Single and combined enzymes were tested in Pseudomonasfluorescens biofilm removal, evaluating both the cell and biomass removal yields. Amongst the three single polysaccharide-degrading enzymes tested, pectin esterase was the most effective as it removed about three quarters of the biofilm cells. Soluble Pronase removed about 30% of the biofilm biomass and 80% of the cells. Sequential (two-step) treatment with cellulase, pectin esterase or pectin lyase, followed by soluble Pronase, obtained a 2-3 log 10 count reduction and up to 94% biomass removal. One-step treatment was performed using a newly developed form of delayed release encapsulated Pronase (DRE Pro) that allows soluble accompanying enzymes to work for 20-60 min before proteolytic onset. Combinations of the DRE Pro with certain polysaccharide-degrading enzymes achieved up to 4 log(10) reductions in biofilm cells and 96% removal of the biofilm mass, 2 h after a single product application at 25 degrees C. (c) 2006 Elsevier Inc. All rights reserved.