococcus hirae, an unusual pathogen in humans causing urinary tract infection in a patient with benign prostatic hyperplasia: First case report in Algeria,” New Microbes and New Infections, vol. 8, pp. 7–9, Nov. 2015.
[10] P. V. Dicpinigaitis, M. D. Aguirre, and J. Divito, “Enterococcus hirae bacteremia associated with acute pancreatitis and septic shock. Case reports in infectious diseases,” Case Reports in Infectious Diseases, vol. 2015, pp. 1–3, 2015.
[11] L. Devriese, M. Vancanneyt, P. Descheemaeker, M. Baele, H. Van Landuyt, B. Gordts, P. Butaye, J. Swings, and F. Haesebrouck, “Differentiation and identification of Enterococcus durans, E. hirae and E. villorum,” Journal of Applied Microbiology, vol. 92, no. 5, pp. 821–827, Apr. 2002.
[12] V. Cavicchioli, A. Camargo, S. Todorov, and L. Nero, “Novel bacteriocinogenic Enterococcus hirae and Pediococcus pentosaceus strains with antilisterial activity isolated from Brazilian artisanal cheese,” Journal of Dairy Science, vol. 100, no. 4, pp. 2526–2535, Apr. 2017.
[13] S. Ennahar, Y. Asou, T. Zendo, K. Sonomoto, and A. Ishizaki, “Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81,” International Journal of Food Microbiology, vol. 70, no. 3, pp. 291–301, Nov. 2001.
[14] K. Rumjuankiat, R. H. Perez, K. Pilasombut, S. Keawsompong, T. Zendo, K. Sonomoto, and S. Nitisinprasert, “Purification and characterization of a novel plantaricin, KL-1Y, from Lactobacillus plantarum KL-1,” World Journal of Microbiology and Biotechnology, vol. 31, no. 6, pp. 983–994, Apr. 2015.
[15] M. M. Bradford, “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding,” Analytical Biochemistry, vol. 72, no. 1–2, pp. 248–254, 1976.
[16] P. Therdtatha, C. Tandumrongpong, K. Pilasombut, H. Matsusaki, S. Keawsompong, and S. Nitisinprasert, “Characterization of antimicrobial substance from Lactobacillus salivarius KL-D4 and its application as biopreservative for creamy filling,” SpringerPlus, vol. 5, pp. 1–21, 2016.
[17] S. Elayaraja, N. Annamalai, P. Mayavu, and T. Balasubramanian, “Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum,” Asian Pacific Journal of Tropical Biomedicine, vol. 4, pp. 305– 311, May 2014.
[18] M. Farías, R. Farías, A. D. R. Holgado, and F. Sesma, “Purification and N‐terminal amino acid sequence of Enterocin CRL 35, a ‘pediocinlike’bacteriocin produced by Enterococcus faecium CRL 35,” Letters in Applied Microbiology, vol. 22, no. 6, pp. 417–419, Jun. 1996.
[19] H. Tomita, S. Fujimoto, K. Tanimoto, and Y. Ike, “Cloning and genetic organization of the bacteriocin 31 determinant encoded on the Enterococcus faecalis pheromone-responsive conjugative plasmid pYI17,” Journal of Bacteriology, vol. 178, no. 12, pp. 3585–3593, 1996.
[20] P. Casaus, T. Nilsen, L. M. Cintas, I. F. Nes, P. E. Hernández, and H. Holo, “Enterocin B, a new bacteriocin from Enterococcus faecium T136 which can act synergistically with enterocin A,” Microbiology, vol. 143, no. 7, pp. 2287–2294, Jul. 1997.
[21] R. D. Campo, C. Tenorio, R. Jiménez-Dı́az, C. Rubio, R. Gómez-Lus, F. Baquero, and C. Torres, “Bacteriocin production in vancomycin-resistant and vancomycin-susceptible Enterococcus isolates of different origins,” Antimicrobial Agents and Chemotherapy, vol. 45, no. 3, pp. 905–912, 2001.
[22] T. Eguchi, K. Kaminaka, J. Shima, S. Kawamoto, K. Mori, S.-H. Choi, K. Doi, S. Ohmomo, and S. Ogata, “Isolation and characterization of enterocin SE-K4 produced by thermophilic enterococci, Enterococcus faecalis K-4,” Bioscience, Biotechnology and Biochemistry, vol. 65, no. 2, pp. 247–253, 2001.
