Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Salmonella typhimurium LPS Confers Its Resistance to Antibacterial Agents of Baicalin of Scutellaria baicalensis George and Novobiocin: Complementation of the rfaE Gene Required for ADP-L-glycero-D-manno-heptose Biosynthesis of Lipopolysaccharide

  • Chung, Tae-Wook (National Research Laboratory for Glycobiology, Ministry of Science and Technology of Korean Government and Department of Biochemistry and Molecular Biology, Dongguk University COM) ;
  • Jin, Un-Ho (National Research Laboratory for Glycobiology, Ministry of Science and Technology of Korean Government and Department of Biochemistry and Molecular Biology, Dongguk University COM) ;
  • Kim, Cheorl-Ho (National Research Laboratory for Glycobiology, Ministry of Science and Technology of Korean Government and Department of Biochemistry and Molecular Biology, Dongguk University COM)
  • Published : 2003.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

The antibacterial mechanism of enterobacter Salmonella typhimurium was studied. The rfa (Waa) gene cluster of S. typhimurium encodes the core oligosaccharide biosynthesis of lipopolysaccharide (LPS). Among the rfa gene cluster, we recently cloned the rfaE gene, which is involved in ADP-L-glycero-D-manno-heptose biosynthesis. The rfaE mutant synthesizes heptose-deficient LPS, which consists of only lipid A and 3-deoxy-D-manno-octulosonic acid (KDO), thus making an incomplete LPS and a rough phenotype mutant. S. typhimurium deep-rough mutants with the heptose region of the inner core show a reduced growth rate, sensitivity to high temperature, and hypersensitivity to hydrophobic antibiotics such as baicalin isolated from the medicinal herb of Scutellaria baicalensis Georgi. Thus, in this study, the cloned rfaE gene was added to the S. typhimurium rfaE mutant strain SL1102 (rfaE543), which makes heptose-deficient LPS and has a deep-rough phenotype. The complementation created a smooth phenotype in the SL1102 strain. The sensitivity of SL1102 to bacteriophages was also recovered to that of wild-type strain, indicating that LPS is used as the receptor for bacteriophage infection. The permeability barrier of SL1102 to hydrophobic antibiotics such as novobiocin and baicalin was restored to that of the wild-type, suggesting that antibiotic resistance of the wild-type strain is highly correlated with their LPS. Through an agar diffusion assay, the growth-inhibition activity of baicalin was fully observed in the mutant SL1102 strain. However, only a half of the inhibitory activity was detected in the rfaE complemented SL1102 strain. Furthermore, the LPS produced by the rfaE-complemented SL1102 strain was indistinguishable from LPS biosynthesis of smooth strains.

