Polypyrrole nanoparticles were successfully synthesized in cetyltrimethyl ammonium bromide (CTAB)/hexanol/water reverse micelle. The morphology and particle size of the obtained nanoparticles were characterized with transmission electron microscope (TEM) and scanning electron microscopy (SEM). Glucose biosensors were formed with glucose oxidase (GOx) immobilized in conducting composite material consisting of polypyrrole nanoparticles and ethyl cellulose. The effects of reaction conditions such as molar ratio of polypyrrole nanoparticles to ethyl cellulose, working voltage, glucose concentration, temperature and solution pH on the electrochemical response of the GOx electrode were studied. Experimental results showed that the linear range of GOx electrode was 1.0 x 10(-6)-6 x 10(-3) mol/L and the detection limit was 1.0 x 10(-7) mol/L. The electrode exhibited fine repeatability and selectability, and its lifetime was greater than one month. AFM showed that the surface of conducting composite material-glucose oxidase electrode's presents uniform granular after washing paraffin wax with cyclohexane, which was favorable for enzyme-catalyzed reaction.