Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1978 Dec;22(3):842–851. doi: 10.1128/iai.22.3.842-851.1978

Extractability of Cell Wall Polysaccharide from Lactobacilli and Streptococci by Autoclaving and by Dilute Acid

Lindy K Campbell 1, Kenneth W Knox 1, Anthony J Wicken 2
PMCID: PMC422236  PMID: 32140

Abstract

Autoclaving cell wall of Streptococcus mutans Ingbritt for 15 min under the Rantz and Randall conditions released one-tenth of the total cell wall carbohydrate, whereas two-thirds was extracted after autoclaving for 180 min. The extract contained the serotype c-specific antigen but lacked the lipoteichoic acid component extracted when whole cells were autoclaved. Autoclaving cell wall preparations from other strains of S. mutans and also Streptococcus salivarius and Streptococcus mitis in 0.85% NaCl for 180 min released the major proportion of the wall polysaccharide fraction. Approximately 50 to 90% of wall carbohydrate of Lactobacillus fermentum and Lactobacillus casei was released when cell wall preparations were autoclaved in 0.85% NaCl for 180 min. For wall preparations from several strains of S. mutans, autoclaving for 60 min at pH 3.75 released only 39 to 62% of wall carbohydrate, whereas almost total release could be achieved with the lactobacilli. Heating S. mutans Ingbritt cell wall for 24 h at 60°C in 0.1 N H2SO4 released only two-thirds of the wall carbohydrate; by comparison nearly all of the wall carbohydrate was released in 3 h from L. casei and L. fermentum. Autoclaving L. casei cell wall and purified soluble wall fractions hydrolyzed the phosphodiester bond between the polysaccharide and peptidoglycan. This was shown by the release of reactive N-acetylhexosamine in both cases and the presence of a phosphomonoester in the autoclaved soluble wall fractions. The results indicate that autoclaving can hydrolyze covalent linkages, and this must be considered when the Rantz and Randall procedure is used to obtain antigen preparations.

