Due to water scarcity, alternative water management strategies are now being implemented. To control this, general toxicity bacterial sensors are being developed for the timely detection of toxics through their side effects on bacterial proliferation. Main limitation of these biosensors is on the composition of the biorecognition membranes, since traditional synthetic polymers rely on poorly biocompatible polymerization processes and natural biomaterials present low stability. Silicate-rich inorganic materials present excellent bioaffinity and long-term stability, as well as low production costs and ease functionalization but, until now, This work describes the use of Montmorillonite clay (MMT) based ionic liquids in the development of a low-cost amperometric microbial biosensors for the rapid detection of chemical pollutants, here demonstrated by the pesticide 3.5-Diclorophenol (DCP). In the biosensor, Escherichia coli (E.coli) are immobilized in the clay matrix doped with ionic liquids as sensing element and optimized to guarantee maximal bacterial retention and metabolic activity. The metabolic activity is determined using ferricyanide, which reduced by bacteria into ferrocyanide, thus producing an electrochemically quantifiable signal. The toxicity assessment is completed in 30 minutes by incubating the sample under study with the sensor and detecting ferrocyanide production at 37°C. Under optimal conditions, the results showed the inhibitory effect of the DCP pesticide on bacterial growth at concentrations of 8.25 ppm, then finding a maximum mean effective concentration (CE 50 ) of 8.0 ppm. Biosensor developed is presented as an effective alternative in the development of low cost and high sensitivity portable systems for preventive monitoring of environmental pollution.