[23] S. Kawamoto, J. Shima, R. Sato, T. Eguchi, S. Ohmomo, J. Shibato, N. Horikoshi, K. Takeshita, and T. Sameshima, “Biochemical and genetic characterization of mundticin KS, an antilisterial peptide produced by Enterococcus mundtii NFRI 7393,” Applied and Environmental Microbiology, vol. 68, no. 8, pp. 3830–3840, 2002.
[24] M. D. Kwaadsteniet, T. Fraser, C. V. Reenen, and L. Dicks, “Bacteriocin T8, a novel class IIa secdependent bacteriocin produced by Enterococcus faecium T8, isolated from vaginal secretions of children infected with human immunodeficiency virus,” Applied and Environmental Microbiology, vol. 72, no. 7, pp. 4761–4766, 2006.
[25] J. Sanchez, D. B. Diep, C. Herranz, I. F. Nes, L. M. Cintas, and P. E. Hernández, “Amino acid and nucleotide sequence, adjacent genes, and heterologous expression of hiracin JM79, a secdependent bacteriocin produced by Enterococcus hirae DCH5, isolated from Mallard ducks (Anas platyrhynchos),” FEMS Microbiology Letters, vol. 270, no. 2, pp. 227–236, May 2007.
[26] A. Gupta and S. K. Tiwari, “Probiotic potential of bacteriocin-producing Enterococcus hirae strain LD3 isolated from dosa batter,” Annals of Microbiology, vol. 65, no. 4, pp. 2333–2342, Dec. 2015.
[27] G. R. Siragusa, “Production of bacteriocin inhibitory to Listeria species by Enterococcus hirae,” Applied and Environmental Microbiology, vol. 58, no. 11, pp. 3508–3513, 1992.
[28] F. Achemchem, R. Cebrián, J. Abrini, M. Martínez- Bueno, E. Valdivia, and M. Maqueda, “Antimicrobial characterization and safety aspects of the bacteriocinogenic Enterococcus hirae F420 isolated from Moroccan raw goat milk,” Canadian Journal of Microbiology, vol. 58, no. 5, pp. 596– 604, Apr. 2012.
[29] A. Gálvez, E. Valdivia, H. Abriouel, E. Camafeita, E. Mendez, M. Martínez-Bueno, and M. Maqueda, “Isolation and characterization of enterocin EJ97, a bacteriocin produced by Enterococcus faecalis EJ97,” Archives of Microbiology, vol. 171, no. 1, pp. 59–65, Dec. 1998.
[30] E. Balla, L. Dicks, V. D. M. D. Toit, M. V. D. Merwe, and W. Holzapfel, “Characterization and cloning of the genes encoding enterocin 1071A and enterocin 1071B, two antimicrobial peptides produced by Enterococcus faecalis BFE 1071,” Applied and Environmental Microbiology, vol. 66, no. 4, pp. 1298–1304, 2000.
[31] T. Inoue, H. Tomita, and Y. Ike, “Bac 32, a novel bacteriocin widely disseminated among clinical isolates of Enterococcus faecium,” Antimicrobial Agents and Chemotherapy,” vol. 50, no. 4, pp. 1202–1212, 2006.
[32] R. Aunpad and K. Na-Bangchang, “Pumilicin 4, a novel bacteriocin with anti-MRSA and anti- VRE activity produced by newly isolated bacteria Bacillus pumilus strain WAPB4,” Current Microbiology, vol. 55, no. 4, pp. 308–313, 2007.
[33] K. Saelim, S. Kaewsuwan, A. Tani, and S. Maneerat, “Physical, biochemical and genetic characterization of enterocin CE5-1 produced by Enterococcus faecium CE5-1 isolated from Thai indigenous chicken intestinal tract,” Songklanakarin Journal of Science & Technology, vol. 37, no. 3, pp. 299–307, 2015.
[34] M. Galvin, C. Hill, and R. P. Ross, “Lacticin 3147 displays activity in buffer against Gram‐positive bacterial pathogens which appear insensitive in standard plate assays,” Letters in Applied Microbiology, vol. 28, no. 5, pp. 355–358, 1999.
