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. 1985 Aug;50(2):298–303. doi: 10.1128/aem.50.2.298-303.1985

Destruction of the outer membrane permeability barrier of Escherichia coli by heat treatment.

T Tsuchido, N Katsui, A Takeuchi, M Takano, I Shibasaki
PMCID: PMC238619  PMID: 3901917

Abstract

Heat treatment of a wild-type Escherichia coli strain at 55 degrees C in 50 mM Tris-hydrochloride buffer with or without 10 mM magnesium sulfate or HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer at pH 8.0 caused an increase in cell surface hydrophobicity. By determining the location of n-hexadecane droplets attached to cells by phase-contrast microscopy, the septal and polar regions of heated cells appeared to become the most frequently hydrophobic. Some of the lipopolysaccharide molecules in the outer membrane were released from heated cells, and the cells became susceptible to the hydrolytic action of added phospholipase C. Heat-treated cells also became permeable to the hydrophobic dye crystal violet, which was added externally. The release of part of the outer membrane by heat treatment appeared to bring about the disorganization of the outer membrane structure and, as a consequence, to result in the partial disruption of the permeability barrier function of the outer membrane. Tris was found to enhance damage to the outer membrane by heat.

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Selected References

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  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Burdett I. D., Murray R. G. Septum formation in Escherichia coli: characterization of septal structure and the effects of antibiotics on cell division. J Bacteriol. 1974 Jul;119(1):303–324. doi: 10.1128/jb.119.1.303-324.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Coleman W. G., Jr, Leive L. Two mutations which affect the barrier function of the Escherichia coli K-12 outer membrane. J Bacteriol. 1979 Sep;139(3):899–910. doi: 10.1128/jb.139.3.899-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Costerton J. W., Ingram J. M., Cheng K. J. Structure and function of the cell envelope of gram-negative bacteria. Bacteriol Rev. 1974 Mar;38(1):87–110. doi: 10.1128/br.38.1.87-110.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fung J., MacAlister T. J., Rothfield L. I. Role of murein lipoprotein in morphogenesis of the bacterial division septum: phenotypic similarity of lkyD and lpo mutants. J Bacteriol. 1978 Mar;133(3):1467–1471. doi: 10.1128/jb.133.3.1467-1471.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grau F. H. Significance of the inactivation of transport in thermal death of Escherichia coli. Appl Environ Microbiol. 1978 Aug;36(2):230–236. doi: 10.1128/aem.36.2.230-236.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gustafsson P., Nordström K., Normark S. Outer penetration barrier of Escherichia coli K-12: kinetics of the uptake of gentian violet by wild type and envelope mutants. J Bacteriol. 1973 Nov;116(2):893–900. doi: 10.1128/jb.116.2.893-900.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hitchener B. J., Egan A. F. Outer-membrane damage in sublethally heated Escherichia coli K-12. Can J Microbiol. 1977 Mar;23(3):311–318. doi: 10.1139/m77-046. [DOI] [PubMed] [Google Scholar]
  9. Hurst A., Hughes A., Collins-Thompson D. L., Shah B. G. Relationship between loss of magnesium and loss of salt tolerance after sublethal heating of Staphylococcus aureus. Can J Microbiol. 1974 Aug;20(8):1153–1158. doi: 10.1139/m74-178. [DOI] [PubMed] [Google Scholar]
  10. Iandolo J. J., Ordal Z. J. Repair of thermal injury of Staphylococcus aureus. J Bacteriol. 1966 Jan;91(1):134–142. doi: 10.1128/jb.91.1.134-142.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Irvin R. T., MacAlister T. J., Costerton J. W. Tris(hydroxymethyl)aminomethane buffer modification of Escherichia coli outer membrane permeability. J Bacteriol. 1981 Mar;145(3):1397–1403. doi: 10.1128/jb.145.3.1397-1403.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kamio Y., Nikaido H. Outer membrane of Salmonella typhimurium: accessibility of phospholipid head groups to phospholipase c and cyanogen bromide activated dextran in the external medium. Biochemistry. 1976 Jun 15;15(12):2561–2570. doi: 10.1021/bi00657a012. [DOI] [PubMed] [Google Scholar]
  13. Karkhanis Y. D., Zeltner J. Y., Jackson J. J., Carlo D. J. A new and improved microassay to determine 2-keto-3-deoxyoctonate in lipopolysaccharide of Gram-negative bacteria. Anal Biochem. 1978 Apr;85(2):595–601. doi: 10.1016/0003-2697(78)90260-9. [DOI] [PubMed] [Google Scholar]
