Amperometric bienzyme sensors were constructed based on solid graphite electrodes modified with polymer redox mediator, horseradish peroxidase and a variety of oxidases and covered with a bipolar electrostatic protective layer. Glucose, ethanol, choline, glycero-3-phosphate and cholesterol sensors were designed and operated at 0 mV vs. Ag/AgCl using single potential step chronoamperometry. The protection of the sensor against interference from ascorbic acid and linearity of response were provided by a bipolar electrostatic layer. Sensitivities as high as 0.74 mu A cm(-2) mM(-1) of glucose, 0.71 mu A cm(-2) mM(-1) of ethanol 5.7 mu A cm(2) mM(-1) of glycerol-1-phosphate, and 28.3 nA cm(-2) per 10 mu M of choline, were achieved.