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Antimicrobial and Antioxidant Peptide from Bacillus Strain CBS73 Isolated from Korean Food

  • Received : 2017.07.11
  • Accepted : 2017.09.25
  • Published : 2017.09.30

Abstract

An antimicrobial peptides-producing Bacillus strain CBS73 was isolated from fermented food (kimchi) that produces low-molecular-weight proteins with broad-spectrum antimicrobial activity. Our goal was to explore the therapeutic potential of antimicrobial substances produced by Bacillus species. Peptide CBS73 was purified from Bacillus subtilis subsp. subtilis with identity of 99.79%. It was found to be stable at pH 4.0-10.0 and temp $20-60^{\circ}C$. A protein band around 5.2 kDa was detected in tricine-SDS-PAGE and band was confirmed by MALDI-TOF test. Peptide CBS73 showed antimicrobial activity against MDR bacteria. The minimal inhibitory concentration (MIC) of peptide CBS73 for vancomycin-resistant S. aureus (VRSA), vancomycin resistant Enterococci (VRE) and Salmonella typhimurium ranged from $10-40{\mu}g/mL$. The antioxidant activity of peptide CBS73 was measured by DPPH scavenging, reducing power activity and total phenolic content. Cell viability and NO production result showed less cytotoxic effect upto $12{\mu}g/mL$. Peptide CBS73 could be a promising antimicrobial agent for clinical application.

