Multi-targets detection via combination of multi-stimulus-response engineered bacteria and hydrogel-based separation platform.

Establishing a method similar to ICP-MS that can quantitatively analyze multiple heavy metals simultaneously, conveniently, and in situ is highly anticipated. In this study, we integrated the sensing elements of multiple targets and different fluorescence reporting elements to construct an engineered Escherichia coli. When these targets are present, the engineered bacteria can emit a fluorescent signal at the corresponding wavelength. To avoid the inability to accurately distinguish and quantify the content of each target due to the overlap of fluorescence signals when multiple targets coexist, a hydrogel-based separation platform similar to a separation column was constructed. The hydrogel platform can change the detection limit (LOD) and sensitivity by adjusting the adsorption strength towards different targets, so as to realize the differentiation and recognition of their respective detection signals. The LODs of this new detection method for Cd(II), Hg(II), As(III), and Pb(II) are 1.249, 0.380, 3.917, and 0.755 μg/L, respectively. In addition, this biosensor system was applied to detect coexisting Cd(II), Hg(II), As(III), and Pb(II) in actual samples with a recovery rate of 85.61-110.30 %, which is consistent with the classical ICP-MS detection results, confirming the accuracy and reliability of the method for detecting multiple heavy metal coexisting samples.