Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains.

Journal of Bacteriology
R P Darveau, R E Hancock

Abstract

Lipopolysaccharide (LPS) is a major component of the outer membrane of gram-negative bacteria. It is now well established that within a single organism, size heterogeneity of this molecule can exist. We have developed a LPS isolation procedure which is effective in extracting both smooth and rough LPS in high yields (51 to 81% of the LPS present in whole cells as quantitated by using hydroxy fatty acid, heptose, and 2-keto-3-deoxyoctonate yields) and with a high degree of purity. The contamination by protein (0.1% by weight of LPS), nucleic acids (1%), lipids (2 to 5%), and other bacterial products was low. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the LPS demonstrated the presence of a high degree of size heterogeneity in the isolated smooth LPS as well as the presence of significant amounts of rough-type LPS. The Pseudomonas aeruginosa LPS interacted well with a monoclonal antibody in a variety of immunochemical analyses. The usefulness of the procedure was demonstrated by comparing LPS preparations obtained from wild-type and mutant strains of P. aeruginosa and Salmonella typhimurium. For example, it was shown that the LPS of an antibiotic supersusceptible mutant Z61 of P. aeruginosa, which was previously ...Continue Reading

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Citations

Jan 1, 1989·Archives of Microbiology·A SonessonJ Eng
Aug 15, 1994·Annals of the New York Academy of Sciences·R F RestD Simon
Jan 1, 1994·Antonie van Leeuwenhoek·J P Maskell
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Sep 24, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Stephen R CoatsRichard P Darveau
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May 28, 2008·Journal of Periodontal Research·C BodetD Grenier
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May 31, 2014·BioMed Research International·James LiaoDaniel Grenier
Nov 12, 2014·Journal of Basic Microbiology·Periyanaina KesikaKrishnaswamy Balamurugan
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Aug 25, 2020·Journal of Biomedical Science·Tzu-Wei ChiuYuan Chuan Lee
Jan 1, 1990·Critical Reviews in Microbiology·Y A Knirel

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