The effect of enzyme pre-treatment on dewaterability of anaerobically digested sludge was investigated at both laboratory and pilot scale. Our results revealed a significant increase in cake solid content (27% cake solids compared to 18% without enzyme pre-treatment), using an enzyme dose of only 20 mg/L. In order to assess practical application, enzyme pre-treatment was applied at the Wilmington, Delaware (U.S.) wastewater treatment plant, using a pilot-scale centrifuge. However, the efficiency reached in laboratory scale could not be obtained in pilot scale, where the final cake solids content did not exceed 20%. Centrifuge and belt filter press (simulated by Crown Press) dewatering were compared in terms of the process efficiencies in the absence and presence of enzyme pre-treatment. Possible factors that might cause the differences were tested by experimental and statistical comparisons. Results indicated that the higher shear applied in centrifugation is responsible for the lack of improved cake solids. The network strength of sludge determined by rheological measurements revealed that enzymatic treatment weakens the gel structure of the sludge floc through the hydrolysis of extracellular polymeric substances; this allows improved dewatering by filtration processes, but leads to floc deterioration when subjected to high shear during centrifugation.