[35] D. Shokri, S. Zaghian, F. Khodabakhsh, H. Fazeli, S. Mobasherizadeh, and B. Ataei, “Antimicrobial activity of a UV-stable bacteriocin-like inhibitory substance (BLIS) produced by Enterococcus faecium strain DSH20 against vancomycinresistant Enterococcus (VRE) strains,” Journal of Microbiology, Immunology and Infection, vol. 47, no. 5, pp. 371–376, Jan. 2014.
[36] H. Khan, S. Flint, and P. L. Yu, “Development of a chemically defined medium for the production of enterolysin A from Enterococcus faecalis B 9510,” Journal of applied microbiology, vol. 114, no. 4, pp. 1092–1102, Dec. 2013.
[37] K. Venema, M. L. Chikindas, J. Seegers, A. J. Haandrikman, K. J. Leenhouts, G. Venema, and J. Kok, “Rapid and efficient purification method for small, hydrophobic, cationic bacteriocins: Purification of lactococcin B and pediocin PA-1,” Applied and Environmental Microbiology, vol. 63, no. 1, pp. 305–309, 1997.
[38] L. Burianek and A. Yousef, “Solvent extraction of bacteriocins from liquid cultures,” Letters in Applied Microbiology, vol. 31, no. 3, pp. 193–197, 2000.
[39] M. S. Kim, J. Zhong, and A. Pandey, “Common errors in mass spectrometry‐based analysis of post‐translational modifications,” Proteomics, vol. 16, no. 5, pp. 700–714, Dec. 2016.
[40] C. M. Franz, M. J. V. Belkum, W. H. Holzapfel, H. Abriouel, and A. Gálvez, “Diversity of enterococcal bacteriocins and their grouping in a new classification scheme,” FEMS Microbiology Reviews, vol. 31, no. 3, pp. 293–310, Apr. 2007.
[41] A. Gupta, S. K. Tiwari, V. Netrebov, and M. L. Chikindas, “Biochemical properties and mechanism of action of enterocin LD3 purified from Enterococcus hirae LD3,” Probiotics and Antimicrobial Proteins, vol. 8, no. 3, pp. 161–169, Sep. 2016.
[42] C. Gabrielsen, D. A. Brede, I. F. Nes, and D. B. Diep, “Circular bacteriocins: Biosynthesis and mode of action,” Applied and Environmental Microbiology, vol. 80, no. 22, pp. 6854–6862, 2014.
[43] S. Iwatani, T. Zendo, F. Yoneyama, J. Nakayama, and K. Sonomoto, “Characterization and structure analysis of a novel bacteriocin, lacticin Z, produced by Lactococcus lactis QU14,” Bioscience, Biotechnology, and Biochemistry, vol. 71, no. 8, pp. 1984–1992, 2007.
[44] S. El-Ghaish, A. El-Baz, N. Hwanhlem, M. Zommara, E. Ayad, Y. Choiset, T. Haertlé, and J.-M. Chobert, “Bacteriocin production and safety evaluation of non-starter Enterococcus faecium IM1 and Enterococcus hirae IM1 strains isolated from homemade Egyptian dairy products,” European Food Research and Technology, vol. 240, no. 6, pp. 1211–1223, Feb. 2015.
[45] Y. Gao, B. Li, D. Li, and L. Zhang, “Purification and characteristics of a novel bacteriocin produced by Enterococcus faecalis L11 isolated from Chinese traditional fermented cucumber,” Biotechnology Letters, vol. 38, no. 5, pp. 871– 876, May 2016.
[46] S. Maisnier-Patin, E. Forni, and J. Richard, “Purification, partial characterisation and mode of action of enterococcin EFS2, an antilisterial bacteriocin produced by a strain of Enterococcus faecalis isolated from a cheese,” International Journal of Food Microbiology, vol. 30, no. 3, pp. 255–270, Jul. 1996.
[47] G. Liu, L. Ren, Z. Song, C. Wang, and B. Sun, “Purification and characteristics of bifidocin A, a novel bacteriocin produced by Bifidobacterium animals BB04 from centenarians’ intestine,” Food Control, vol. 50, pp. 889–895, Apr. 2015.
DOI: 10.14416/j.asep.2020.04.004
Refbacks
- There are currently no refbacks.
.png){kind=logo}
