Chromosomal genes conferring tolerance to heavy metal (Ag) toxicity

Bacterial strains were isolated from sediment samples from the Thames River. Successive transfer growth of the various strains on nutrient agar containing increasing concentrations of AgNO3 revealed that three of the bacterial isolates were found to be capable of tolerating high concentrations of AgNO3 ranging from 20 to 80 mM on a solid medium and up to 10 mM AgNO3 in liquid medium. Molecular characterization and identification based on 16S rDNA gene sequencing of three strains of bacteria that are tolerant to silver nitrate showed that the major tolerant strains include the superbug, Shewanella oneidensis, Pseudomonas sp. and Bacillus sp. Protein extraction and two-dimensional (2D) sodium dodecyl sulfate SDS-polyacrylamide gel electrophoresis (PAGE) of the protein extracts in bacteria exposed to very high concentrations of AgNO3 revealed a general reduction in the number of expressed proteins, although two protein spots were conspicuously over expressed in the exposed bacteria compared to control. The N-terminal amino acid sequence analysis of the protein spots identified the major up-regulated proteins as the outer membrane protein To1C (45.2 kDa) and the structural protein of the flagellar filament, flagellin (28.34 kDa), encoded for by the to1C and fliC genes, respectively. The roles of these genes in a number of multi-drug resistant pathogen and potentials for biotechnological applications in toxic metal control for treatment of contaminated ecosystems and biomining were discussed. © 2008 Springer Science+Business Media, LLC.