Glucose oxidase (GOx), a flavoprotein enzyme with the nomenclature of E.C.1.1.3.4, is widely employed in the different sectors such as an implantable fuel cell. In this paper, optimal conditions for the size of carbon paste, GOx volume, and glutaraldehyde concentrations for fabrication of polyaniline nanofiber modified carbon paste electrodes (GOx/MCPE) were investigated. The optimum parameters of the carbon paste size, GOx volume, and glutaraldehyde concentration were found to be 3 mm x 3 mm, 5 µL and 0.5%, respectively. The maximum current in carbon paste size (3 mm x 3 mm), GOx volume (5 µL) and glutaraldehyde concentration (0.5%) were estimated to be 5.39 mA, 4.12 mA, and 7.10 mA, respectively. This GOx/MCPE showed great potential in fuel cell application.