The electrophoretic deposition of glucose oxidase from water using asymmetrical alternating voltages is investigated. Using asymmetric voltages, glucose oxidase layers with a thickness of 7 mu m could be deposited on a platinum electrode in 20 min time as verified with a microbalance, carbon analysis and scanning electron microscopy. In contrast, if a symmetrical alternating signal is used under the same conditions, a layer of 0.5 mu m is formed. We believe the deposition is due to two effects: the electrophoretic migration of the enzyme towards the deposition electrode and the pH induced precipitation of the enzyme near the deposition electrode. The electrophoretic migration is due to the non-linear dependence of the electrophoretic mobility on the electric field caused by the asymmetry of the applied alternating current signal. In addition, pH changes near the deposition electrode drive the enzyme towards its point of zero charge (PZC), perhaps causing the precipitation of GOx on the substrate. The effect of amplitude, frequency, deposition time and GOx concentration on the deposition rate was studied. An amplitude of 160 Vp-p and a frequency of 30 Hz was found to be optimal for the formation of thick enzyme layers, which excludes a big part of the interferences. (C) 2009 Elsevier B.V. All rights reserved.