Structure elucidation and gene cluster annotation of the O-antigen of Escherichia coli O39; application of anhydrous trifluoroacetic acid for selective cleavage of glycosidic linkages

•Structure of the O-polysaccharide of Shiga toxin-producing E. coli O39 was established.•Solvolysis with trifluoroacetic acid was applied to cleave selectively the rhamnosidic linkage.•The O-antigen gene cluster of E. coli O39 has been sequenced.

O-Polysaccharide (O-antigen) accompanied by a minor mannan was isolated from the lipopolysaccharide of Escherichia coli O39 and studied by component analyses, methylation, Smith degradation, mass spectrometry, and 1D and 2D NMR spectroscopy. In addition, a new approach, solvolysis with anhydrous trifluoroacetic acid, was applied to cleave selectively the rhamnosidic linkage. The following structure of the O-polysaccharide was established:Figure optionsDownload full-size imageDownload as PowerPoint slide where d-Qui4N(R3Hb) indicates 4,6-dideoxy-4-[(R)-3-hydroxybutanoylamino]-d-glucose. The O-antigen gene cluster of E. coli O39 has been sequenced. The gene functions were tentatively assigned by a comparison with sequences in the available databases and found to be in agreement with the O-polysaccharide structure.

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