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. 1993 May;175(9):2490–2500. doi: 10.1128/jb.175.9.2490-2500.1993

Sequential assembly and polymerization of the polyprenol-linked pentasaccharide repeating unit of the xanthan polysaccharide in Xanthomonas campestris.

L Ielpi 1, R O Couso 1, M A Dankert 1
PMCID: PMC204549  PMID: 7683019

Abstract

Lipid-linked intermediates are involved in the synthesis of the exopolysaccharide xanthan produced by the bacterium Xanthomonas campestris (L. Ielpi, R. O. Couso, and M. A. Dankert, FEBS Lett. 130:253-256, 1981). In this study, the stepwise assembly of the repeating pentasaccharide unit of xanthan is described. EDTA-treated X. campestris cells were used as both enzyme preparation and lipid-P acceptor, and UDP-Glc, GDP-Man, and UDP-glucuronic acid were used as sugar donors. A linear pentasaccharide unit is assembled on a polyprenol-P lipid carrier by the sequential addition of glucose-1-P, glucose, mannose, glucuronic acid, and mannose. The in vitro synthesis of pentasaccharide-P-P-polyprenol was also accompanied by the incorporation of radioactivity into a polymeric product, which was characterized as xanthan, on the basis of gel filtration and permethylation studies. Results from two-stage reactions showed that essentially pentasaccharide-P-P-polyprenol is polymerized. In addition, the direction of chain elongation has been studied by in vivo experiments. The polymerization of lipid-linked repeat units occurs by the successive transfer of the growing chain to a new pentasaccharide-P-P-polyprenol. The reaction involves C-1 of glucose at the reducing end of the polyprenol-linked growing chain and C-4 of glucose at the nonreducing position of the newly formed polyprenol-linked pentasaccharide, generating a branched polymer with a trisaccharide side chain.

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Selected References

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  1. Cadmus M. C., Rogovin S. P., Burton K. A., Pittsley J. E., Knutson C. A., Jeanes A. Colonial variation in Xanthomonas campestris NRRL B-1459 and characterization of the polysaccharide from a variant strain. Can J Microbiol. 1976 Jul;22(7):942–948. doi: 10.1139/m76-136. [DOI] [PubMed] [Google Scholar]
  2. Chapman A., Li E., Kornfeld S. The biosynthesis of the major lipid-linked oligosaccharide of Chinese hamster ovary cells occurs by the ordered addition of mannose residues. J Biol Chem. 1979 Oct 25;254(20):10243–10249. [PubMed] [Google Scholar]
  3. Couso R. O., Ielpi L., Garcia R. C., Dankert M. A. Biosynthesis of polysaccharides in Acetobacter xylinum. Sequential synthesis of a heptasaccharide diphosphate prenol. Eur J Biochem. 1982 Apr;123(3):617–627. doi: 10.1111/j.1432-1033.1982.tb06577.x. [DOI] [PubMed] [Google Scholar]
  4. Couso R. O., Ielpi L., García R. C., Dankert M. A. Synthesis of mannosyl cellobiose diphosphate prenol in Acetobacter xylinum. Arch Biochem Biophys. 1980 Oct 15;204(2):435–443. doi: 10.1016/0003-9861(80)90054-5. [DOI] [PubMed] [Google Scholar]
  5. García R. C., Recondo E., Dankert M. Polysaccharide biosynthesis in Acetobacter xylinum. Enzymatic synthesis of lipid diphosphate and monophospate sugars. Eur J Biochem. 1974 Mar 15;43(1):93–105. doi: 10.1111/j.1432-1033.1974.tb03389.x. [DOI] [PubMed] [Google Scholar]
  6. Harding N. E., Cleary J. M., Cabañas D. K., Rosen I. G., Kang K. S. Genetic and physical analyses of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris. J Bacteriol. 1987 Jun;169(6):2854–2861. doi: 10.1128/jb.169.6.2854-2861.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hassler R. A., Doherty D. H. Genetic engineering of polysaccharide structure: production of variants of xanthan gum in Xanthomonas campestris. Biotechnol Prog. 1990 May-Jun;6(3):182–187. doi: 10.1021/bp00003a003. [DOI] [PubMed] [Google Scholar]
  8. Hötte B., Rath-Arnold I., Pühler A., Simon R. Cloning and analysis of a 35.3-kilobase DNA region involved in exopolysaccharide production by Xanthomonas campestris pv. campestris. J Bacteriol. 1990 May;172(5):2804–2807. doi: 10.1128/jb.172.5.2804-2807.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ielpi L., Couso R. O., Dankert M. A. Pyruvic acid acetal residues are transferred from phosphoenolpyruvate to the pentasaccharide-P-P-lipid. Biochem Biophys Res Commun. 1981 Oct 30;102(4):1400–1408. doi: 10.1016/s0006-291x(81)80167-2. [DOI] [PubMed] [Google Scholar]
  10. Ielpi L., Couso R., Dankert M. Lipid-linked intermediates in the biosynthesis of xanthan gum. FEBS Lett. 1981 Aug 3;130(2):253–256. doi: 10.1016/0014-5793(81)81132-5. [DOI] [PubMed] [Google Scholar]
  11. Jansson P. E., Kenne L., Lindberg B. Structure of extracellular polysaccharide from Xanthomonas campestris. Carbohydr Res. 1975 Dec;45:275–282. doi: 10.1016/s0008-6215(00)85885-1. [DOI] [PubMed] [Google Scholar]
  12. Köplin R., Arnold W., Hötte B., Simon R., Wang G., Pühler A. Genetics of xanthan production in Xanthomonas campestris: the xanA and xanB genes are involved in UDP-glucose and GDP-mannose biosynthesis. J Bacteriol. 1992 Jan;174(1):191–199. doi: 10.1128/jb.174.1.191-199.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Li E., Tabas I., Kornfeld S. The synthesis of complex-type oligosaccharides. I. Structure of the lipid-linked oligosaccharide precursor of the complex-type oligosaccharides of the vesicular stomatitis virus G protein. J Biol Chem. 1978 Nov 10;253(21):7762–7770. [PubMed] [Google Scholar]
  14. Marzocca M. P., Harding N. E., Petroni E. A., Cleary J. M., Ielpi L. Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris. J Bacteriol. 1991 Dec;173(23):7519–7524. doi: 10.1128/jb.173.23.7519-7524.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Melton L. D., Mindt L., Rees D. A., Sanderson G. R. Covalent structure of the extracellular polysaccharide from Xanthomonas campestris: evidence from partial hydrolysis studies. Carbohydr Res. 1976 Feb;46(2):245–257. doi: 10.1016/s0008-6215(00)84296-2. [DOI] [PubMed] [Google Scholar]
  16. Robbins P. W., Bray D., Dankert B. M., Wright A. Direction of chain growth in polysaccharide synthesis. Science. 1967 Dec 22;158(3808):1536–1542. doi: 10.1126/science.158.3808.1536. [DOI] [PubMed] [Google Scholar]
  17. Thorne L., Tansey L., Pollock T. J. Clustering of mutations blocking synthesis of xanthan gum by Xanthomonas campestris. J Bacteriol. 1987 Aug;169(8):3593–3600. doi: 10.1128/jb.169.8.3593-3600.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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