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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Jan;129(1):225–236. doi: 10.1128/jb.129.1.225-236.1977

Role of a sugar-lipid intermediate in colanic acid synthesis by Escherichia coli.

J G Johnson, D B Wilson
PMCID: PMC234919  PMID: 318640

Abstract

Membrane fractions from a lon strain of Escherichia coli but not a wild-type strain catalyze the incorporation of fucose from guanosine 5'-diphosphate-fucose into a lipid and into polymeric material. Both incorporation reactions specifically require only uridine 5'-diphosphate (UDP)-glucose. The sugar lipid was shown to be an intermediate in the synthesis of the polymer which was related to colanic acid. The sugar lipid had the structure (fucose3, glucose2)-glucose P-P-lipid. Its behavior on column and thin-layer chromatography, the rates of its hydrolysis in acid and base, and the response of its synthesis to inhibitors are all identical to the other sugar-lipid intermediates which have been shown to contain sugars attached to the C55-polyisoprenol, undecaprenol, by a pyrophosphate linkage. The membrane fractions from both the lon strain and the wild-type strain also catalyzed the incorporation of either glucose from UDP-glucose or galactose from UDP-galactose into a lipid fraction which was shown to contain the free sugar attached by a monophosphate linkage to an undecaprenol-like lipid. This lipid was isolated and its nuclear magnetic resonance spectra was identical to undecaprenol. The membrane fractions from both strains also incorporated glucose from UDP-glucose into glycogen and into a polymer that behaved like Escherichia coli lipopolysaccharide. Conditions were found where the incorporation of glucose could be directed specifically into each compound by adding the appropriate inhibitors.

