The electrophoretic deposition of carbon nanotubes (CNT) on glassy carbon electrode was used here for the development of porous matrix with enhanced electroactive surface area. The prepared layer displayed a catalytic activity toward NADH oxidation and was used as a support for the d-sorbitol dehydrogenase immobilization. The electrochemically assisted deposition (EAD) method was performed to generate around carbon nanotubes sol–gel biocomposite films containing the enzyme and a cationic polyelectrolyte. The response of the immobilized enzyme strongly depended on the parameters of sol–gel deposition as well as the thickness of carbon nanotubes layer. The electrocatalytic oxidation of d-sorbitol using such modified electrode was used here as a model to discuss the interest of such silica–carbon nanotubes composite in the field of bioelectrochemistry and biosensors.