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. 1979 May;138(2):361-9.
doi: 10.1128/jb.138.2.361-369.1979.

Purification and characterization of smooth and rough lipopolysaccharides from Brucella abortus

E Moreno, M W Pitt, L M Jones, G G Schurig, D T Berman

Purification and characterization of smooth and rough lipopolysaccharides from Brucella abortus

E Moreno et al. J Bacteriol. 1979 May.

Abstract

In an attempt to obtain pure and well characterized smooth lipopolysaccharide (S-LPS) and rough lipopolysaccharide (R-LPS), smooth and rough strains of Brucella abortus were extracted by two different modifications of the phenol-water method. S-LPS was obtained in the phenol phase, and R-LPS was obtained in the aqueous phase. Further purification was accomplished by treatment with enzymes, detergents, NaI as a chaotropic agent to separate non-covalently bound contaminants, and by gel filtration. The degree of purity of the molecules was determined by chemical and immunological analysis and by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. Lipid identification by gas-liquid chromatography showed seven major fatty acids. Palmitic acid accounts for about 50%, stearic acid accounts for about 10%, and hydroxylated fatty acids account for less than 5% of total fatty acids. 2-Keto-3-deoxyoctonate but not heptose was detected in the sugar analysis. Protein was found to be firmly bound to S-LPS but not to R-LPS.

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