PARTIAL PURIFICATION AND CHARACTERIZATION OF BACTERIOCIN-LIKE PEPTIDE PRODUCED BY STAPHYLOCOCCUS XYLOSUS

Veslava Matikeviciene, Saulius Grikiškis, Erika Lubyte, Gervydas Dienys

Abstract


The extensive use of antibiotics leads to an increasing number of antibiotic-resistant pathogenic microorganisms. The development of new antimicrobials is needed for clinical, veterinary, and food applications. Bacteriocins are small peptides with antimicrobial activity ribosomally synthesized by bacteria and could be applied as an alternative to classical antibiotics. In this study, the bacteriocin-like (BLIS) peptide, produced by Staphylococcus xylosus was partially purified and main characteristics (pH, thermal stability, resistance to some protease enzymes and molecular weight) were evaluated. Ammonium sulfate precipitation, acetone extraction and ion-exchange chromatography methods were applied for purification of bacteriocin. The activity of bacteriocin was detected using a well diffusion assay method and the amount of protein concentration was estimated by Lowry method. Molecular weight (~ 6 kDa) of purified bacteriocin was determined by sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS PAGE) method. The highest purification yield (80 %) was obtained using ion-exange chromatography and SP-sepharose as sorbent. The purified bacteriocin remained stable at pH values between 2.0 and 12.0 for 4 h. No decrease in antibacterial activity was estimated after 30 min at 121º C temperature. The purified bacteriocin was resistant to papain, pepsin and trypsin action. The BLIS inhibits a growth of Listeria monocytogenes (93 ± 3.0 %), Bacillus subtilis (85 ± 4.0 %), Pediococcus pentosaceus (79 ± 4.0 %), Staphylococcus aureus (51 ± 5.0 %) and Propionibacterium acnes (70 ± 5 %) up to 24 hours. Such bacteriocin preparation could be applied as antimicrobial agent in medical and food industry.

Keywords


antimicrobial agent; bacteriocin, purification; Staphylococcus xylosus

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References


J. L. Parada, C. R. Caron, A. P. Medeiros and C. R. Soccol, “Bacteriocins from Lactic Acid Bacteria: Purification, Properties and use as Biopreservatives”, Brazilian Archives of Biology and Technology, vol.50, n. 3, pp.521-542, May 2007.

Alvarez-Sieiro, P., Montalbán-López, M., Mu, D., & Kuipers, O. P. (2016). Bacteriocins of lactic acid bacteria: extending the family. Appl. Microbiol. Biotechnol., 100 (7), pp. 2939-2951.

Martínez, B., Rodríguez, A., & Suárez, E. (2016). Antimicrobial Peptides Produced by Bacteria: The Bacteriocins. In New Weapons to Control Bacterial Growth (pp. 15-38). Springer International Publishing.

H. Holo, T. Fayec, D. Bredea, T. Nilsena, I. Ødegårdb, Th. Langsrudc, J. Brendehaugb, I. F. Nes, Bacteriocins of propionic acid bacteria. Lait 82 (2002) 59-68, Oral communication at the 3rd International Symposium on Propionibacteria, Zurich, Switzerland, July 8–11, 2001.

S. Elayaraja, N. Annamalai, P. Mayavu, T. Balasubramanian, “Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum”, Asian Pac J Trop Biomed, vol. 4(Suppl 1), pp. S305-S311, 2014.

V. Kaškonienė, M. Stankevičius, K.Bimbiraitė-Survilienė, G. Naujokaitytė, L. Šernienė, K. Mulkytė, M. Malakauskas, A. Maruška, “Current state of purification, isolation and analysis of bacteriocins produced by lactic acid bacteria”, Appl Microbiol Biotechnol, vol. 101, pp. 1323–1335, 2017.

D. Cizeikiene, G. Juodeikiene, A. Paskevicius, E, Bartkiene, “Antimicrobial activity of lactic acid bacteria against pathogenic and spoilage microorganism isolated from food and their control in wheat bread”, Food Control, vol. 31(2), pp. 539-545, 2013.

J. H. Waterborg, The Lowry Method for Protein Quantitation. The Protein Protocols Handbook, 2nd Edition. Humana Press, 2002, pp. 7-9.

Guyonnet, D.; Fremaux, C.; Cenatiempo, Y. and Berjeaud, J. M., “Method for rapid purification of class IIa bacteriocins and comparison of their activities. Appl. Environ. Microbiol., vol. 66, pp. 1744-1748, 2000.

J. W. Hastings, M. Sailer, K. Johnson, K. L. Roy, J. C. Vederas, and M. E. Stiles, “Characterization of leucocin A-UAL 187 and cloning of the bacteriocin gene from Leuconostoc gelidum”, J. Bacteriol., 173, pp. 7491-7500, 1991.

S. D. Torodov, L. M. T. Dicks, “Lactobacillus plantarum isolated from molasses produces bacteriocins active against Gram-negative bacteria”. Enzyme and Microbial. Technol., vol. 36, pp. 318-326, 2005.

M. Papagianni, E. M. Papamichael, “Purification, amino acid sequence and characterization of the class IIa bacteriocin weissellin A, produced by Weissella paramesenteroides DX”, Bioresource Technology, vol. 102, pp. 6730–6734, 2011.

H. F. Goh, K.Philip, “Purification and Characterization of Bacteriocin Produced by Weissella confusa A3 of Dairy Origin”, PLoS ONE 10(10): e0140434. doi:10.1371/journal.pone.0140434




DOI: http://dx.doi.org/10.17770/etr2017vol3.2586

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