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. 1977 Oct 1;167(1):147–154. doi: 10.1042/bj1670147

Characterization of lipid A and polysaccharide moieties of the lipopolysaccharides from Vibrio cholerae.

S Raziuddin
PMCID: PMC1183631  PMID: 588246

Abstract

Lipid A and polysaccharide moieties obtained by mild acid hydrolysis of the lipopolysaccharides from Vibrio cholerae 569 B (Inaba) and Vibrio el-tor (Inaba) were characterized. Heterogeneity of lipid A fractions was indicated by t.l.c. and by gel filtration of the de-O-acylated products from mild alkaline methanolysis of the lipids. Presumably lipid A contains a glucosamine backbone, and the fatty acids are probably bound to the hydroxyl and amino groups of glucosamine residues. Approximately equal amounts of fatty acids C16:0, C18:1 and 3-hydroxylauric acid were involved in ester linkages, but 3-hydroxymyristic acid was the only amide-linked fatty acid. Sephadex chromatography of the polysaccharide moiety showed the presence of a high-molecular-weight heptose-free fraction and a low-molecular-weight heptose-containing fraction. Haemagglutination-inhibition assays of these fractions showed the heptose-free fraction to be an O-specific side-chain polysaccharide, whereas the heptose-containing fraction was the core polysaccharide region of the lipopolysaccharides. Identical results were obtained for both organisms.