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Selected References

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  1. ANDERSON J. S., MATSUHASHI M., HASKIN M. A., STROMINGER J. L. LIPID-PHOSPHOACETYLMURAMYL-PENTAPEPTIDE AND LIPID-PHOSPHODISACCHARIDE-PENTAPEPTIDE: PRESUMED MEMBRANE TRANSPORT INTERMEDIATES IN CELL WALL SYNTHESIS. Proc Natl Acad Sci U S A. 1965 Apr;53:881–889. doi: 10.1073/pnas.53.4.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bukhari A. I., Zipser D. Mutants of Escherichia coli with a defect in the degradation of nonsense fragments. Nat New Biol. 1973 Jun 20;243(129):238–241. doi: 10.1038/newbio243238a0. [DOI] [PubMed] [Google Scholar]
  3. Dankert M., Wright A., Kelley W. S., Robbins P. W. Isolation, purification, and properties of the lipid-linked intermediates of O-antigen biosynthesis. Arch Biochem Biophys. 1966 Sep 26;116(1):425–435. doi: 10.1016/0003-9861(66)90049-x. [DOI] [PubMed] [Google Scholar]
  4. GOEBEL W. F. Colanic acid. Proc Natl Acad Sci U S A. 1963 Apr;49:464–471. doi: 10.1073/pnas.49.4.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. HENRIKSEN S. D. Studies on the Klebsiella group (Kauffmann). III. Demonstration of the M-antigen of Escherichia coli in Klebsiella (Aerobacter). Acta Pathol Microbiol Scand. 1954;34(3):266–270. [PubMed] [Google Scholar]
  6. Higashi Y., Strominger J. L., Sweeley C. C. Biosynthesis of the peptidoglycan of bacterial cell walls. XXI. Isolation of free C55-isoprenoid alcohol and of lipid intermediates in peptidoglycan synthesis from Staphylococcus aureus. J Biol Chem. 1970 Jul 25;245(14):3697–3702. [PubMed] [Google Scholar]
  7. Hua S. S., Markovitz A. Multiple regulator gene control of the galactose operon in Escherichia coli K-12. J Bacteriol. 1972 Jun;110(3):1089–1099. doi: 10.1128/jb.110.3.1089-1099.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Lahav M., Chiu T. H., Lennarz W. J. Studies on the biosynthesis of mannan in Micrococcus lysodeikticus. II. The enzymatic synthesis of mannosyl-l-phosphoryl-undecaprenol. J Biol Chem. 1969 Nov 10;244(21):5890–5898. [PubMed] [Google Scholar]
  10. Lennarz W. J., Scher M. G. Metabolism and function of polyisoprenol sugar intermediates in membrane-associated reactions. Biochim Biophys Acta. 1972 Aug 4;265(3):417–441. doi: 10.1016/0304-4157(72)90015-9. [DOI] [PubMed] [Google Scholar]
  11. MARKOVITZ A. REGULATORY MECHANISMS FOR SYNTHESIS OF CAPSULAR POLYSACCHARIDE IN MUCOID MUTANTS OF ESCHERICHIA COLI K12. Proc Natl Acad Sci U S A. 1964 Feb;51:239–246. doi: 10.1073/pnas.51.2.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mackie G., Wilson D. B. Regulation of the gal operon of Escherichia coli by the capR gene. J Biol Chem. 1972 May 25;247(10):2973–2978. [PubMed] [Google Scholar]
  13. Markovitz A., Rosenbaum N. A regulator gene that is dominant on an episome and recessive on a chromosome. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1084–1091. doi: 10.1073/pnas.54.4.1084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nikaido H., Nikaido K., Nakae T., Mäkelä P. H. Glucosylation of lipopolysaccharide in Salmonella: biosynthesis of O antigen factor 12 2 . I. Over-all reaction. J Biol Chem. 1971 Jun 25;246(12):3902–3911. [PubMed] [Google Scholar]
  15. Nikaido K., Nikaido H. Glucosylation of lipopolysaccharide in Salmonella: biosynthesis nof O antigen factor n12 2 . II. Structure of the lipid intermediate. J Biol Chem. 1971 Jun 25;246(12):3912–3919. [PubMed] [Google Scholar]
  16. 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]
  17. Osborn M. J., Weiner I. M. Biosynthesis of a bacterial lipopolysaccharide. VI. Mechanism of incorporation of abequose into the O-antigen of Salmonella typhimurium. J Biol Chem. 1968 May 25;243(10):2631–2639. [PubMed] [Google Scholar]
  18. SAPELLI R. V., GOEBEL W. F. THE CAPSULAR POLYSACCHARIDE OF A MUCOID VARIANT OF E. COLI K 12. Proc Natl Acad Sci U S A. 1964 Aug;52:265–271. doi: 10.1073/pnas.52.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SCOTT J. E. Aliphatic ammonium salts in the assay of acidic polysaccharides from tissues. Methods Biochem Anal. 1960;8:145–197. doi: 10.1002/9780470110249.ch4. [DOI] [PubMed] [Google Scholar]
  20. Scher M., Lennarz W. J. Studies on the biosynthesis of mannan in Micrococcus lysodeikticus. I. Characterization of mannan-14C formed enzymatically from mannosyl-1-phosphoryl-undecaprenol. J Biol Chem. 1969 May 25;244(10):2777–2789. [PubMed] [Google Scholar]
  21. Scher M., Lennarz W. J., Sweeley C. C. The biosynthesis of mannosyl-1-phosphoryl-polyisoprenol in Micrococcus lysodeikticus and its role in mannan synthesis. Proc Natl Acad Sci U S A. 1968 Apr;59(4):1313–1320. doi: 10.1073/pnas.59.4.1313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stone K. J., Strominger J. L. Mechanism of action of bacitracin: complexation with metal ion and C 55 -isoprenyl pyrophosphate. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3223–3227. doi: 10.1073/pnas.68.12.3223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Struve W. G., Sinha R. K., Neuhaus F. C. On the initial stage in peptidoglycan synthesis. Phospho-N-acetylmuramyl-pentapeptide translocase (uridine monophosphate). Biochemistry. 1966 Jan;5(1):82–93. doi: 10.1021/bi00865a012. [DOI] [PubMed] [Google Scholar]
  24. Sutherland I. W. Enzymic hydrolysis of colanic acid. Eur J Biochem. 1971 Dec 10;23(3):582–587. doi: 10.1111/j.1432-1033.1971.tb01657.x. [DOI] [PubMed] [Google Scholar]
  25. Sutherland I. W. Structural studies on colanic acid, the common exopolysaccharide found in the enterobacteriaceae, by partial acid hydrolysis. Oligosaccharides from colanic acid. Biochem J. 1969 Dec;115(5):935–945. doi: 10.1042/bj1150935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Troy F. A., Frerman F. E., Heath E. C. The biosynthesis of capsular polysaccharide in Aerobacter aerogenes. J Biol Chem. 1971 Jan 10;246(1):118–133. [PubMed] [Google Scholar]
  27. Umbreit J. N., Strominger J. L. Isolation of the lipid intermediate in peptidoglycan biosynthesis from Escherichia coli. J Bacteriol. 1972 Dec;112(3):1306–1309. doi: 10.1128/jb.112.3.1306-1309.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wright A., Dankert M., Fennessey P., Robbins P. W. Characterization of a polyisoprenoid compound functional in O-antigen biosynthesis. Proc Natl Acad Sci U S A. 1967 Jun;57(6):1798–1803. doi: 10.1073/pnas.57.6.1798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wright A. Mechanism of conversion of the salmonella O antigen by bacteriophageepsilon 34. J Bacteriol. 1971 Mar;105(3):927–936. doi: 10.1128/jb.105.3.927-936.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yuasa R., Levinthal M., Nikaido H. Biosynthesis of cell wall lipopolysaccharide in mutants of Salmonella. V. A mutant of Salmonella typhimurium defective in the synthesis of cytidine diphosphoabequose. J Bacteriol. 1969 Oct;100(1):433–444. doi: 10.1128/jb.100.1.433-444.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]

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