Keywords

References

  1. S. Wu, S. Jia, D. Sun, M. Chen, X. Chen, J. Zhong, and L. Huan, "Purification and characterization of two novel antimicrobial peptides subpeptin JM4-A and subpeptin JM4-B produced by Bacillus subtilis JM4", Curr. Microbiol., Vol. 51, pp. 292-296, 2005. https://doi.org/10.1007/s00284-005-0004-3
  2. H. Jenssen, P. Hamill, and R. E. Hancock, "Peptide antimicrobial agents", Clin. Microbiol. Rev., Vol. 19, pp. 491-511, 2006. https://doi.org/10.1128/CMR.00056-05
  3. T. R. Klaenhammer, "Genetics of bacteriocins produced by lactic acid bacteria", FEMS Microbiol. Rev., Vol. 12, pp. 39-85, 1993.
  4. L. Rodrigues, I. M. Banat, J. Teixeira, and R. Oliveira, "Biosurfactants: potential applications in medicine", J. Antimicrob. Chemother., Vol. 57, pp. 609-618, 2006. https://doi.org/10.1093/jac/dkl024
  5. S. Mukherjee, P. Das, and R. Sen, "Towards commercial production of microbial surfactants", TRENDS Biotechnol., Vol. 24, pp. 509-515, 2006. https://doi.org/10.1016/j.tibtech.2006.09.005
  6. N. A. Logan and P. D. Vos, "Bacillus", Bergey's Manual of Systematics of Archaea and Bacteria, pp. 1-163, 2015.
  7. D. M. Bollag, S. J. Edelstein, and M. D. Rozycki, "Proteins methods", No. 547.96 BOL., New York: Wiley, 1996.
  8. M. M. Bradford, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem., Vol. 72, pp. 248-254, 1976. https://doi.org/10.1016/0003-2697(76)90527-3
  9. I. Wiegand, K. Hilpert, and R. E. W. Hancock, "Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances", Nat. protoc., Vol. 3, pp. 163-175, 2008. https://doi.org/10.1038/nprot.2007.521
  10. S. Regmi, Y. H. Choi, Y. S. Choi, M. R. Kim, and J. C. Yoo, "Antimicrobial peptide isolated from Bacillus amyloliquefaciens K14 revitalizes its use in combinatorial drug therapy", Folia Microbiol., Vol. 62, pp. 127-138, 2017. https://doi.org/10.1007/s12223-016-0479-2
  11. H. Schagger, "Tricine-sds-page", Nat. protoc., Vol. 1, pp. 16-22, 2006. https://doi.org/10.1038/nprot.2006.4
  12. X. Zhang, J. Koo, and J.-B. Eun., "Antioxidant activities of methanol extracts and phenolic compounds in Asian pear at different stages of maturity", Food Sci. Biotechnol., Vol. 15, pp. 44-50, 2006.
  13. S. Regmi, S. Adhikari, and S. Panthi, "Antioxidant, antimicrobial and cytotoxic characteristics of a bioactive compound isolated from the aerial part of Dicksonia antarctica labb", World Journal of Pharmaceutical Research, Vol. 5, pp. 14-29, 2015.
  14. H. C. Wu, H.-M. Chen, and C.-Y. Shiau, "Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus)", Food Res. Int., Vol. 36, pp. 949-957, 2003. https://doi.org/10.1016/S0963-9969(03)00104-2
  15. V. Klompong, S. Benjakul, D. Kantachote, and F. Shahidi, "Antioxidative activity and functional properties of protein hydrolysate of yellow stripe trevally (Selaroides leptolepis) as influenced by the degree of hydrolysis and enzyme type", Food Chem., Vol. 102, pp. 1317-1327, 2007. https://doi.org/10.1016/j.foodchem.2006.07.016
  16. I. Gulcin, G. Sat, S. Beydemir, M. Elmastas, and O. I. Kufrevio?lu, "Comparison of antioxidant activity of clove (Eugenia caryophylata Thunb) buds and lavender (Lavandula stoechas L.)", Food Chem., Vol. 87, pp. 393-400, 2004. https://doi.org/10.1016/j.foodchem.2003.12.008
  17. M. S. Rahman, Y. H. Choi, Y. S. Choi, M. B. Alam, S. H. Lee, and J. C. Yoo, "A novel antioxidant peptide, purified from Bacillus amyloliquefaciens, showed strong antioxidant potential via Nrf-2 mediated heme oxygenase-1 expression", Food Chem., Vol. 239, pp. 502-510, 2018. https://doi.org/10.1016/j.foodchem.2017.06.106
  18. S. Regmi, G. C. Pradeep, Y. H. Choi, Y. S. Choi, J. E. Choi, S. S. Cho, and J. C. Yoo, "A multi-tolerant low molecular weight mannanase from Bacillus sp. CSB39 and its compatibility as an industrial biocatalyst", Enzyme Microb. Technol., Vol. 92, pp. 76-85, 2016. https://doi.org/10.1016/j.enzmictec.2016.06.018
  19. S. Regmi, Y. S. Choi, Y. H. Choi, Y. K. Kim, S. S. Cho, J. C. Yoo, and J.-W. Suh, "Antimicrobial peptide from Bacillus subtilis CSB138: characterization, killing kinetics, and synergistic potency", Int. Microbiol., Vol. 20, pp. 43-53, 2017.
  20. S. Regmi, Y. H. Yoo, Y. H. Choi, Y. S. Choi, J. C. Yoo, and S. W. Kim, "Prospects for bio-industrial application of an extremely alkaline mannanase from Bacillus subtilis subsp. inaquosorum CSB31", Biotechnol. J., Accessed 26 August, 2017, doi: 10.1002/biot.201700113.
  21. B. Alkotaini, N. Anuar, and A. A. H. Kadhum, "The effects of temperature, pH and carbon sources on antimicrobial peptide AN5-1 production using Paenibacillus alvei AN 5", Asian J. Microbiol. Biotechnol. Environ. Sci., Vol. 15, pp. 195-201, 2013.
  22. D. Bizani and A. Brandelli, "Characterization of a bacteriocin produced by a newly isolated Bacillus sp. Strain 8 A", J. Appl. Microbiol., Vol. 93, pp. 512-519, 2002. https://doi.org/10.1046/j.1365-2672.2002.01720.x
  23. C.-I. Cheigh, H.-J. Choi, H. Park, S.-B. Kim, M.-C. Kook, T.-S. Kim, J.-k. Hwang, and Y.-R. Pyun, "Influence of growth conditions on the production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi", Journal of Biotechnology, Vol. 95, pp. 225-235, 2002. https://doi.org/10.1016/S0168-1656(02)00010-X
  24. F. Kamoun, H. Mejdoub, H. Aouissaoui, J. Reinbolt, A. Hammami, and S. Jaoua, "Purification, amino acid sequence and characterization of Bacthuricin F4, a new bacteriocin produced by Bacillus thuringiensis", J. Appl. Microbiol., Vol. 98, pp. 881-888, 2005. https://doi.org/10.1111/j.1365-2672.2004.02513.x
  25. M. S. Shin, S. k. Han, J. S. Ryu, K. S. Kim, and W. K. Lee, "Isolation and partial characterization of a bacteriocin produced by Pediococcus pentosaceus K23-2 isolated from Kimchi", J. Appl. Microbiol., Vol. 105, pp. 331-339, 2008. https://doi.org/10.1111/j.1365-2672.2008.03770.x