Keywords

References

  1. Current Protocols in Molecular Biology Ausubel,F.M.;R.Brent;R.E.Kingston;D.D.Moore;J.G.Seidman;J.A.Smith;K.Struhl
  2. J. Infect. Dis. v.165 Inhibition of human T cell leukemia virus by the plant flavonoid baicalin (7-glucuronic acid, 5,6-dihydroxyflavone) Baylor,N.W.;T.Fu;Y.D.Yan;F.W.Ruscetti https://doi.org/10.1093/infdis/165.3.433
  3. Can. J. Microbiol. v.22 Leakage of periplasmic enzymes from lipopolysacharide-defectiv mutants of Salmonella typhimurium Chatterjee,A.K.;K.E.Sanderson;H.Ross;S.Schlecht;O.Luderitz https://doi.org/10.1139/m76-226
  4. Biochim. Biophys. Acta v.1571 Flavones from Scutellaria baicalensis Georgi attenuate apoptosis and protein oxidation in neuronal cell lines Choi,J.;C.C.Conrad;C.A.Malakowsky;J.M.Talent;C.S.Yuan;R.W.Gracy https://doi.org/10.1016/S0304-4165(02)00217-9
  5. J. Biol. Chem. v.258 The rfaD gene codes for ADP-L- glycero-D-mannoheptose-6-epimerase. An enzyme required for lipopolysaccharide core biosynthesis Coleman,W.G.
  6. Biochem. Biophys. Res. Commun. v.266 Lipoxygenase inhibition induced apoptosis, morphological changes, and carbonic anhydrase expression in human pancreatic cancer cells Ding,X.Z.;C.A.Charles;T.H.Kuszynski;T.H.El-Metwally;T.E.Adrian https://doi.org/10.1006/bbrc.1999.1824
  7. J. Clin. Microbiol. v.12 Single-disk diffusion testing (Kirby-Bauer) of susceptibility of Proteus mirabilis to chloramphenicol:Significance of the intermediate category Furtado,G.L.;A.A.Medeiros
  8. J. Mol. Biol. v.166 Studies on transformation of Escherichia coli with plasmids Hanahan,D. https://doi.org/10.1016/S0022-2836(83)80284-8
  9. Mol. Microbiol. v.30 Molecular basis for structural diversity in the core regions of the lipopolysaccharides of Escherichia coli and Salmonella enterica Heinrichs,D.E.;J.A.Yethon;C.Whitfield https://doi.org/10.1046/j.1365-2958.1998.01063.x
  10. J. Bacteriol. v.154 Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels Hitchcock,P.J.;T.M.Brown
  11. Glycoconjugate J. v.18 Molecular cloning and functional expression of the rfaE gene required for lipopolysaccharide biosynthesis in Salmonella typhimurium Jin,U.H.;T.W.Chung;Y.C.Lee;S.D.Ha;C.H.Kim https://doi.org/10.1023/A:1021103501626
  12. J. Biol. Chem. v.273 Enzymatic synthesis of lipopolysaccharide in Escherichia coli. Purification and properties of heptosyltransferase i Kadrmas,J.L.;C.R.H.Raetz https://doi.org/10.1074/jbc.273.5.2799
  13. Nature(London) v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli,U.K. https://doi.org/10.1038/227680a0
  14. Infect. Immun. v.63 Molecular cloning and characterization of the nontypeable Haemophilus influenzae 2019 rfaE gene required for lipopolysacharide biosynthesis Lee,N.G.;M.G.Sunshine;M.A.Apicella
  15. Bacterial Endotoxic Lipopolysaccharides v.1 Morrison,D.C.;J.L.Ryan
  16. J. Bacteriol. v.174 Role of the rfaG and rfaP genes in determining the lipopolysaccharide core structure and cell surface properties of Escherichia coli K-12 Parker,C.T.;A.W.Kloser;C.A.Schnaitman;M.A.Stein;S.Gottesman;B.W.Gibson
  17. Escherichia coli and salmonella. Cellular and Molecular Biology Bacterial lipopolysacharides:A remarkable family of bioactive macroamphiphiles Raetz,C.R.H;Neidhardt,F.C.(ed.);R.Curtiss(ed.);J.L.Ingraham(ed.);E..C.Lin(ed.);K.B.Low(ed.)
  18. Escherichia coli and salmonella typhimurium: Cellular and Molecular Biology Lipopolysaccharide biosynthesis Rick,P.D.;F.C.Neidhardt(ed.);J.L.Ingraham(ed.);K.B.Low(ed.);B.Magasanik(ed.);M.A.Schaechter(ed.);H.E.Umbarger(ed.)
  19. Molecular Cloning:A Laboratory Manual(Cold Spring Harbor Laboratory) Sambrook,J.;E.F.Fritsch;T.Maniatis
  20. Microbiol. Rev. v.57 Genetics of lipopolysaccharide biosynthesis in enteric bacteria Schnaitman,C.A.;J.D.Klena
  21. J. Microbiol. Biotechnol. v.11 Characterization of a cell line HFH-T2, proucing viral particles, from primary human fetal hepatocytes infected with hepatitis B virus Shim,J.K.;D.W.Kim;T.H.Chung;J.K.Kim;J.I.Suh;C.Park;Y.C.Lee;T.W.Chung;E.Y.Song;C.H.Kim
  22. J. Microbiol. Biotechnol. v.12 Intestinal colonization characteristics of lactobacillus spp. isolated from chicken cecum and competitive inhibition against Salmonella typhimurium Shin,J.W.;J.K.Kang;K.I.Jang;K.Y.Kim
  23. J. Biol. Chem. v.267 The rfaC gene of salmonella typhimurium. Cloning, sequencing, and enzymatic function in heptose transfer to lipopolysaccharide Sirisena,D.M.;K.A.Brozek;P.R.MacLachlan;K.E.Sanderson;C.R.H.Raetz
  24. Anal. Biochem. v.119 A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels Tsai,C.M.;C.E.Frasch https://doi.org/10.1016/0003-2697(82)90673-X
  25. J. Gen. Microbiol. v.70 Non-smooth mutants of Salomonella typhimurium: Differentiation by phage sensitivity and genetic mapping Wilkinson,R.G.;P.Gemski;B.A.D.Stocker https://doi.org/10.1099/00221287-70-3-527
  26. J. Microbiol. Biotechnol. v.11 Prediction of the secondary structure of the afgA subunit of Salomonella enteritidis overexpressed as an MBP-fused protein Won,M.S.;S.Y.Kim;S.H.Lee;C.J.Kim;H.S.Kim;M.H.Jun;K.B.Song