Antimicrobial complex Enterococcus mundtii PPHS-5/13 of peptide nature
An irregular application of antibiotics in medicine, veterinary and agriculture is known to result in a wide spread of drug-resistant microorganisms worldwide. Today new natural antimicrobials that would differ from antibiotics are urgently needed to control such microorganisms. Probiotic bacteria, bacteriophages and bacteriocins are often considered as candidates to play the role. In this respect nonhemolytic Enterococcus species, such as E. faecium and E. mundtii, as well as their poorly investigated species producing Class IIa bacteriocins are especially attractive. The aim of our research was to determine the most significant factors that may influence the ability of our E. mundtii PPHS-5/13 strain to produce bacteriocidal substance during submerged cultivation, to elaborate a method of purification of the substance and to study its nature and the main properties. We showed that this strain can produce bactericidal substance in concentrations to 12.800 AU/ml on simple media and under conventional aerobic growth conditions. This allows microorganisms to be technologically applicable as a producer of a new antimicrobial remedy. Our research allowed us: to test a low-cost technology of isolation of the bacteriocidal complex from the culture fluid; to determine the peptide nature of the active agent; to determine what fraction inhibiting the growth of gram-positive bacteria; to evaluate tolerance of the peptide to pH, heat and proteolytic enzymes. The yield of the antimicrobial substance from the cell-free supernatant E. mundtii PPHS-5/13 strain with a phase separation method using dichloromethane 2-8 times higher than the salt precipitation method with ammonium sulfate or adsorption on solid media, while reducing the separation time from 24 to 3 hours. We found out that the antimicrobial complex E. mundtii PPHS-5/13 is of peptide nature, consists mainly of substances with molecular mass 5,762 kDa and its main properties, including and activity against Listeria spp., are similar to subclass IIA bacteriocins. By the example of broiler poultry skin we demonstrated the possibility of practical application of BLIS Enterococcus mundtii PPHS-5/13 as a means to control the level of microbial contamination of certain food stuff.
Keywords
antimicrobial complex,
BLIS,
bacteriocins,
Enterococcus,
антимикробный комплекс,
BLIS,
бактериоцины,
EnterococcusAuthors
| Khramov Vladimir M. | State Research Center for Applied Microbiology and Biotechnology (Obolensk, Moscow Region) | Khramov-vladimir@yandex.ru |
| Kalmantaev Timur A. | State Research Center for Applied Microbiology and Biotechnology (Obolensk, Moscow Region) | kalmantaev@yandex.ru |
| Sadikova Gulnur T. | State Research Center for Applied Microbiology and Biotechnology (Obolensk, Moscow Region) | Vpokhil003@yandex.ru |
| Perelygin Vladimir V. | State Research Center for Applied Microbiology and Biotechnology (Obolensk, Moscow Region) | perelygin@obolensk.org |
| Pokhilenko Viktor D. | State Research Center for Applied Microbiology and Biotechnology (Obolensk, Moscow Region) | Vpokhil003@yandex.ru |
Всего: 5
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