Amperometric urea sensor is more suitable than optical and potentiometric urea sensor to diagnose hyperammonemia. However, because sensitivity in low concentration decreases remarkably, despite amperometric urea sensor has been studied for a long time it has not been applied for clinical diagnosis. In this paper, a new structure for an amperometric urea sensor was fabricated by MEMS, electrochemical etching, and electrostatic covalent binding techniques. Until now most amperometric urea sensors have had a membrane fixed on top of the transducer. That method often leads to malfunction of the sensor, arising from problems such as inadequate membrane adhesion, insufficient mechanical stability, and low sensitivity. To solve these kinds of problems, urease (Urs) was immobilized by electrostatic covalent binding method on the porous silicon (PSi) substrate coated self-assembled monolayer (SAM). Electrostatic covalent binding method was used to keep anisotropic orientation of urease on SAM.