Metabolites produced by the degradation of polyester polyurethane (PUR) by Comamonas acidovorans strain TB-35 were investigated. GC-MS analysis revealed that they were diethylene glycol, trimethylolpropane and adipic acid. These metabolites were considered to be derived from polyester segments of the PUR as a result of hydrolytic cleavage of ester bonds. In addition, when the culture broth was alkaline treated, a previously undetected product was detected by GC analysis. This product was identified as 2,4-diaminotoluene by GC-MS analysis. This indicates that an additional metabolite exists in the culture broth. This metabolite was considered to have been derived from polyisocyanate segments of the PUR. A preliminary study on a PUR degrading enzyme was also performed. Strain TB-35 produced two different esterases, one which is secreted to the culture broth and one which is bound to the cell surface. Between them, only cell-surface-bound esterase catalyzes the degradation of the polyester PUR.