Antibacterial Activity against Salmonella enteritidis JK-15 and LPS Changes Caused by Rose Flower Extracts

장미꽃 추출물에 의한 식중독 세균 Salmonella enteritidis JK-15에 대한 살균활성 및 그에 따른 LPS 변화

  • Song, You-Jin (Department of Biotechnology, Soonchunhyang University) ;
  • Cho, Yun-Seok (Department of Biotechnology, Soonchunhyang University) ;
  • Oh, Kye-Heon (Department of Biotechnology, Soonchunhyang University)
  • 송유진 (순천향대학교 생명공학과) ;
  • 조윤석 (순천향대학교 생명공학과) ;
  • 오계헌 (순천향대학교 생명공학과)
  • Received : 2009.08.04
  • Accepted : 2009.10.18
  • Published : 2009.12.31

Abstract

The aim of this work was to investigate the antibacterial effect of the food-poisoning bacterium, Salmonella enteritidis JK-15 exposed to rose extracts. Initially, the isolate S. enteritidis JK-15 was enriched and isolated from stale food. BIOLOG and 16S rRNA analyses revealed that strain S. enteritidis JK-15 was 98% similar to the S. enteritidis species cluster; therefore we have designated this strain as S. enteritidis JK-15. Bactericidal effects of S. enteritidis JK-15 exposed to rose extracts ranging from 5 mg/ml to 100 mg/ml were monitored, and complete bactericidal effects were achieved within 6 h at 100 mg/ml and 12 h at 50 mg/ml, respectively. SDSPAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain S. enteritidis JK-15 treated to different concentrations and exposing periods of rose extracts in exponentially growing cultures. Scanning electron microscopic analysis, demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with rose extracts.

본 연구는 식중독 원인 세균인 Salmonella enteritidis JK-15를 장미추출물에 노출시켜 나타나는 살균효과를 조사하기 위하여 실시하였다. 분리세균인 S. enteritidis JK-15는 장시간 노출된 음식으로부터 농화, 분리되었다. 16S rRNA PCR을 수행하여 S. enteritidis JK-15의 유전학적 계통수를 작성하였다. BIOLOG 분석 시스템과 16S rRNA 염기서열 분석을 통하여 S. enteritidis 종(species)과 98% 유사성을 가졌으며, 이 균주를 S. enteritidis JK-15로 명명 하였다. 5~100 mg/ml 범위의 장미추출물에 노출된 S. enteritidis JK-15의 살균효과를 조사하였고, 100 mg/ml의 장미추출물에서 6시간, 그리고 50 mg/ml에서 12시간 이내에 완전히 살균되었다. 지수생장기의 S. enteritidis JK-15 균주는 노출된 장미추출물의 농도와 시간에 따라 lipopolysaccharide의 양이 변화하는 것을 SDS-PAGE와 silver staining을 통하여 관찰하였다. 또한, 주사전자현미경을 이용하여 장미추출물에 노출시킨 세균의 외부 형태를 분석한 결과, 움푹 패이고 불규칙적인 간균 형태로 관찰되었다.

