E. coli TG1 pBS TOM Green was cultured to produce toluene-o-monooxygenase (TOM). A biosensor system was successfully constructed using purified TOM to effectively detect trichloroethene (TCE) and tetrachloroethene (PCE), which represent some of the major contaminants in groundwater and soil. In order to utilize TOM as a sensor, NADH, a biological oxidizer, was replaced with hydrogen peroxide which is a chemical oxidizing agent. A three-layered sandwich-type sensing tip was fabricated on the outside of the hydrophilic polyvinylidene fluoride membrane. TCE and PCE were applied to the sensor and the hydrogen ions were measured by a fiber optic fluorometer using fluoresceinamine. Calibration curves were obtained for TCE and PCE in the concentration range of 0.2-100 mg/l, and the detection limit of the system was $10{\mu}g/l$ for TCE and PCE.

Keywords: TCE;PCE;Biosensor;Shunting;Fiber optic;