A novel technique for enzyme immobilization on the surface of the Au electrode for designing amperometric biosensor by electrospinning poly(vinyl alcohol) (PVA) and glucose oxidase (GOD) is presented in this paper. The immobilized GOD remained active inside the electrospun PVA fibrous membranes. The membranes are promising candidates for immobilization of enzymes because of their high specific surface area and porous structure. The infrared spectrum, the UV-Vis spectrum, and the scanning electronic microscope of the membranes showed that the enzyme had been immobilized inside the PVA membranes. Chronoamperometric measurements demonstrated that electrospun fibrous enzymatic electrodes exhibited a rapid response (1 s) and a higher response current (mu A level) to glucose in the normal and diabetic level. The linear response range (from 1 to 10 mM) and the lower detection limit (0.05 mM) of the sensor are satisfying. The electrospun method makes it convenient and efficient to prepare the enzymatic electrode for biosensors. (c) 2006 Elsevier B.V. All rights reserved.