Polycaprolactone films (PCL) with different degree of crystallinity were prepared by solvent casting and the effect of microstructure on enzymatic degradation by a lipase type enzyme was studied in vitro. The extent of degradation of PCL films was examined by weight loss, differential scanning calorimetry (DSC), FTIR spectroscopy and optical microscopy. Weight loss during enzymatic degradation of PCL films having different crystallinity as a function of time suggested that that the degradation depends on crystallinity and morphology of starting PCL films. PCL film with lower crystallinity (24%) degrades in a way of layer-by-layer. Enzymatic degradation of low crystallinity PCL film usually starts from the amorphous region where the degradation rate is much higher than that in the crystalline region while degradation of PCL film with higher crystallinity (45%) occurs in the crystalline region as well in the amorphous region. Effect of degradation on crystallinity, melting temperature and conformational changes if any were investigated using FTIR and DSC. These results will be discussed.