Аннотация

In recent years, we have witnessed a considerable growth in number of strains resistant to antibiotics. Therefore, research on new antimicrobial components that might be used for development of new-generation drugs is currently very important. We have studied antibiotic activity of Bacillus safensis , Bacillus endopheticus, Bacillus subtilis strains, isolated their bacteriocins, and evaluated their properties. The study was carried out in the scientific research institute for biotechnology, Kemerovo Institute of Food Science and Technology, in the city of Kemerovo. Strains of microorganisms were isolated from vegetables grown in Krasnodar region, namely, samples of Manas onions, Big Beef tomatoes, and Capia bell peppers. Antibiotic activity of the strains was evaluated in liquid nutrient medium. All test strains demonstrated some level of antimicrobial activity which varied from 18 to 91%. We established minimum inhibitory concentrations for the isolated strains based on measured optical density; MIC for Bacillus safensis was 1.5*106 CFU/сm3, for Bacillus endopheticus , 1.5*106 CFU/сm3, for Bacillus subtilis , 1.5*108 CFU/сm3. We then isolated respective bacteriocins and purified them by HPLC method. During disk diffusion tests, bacteriocin preparations proved active against Micrococcus luteus strain. Molecular weight was determined by PAGE electrophoresis. Molecular weight of bacteriocins varied from 3.6 through 4.21 kDа. Isolated bacteriocins were proved to belong to the lantibiotics class.

