Construction of two lux-tagged Hg2+-specific biosensors and their luminescence performance

Two Hg(2+)-specific biosensors were constructed using bacterial luciferase as reporter gene and plasmid-free Pseudomonas putida X4 and Enterobacter aerogenes NTG-01 as host strains. The performance of X4 biosensor was compared with that of NTG-01 biosensor in the same assay conditions. The maximum bioluminescence for X4 (pmerRluxCDABE-Kan) biosensor was found during the midexponential phase and that for NTG-01 (pmerRluxCDABE-Kan) was at the late exponential phase. The shortest induction time of two biosensors was 30 min. The maximum light signal output for NTG-01 and X4 sensors was observed at the incubation time of 5 and 4 h, respectively. The lowest detectable concentration of mercury by the two biosensors were both of 100 pM at 28 degrees C, pH 7 and an initial cell number of 10(6) CFU ml(-1). Cd(2+), Zn(2+), Co(2+), Cu(2+), and Pb(2 +) ions at nanomolar level did not interfere with the measurement by the biosensors. These results show that the sensitivity of the two biosensors is sufficient for the detection of Hg(2+) under most contaminated environments.