A method of irreversibly immobilizing an enzyme in a polyurethane and an enzyme-containing polyurethane having a degree of immobilization of the enzyme of approximately 100%. The synthesis of waterborne polyurethanes in the presence of enzyme has enabled the irreversible attachment of the enzyme to the polymeric matrix. The distribution of immobilized enzyme as well as activity retention are homogeneous within the polyurethane. Decreasing ECC hydrophobicity, via the use of a less hydrophobic polyisocyanate prepolymer during polymerization, significantly enhanced the intrinsic activity of the ECC.