Structural characterization of the lipid A component of Bacteroides fragilis strain NCTC 9343 lipopolysaccharide

Abstract

The chemical structure of Bacteroides fragilis NCTC 9343 lipid A was characterized by using conventional chemical procedures, methylation analysis, and laser desorption mass spectrometry. It was found that B. fragilis lipid A consists of a β‐D‐glucosaminyl‐(1–6)‐D‐glucosaminyl‐1‐O‐phosphate backbone whose hydroxyl groups in positions 4, 4′ and 6′ are free, the latter serving as the attachment site for the polysaccharide component in lipopolysaccharide. This backbone molecule carries up to of five molecules of ester‐ and amide‐bound long chain non‐hydroxylated and (R)‐3‐hydroxy fatty acids. With regard to the distribution of the fatty acids on the lipid A backbone, a considerable heterogeneity was revealed by laser desorption mass spectrometry. Despite this heterogeneity, a major species of B. fragilis lipid A could be defined in which the hydroxyl group at position 3′ of the distal GlcN carries (R)‐3‐hydroxyhexadecanoic acid and the hydroxyl group at position 3 of the reducing GlcN is acylated by (R)‐3‐hydroxypentadecanoic acid. The amino group of the distal GlcN residue carries (R)‐3‐(13‐methyltetradecanoyloxy)‐15‐methylhexadecanoic acid and that of the reducing GlcN group (R)‐3‐hydroxyhexadecanoic acid. The absence of ester‐bound phosphate and ester‐linked 3‐acyloxyacyl groups, the presence of not more than five acyl residues and the predominance of fatty acids possessing 15–17 carbon atoms are unique features of B. fragilis lipid A which differentiate it from enterobacterial and other lipids A and which are likely to be related to its low endotoxic activity.