An amperometric biosensor has been developed for the measurement of urea. To construct the proposed biosensor urease enzyme was immobilized onto poly(N-glycidylpyrrole-co-pyrrole) conducting film by direct covalent attachment. The biosensor surface was characterized by FTIR spectroscopy and atomic force microscopy (AFM). The response studies were carried out as a function of urea concentration with amperometric measurements. The biosensor showed a linear current response to the urea concentration ranging from 0.1 to 0.7 mM. The urea biosensor exhibited a sensitivity of 4.5 μA/mM with a response time of 4 s. The factors influencing on the performance of the resulting biosensor were also studied in detail.