  14. Katsui N., Tsuchido T., Hiramatsu R., Fujikawa S., Takano M., Shibasaki I. Heat-induced blebbing and vesiculation of the outer membrane of Escherichia coli. J Bacteriol. 1982 Sep;151(3):1523–1531. doi: 10.1128/jb.151.3.1523-1531.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Katsui N., Tsuchido T., Takano M., Shibasaki I. Effect of preincubation temperature on the heat resistance of Escherichia coli having different fatty acid compositions. J Gen Microbiol. 1981 Feb;122(2):357–361. doi: 10.1099/00221287-122-2-357. [DOI] [PubMed] [Google Scholar]
  16. Leive L. The barrier function of the gram-negative envelope. Ann N Y Acad Sci. 1974 May 10;235(0):109–129. doi: 10.1111/j.1749-6632.1974.tb43261.x. [DOI] [PubMed] [Google Scholar]
  17. MALAMY M., HORECKER B. L. The localization of alkaline phosphatase in E. coli K12. Biochem Biophys Res Commun. 1961 Jun 2;5:104–108. doi: 10.1016/0006-291x(61)90020-1. [DOI] [PubMed] [Google Scholar]
  18. Mao J. C., Putterman M. Accumulation in gram-postive and gram-negative bacteria as a mechanism of resistance to erythromycin. J Bacteriol. 1968 Mar;95(3):1111–1117. doi: 10.1128/jb.95.3.1111-1117.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nikaido H., Nakae T. The outer membrane of Gram-negative bacteria. Adv Microb Physiol. 1979;20:163–250. doi: 10.1016/s0065-2911(08)60208-8. [DOI] [PubMed] [Google Scholar]
  20. Nikaido H. Outer membrane of Salmonella typhimurium. Transmembrane diffusion of some hydrophobic substances. Biochim Biophys Acta. 1976 Apr 16;433(1):118–132. doi: 10.1016/0005-2736(76)90182-6. [DOI] [PubMed] [Google Scholar]
  21. Russell A. D., Harries D. Some aspects of thermal injury in Escherichia coli. Appl Microbiol. 1967 Mar;15(2):407–410. doi: 10.1128/am.15.2.407-410.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. STRANGE R. E., SHON M. EFFECTS OF THERMAL STRESS ON VIABILITY AND RIBONUCLEIC ACID OF AEROBACTER AEROGENES IN AQUEOUS SUSPENSION. J Gen Microbiol. 1964 Jan;34:99–114. doi: 10.1099/00221287-34-1-99. [DOI] [PubMed] [Google Scholar]
  23. Schindler M., Osborn M. J. Interaction of divalent cations and polymyxin B with lipopolysaccharide. Biochemistry. 1979 Oct 2;18(20):4425–4430. doi: 10.1021/bi00587a024. [DOI] [PubMed] [Google Scholar]
  24. Sheu C. W., Freese E. Lipopolysaccharide layer protection of gram-negative bacteria against inhibition by long-chain fatty acids. J Bacteriol. 1973 Sep;115(3):869–875. doi: 10.1128/jb.115.3.869-875.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smit J., Kamio Y., Nikaido H. Outer membrane of Salmonella typhimurium: chemical analysis and freeze-fracture studies with lipopolysaccharide mutants. J Bacteriol. 1975 Nov;124(2):942–958. doi: 10.1128/jb.124.2.942-958.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stárková Z., Bonnaveiro N., Stárka J. Hydrolysis of phospholipids by phospholipase C in intact cells of wild-type and envelope mutants of Escherichia coli K12. FEBS Lett. 1981 Aug 3;130(2):261–264. doi: 10.1016/0014-5793(81)81134-9. [DOI] [PubMed] [Google Scholar]
  27. Suzuki H., Nishimura Y., Yasuda S., Nishimura A., Yamada M., Hirota Y. Murein-lipoprotein of Escherichia coli: a protein involved in the stabilization of bacterial cell envelope. Mol Gen Genet. 1978 Nov 16;167(1):1–9. doi: 10.1007/BF00270315. [DOI] [PubMed] [Google Scholar]
  28. TORRIANI A., ROTHMAN F. Mutants of Escherichia coli constitutive for alkaline phosphatase. J Bacteriol. 1961 May;81:835–836. doi: 10.1128/jb.81.5.835-836.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tamaki S., Sato T., Matsuhashi M. Role of lipopolysaccharides in antibiotic resistance and bacteriophage adsorption of Escherichia coli K-12. J Bacteriol. 1971 Mar;105(3):968–975. doi: 10.1128/jb.105.3.968-975.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Voss J. G. Effects of organic cations on the gram-negative cell wall and their bactericidal activity with ethylenediaminetetra-acetate and surface active agents. J Gen Microbiol. 1967 Sep;48(3):391–400. doi: 10.1099/00221287-48-3-391. [DOI] [PubMed] [Google Scholar]
  31. van Alphen L., Verkleij A., Leunissen-Bijvelt J., Lugtenberg B. Architecture of the outer membrane of Escherichia coli. III. Protein-lipopolysaccharide complexes in intramembraneous particles. J Bacteriol. 1978 Jun;134(3):1089–1098. doi: 10.1128/jb.134.3.1089-1098.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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