An optical biosensor using an electrically controlled-release system was developed for the measurement of peroxide concentration. The electrically controlled-release system consisted of a current-supplying system and a polymer complex by hydrogen bonding between the carboxylic and oxazoline group. The polymer complex was formed below pH 5.0 and was degraded above pH 5.4. The local pH change near the surface of the polymer complex could be controlled by applying the electric current to release an enzyme reaction reagent, 4-hydroxyphenylacetic acid (HPA), in the polymer complex. The releasing rate of HPA was proportional to the electric current applied to the polymer complex. The model of the controlled-release system was proposed to predict the degradation velocity of the polymer complex, which is equivalent to the releasing rate of HPA. The released HPA and analyte, peroxide, flowed into the reactor with the immobilized enzyme and then reacted with the enzyme. The peroxide concentration was measured based on the fluorescence detection of enzyme reaction product, 6,6'-dihydroxy (1,1'-biphenyl) 3,3'-diacetic acid (DBDA). The proposed biosensor had the linear analytical range of 0.025 approximately 1.0 mM with a response time of 20 min, good repeatability, and reproducibility.