Enzymatic degradation of a series of polyesters prepared from 1,4:3.6-dianhydro-D-glucitol (1) and aliphatic dicarboxylic acids of the methylene chain length ranging from 2 to 10 were examined using seven different enzymes. Enzymatic degradability of these polyesters as estimated by water-soluble total organic carbon (TOC) measurement is dependent on the methylene chain length (m) of the dicarboxylic acid component for most of the enzymes examined. The most remarkable substrate specificity was observed for Rhizopus delemar lipase, which degraded polyester derived from 1 and suberic acid (m = 6) most readily. In contrast, degradation by Porcine liver esterase was nearly independent of the structure of the polyesters. Enzymatic degradability of the polyesters based on three isomeric 1,4:3.6-dianhydrohexitols and sebacic acid was found to decrease in the order of 1, 1,4:3.6-dianhydro-D-mannitol (2), and 1,4:3.6-dianhydro-L-iditol (3). Structural analysis of water-soluble degradation products formed during the enzymatic hydrolysis of polyester 5g derived from 1 and sebacic acid has shown that the preferential ester cleavage occurs at the O(5) position of 1,4:3.6-dianhydro-D-glucitol moiety in the polymer chain by enzymes including Porcine pancreas lipase, Rhizopus delemar lipase, and Pseudomonas sp. lipase.