Structural investigation of the antagonist LPS from the cyanobacterium Oscillatoria planktothrix FP1

Cyanobacteria are aquatic and photosynthetic microorganisms, which contribute up to 30% of the yearly oxygen production on the earth. They have the distinction of being the oldest known fossils, more than 3.5 billion years old, and are one of the largest and most important groups of bacteria on earth. Cyanobacteria are an emerging source of potentially pharmacologically active products and, among these, there are the lipopolysaccharides. Despite their significant and well documented activity, very little is known about the cyanobacteria lipopolysaccharides (LPS) structure. The aim of this work is to investigate the structure of the highly TLR4-antagonist lipopolysaccharide from the cyanobacterium Oscillatoria plankthotrix FP1. The LPS was purified and analysed by means of chemical analysis and H-1 and C-13 NMR spectroscopy. The LPS was then degraded by Smith degradation, HF and acetic acid hydrolyses. All the obtained products were investigated in detail by chemical analysis, NMR spectroscopy and by mass spectrometry. The LPS consists of a high molecular mass and very complex molecule lacking Kdo and heptose residues, where the polysaccharide chain is mainly constituted by a backbone of 3-substituted alpha-L-rhamnose units. The core region is rich in galacturonic acid and mannose residues. Moreover a glycolipid portion, similar to Gram-negative lipid A, was identified. This was built up of a non phosphorylated (1'-> 6) linked glucosamine disaccharide, acylated with 3-hydroxylated fatty acids. In particular 3-hydroxypentadecanoic and 3-hydroxyesadecanoic acids were found, together with esadecanoic and tetradecanoic ones. Finally the presence of a galacturonic acid residue at 6-position of the distal glucosamine in place of the Kdo residue is suggested. (C) 2013 Elsevier Ltd. All rights reserved.