Fluorescence Enhancement from Nitro-Compound-Sensitive Bacteria within Spherical Hydrogel Scaffolds

Abstract

For the safety of both production and life, it is a very significant issue to detect explosive nitro compounds in a remote way or over a long distance. Here, we report that nitro compounds were detected by the bacterial sensor based on hydrogel microbeads as a platform. Green fluorescent protein-producing Escherichia coli, which was genetically engineered to be sensitive to nitro compounds, was loaded within poly(2-hydroxyethyl methacrylate) [poly(HEMA)]-based hydrogel beads, in which fluorescent signals from bacteria were concentrated and strong enough to be easily detected. For efficient loading of negatively charged bacteria, the surface charge of poly(HEMA)-based beads was controlled by copolymerization with 2-(methacryloyloxy)-ethyltrimethylammonium chloride (MAETC) as a cationic monomer. With the addition of MAETC, the cell affinity was nine times enhanced by the interaction between the positively charged poly(HEMA-co-MAETC) beads and negatively charged bacteria. The increased cell affinity resulted in an enhancement of a sensing signal. After exposure to 2,4,6-trinitrotoluene, a typical explosive nitro compound, the fluorescence intensity of bacterial sensors using poly(HEMA-co-MAETC) beads having 80 wt % MAETC was five times increased compared to those based on poly(HEMA) beads. This amplification of the fluorescent signal enables easier detection of explosives efficiently by a remote detection, even over a long distance.