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Selected References

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  1. Adams G. A., Singh P. P. Structural features of lipid A preparations isolated from Escherichia coli and Shigella flexneri. Biochim Biophys Acta. 1970 May 5;202(3):553–555. doi: 10.1016/0005-2760(70)90128-1. [DOI] [PubMed] [Google Scholar]
  2. Adams G. A., Singh P. P. The chemical constitution of lipid A from Serratia marcescens. Can J Biochem. 1970 Jan;48(1):55–62. doi: 10.1139/o70-010. [DOI] [PubMed] [Google Scholar]
  3. Armstrong I. L., Redmond J. W. The fatty acids present in the lipopolysaccharide of Vibrio cholerae 569B (Inaba). Biochim Biophys Acta. 1974 May 29;348(2):302–305. doi: 10.1016/0005-2760(74)90242-2. [DOI] [PubMed] [Google Scholar]
  4. Bishop D. G., Hewett M. J., Knox K. W. Biochemical studies on lipopolysaccharides of Veillonella. Eur J Biochem. 1971 Mar 11;19(2):169–175. doi: 10.1111/j.1432-1033.1971.tb01301.x. [DOI] [PubMed] [Google Scholar]
  5. Chester I. R., Meadow P. M., Pitt T. L. The relationship between the O-antigenic lipopolysaccharides and serological specificity in strains of Pseudomonas aeruginosa of different O-serotypes. J Gen Microbiol. 1973 Oct;78(2):305–318. doi: 10.1099/00221287-78-2-305. [DOI] [PubMed] [Google Scholar]
  6. Drewry D. T., Gray G. W., Wilkinson S. G. Low-molecular-weight solutes released during mild acid hydrolysis of the lipopolysaccharide of Pseudomonas aeruginosa. Biochem J. 1972 Nov;130(1):289–295. doi: 10.1042/bj1300289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drewry D. T., Lomax J. A., Gray G. W., Wilkinson S. G. Studies of lipid A fractions from the lipopolysaccharides of Pseudomonas aeruginosa and Pseudomonas alcaligenes. Biochem J. 1973 Jul;133(3):563–572. doi: 10.1042/bj1330563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fensom A. H., Gray G. W. The chemical composition of the lipopolyacarideof Pseudomonas aeruginosa. Biochem J. 1969 Sep;114(2):185–196. doi: 10.1042/bj1140185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fensom A. H., Meadow P. M. Evidence for two regions in the polysaccharide moiety of the lipopolysaccharide of Pseudomonas aeruginosa 8602. FEBS Lett. 1970 Jul 29;9(2):81–84. doi: 10.1016/0014-5793(70)80318-0. [DOI] [PubMed] [Google Scholar]
  10. Galanos C., Rietschel E. T., Lüderitz O., Westphal O., Kim Y. B., Watson D. W. Biological activities of lipid A complexed with bovine-serum albumin. Eur J Biochem. 1972 Dec 4;31(2):230–233. doi: 10.1111/j.1432-1033.1972.tb02524.x. [DOI] [PubMed] [Google Scholar]
  11. Gmeiner J., Lüderitz O., Westphal O. Biochemical studies on lipopolysaccharides of Salmonella R mutants. 6. Investigations on the structure of the lipid A component. Eur J Biochem. 1969 Jan;7(3):370–379. doi: 10.1111/j.1432-1033.1969.tb19618.x. [DOI] [PubMed] [Google Scholar]
  12. Gmeiner J., Simon M., Lüderitz O. The linkage of phosphate groups and of 2-keto-3-deoxyoctonate to the lipid A component in a Salmonella minnesota lipopolysaccharide. Eur J Biochem. 1971 Aug 16;21(3):355–356. doi: 10.1111/j.1432-1033.1971.tb01476.x. [DOI] [PubMed] [Google Scholar]
  13. HAKOMORI S. A RAPID PERMETHYLATION OF GLYCOLIPID, AND POLYSACCHARIDE CATALYZED BY METHYLSULFINYL CARBANION IN DIMETHYL SULFOXIDE. J Biochem. 1964 Feb;55:205–208. [PubMed] [Google Scholar]
  14. Hancock I. C., Humphreys G. O., Meadow P. M. Characterisation of the hydroxy acids of Pseudomonas aeruginosa 8602. Biochim Biophys Acta. 1970 Mar 10;202(2):389–391. doi: 10.1016/0005-2760(70)90204-3. [DOI] [PubMed] [Google Scholar]
  15. Hämmerling G., Lehmann V., Lüderitz O. Structural studies on the heptose region of Salmonella lipopolysaccharides. Eur J Biochem. 1973 Oct 18;38(3):453–458. doi: 10.1111/j.1432-1033.1973.tb03079.x. [DOI] [PubMed] [Google Scholar]
  16. Hämmerling G., Lüderitz O., Westphal O., Mäkelä P. H. Structural investigations on the core polysaccharide of Escherichia coli 0100. Eur J Biochem. 1971 Oct 14;22(3):331–344. doi: 10.1111/j.1432-1033.1971.tb01549.x. [DOI] [PubMed] [Google Scholar]
  17. Jackson G. D.F., Redmond J. W. Immunochemical studies of the O-antigens of Vibrio cholerae. The constitution of a lipopolysaccharide from V. cholerae 569B (Inaba). FEBS Lett. 1971 Feb 19;13(2):117–120. doi: 10.1016/0014-5793(71)80213-2. [DOI] [PubMed] [Google Scholar]
  18. Jann B., Jann K., Beyaert G. O. 2-Amino-2,6-dideoxy-d-glucose (D-quinovosamine): a constituent of the lipopolysaccharides of Vibrio cholerae. Eur J Biochem. 1973 Sep 3;37(3):531–534. doi: 10.1111/j.1432-1033.1973.tb03015.x. [DOI] [PubMed] [Google Scholar]
  19. Kasai N., Nowotny A. Endotoxic glycolipid from a heptoseless mutant of Salmonella minnesota. J Bacteriol. 1967 Dec;94(6):1824–1836. doi: 10.1128/jb.94.6.1824-1836.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lacave C., Asselineau J., Serre A., Roux J. Comparaison de la composition chimique d'une fraction lipopolysaccharidique et d'une fraction polysaccharidique isolées de Brucella melitensis. Eur J Biochem. 1969 Jun;9(2):189–198. doi: 10.1111/j.1432-1033.1969.tb00594.x. [DOI] [PubMed] [Google Scholar]
