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Detection of 2-keto-3-deoxyoctonate in endotoxins isolated from six reference strains of the Bacteroides fragilis group
Authors:I Beckmann  H G van Eijk  F Meisel-Miko?ajczyk  H C Wallenburg
Institution:Department of Obstetrics and Gynecology, Erasmus University, Rotterdam, The Netherlands.
Abstract:1. Endotoxins isolated from six serotype specific reference strains of the Bacteroides fragilis group were dephosphorylated by treatment with aqueous 50% hydrofluoric acid. 2. Mild acidic hydrolysis of the dephosphorylated endotoxins released 2-keto-3-deoxyaldonic acid, the presence of which was demonstrated by the colorimetric thiobarbituric acid assay (TBA). 3. Thin layer chromatography of the dephosphorylated lipopolysaccharide of B. fragilis IPL E 323 (serotype E2), after acidic hydrolysis, revealed a TBA-positive substance with the same Rf-value as authentical 2-keto-3-deoxyoctolusonic acid (KDO). 4. Quantification of 2-keto-3-deoxyoctonate-in the lipopolysaccharide of B. fragilis IPL E 323 by means of the TBA resulted in a KDO content of 15 nM mg-1 lipopolysaccharide.
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