An amperometric biosensor for glucose based on a novel polymeric matrix is described. The sensor was prepared by electrochemical polymerization of p-aminophenol in the presence of glucose oxidase on Pt substrates in a KCl solution at a potential of 0.6 V (vs. Ag/AgCl). The chronoamperometric response of poly(p-aminophenol)-glucose oxidase biosensor to hydrogen peroxide formed as the result of enzymatic reaction was measured at a potential of 0.7 V (vs. Ag/AgCl) in PBS solution. It was shown that a biosensor prepared by this onestep procedure has a linear range up to 6 mM glucose, a response time less than 5 s and a lifetime around 15 days. The sensitivity of the electrode was superior to that of much publicized polypyrrole based glucose oxidase electrode. Also, the substrate selectivity of PAP/GOx electrode was tested for coexisting substances such as lactose, sucrose, urea, paracetamol, uric and ascorbic acid and no discernible anodic response was observed above the background current.