A new conductometric biosensor has been developed for the determination of short chain primary aliphatic alcohols. The biosensor assembly was prepared through immobilization of alcohol oxidase from Hansenula sp. and bovine liver catalase in a photoreticulated poly(vinyl alcohol) membrane at the surface of interdigitated microelectrodes. The local conductivity increased rapidly after alcohol addition, reaching steady-state within 10min. The sensitivity was maximal for methanol (0.394±0.004μSμM −1 , n=5) and decreased by increasing the alcohol chain length. The response was linear up to 75μM for methanol, 70μM for ethanol and 65μM for 1-propanol and limits of detection were 0.5μM, 1μM and 3μM, respectively (S/N=3). No significant loss of the enzyme activities was observed after 3 months of storage at 4°C in a 20mM phosphate buffer solution pH 7.2 (two or three measurements per week). After 4 months, 95% of the initial signal still remained. The biosensor response to ethanol was not significantly affected by acetic, lactic, ascorbic, malic, oxalic, citric, tartaric acids or glucose. The bi-enzymatic sensor was successfully applied to the determination of ethanol in different alcoholic beverages.