  21. Lomax J. A., Gray G. W., Wilkinson S. G. Studies of the polysaccharide fraction from the lipopolysaccharide of Pseudomonas alcaligenes. Biochem J. 1974 Jun;139(3):633–643. doi: 10.1042/bj1390633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lüderitz O., Staub A. M., Westphal O. Immunochemistry of O and R antigens of Salmonella and related Enterobacteriaceae. Bacteriol Rev. 1966 Mar;30(1):192–255. doi: 10.1128/br.30.1.192-255.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. NOWOTNY A. Relation of structure to function in bacterial O antigens. II. Fractionation of lipids present in Boivin-type endotoxin of Serratia marcescens. J Bacteriol. 1963 Feb;85:427–435. doi: 10.1128/jb.85.2.427-435.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Raziuddin S. R. Effect of growth temperature and culture age on the lipid composition of Vibrio cholerae 569B (Inaba). J Gen Microbiol. 1976 Jun;94(2):367–372. doi: 10.1099/00221287-94-2-367. [DOI] [PubMed] [Google Scholar]
  26. Raziuddin S., Ambegaokar S. D. Chemical composition & fatty acid pattern of lipopolysaccharides from Vibrio cholerae. Indian J Biochem Biophys. 1976 Mar;13(1):57–61. [PubMed] [Google Scholar]
  27. Raziuddin S., Kawasaki T. Biochemical studies on the cell wall lipopolysaccharides (O-antigens) of Vibrio cholerae 569 B (Inaba) and El-tor (Inaba). Biochim Biophys Acta. 1976 Apr 22;431(1):116–126. doi: 10.1016/0005-2760(76)90265-4. [DOI] [PubMed] [Google Scholar]
  28. Redmond J. W. 4-Amino-4,6-dideoxy-D-mannose (D-perosamine): a component of the lipopolysaccharide of Vibrio cholerae 569B (Inaba). FEBS Lett. 1975 Feb 1;50(2):147–149. doi: 10.1016/0014-5793(75)80476-5. [DOI] [PubMed] [Google Scholar]
  29. Rietschel E. T., Gottert H., Lüderitz O., Westphal O. Nature and linkages of the fatty acids present in the lipid-A component of Salmonella lipopolysaccharides. Eur J Biochem. 1972 Jul 13;28(2):166–173. doi: 10.1111/j.1432-1033.1972.tb01899.x. [DOI] [PubMed] [Google Scholar]
  30. Rietschel E. T., Palin W. J., Watson D. W. Nature and linkages of the fatty acids present in lipopolysaccharides from Vibrio metchnikovii and Vibrio parahemolyticus. Eur J Biochem. 1973 Aug 1;37(1):116–120. doi: 10.1111/j.1432-1033.1973.tb02965.x. [DOI] [PubMed] [Google Scholar]
  31. Rooney S. A., Goldfine H. Isolation and characterization of 2-keto-3-deoxyoctonate-lipid A from a heptose-deficient mutant of Escherichia coli. J Bacteriol. 1972 Aug;111(2):531–541. doi: 10.1128/jb.111.2.531-541.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rooney S. A., Goldfine H., Sweeley C. C. The identification of trans-2-tetradecenoic acid in hydrolysates of lipid A from Escherichia coli. Biochim Biophys Acta. 1972 Jul 7;270(3):289–295. doi: 10.1016/0005-2760(72)90192-0. [DOI] [PubMed] [Google Scholar]
  33. Russa R., Lorkiewicz Z. Fatty acids present in the lipopolysaccharide of Rhizobium trifolii. J Bacteriol. 1974 Sep;119(3):771–775. doi: 10.1128/jb.119.3.771-775.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schmidt G., Jann B., Jann K. Immunochemistry of R lipopolysaccharides of Escherichia coli. Different core regions in the lipopolysaccharides of O group 8. Eur J Biochem. 1969 Oct;10(3):501–510. doi: 10.1111/j.1432-1033.1969.tb00717.x. [DOI] [PubMed] [Google Scholar]
  35. Weckesser J., Drews G., Fromme I. Chemical analysis of and degradation studies on the cell wall lipopolysaccharide of Rhodopseudomonas capsulata. J Bacteriol. 1972 Mar;109(3):1106–1113. doi: 10.1128/jb.109.3.1106-1113.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wilkinson S. G., Galbraith L., Lightfoot G. A. Cell walls, lipids, and lipopolysaccharides of Pseudomonas species. Eur J Biochem. 1973 Feb 15;33(1):158–174. doi: 10.1111/j.1432-1033.1973.tb02666.x. [DOI] [PubMed] [Google Scholar]
  37. Wilkinson S. G., Galbrath L. Studies of lipopolysaccharides from Pseudomonas aeruginosa. Eur J Biochem. 1975 Mar 17;52(2):331–343. doi: 10.1111/j.1432-1033.1975.tb04001.x. [DOI] [PubMed] [Google Scholar]
  38. Wober W., Alaupović P. Studies on the protein moiety of endotoxin from gram-negative bacteria. Characterization of the protein moiety isolated by acetic acid hydrolysis of endotoxin from Serratia marcescens 08. Eur J Biochem. 1971 Apr;19(3):357–367. doi: 10.1111/j.1432-1033.1971.tb01324.x. [DOI] [PubMed] [Google Scholar]
  39. Wober W., Alaupović P. Studies on the protein moiety of endotoxin from gram-negative bacteria. Characterization of the protein moiety isolated by phenol treatment of endotoxin from Serratia marcescens 08 and Escherichia coli 0 141:K85(B). Eur J Biochem. 1971 Apr;19(3):340–356. doi: 10.1111/j.1432-1033.1971.tb01323.x. [DOI] [PubMed] [Google Scholar]

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