Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
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Effects of Sulforaphane, Grapefruit Seed Extracts, and Reuterin on Virulence Gene Expression Using hilA and invF Fusion Strains of Salmonella typhimurium

  • Kim, Ji-Yeun (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University) ;
  • Ryu, Sang-Ryul (Department of Food Science and Technology, Seoul National University) ;
  • Ji, Geun-Eog (Department of Food and Nutrition, Research Institute of Human Ecology, Seoul National University)
  • Published : 2007.10.31
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Abstract

This study assessed the effects of the antimicrobial substances sulforaphane, grapefruit seed extracts (GSE), and reuterin on the expression of Salmonella HilA and InvF virulence gene using a LacZY assay (${\beta}$-galactosidase assay) with hilA:lacZY and invF:lacZY fusion strains of Salmonella typhimurium SL1344. Salmonella was grown for 8 hr at $37^{\circ}C$ in the presence of diluted antimicrobial substances ($2\;{\mu}g/mL$ sulforaphane, $20\{\mu}g/mL$ GSE, and 0.26 mM reuterin) at concentrations that did not inhibit the cellular growth of Salmonella. Sulforaphane inhibited the expression of HilA and InvF by 50-90 and 20-80%, respectively. GSE also inhibited the expression of both genes, but to a lesser degree. Among the 3 antimicrobial substances, reuterin showed the least inhibition, which was abolished after 3-4 hr. None of the antimicrobial substances inhibited the ${\beta}$-galactosidase enzyme activity of S. typhimurium. The assay used in this study represents a very sensitive method for screening bioactive substances that inhibit the expression of virulence genes in Salmonella.

Keywords

References

  1. Bajaj V, Hwang C, Lee CA. HilA is a novel ompR/toxR family member that activates the expression of Salmonella typhimurium invasion genes. Mol. Microbiol. 18: 715-727 (1995) https://doi.org/10.1111/j.1365-2958.1995.mmi_18040715.x
  2. Kaniga K, Bossi JC, Galan JE. The Salmonella typhimurium invasion genes invF and invG encode homologues of the AraC and PulD family of proteins. Mol. Micobiol. 13: 555-568 (1994) https://doi.org/10.1111/j.1365-2958.1994.tb00450.x
  3. Jones BD, Ghori N, Falkow S. Salmonella typhimurium initiates murine infection by penetrating and destroying the specialized epithelial M cells of the Peyer's patches. J. Exp. Med. 180: 15-23 (1994) https://doi.org/10.1084/jem.180.1.15
  4. Shin DH, An DS, Koh JS, Lee DS. Wax-coating combined with grapefruit seed extract to reduce physiological quality changes and decay of Satsuma mandarin. Food Sci. Biotechnol. 7: 214-220 (1998)
  5. Park WP, Park KD, Cho SH. Effect of grapefruit seed extract on kimchi fermentation. Food Sci. Biotechnol. 5: 91-93 (1996)
  6. Cvetnic Z, Vladimir-Knezevic S. Antimicrobial activity of grapefruit seed and pulp ethanolic extract. Acta Pharm. 54: 243-250 (2004)
  7. Ortunõ A, García-Puig D, Fuster MD, Pérez ML, Sabater F, Porras I, García-Lidón A, Del Río JA. Flavanone and nootkatone levels in different varieties of grapefruit and pummelo. J. Agr. Food Chem. 43: 1-5 (1995) https://doi.org/10.1021/jf00049a001
  8. Sakamoto S, Sato K, Maitani T, Yamada T. Analysis of components in natural food additive 'grapefruit seed extract' by HPLC and LC/ MS. Bull. Nat. Institute Hyg. Sci. 114: 38-42 (1996)
  9. Takeoka GR, Dao LT, Wong RY, Harden LA. Identification of benzalkonium chloride in commercial grapefruit seed extracts. J. Agr. Food Chem. 53: 7630-7636 (2005) https://doi.org/10.1021/jf0514064
  10. von Woedtke T, Schlüter B, Pflegel P, Lindequist U, Jülich WD. Aspects of the antimicrobial efficacy of grapefruit seed extract and its relation to preservative substances contained. Pharmazie 54: 452- 456 (1999)
  11. Kim JT, Jung HY, Lee NK, Rhim SL, Paik HD. Isolation, identification, and probiotic properties of Lactobacillus reuteri HY701 from human feces. Food Sci. Biotechnol. 15: 677-682 (2006)
  12. Vollenweider S, Lacroix C. 3-Hydroxy-propionaldehyde: Applications and perspectives of biotechnological production. Appl. Microbiol. Biotechnol. 64: 16-27 (2004) https://doi.org/10.1007/s00253-003-1497-y
  13. Dornberger K, Bockel V, Heyer J, Schonfeld C, Tonew M, Tonew E. Investigations of the isothiocyanates erysolin and sulforaphan of Cardaria draba L. Pharmazie 30: 792-796 (1975)
  14. Fahey JW, Zalcmann AT, Talalay P. The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. Phytochemistry 56: 5-51 (2001) https://doi.org/10.1016/S0031-9422(00)00316-2
  15. Nutt JD, Pillai SD, Woodward CL, Sternes KL, Zabala-Diaz IB, Kwon YM, Ricke SC. Use of a Salmonella typhimurium hilA fusion strain to assess effects of environmental fresh water sources on virulence gene expression. Water Res. 37: 3319-3326 (2003) https://doi.org/10.1016/S0043-1354(03)00244-6
  16. Dobrogosz WJ, Lindgren SE. Method of determining the presence of an antibiotic produced by Lactobacillus reuteri. U.S. patent 5, 352, 586 (1994)
  17. Luthi-Peng Q, Scharer S, Puhan Z. Production and stability of 3- hydroxypropionaldehyde in Lactobacillus reuteri. Appl. Microbiol. Biotechnol. 60: 73-80 (2002) https://doi.org/10.1007/s00253-002-1099-0
  18. Miller JH. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, New York, NY, USA (1972)
  19. EichelbergK, Galan JE. Differential regulation of Salmonella typhimurium type III secreted proteins by pathogenicity island 1 (SPI-1)-encoded transcriptional activators InvF and HilA. Infect. Immun. 67: 4099-4105 (1999)
  20. Zhang Y. Molecular mechanism of rapid cellular accumulation of anticarcinogenic isothiocyanates. Carcinogenesis 22: 425-431 (2001) https://doi.org/10.1093/carcin/22.3.425
  21. Stoner GD, Morrissey DT, Heur HY, Daniel M, Galati AJ, Wagner SA. Inhibitory effects of phenethyl isothiocyanate on N-nitro-sobenzylmethylamine carcinogenesis in the rat esophagus. Cancer Res. 51: 2063-2068 (1991)
  22. Heggers JP, Cottingham J, Gusman J, Reagor L, McCoy L, Carino E, Cox R, Zhao JG. The effectiveness of processed grapefruit-seed extract as an antibacterial agent: II. Mechanism of action and in vitro toxicity. J. Altern. Complem. Med. 8: 333-340 (2002) https://doi.org/10.1089/10755530260128023
  23. Brzozowski T, Konturek PC, Drozdowicz D, Konturek SJ, Zayachivska O, Pajdo R, Kwiecien S, Pawlik WW, Hahn EG. Grapefruit-seed extract attenuates ethanol- and stress-induced gastric lesions via activation of prostaglandin, nitric oxide, and sensory nerve pathways. World J. Gastroentero. 11: 6450-6458 (2005) https://doi.org/10.3748/wjg.v11.i41.6450