The characteristics of a glucose sensor based on an ion-sensitive TiO2/Ti extended gate electrode field-effect transistor (EGFET) are reported. A glucose oxidase-containing silk fibroin membrane was immobilized on a TiO2/Ti surface as the bio-sensing component. This EGFET-type biosensor was estimated to be able to detect a glucose concentration as low as 0.001 mg/mL in an aqueous electrolyte, which enables the sensing of glucose in the saliva and sweat. The endurance of this sensor was also examined, and it was found that the retention time of the original sensitivity for repeated use at room temperature was more than 30 days, with a high heat tolerance temperature close to 60 °C.