Ключевые слова

antibacterial activity, Bacillus strains, bacteriocins, pathogenic strains

СПИСОК ЛИТЕРАТУРЫ

1. Klaenhammer T.R. Bacteriocins of lactic acid bacteria. Biochimie, 1988, vol. 70, iss. 3, pp. 337-349. DOI: 10.1016/ 0300-9084(88)90206-4.
2. Messaoudi S., Manai M., Kergourlay G., et al. Lactobacillus salivarius: bacteriocin and probiotic activity. Food Microbiology, 2013, vol. 32, iss. 2, pp. 296-304. DOI: http://dx.doi.org/10.1016/j.fm.2013.05.010.
3. Maldonado-Barragán A., Caballero-Guerrero B., Lucena-Padrós H., Ruiz-Barba J.L. Induction of bacteriocin production by coculture is widespread among plantaricin-producing Lactobacillus plantarum strains with different regulatory operons. Food Microbiology, 2013, vol. 33, iss. 1, pp. 40-47. DOI: http://dx.doi.org/ 10.1016/ j.fm.2012.08.009.
4. Khalisanni K. An overview of lactic acid bacteria. International Journal of Biosciences, 2011, vol. 1, no. 3,pp. 1-13.
5. VelhoL R.V., Medina F.C., Segalin J., and Brandelli A. Production of lipopeptides among Bacillus strains showing growth inhibition of phytopathogenic fungi. Folia Microbiologica, 2011, vol. 56, pp. 297-303. DOI: 10.1007/ s12223-011-0056-7.
6. Katz E. and Demain A.L. The peptide antibiotics of Bacillus: chemistry, biogenesis, and possible role. Bacteriological Reviews, 1977, vol. 41, pp. 449-474.
7. Shoji J. Recent chemical studies on peptide antibiotics from the genus Bacillus. Advances in Applied Microbiology, 1978, vol. 24, pp. 187-214.
8. Smirnov V.V., Reznik S.R. and Vasilievskaya I.A. Aerobe Endospore-forming Bacteria. Budapest: Medicina KoÈnyvkiadoÂ, 1986. (In Hungarian).
9. Stein T., Heinzmann S., Dusterhus S., and Borchert S. Expression and functional analysis of the subtilin immunity genes spaIFEG in the subtilin-sensitive host Bacillus subtilis MO1099. Journal of Bacteriology, 2005, vol. 187, no. 3, pp. 822-828. DOI: 10.1128/JB.187.3.822-828.2005.
10. LoefflerW., Tschen J., Nongnuch M., et al. Effects of bacilysin and fengymycin from Bacillus suhtilis F-29-3 a comparison with activities of other bacillus antihiotics. Journal of Phytopathology, 1986, vol. 115, iss. 3, pp. 204-213. DOI: 10.1111/j.1439-0434.1986.tb00878.x.
11. Tsuge K., Ano T., and Shoda M. Isolation of a gene essential for biosynthesis of the lipopeptide antibiotics plipastatin B1 and surfactin in Bacillus subtilis YB8. Archives of Microbiology, 1996, vol. 165, no. 4, pp. 243-251. DOI: 10.1007/s002030050322.
12. Hyronimus B., Marrec C., and Urdaci C. Coagulin, a bacteriocin-like inhibitory substance produced by Bacillus coagulans I4. Journal of Applied Microbiology, 1998, vol. 85, iss. 1, pp. 42-50. DOI: 10.1046/j.1365-2672.1998.00466.x.
13. Paik H.D., Bae S.S., Park S.H., and Pan J.G. Identification and partial characterization of tochicin, a bacteriocin produced by Bacillus thuringiensis subsp tochigiensis. Journal of Industrial Microbiology & Biotechnology, 199, vol. 19, iss. 4, pp. 294-298. DOI: 10.1038/sj.jim.2900462.
14. Simha B.V., Sood S.K., Kumariya R., and Garsa A.K. Simple and rapid purification of pediocin PA-1 from Pediococcus pentosaceous NCDC 273 suitable for industrial application. Microbiological Research, 2012, vol. 167, iss. 9, pp. 544-549. DOI: http://dx.doi.org/10.1016/j.micres.2012.01.001.
15. Halimi B., Dortu C., Arguelles-Arias A., et al. Antilisterial Activity on Poultry Meat of Amylolysin, a Bacteriocin from Bacillus amyloliquefaciens GA1. Probiotics and Antimicrobial Proteins, 2010, vol. 2, iss. 2, pp. 120-125. DOI: 10.1007/s12602-010-9040-9.
16. Abrioue H., Franz C., Galvez A., et al. Diversity and applications of Bacillus bacteriocins. FEMS Microbiology Reviews, 2011, vol. 35, iss. 1, pp. 201-232.
17. Alisky J., Iczkowski K., Rapoport A., and Troitsky N. Bacteriophages show promise as antimicrobial agents. Journal of Infection, 1998, vol. 36, iss. 1, pp. 5-15. DOI: http://dx.doi.org/10.1016/S0163-4453(98)92874-2
18. Macfarlane G.T. and Cummings J.H. Probiotics, infection and immunity. Current Opinion in Infectious Diseases, 2002, vol. 15, no. 5, pp. 501-506.
19. Joerger R.D. Alternatives to antibiotics: bacteriocins, antimicrobial peptides and bacteriophages. Poultry Science, 2003, vol. 82, pp. 640-647.
20. Twomey D., Ross R.P., Ryan M., et al. Lantibiotics produced by lactic acid bacteria: structure, function and applications. Antonie Van Leeuwenhoek, 2002, vol. 82, iss. 1, pp. 165-185. DOI: 10.1023/A:1020660321724.
21. Foèldes T., Baânhegyi I., Herpai Z., et al. Isolation of Bacillus strains from the rhizosphere of cereals and in vitro screening for antagonism against phytopathogenic, food-borne pathogenic and spoilage micro-organisms. Journal of Applied Microbiology, 2000, vol. 89, iss. 5, pp. 840-846. DOI: 10.1046/j.1365-2672.2000.01184.x.
22. Kayalvizhi N. and Gunasekaran P. purification and characterization of a novel broad-spectrum bacteriocin from Bacillus licheniformis MKU3. Biotechnology and Bioprocess Engineering, 2010, vol. 15, no. 2, pp. 365-370.
23. Hernández D., Cardell E., and Zárate V. Antimicrobial activity of lactic acid bacteria isolated from Tenerife cheese: initial characterization of plantaricin TF711, a bacteriocin-like substance produced by Lactobacillus plantarum TF711. Journal of Applied Microbiology, vol. 99, iss. 1, pp. 77-84. DOI: 10.1111/j.1365-2672.2005.02576.x.
24. Motta A.S. and Brandelli A. Characterization of an antimicrobial peptide produced by Brevibacterium linens. Journal of Applied Microbiology, 2002, vol. 92, iss. 1, pp. 63-70. DOI: 10.1046/j.1365-2672.2002.01490.x.
25. De Oliveira S.S., Abrantes J., Cardoso M., Sordelli D., and Bastos M.C.F. Staphylococcal strains involved in bovine mastitis are inhibited by Staphylococcus aureus antimicrobial peptides. Letters in Applied Microbiology, 1998, vol. 27, iss. 5, pp. 287-291. DOI: 10.1046/j.1472-765X.1998.00431.x.
26. Kurbanov M.G. and Razumnikova J.S. Protein hydrolysates with biologically active peptides. Dairy Industry, 2010, vol. 9, pp.13-17.
27. Prosekov A.Yu., Dyshlyuk L.S., Milentieva I.S., et al. Study of biocompatibility and antitumor activity of lactic acid bacteria isolated from the human gastrointestinal tract. International Journal of Pharmacy & Technology, 2016, vol. 8, iss. 2, pp. 13647-13661.
28. Prosekov A., Milenteva I., Sukhikh S., et al. Identification of probiotic strains isolated from human gastrointestinal tract and investigation of their antagonistic, antioxidant and antiproliferative properties. Biology and Medicine, 2017, vol. 7, iss. 5, pp. 1-5.
29. Mayo B., Aleksandrzak-Piekarczyk T., Fernandez M., et al. Updates in the metabolism of lactic acid bacteria. In: Mozzi F., Raya R.R., and Vignolo G.M. (eds.). Biotechnology of lactic acid bacteria-novel applications. Iowa, USA: Wiley Blackwell Publ., 2010, pp. 3-34.
30. Oguntoyinbo F.A. and Narbad A. Molecular characterization of lactic acid bacteria and in situ amylase expression during traditional fermentation of cereal foods. Food Microbiology, 2012, vol. 31, iss. 2, pp. 254-262. DOI: http://dx.doi.org/ 10.1016/j.fm.2012.03.004.
31. Klaenhammer T.R. Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiology Reviews, 1993, vol. 12, iss. 1-2, pp. 39-85. DOI: 10.1111/j.1574-6976.1993.tb00012.x.