Purification and characterization of a soluble polyurethane degrading enzyme from Comamonas acidovorans

A soluble esterase involved in the biodegradation of polyester polyurethane (PU) was purified to apparent electrophoretic homogeneity in high yield, s imilar to 83%. The enzyme displayed a single band on both non-denaturing (N D-) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE ) with an apparent molecular mass of 42 kDa. Using rho-nitrophenylacetate a s the substrate, the enzyme displayed steady-state kinetic parameters K-m a nd V-max of 51.5 mM and 180 U mg(-1) respectively. Esterase activity was th ermally stable and could be inhibited with phenylmethylsulfonylfluoride (PM SF) and soybean trypsin inhibitor (STI). (C) 1999 Elsevier Science Ltd. All rights reserved.