The fabrication of novel uranyl (UO(2)(2+)) binding protein based sensors is reported. The new biosensor responds to picomolar levels of aqueous uranyl ions within minutes using Lysinibacillus sphaericus JG-A12 S-layer protein tethered to gold electrodes. In comparison to traditional self assembled monolayer based biosensors the porous bioconjugated layer gave greater stability, longer electrode life span and a denser protein layer. Biosensors responded specifically to UO(2)(2+) ions and showed minor interference from Ni(2+), Cs(+), Cd(2+) and Co(2+). Chemical modification of JG-A12 protein phosphate and carboxyl groups prevented UO(2)(2+) binding, showing that both moieties are involved in the recognition to UO(2)(2+).
Keywords: S-layer; impedance spectroscopy; metal ion; protein biosensor; sequestering; surface layer; uranium; uranyl.