Composition and molar mass characterisation of bacterial extracellular polymeric substances by using chemical, spectroscopic and fractionation techniques

The chemical composition and molar-mass distribution of extracellular polymeric substances (EPS) produced by the bacterium Sinorhizobium meliloti have been characterised by combining asymmetrical flow field-flow fractionation (AFlFFF), chemical and spectroscopic techniques. The relationship between the EPS composition and molar-mass distribution has been studied by comparing the characteristics of EPS excreted by the wild type S. meliloti and by a mutant deficient in the production of high-molar-mass EPS, as well as by the analysis of total protein content in the collected AFlFFF fractions. Total organic carbon, protein and polysaccharide contents of the EPS were also determined. Obtained results demonstrate the existence of two major populations with weight-average molar masses of 1.40 x 10(5) and 4.57 x 10(5) g mol(-1) respectively. The lower molar-mass population contained predominantly protein-like substances, detectable by UV-VIS spectroscopy, whereas the higher molar-mass population was rich in exopolysaccharides and exoproteins. These findings are in general agreement with the size distributions and chemical heterogeneity observed by nanoparticle tracking analysis, and the characterisation of the composition of all the EPS by different analytical techniques.