Penicillin sensor was manufactured by immobillizing penicillinase with glutaraldehyde on the $H^+$-selective membrane based on PVC-COOH-TDDA. This membrane was not inter fered by $K^+$ ion in Pc-G potassium salt. When enzyme was immobilized with glutaraldehyde, the PVC-COOH matrix was more effective than PVC matrix. Calibration curve calculated from Nernst equation was not linear. But potential was relative to concentration of Pc-G. And maximal potentiometric velocity was also relative to concentration of Pc-G. Therefore, it may be applied to Michaelis-Menten equation. The penicillin sensor was useful for determination of Pc-G at concentration of 0.1~10mM level.