Keywords

References

  1. Ahn, D.K., T.W. Han, H.Y. Shin, I.N. Jin, and S.Y. Ghim. 2003. Diversity and antibacterial activity of lactic acid bacteria isolated from kimchi. Kor. J. Microbiol. Biotechnol. 31, 191-196
  2. Bollag, D.M., M.D. Rozycki, and S.J. Edelstein. 1996. Protein methods. Wiley-Liss. 2, 108-128
  3. Cho, Y.S., N.L. Schiller, H.Y. Kahng, and K.H. Oh. 2007. Cellular responses and proteomic analysis of Escherichia coli exposed to green tea polyphenols. Curr. Microbiol. 53, 501-506 https://doi.org/10.1007/s00284-007-9021-8
  4. Doerrler, W.T. 2006. Lipid trafficking to the outer membrane of Gram-negative bacteria. Mol. Microbiol. 60, 542-552 https://doi.org/10.1111/j.1365-2958.2006.05130.x
  5. Dusch, H. and M. Altwegg. 1995. Evaluation of five new plating media for isolation of Salmonella species. J. Clin. Microbiol. 33, 802-804
  6. Fomsgaard, A., M.A. Freudenberg, and C. Galanos. 1990. Modification of the silver staining technique to detect lipopolysaccharide in polyacrylamide gels. J. Clin. Microbiol. 28, 2627-2631
  7. Hajime, I., T. Nakae, Y. Hara, and T. Shimamura. 1993. Bactericidal catechins damage the lipid bilayer. Biochim. Biophys. Acta. 1147, 132-136 https://doi.org/10.1016/0005-2736(93)90323-R
  8. Hitchcock, P.J. and T.M. Brown. 1983. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silverstained polyacrylamide gels. J. Bacteriol. 154, 269-277
  9. Jay, M.T., V. Garrett, J.C. Mohle-Boetani, M. Barros, J.A. Farrar, R. Rios, S. Abbott, R. Sowadsky, K. Komatsu, R. Mandrell, J. Sobel, and S.B. Werner. 2004. A multistate outbreak of Escherichia coli O157:H7 infection linked to consumption of beef tacos at a fast-food restaurant chain. Clin. Infec. Dis. 39, 1-7 https://doi.org/10.1086/421088
  10. Johnson, K.G. and M.B. Perry. 1976. Improved techniques for the preparation of bacterial lipopolysaccharides. Can. J. Microbiol. 22, 29-34 https://doi.org/10.1139/m76-004
  11. Jones, J.B., A.R. Chase, and G.K. Harris. 1993. Evaluation of the biolog GN microplate system for identification of some plantpathogenic bacteria. Plant Disease 77, 553-558 https://doi.org/10.1094/PD-77-0553
  12. Li, A., J. Chen, W. Zhu, T. Jiang, X. Zhang, and Q. Gu. 2007. Antibacterial activity of gallic acid from the flowers of Rosa chinensis jacq. against fish pathogens. Aquacul. Res. 38, 1110-1112 https://doi.org/10.1111/j.1365-2109.2007.01745.x
  13. Lim, Y.J., Y.S. Cho, and K.H. Oh. 2008. Antibacterial synergic effect and cellular responses of nalidixic acid-resistant Salmonella typhimurium exposed to tea polyphenols and nalidixic acid. Kor. J. Microbiol. 44, 122-129
  14. Lin, C.M., S.Y. Fernando, and C.I. Wei. 1996. Occurrence of Listeria monocytogenes, Salmonella spp., Escherichia coli and E. coli O157:H7 in vegetable salads. Food Control. 7, 135-140 https://doi.org/10.1016/0956-7135(96)00019-9
  15. Ng, T.B., W. Gao, L. Li, S.M. Niu, L. Zhao, J. Liu, L.S. Shi, M. Fu, and F. Liu. 2005. Rose (Rosa rugosa)-flower extract increases the activities of antioxidant enzymes and their gene expression and reduces lipid peroxidation. Biochem. Cell Biol. 83, 78-85 https://doi.org/10.1139/o04-100
  16. $\ddot{O}$zkan, G., O. Sagdi$\c{c}$, N.G. Baydar, and H. Baydar. 2004. Antioxidant and antibacterial activities of Rosa damascena flower extracts. Food Sci. Tech. Int. 10, 277-281 https://doi.org/10.1177/1082013204045882
  17. Pan, T.M., T.K. Wang, C.L. Lee, S.W. Chien, and C.B. Horng. 1997. Food-borne disease outbreaks due to bacteria in Taiwan, 1986 to 1995. J. Clin. Microbiol. 35, 1260-1262
  18. Park, S.H., K.H. Oh, and C.K. Kim. 2001. Adaptive and cross-protective responses of Pseudomonas sp. DJ-12 to several aromatics and other stress shocks. Curr. Microbiol. 43, 176-181 https://doi.org/10.1007/s002840010283
  19. Robers, D. 2003. Food poisoning / classification. Encycl. Food Sci. Nutri. 2654-2658
  20. Shimamura, T., W.H. Zhao, and Z.Q. Hu. 2007. Mechanism of action and potential for use of tea catechin as an anti-infective agent. Medi. Chem. 6, 57-62 https://doi.org/10.2174/187152107779314124
  21. Tsen, H.Y. and J.S. Lin. 2001. Analysis of Salmonella enteritidis strains isolated from food-poisoning cases in Taiwan by pulsed field gel electrophoresis, plasmid profile and phage typing. J. Appl. Microbiol. 91, 72-79 https://doi.org/10.1046/j.1365-2672.2001.01343.x
  22. Vinokur, Y., V. Rodov, N. Reznick, G. Goldman, B. Horev, N. Umiel, and H. Friedman. 2006. Rose petal tea as an antioxidantrich beverage: cultivar effects. J. Food Sci. 71, 42-47 https://doi.org/10.1111/j.1365-2621.2006.tb12404.x