A poly(ethylene succinate) (PESu) hydrolase was purified from the culture supernatant of the PESu-degrading fungus Aspergillus clavatus NKCM 1003, which was cultivated on a mineral medium containing PESu as the sole carbon source. The enzyme had a molecular mass of 57,000 Da and an isoelectric point of 5.6, and it was stable in the rage pH5-8; the optimum pH was 7.0. Its PESu hydrolytic activity remained constant below 40 8C after incubation for 30 min. Liquid chromatography electrospray mass spectrometry analysis of enzymatic hydrolysates revealed that the major products are ES, ESE, SES, and ESES(S; succinate unit, E; ethylene glycol unit). This enzyme degraded not only PESu but also poly(3-hydroxybutyrate) (P(3HB)) and its copolymers. Further, it showed high binding affinity to P(3HB). It was concluded that PESu hydrolase belongs to the class of P(3HB) depolymerases.