The structure of the heptose-3-deoxy-D-mannooctulosonic-acid region in a mutant form of Aeromonas salmonicida lipopolysaccharide.

Lipopolysaccharide was isolated from a phage-selected mutant of a wild strain of Aeromonas salmonicida by the aqueous phenol method. The lipopolysaccharide consisted of the R form, containing per mole, three moles of L-glycero-D-manno-heptopyranose, one mole of 3-deoxy-D-manno-2-octulosonic acid (dOclA) and lipid A. The dOclA was not fully assayable by the thiobarbituric acid methods usually used, but its degradation product was detected, after Smith degradation of the lipopolysaccharide, either as free 3-deoxy-2-heptulosonic acid (after hydrolysis) or substituted by a mannopyranosyl residue derived from heptose. Mass spectrometry indicated that the dOclA existed in the furanose form and was substituted by the heptose trisaccharide through position six. Methylation analysis, chemical degradation, chromium trioxide oxidation and nuclear magnetic resonance spectroscopy were used to identify the structure of the core oligosaccharide as: L alpha DHepp(1----2)L alpha DHepp(1----3)L alpha DHepp(1----6)dOclAf(2----.

Structural studies on the O-antigen of Aeromonas salmonicida.

Lipopolysaccharide from a strain of Aeromonas salmonicida salmonicida was isolated from cells by the aqueous phenol method in 2.3% yield (based on dry weight of bacteria). Hydrolysis of the lipopolysaccharide in 1% acetic acid afforded O-polysaccharide (19% by weight), core-oligosaccharide (12.2%) and lipid A (44.6%). Analysis indicated that 3-deoxy-D-manno-2-octulosonic acid was absent from the lipopolysaccharide and that no low-molecular-weight compounds were released by the mild hydrolysis. The O-polysaccharide had the monosaccharide composition of rhamnose, glucose and N-acetylmannosamine in molar ratio of 1.0:1.58:0.83. 75% of the N-acetylmannosamine residues were substituted at position 4 by O-acetyl groups. Hydrolysis of the methylated polysaccharide proved to be both difficult and dependent on the method of hydrolysis chosen, in all cases a partially methylated disaccharide of rhamnose and N-acetylmannosamine was identified in the hydrolysate. Methylation analysis, periodate oxidation and proton magnetic resonance analysis were used to confirm the structure of the repeating unit as: (formula; see text).

Optional production and utilization of polysaccharide by Aeromonas hydrophila.

Glucans from the fish pathogen Aeromonas hydrophila have been extracted and purified by a method utilizing phenol/water followed by sodium deoxycholate rather than the traditional sodium hydroxide extraction. Presence of substantial amounts of these glucans was shown to be dependant on whether or not the substrate contained dextrose, a point which had import because of the low carbohydrate environment in which this species must survive and multiply. These glucans, produced in the log phase, were utilized during the growth period. The structures of the two purified glucans were examined by methylation analysis, periodate oxidation, and enzymatic degradation. The results indicated that A. hydrophila under low-carbohydrate growth conditions produced two similar but distinguishable alpha 1 leads to 4 linked glucans substituted alpha 1 leads to 6 by single monosaccharide residues or short chains to give an amylopectinglucogen type of polysaccharide.