Growth of Pseudomonas fluorescens on a polyester-polyurethane and the purification and characterization of a polyurethanase-protease enzyme

A Pseudomonas fluorescens was found to degrade and utilize a polyester poly urethane as a sole carbon and energy source. Polyurethane utilization by P. fluorescens, followed simple Michaelis-Menten kinetics. The K-s and mu(max ) values were 0.9 mg ml(-1) and 1.61 doublings . h(-1) respectively. The en zymes from P. fluorescens responsible for polyurethane degradation were fou nd to be extracellular. Analysis of the polyurethane degrading proteins usi ng non-denaturing polyacrylamide gel electrophoresis revealed one active pr otein band with an R-f value of 0.083. A polyurethane degrading enzyme was purified and displayed protease activity. This enzyme was inhibited by phen ylmethylsulfonyl fluoride and had a molecular weight of 29,000 daltons. (C) 1999 Elsevier Science Ltd. All rights reserved.