Was Mineral Surface Complexity and Toxicity an Impetus for Evolution of Microbial Extracellular Polymeric Substances?
Modern ecological niches are teeming with an astonishing diversity of microbial life closely associated with mineral surfaces, highlighting the remarkable success of microorganisms in conquering the challenges and capitalizing on the benefits presented by the mineral-water interface. Such community-living is enabled by an extracellular, polymeric, biofilm matrix developed at cell surfaces. Despite the energetic penalties, biofilm formation capability likely evolved on early Earth because of crucial cell survival functions, of which recognized roles include facilitating cell-attachment at mineral surfaces, intercellular signaling and lateral gene transfer, protection from dessication in tidal pools, and screening toxic UV light and toxic soluble metals. Cell-attachment to mineral surfaces was likely critical for cell survival and function, but the potential toxicity of mineral surfaces towards cells and the complexities of the mineral-water-cell interface in promoting biofilm formation, have not been fully appreciated. We examined the effects of nanoparticulate oxides (amorphous SiO2, anatase beta-TiO2, and gamma-Al2O3) on EPS- and biofilm-producing wild-type strains and their isogenic knock-out mutants which are defective in EPS-producing ability. In detail, we used Gram-negative wild-type Pseudomonas aeruginosa PAO1 and its EPS knock-out mutant Delta-psl, and the Gram-positive wild-type Bacillus subtilis NCIB3610 and its EPS-knock-out mutant yhxBDelta. We conducted bacterial growth experiments in the presence of each oxide in order to determine the viability of each cell type relative to oxide-free controls. The amount of EPS generated in the presence of oxides was also quantified and qualitatively analyzed by fluorescent stains. The results indicated a previously unrecognized role for microbial extracellular polymeric substances (EPS) in shielding both Gram-negative and Gram-positive cells against the toxic effects of mineral surfaces. This role is distinct from the protection provided against toxic soluble metals. Furthermore, we found that mineral toxicity is specific to the surface chemistry and particle size of the mineral, and that EPS protect against this mineral-specific toxicity via different mechanisms. Most intriguingly, we determined that EPS production is mineral-induced. By addressing the mechanistic detailed interactions at the mineral-water-cell interface, we provide insight to the potential impact of nanoparticulate mineral surfaces in promoting increased complexity of cell surfaces, including EPS and biofilm formation, on early Earth.