Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1966 Apr;99(1):53–61. doi: 10.1042/bj0990053

Chemical and biological properties of an extracellular lipopolysaccharide from Escherichia coli grown under lysine-limiting conditions

Alina Taylor 1,*, K W Knox 1,, Elizabeth Work 1
PMCID: PMC1264956  PMID: 5337757

Abstract

Lipopolysaccharide was prepared from the extracellular lipoglycopeptide produced by the lysine-requiring mutant Escherichia coli A.T.C.C. 12408 grown under lysine-limiting conditions. The lipid moiety, containing glucosamine phosphate and four fatty acids (lauric acid, myristic acid, β-hydroxymyristic acid and palmitic acid) corresponded in composition to lipid A of known bacterial lipopolysaccharides. The components of the polysaccharide moiety were d-glucose, d-galactose, l-glycero-d-manno-heptose, 3-deoxy-2-oxo-octonic acid, ethanolamine and phosphate. These are the constituents of the polysaccharide of the cell-wall antigens from rough strains of E. coli. Lipopolysaccharides were also prepared from whole cells of E. coli 12408 grown with excess or limited amounts of lysine; they were identical in carbohydrate composition with the extracellular lipopolysaccharide. The biological properties of this material also resembled those of known lipopolysaccharides; it was antigenic, pyrogenic, toxic and had adjuvant activity.

