Studies on the structure and the solution conformation of an acidic extracellular polysaccharide isolated from Bradyrhizobium

The structure of an acidic extracellular polysaccharide isolated from Bradyrhizobium (Chamaecytisus proliferus) has been elucidated by hydrolysis, methylation analysis, and 1D and 2D 1H- and 13C-NMR spectroscopy of the complete polysaccharide. The NMR spectrum showed that microheterogeneity was present due to the minor existence of a variety of O-acetyl groups. Thus, a deacetylated sample was prepared by alkaline treatment which was then fully analysed. The deacetylated polysaccharide has the following sequence: The sample is partially O-methylated at position 4 of the α-d-Galp-(1 → 6) unit. In addition, the same moiety of the native sample is also partially and heterogeneously O-acetylated. The conformational features of the deacetylated sample have been evaluated by molecular mechanics and dynamics calculations and NOE spectroscopy. The results indicate that the polysaccharide may adopt a variety of three dimensional shapes, and that there is a fair agreement between the NMR-derived distances and those provided by the calculations.