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Isolation and purification of cell wall polysaccharide of Bacillus anthracis (Δ Sterne)

F.S. Ekwunife, J. Singh, K.G. Taylor, R.J. Doyle
DOI: http://dx.doi.org/10.1111/j.1574-6968.1991.tb04891.x 257-262 First published online: 1 August 1991

Summary

A polysaccharide fraction was isolated from sodium-dodecyl-sulfate (SDS) treated cell walls of Bacillus anthracis (Δ Sterne) by hydrofluoric acid (HF) hydrolysis and ethanolic precipitation. The polysaccharide fraction was subsequently purified by several washings with absolute ethanol. Purity of the isolated polysaccharide was tested using the anthrone assay and amino acid analyzer. The molecular mass of the polysaccharide fraction as determined by gel filtration chromatography was about 12 000 Da. Preliminary analyses of the polysaccharide was done using thin layer chromatography and amino acid analyzer, and results obtained from these analyses were further confirmed by gas liquid chromatography and 13CNMR spectroscopy. Results showed that the polysaccharide moiety contained galactose, N-acetylglucosamine, and N-acetylmannosamine in an approximate molar ratio of 3:2:1. This moiety was devoid of muramic acid, alanine, diaminopimelic acid, glutamic acid, and lipid, thus indicating that the isolated polysaccharide was of pure quality.

Key words
  • Cell wall polysaccharide
  • HF-extracted polysaccharide
  • Ethanolic precipitation
  • Bacillus anthracis

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