The effect of degradation of polymeric substrates (starch and soy proteins mixture) on the structure of biofilms has been studied. The characteristics of the obtained biofilms were compared to those obtained on corresponding monomeric substrates (glucose and aspartic acid). Based on literature suggestions it was hypothesized that the polymeric substrates, which have a low diffusion rate in the biofilm matrix, would affect the biofilm structure if hydrolytic activity occurs in the biofilm. The obtained biofilm could be expected to present properties like low density and rough surface, facilitating transport and conversion of large polymeric molecules. From the present study it was concluded that the structure of the formed biofilms was influenced by the substrate degraded, however no unequivocal effect of degradation of a polymer on the biofilm structure could be observed. The hydrolytic activity with soy protein and starch as substrate was under stable conditions found to be mainly associated to the biofilm (more than 95% of the total activity). During unstable conditions or start-up significant hydrolytic activity occurred outside the biofilm. © 2002 Elsevier Science Ltd. All rights reserved.