Characterization of the extracellular polymeric substances (EPS) of Virgibacillus strains capable of mediating the formation of high Mg-calcite and protodolomite

Abstract

The origin of dolomite –a common mineral in the geological record– is the subject of an ongoing debate. Among different hypotheses, it has been proposed that extracellular polymeric substances (EPS) excreted by microbes include organic molecules that catalyze the incorporation of Mg in the carbonate mineral. However, limited information exists on the composition of the EPS produced by CaMg carbonate-forming microbes, which in turn hampers a precise understanding of their role in the mineralization mechanism. Here, we present the results of laboratory experiments in which we cultured different strains of microbes, characterized their EPS, and identified components associated with carbonate minerals with high mol% Mg. Two Virgibacillus strains known to mediate the formation of Mg-rich carbonates, as well as a strain of Bacillus licheniformis –a negative control that does not mediate mineral formation but produces EPS, were grown under different salinities and temperatures, which caused them to produce EPS with different compositions. The EPS were subsequently characterized by measuring total carbohydrate (TCHO) and total protein (TP) contents, as well as by Fourier-transform infrared spectroscopy (FTIR). At the tested conditions, we found that Mg-carbonates with a mol% Mg higher than 40% (i.e., potential dolomite precursor phases) formed exclusively in association with EPS rich in carbohydrates (TCHO > than 75% of the total mass). FTIR spectra of CaMg carbonate-forming strains were distinct from those of the non-mineral-forming strain in areas associated with the protein structures responsible for the formation of hydrogels, which contribute to hydration or dehydration of ionic clusters; further differences have been observed in the regions of phosphoryl functional groups. These results provide insight on which fraction of organic molecules and specific functional groups are, among the many constituents of EPS, important for mineral nucleation and incorporation of Mg into carbonate minerals, a crucial step for the formation of dolomite in natural environments.