Quorum sensing, in
which bacteria communities use signaling molecules
for inter- and intracellular communication, has been intensively studied
in recent decades. In order to fabricate highly sensitive easy-to-handle
point of care biosensors that detect quorum sensing molecules, we
have developed, as is reported here, reporter bacteria loaded alginate–methacrylate
(alginate-MA) hydrogel beads. The alginate-MA beads, which were obtained
by electrostatic extrusion, were reinforced by photo-cross-linking
to increase stability and thereby to reduce bacteria leaching. In
these beads the genetically engineered fluorescent reporter bacterium Escherichia coli pTetR-LasR-pLuxR-GFP (E. coli pLuxR-GFP) was encapsulated, which responds to the autoinducer N-(3-oxododecanoyl)homoserine lactone secreted by Pseudomonas aeruginosa. After encapsulation in alginate-MA
hydrogel beads with diameters in the range of 100–300 μm
that were produced by an electrostatic extrusion method and rapid
photo-cross-linking, the E. coli pLuxR-GFP were found
to possess a high degree of viability and sensing activity. The encapsulated
bacteria could proliferate inside the hydrogel beads, when exposed
to bacteria culture medium. In media containing the autoinducer N-(3-oxododecanoyl)homoserine lactone, the encapsulated
reporter bacteria responded with a strong fluorescence signal due
to an increased green fluorescent protein (GFP) expression. A prototype
dipstick type sensor developed here underlines the potential of encapsulation
of viable and functional reporter bacteria inside reinforced alginate–methacrylate
hydrogel beads for whole cell sensors for bacteria detection.