Identification of Molecular and Cellular Responses of Desulfovibrio vulgaris Biofilms under Culture Conditions Relevant to Field Conditions for Bioreduction

Desulfovibrio vulgaris ATCC29579 is a sulfate- reducing bacterium (SRB) that is commonly used as a model for direct and indirect heavy metal reduction, and can also be a causitative agent of metal corrosion. During growth with lactate and sulfate, internal carbohydrate levels increased throughout exponential-phase, and peaked as the cells transitioned to stationary-phase. The carbohydrate to protein ratio (C:P) peaked at 0.05 ug/ug as the cells transitioned to stationary-phase, and then declined to 0.02 ug/ug during extended stationary-phase. In contrast, a strain of D. vulgaris that does not contain the megaplasmid, maintained higher internal carbohydrate levels and the C:P ratio peaked at 0.1 ug/ug (2-fold increase compared to wild-type). Under the tested growth conditions, we observed biofilm formation in wild-type cells, but the plasmid-less strain formed less biofilm (2-fold decrease). We hypothesized that carbohydrate was re-allocated to the external cell proper for biofilm formation. However, biofilm contained relatively little carbohydrate (0.6 to 1.0 ug/ml) and had a similar C:P ratio compared to wild-type early stationary-phase cells. Staining with calcafluor white also indicated the presence of little external carbohydrate in D. vulgaris biofilms. Less biofilm was formed in the presence of protinease K, trypsin, and chymotrypsin, however, the growth of planktonic cells was not affected. In addition, when D. vulgaris biofilm was treated with a protease, less biofilm was observed. Electron micrographs suggested the presence of filaments between the biofilm cells, and filaments appeared to be susceptible to protease treatment. Biofilm filtrates contained soluble protein, and SDS-PAGE analysis suggested different polypeptide profiles between a filtrate, a planktonic, and a biofilm sample