Full text

PDF
842

Images in this article

845Image
on p.845

Selected References

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

  1. Bleiweis A. S., Taylor M. C., Deepak J., Brown T. A., Wetherell J. R., Jr Comparative chemical compositions of cell walls of Streptococcus mutans. J Dent Res. 1976 Jan;55:A103–A108. doi: 10.1177/002203457605500102011. [DOI] [PubMed] [Google Scholar]
  2. Bratthall D., Pettersson B. M. Common and unique antigens of Streptococcus mutants. J Dent Res. 1976 Jan;55:A60–A64. doi: 10.1177/002203457605500123011. [DOI] [PubMed] [Google Scholar]
  3. DAHLQVIST A. Determination of maltase and isomaltase activities with a glucose-oxidase reagent. Biochem J. 1961 Sep;80:547–551. doi: 10.1042/bj0800547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DISCHE Z. New color reactions for determination of sugars in polysaccharides. Methods Biochem Anal. 1955;2:313–358. doi: 10.1002/9780470110188.ch11. [DOI] [PubMed] [Google Scholar]
  5. Grenier E. M., Gray R. H., Loesche W. J., Eveland W. C. Microcapsules on Streptococcus mutans serotypes by electron microscopy. J Dent Res. 1977 Feb;56(2):166–176. doi: 10.1177/00220345770560021101. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hall E. A., Knox K. W. Properties of the polysaccharide and mucopeptide components of the cell wall of Lactobacillus casei. Biochem J. 1965 Aug;96(2):310–318. doi: 10.1042/bj0960310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hamada S., Gill K., Slade H. D. Chemical and immunological properties of the type f polysaccharide antigen of Streptococcus mutans. Infect Immun. 1976 Jul;14(1):203–211. doi: 10.1128/iai.14.1.203-211.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hamada S., Masuda N., Ooshima T., Sobue S., Kotani S. Epidemiological survey of Streptococcus mutans among Japanese children. Identification and serological typing of the isolated strains. Jpn J Microbiol. 1976 Feb;20(1):33–44. doi: 10.1111/j.1348-0421.1976.tb00905.x. [DOI] [PubMed] [Google Scholar]
  10. Hamada S., Slade H. D. Adherence of serotype e Streptococcus mutans and the inhibitory effect of Lancefield group E and S mutans type e antiserum. J Dent Res. 1976 Apr;55(Spec No):C65–C74. doi: 10.1177/002203457605500328011. [DOI] [PubMed] [Google Scholar]
  11. Hamada S., Tai S., Slade H. D. Selective adsorption of heterophile polyglycerophosphate antigen from antigen extracts of Streptococcus mutans and other gram-positive bacteria. Infect Immun. 1976 Oct;14(4):903–910. doi: 10.1128/iai.14.4.903-910.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hammond B. F., Steel C. F., Peindl K. S. Antigens and surface components associated with virulence of Actinomyces viscosus. J Dent Res. 1976 Jan;55:A19–A25. doi: 10.1177/002203457605500111011. [DOI] [PubMed] [Google Scholar]
  13. KNOX K. W., HALL E. A. THE ISOLATION OF OLIGOSACCHARIDES FROM THE CELL-WALL POLYSACCHARIDE OF LACTOBACILLUS CASEI, SEROLOGICAL GROUP C. Biochem J. 1965 Mar;94:525–533. doi: 10.1042/bj0940525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. KNOX K. W. Isolation of group specific products from Lactobacillus casei and L.casei var.rhamnosus. J Gen Microbiol. 1963 Apr;31:59–72. doi: 10.1099/00221287-31-1-59. [DOI] [PubMed] [Google Scholar]
  15. Knox K. W., Hall E. A. The linkage between the polysaccharide and mucopeptide components of the cell wall of Lactobacillus casei. Biochem J. 1965 Aug;96(2):302–309. doi: 10.1042/bj0960302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Knox K. W., Holmwood K. J. Structure of the cell wall of Lactobacilli. Role of muramic acid phosphate in Lactobacillus fermenti. Biochem J. 1968 Jul;108(3):363–368. doi: 10.1042/bj1080363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Knox K. W., Markham J. L., Wicken A. J. Formation of cross-reacting antibodies against cellular and extracellular lipoteichoic acid of Streptococcus mutans BHT. Infect Immun. 1976 Mar;13(3):647–652. doi: 10.1128/iai.13.3.647-652.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Knox K. W., Wicken A. J. Serological studies on the teichoic acids of Lactobacillus plantarum. Infect Immun. 1972 Jul;6(1):43–49. doi: 10.1128/iai.6.1.43-49.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Linzer R., Slade H. D. Purification and characterization of Streptococcus mutans group d cell wall polysaccharide antigen. Infect Immun. 1974 Aug;10(2):361–368. doi: 10.1128/iai.10.2.361-368.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Markham J. L., Knox K. W., Wicken A. J., Hewett M. J. Formation of extracellular lipoteichoic acid by oral streptococci and lactobacilli. Infect Immun. 1975 Aug;12(2):378–386. doi: 10.1128/iai.12.2.378-386.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Montague E. A., Knox K. W. Antigenic components of the cell wall of Streptococcus salivarius. J Gen Microbiol. 1968 Dec;54(2):237–246. doi: 10.1099/00221287-54-2-237. [DOI] [PubMed] [Google Scholar]
  22. Mukasa H., Slade H. D. Extraction, purification, and chemical and immunological properties of the Streptococcus mutans group "a" polysaccharide cell wall antigen. Infect Immun. 1973 Aug;8(2):190–198. doi: 10.1128/iai.8.2.190-198.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Olsson J., Glantz P. O., Krasse B. Electrophoretic mobility of oral streptococci. Arch Oral Biol. 1976;21(10):605–609. doi: 10.1016/0003-9969(76)90030-3. [DOI] [PubMed] [Google Scholar]
  24. RANTZ L. A., RANDALL E. Use of autoclaved extracts of hemolytic streptococci for serological grouping. Stanford Med Bull. 1955 May;13(2):290–291. [PubMed] [Google Scholar]
  25. Rosan B., Lai C. H., Listgarten M. A. Streptococcus sanguis: a model in the application in immunochemical analysis for the in situ localization of bacteria in dental plaque. J Dent Res. 1976 Jan;55:A124–A141. doi: 10.1177/002203457605500105011. [DOI] [PubMed] [Google Scholar]
  26. Soprey P., Slade H. D. Immunochemistry of the streptococcal group R cell wall polysaccharide antigen. Infect Immun. 1972 Jan;5(1):91–97. doi: 10.1128/iai.5.1.91-97.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vaught R. M., Bleiweis A. S. Antigens of Streptococcus mutans. II. Characterization of an antigen resembling a glycerol teichoic acid in walls of strain BHT. Infect Immun. 1974 Jan;9(1):60–67. doi: 10.1128/iai.9.1.60-67.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wetherell J. R., Jr, Bleiweis A. S. Antigens of Streptococcus mutans: characterization of a polysaccharide antigen from walls of strain GS-5. Infect Immun. 1975 Dec;12(6):1341–1348. doi: 10.1128/iai.12.6.1341-1348.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wicken A. J., Gibbens J. W., Knox K. W. Comparative studies on the isolation of membrane lipoteichoic acid from Lactobacillus fermenti. J Bacteriol. 1973 Jan;113(1):365–372. doi: 10.1128/jb.113.1.365-372.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wicken A. J., Knox K. W. Lipoteichoic acids: a new class of bacterial antigen. Science. 1975 Mar 28;187(4182):1161–1167. doi: 10.1126/science.46620. [DOI] [PubMed] [Google Scholar]
  31. van Houte J., Saxton C. A. Cell wall thickening and intracellular polysaccharide in microorganisms of the dental plaque. Caries Res. 1971;5(1):30–43. doi: 10.1159/000259730. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES