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. 1991 Sep;173(18):5677–5684. doi: 10.1128/jb.173.18.5677-5684.1991

Identification, isolation, and structural studies of extracellular polysaccharides produced by Caulobacter crescentus.

N Ravenscroft 1, S G Walker 1, G G Dutton 1, J Smit 1
PMCID: PMC208297  PMID: 1885545

Abstract

Caulobacters are adherent prosthecate bacteria that are members of bacterial biofouling communities in many environments. Investigation of the cell surface carbohydrates produced by two strains of the freshwater Caulobacter crescentus, CB2A and CB15A, revealed a hitherto undetected extracellular polysaccharide (EPS) or capsule. Isolation and characterization of the EPS fractions showed that each strain produced a unique neutral EPS which could not be readily removed from the cell surface by washing. Monosaccharide analysis showed that the main CB2A EPS contained D-glucose, D-gulose, and D-fucose in a ratio of 3:1:1, whereas the CB15A EPS fraction contained D-galactose, D-glucose, D-mannose, and D-fucose in approximately equal amounts. Methylation analysis of the main CB2A EPS showed the presence of terminal glucose and gulose groups, 3-linked fucosyl, and two 3,4-linked glucosyl units, thus confirming the pentasaccharide repeating unit indicated by 1H nuclear magnetic resonance analysis. Similar studies of the CB15A EPS revealed a tetrasaccharide repeating unit consisting of terminal galactose, 4-linked fucosyl, 3-linked glucosyl, and 3,4-linked mannosyl residues. EPS was not detectable by thin-section electron microscopy techniques, including some methods designed to preserve or enhance capsules, nor was the EPS readily detected on the cell surface by scanning electron microscopy when conventional fixation techniques were used; however, a structure consistent with EPS was revealed when samples were prepared by cryofixation and freeze-substitution methods.

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Selected References

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  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. Agabian N., Unger B. Caulobacter crescentus cell envelope: effect of growth conditions on murein and outer membrane protein composition. J Bacteriol. 1978 Feb;133(2):987–994. doi: 10.1128/jb.133.2.987-994.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bayer M. E. Visualization of the bacterial polysaccharide capsule. Curr Top Microbiol Immunol. 1990;150:129–157. doi: 10.1007/978-3-642-74694-9_7. [DOI] [PubMed] [Google Scholar]
  4. Beveridge T. J. The bacterial surface: general considerations towards design and function. Can J Microbiol. 1988 Apr;34(4):363–372. doi: 10.1139/m88-067. [DOI] [PubMed] [Google Scholar]
  5. Costerton J. W., Cheng K. J., Geesey G. G., Ladd T. I., Nickel J. C., Dasgupta M., Marrie T. J. Bacterial biofilms in nature and disease. Annu Rev Microbiol. 1987;41:435–464. doi: 10.1146/annurev.mi.41.100187.002251. [DOI] [PubMed] [Google Scholar]
  6. Costerton J. W., Irvin R. T., Cheng K. J. The bacterial glycocalyx in nature and disease. Annu Rev Microbiol. 1981;35:299–324. doi: 10.1146/annurev.mi.35.100181.001503. [DOI] [PubMed] [Google Scholar]
  7. Darveau R. P., Hancock R. E. Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J Bacteriol. 1983 Aug;155(2):831–838. doi: 10.1128/jb.155.2.831-838.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Graham L. L., Beveridge T. J. Effect of chemical fixatives on accurate preservation of Escherichia coli and Bacillus subtilis structure in cells prepared by freeze-substitution. J Bacteriol. 1990 Apr;172(4):2150–2159. doi: 10.1128/jb.172.4.2150-2159.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Graham L. L., Harris R., Villiger W., Beveridge T. J. Freeze-substitution of gram-negative eubacteria: general cell morphology and envelope profiles. J Bacteriol. 1991 Mar;173(5):1623–1633. doi: 10.1128/jb.173.5.1623-1633.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Harris P. J., Henry R. J., Blakeney A. B., Stone B. A. An improved procedure for the methylation analysis of oligosaccharides and polysaccharides. Carbohydr Res. 1984 Apr 2;127(1):59–73. doi: 10.1016/0008-6215(84)85106-x. [DOI] [PubMed] [Google Scholar]
  12. Jann B., Jann K. Structure and biosynthesis of the capsular antigens of Escherichia coli. Curr Top Microbiol Immunol. 1990;150:19–42. doi: 10.1007/978-3-642-74694-9_2. [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. MacRae J. D., Smit J. Characterization of caulobacters isolated from wastewater treatment systems. Appl Environ Microbiol. 1991 Mar;57(3):751–758. doi: 10.1128/aem.57.3.751-758.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  16. Merker R. I., Smit J. Characterization of the adhesive holdfast of marine and freshwater caulobacters. Appl Environ Microbiol. 1988 Aug;54(8):2078–2085. doi: 10.1128/aem.54.8.2078-2085.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mitchell D., Smit J. Identification of genes affecting production of the adhesion organelle of Caulobacter crescentus CB2. J Bacteriol. 1990 Sep;172(9):5425–5431. doi: 10.1128/jb.172.9.5425-5431.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ong C. J., Wong M. L., Smit J. Attachment of the adhesive holdfast organelle to the cellular stalk of Caulobacter crescentus. J Bacteriol. 1990 Mar;172(3):1448–1456. doi: 10.1128/jb.172.3.1448-1456.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Orskov F., Orskov I. The serology of capsular antigens. Curr Top Microbiol Immunol. 1990;150:43–63. [PubMed] [Google Scholar]
  20. POINDEXTER J. S. BIOLOGICAL PROPERTIES AND CLASSIFICATION OF THE CAULOBACTER GROUP. Bacteriol Rev. 1964 Sep;28:231–295. doi: 10.1128/br.28.3.231-295.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Poindexter J. S. The caulobacters: ubiquitous unusual bacteria. Microbiol Rev. 1981 Mar;45(1):123–179. doi: 10.1128/mr.45.1.123-179.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schmidt J. M., Stanier R. Y. The development of cellular stalks in bacteria. J Cell Biol. 1966 Mar;28(3):423–436. doi: 10.1083/jcb.28.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shapiro L. Generation of polarity during Caulobacter cell differentiation. Annu Rev Cell Biol. 1985;1:173–207. doi: 10.1146/annurev.cb.01.110185.001133. [DOI] [PubMed] [Google Scholar]
  24. Smit J., Agabian N. Cell surface patterning and morphogenesis: biogenesis of a periodic surface array during Caulobacter development. J Cell Biol. 1982 Oct;95(1):41–49. doi: 10.1083/jcb.95.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smit J., Agabian N. Cloning of the major protein of the Caulobacter crescentus periodic surface layer: detection and characterization of the cloned peptide by protein expression assays. J Bacteriol. 1984 Dec;160(3):1137–1145. doi: 10.1128/jb.160.3.1137-1145.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Smit J., Grano D. A., Glaeser R. M., Agabian N. Periodic surface array in Caulobacter crescentus: fine structure and chemical analysis. J Bacteriol. 1981 Jun;146(3):1135–1150. doi: 10.1128/jb.146.3.1135-1150.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Whitfield C., Schoenhals G., Graham L. Mutants of Escherichia coli O9:K30 with altered synthesis and expression of the capsular K30 antigen. J Gen Microbiol. 1989 Oct;135(10):2589–2599. doi: 10.1099/00221287-135-10-2589. [DOI] [PubMed] [Google Scholar]

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