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

Abstract

Environmental contextExtracellular polymeric substances (EPS) released by microorganisms are an important component of organic matter in the environment. EPS play an essential role in cell adhesion to surfaces, biofilm and floc formation, soil aggregation and stability and in the activated sludge of waste water treatment plants. EPS are complex mixtures containing components of different chemical nature and molecular size, which make their characterisation difficult. The present work explores the link between chemical composition and molar-mass distribution of the EPS released by the bacterium Sinorhizobium meliloti by using a combination of chemical, spectroscopic and fractionation techniques. AbstractThe 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 × 105 and 4.57 × 105 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.