Full text

PDF
53

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen R. J. The estimation of phosphorus. Biochem J. 1940 Jun;34(6):858–865. doi: 10.1042/bj0340858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BANDURSKI R. S., AXELROD B. The chromatographic identification of some biologically important phosphate esters. J Biol Chem. 1951 Nov;193(1):405–410. [PubMed] [Google Scholar]
  3. BURTON A. J., CARTER H. E. PURIFICATION AND CHARACTERIZATION OF THE LIPID A COMPONENT OF THE LIPOPOLYSACCHARIDES FROM ESCHERICHIA COLI. Biochemistry. 1964 Mar;3:411–418. doi: 10.1021/bi00891a018. [DOI] [PubMed] [Google Scholar]
  4. Bishop D. G., Work E. An extracellular glycolipid produced by Escherichia coli grown under lysine-limiting conditions. Biochem J. 1965 Aug;96(2):567–576. doi: 10.1042/bj0960567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DISCHE Z. Qualitative and quantitative colorimetric determination of heptoses. J Biol Chem. 1953 Oct;204(2):983–997. [PubMed] [Google Scholar]
  6. Edstrom R. D., Heath E. C. Sugar nucleotide transferases in Escherichia coli lipopolysaccharide biosynthesis. Biochem Biophys Res Commun. 1964 Aug 11;16(6):576–581. doi: 10.1016/0006-291x(64)90195-0. [DOI] [PubMed] [Google Scholar]
  7. GHUYSEN J. M., STROMINGER J. L. STRUCTURE OF THE CELL WALL OF STAPHYLOCOCCUS AUREUS, STRAIN COPENHAGEN. I. PREPARATION OF FRAGMENTS BY ENZYMATIC HYDROLYSIS. Biochemistry. 1963 Sep-Oct;2:1110–1119. doi: 10.1021/bi00905a035. [DOI] [PubMed] [Google Scholar]
  8. GROLLMAN A. P., OSBORN M. J. O-PHOSPHORYLETHANOLAMINE: A COMPONENT OF LIPOPOLYSACCHARIDE IN CERTAIN GRAM-NEGATIVE BACTERIA. Biochemistry. 1964 Oct;3:1571–1574. doi: 10.1021/bi00898a031. [DOI] [PubMed] [Google Scholar]
  9. HUGGETT A. S., NIXON D. A. Use of glucose oxidase, peroxidase, and O-dianisidine in determination of blood and urinary glucose. Lancet. 1957 Aug 24;273(6991):368–370. doi: 10.1016/s0140-6736(57)92595-3. [DOI] [PubMed] [Google Scholar]
  10. JOHNSON A. G., GAINES S., LANDY M. Studies on the O antigen of Salmonella typhosa. V. Enhancement of antibody response to protein antigens by the purified lipopolysaccharide. J Exp Med. 1956 Feb 1;103(2):225–246. doi: 10.1084/jem.103.2.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KAUFFMANN F., BRAUN O. H., LUEDERITZ O., STIERLIN H., WESTPHAL O. [Immunochemistry of O-antigens of Enterobacteriaceae. IV. Analysis of the sugar constituents of Escherichia O-antigens]. Zentralbl Bakteriol. 1960 Oct;180:180–188. [PubMed] [Google Scholar]
  12. LARSON C. L., RIBI E., MILNER K. C., LIEBERMAN J. E. A method for titrating endotoxic activity in the skin of rabbits. J Exp Med. 1960 Jan 1;111:1–20. doi: 10.1084/jem.111.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LILLY M. D., CLARKE P. H., MEADOW P. M. THE ACCUMULATION OF NUCLEOTIDES BY ESCHERICHIA COLI STRAIN 26-26. J Gen Microbiol. 1963 Jul;32:103–116. doi: 10.1099/00221287-32-1-103. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. LUDOWIEG J., DORFMAN A. A micromethod for the colorimetric determination of N-acetyl groups in acid mucopolysaccharides. Biochim Biophys Acta. 1960 Feb 26;38:212–218. doi: 10.1016/0006-3002(60)91233-6. [DOI] [PubMed] [Google Scholar]
  16. LUEDERITZ O., RISSE H. J., SCHULTE-HOLTHAUSEN H., STROMINGER J. L., SUTHERLAND I. W., WESTPHAL O. BIOCHEMICAL STUDIES OF THE SMOOTH-ROUGH MUTATION IN SALMONELLA MINNESOTA. J Bacteriol. 1965 Feb;89:343–354. doi: 10.1128/jb.89.2.343-354.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. MACGEE J., DOUDOROFF M. A new phosphorylated intermediate in glucose oxidation. J Biol Chem. 1954 Oct;210(2):617–626. [PubMed] [Google Scholar]
  18. OROSZLAN S. I., MORA P. T. DISSOCIATION AND RECONSTITUTION OF AN ENDOTOXIN. Biochem Biophys Res Commun. 1963 Aug 14;12:345–349. doi: 10.1016/0006-291x(63)90102-5. [DOI] [PubMed] [Google Scholar]
  19. OSBORN M. J., ROSEN S. M., ROTHFIELD L., HORECKER B. L. Biosynthesis of bacterial lipopolysaccharide. I. Enzymatic incorporation of galactose in a mutant strain of Salmonella. Proc Natl Acad Sci U S A. 1962 Oct 15;48:1831–1838. doi: 10.1073/pnas.48.10.1831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. OSBORN M. J., ROSEN S. M., ROTHFIELD L., ZELEZNICK L. D., HORECKER B. L. LIPOPOLYSACCHARIDE OF THE GRAM-NEGATIVE CELL WALL. Science. 1964 Aug 21;145(3634):783–789. doi: 10.1126/science.145.3634.783. [DOI] [PubMed] [Google Scholar]
  21. OSBORN M. J. STUDIES ON THE GRAM-NEGATIVE CELL WALL. I. EVIDENCE FOR THE ROLE OF 2-KETO- 3-DEOXYOCTONATE IN THE LIPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM. Proc Natl Acad Sci U S A. 1963 Sep;50:499–506. doi: 10.1073/pnas.50.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. RONDLE C. J., MORGAN W. T. The determination of glucosamine and galactosamine. Biochem J. 1955 Dec;61(4):586–589. doi: 10.1042/bj0610586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. ROSEN S. M., OSBORN M. J., HORECKER B. L. BIOSYNTHESIS OF BACTERIAL LIPOPOLYSACCHARIDE. 3. CHARACTERIZATION OF THE GALACTOSE INCORPORATION PRODUCT. J Biol Chem. 1964 Oct;239:3196–3200. [PubMed] [Google Scholar]
  24. ROTH H., SEGAL S., BERTOLI D. THE QUANTITATIVE DETERMINATION OF GALACTOSE--AN ENZYMIC METHOD USING GALACTOSE OXIDASE, WITH APPLICATIONS TO BLOOD AND OTHER BIOLOGICAL FLUIDS. Anal Biochem. 1965 Jan;10:32–52. doi: 10.1016/0003-2697(65)90238-1. [DOI] [PubMed] [Google Scholar]
  25. RUEDE E., GOEBEL W. F. Colicine K. V. The somatic antigen of a non-colicinogenic variant of E. coli K235. J Exp Med. 1962 Jul 1;116:73–100. doi: 10.1084/jem.116.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SLEIN M. W., SCHNELL G. W. An aldoheptose phosphate in a polysaccharide isolated from Shigella flexneri. Proc Soc Exp Biol Med. 1953 Apr;82(4):734–738. doi: 10.3181/00379727-82-20231. [DOI] [PubMed] [Google Scholar]
  27. STAUB A. M., WESTPHAL O. ETUDE CHIMIQUE ET BIOCHIMIQUE DE LA SP'ECIFICIT'E IMMUNOLOGIQUE DES POLYOSIDES BACT'ERIENS. Bull Soc Chim Biol (Paris) 1964;46:1647–1684. [PubMed] [Google Scholar]
  28. STOFFYN P. J., JEANLOZ R. W. Identification of amino sugars by paper chromatography. Arch Biochem Biophys. 1954 Oct;52(2):373–379. doi: 10.1016/0003-9861(54)90137-x. [DOI] [PubMed] [Google Scholar]
  29. Sutherland I. W., Lüderitz O., Westphal O. Studies on the structure of lipopolysaccharides of Salmonella minnesota and Salmonella typhimurium R strains. Biochem J. 1965 Aug;96(2):439–448. doi: 10.1042/bj0960439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  31. WARREN L. Thiobarbituric acid spray reaction for deoxy sugars and sialic acids. Nature. 1960 Apr 16;186:237–237. doi: 10.1038/186237a0. [DOI] [PubMed] [Google Scholar]
  32. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]
  33. WORK E., DENMAN R. F. The use of a bacterial culture fluid as a source of alpha-diaminopimelic acid. Biochim Biophys Acta. 1953 Jan;10(1):183–183. doi: 10.1016/0006-3002(53)90226-1. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES