The
ability to detect uranium (U) through environmental monitoring
is of critical importance for informing water resource protection
and nonproliferation efforts. While technologies exist for environmental
U detection, wide-area environmental monitoring, i.e. sampling coverage
over large areas not known to possess U contamination, remains a challenging
prospect that necessitates the development of novel detection approaches.
Herein, we describe the development of a whole-cell U sensor by integrating
two functionally independent, native U-responsive two-component signaling
systems (TCS), UzcRS and UrpRS, within an AND gate circuit in the
bacterium Caulobacter crescentus. Through leverage
of the distinct but imperfect selectivity profiles of both TCS, this
combinatorial approach enabled greater selectivity relative to a prior
biosensor developed with UzcRS alone; no cross-reactivity was observed
with most common environmental metals (e.g, Fe, As, Cu, Ca, Mg, Cd,
Cr, Al) or the U decay-chain product Th, and the selectivity against
Zn and Pb was significantly improved. In addition, integration of
the UzcRS signal amplifier protein UzcY within the AND gate circuit
further enhanced overall sensitivity and selectivity for U. The functionality
of the sensor in an environmental context was confirmed by detection
of U concentrations as low as 1 μM in groundwater samples. The
results highlight the value of a combinatorial approach for constructing
whole-cell sensors for the selective detection of analytes for which
there are no known evolved regulators.