The structure of the antigenic O-polysaccharide (O-PS) of the lipopolysaccharide (LPS) produced by the enterohemorrhagic strain of Escherichia coli O48:H21 (EHEC) has been elucidated. The O-PS obtained by mild acid hydrolysis of the LPS had [alpha](D) +95 (water) and was composed Of L-rhamnose (L-Rha), D-galactose (D-Gal), 2-amino-2-deoxy-D-glucose (D-GlcN), 2-amino-2-deoxy-D-galactose (D-GalN), and D-galacturonic acid (D-GalA) (1: 1: 1: 1: 1). From the results of methylation analysis, mass spectrometry, 2D NMR, and DOC-PAGE, the O-PS was shown to be a high molecular mass polymer of a repeating pentasaccharide unit having the structure: [GRAPHICS] The D-GalpA non-reducing end groups in the O-PS were partially O-acetylated (similar to 30%) at the O-2 and O-3 positions and the degree of acetylation was variable from batch to batch cell production. Crown Copyright (c) 2006 Published by Elsevier Ltd. All rights reserved.