The greatest challenge for biotechnological processes is to have immobilized enzymes acting as good green catalysts with high reusability rates. In this work, we have produced electrospun fibers from poly (lactic acid)/chitosan blends. Further, we evaluated the influence of these materials as support for lipase immobilization. The PLA/chitosan fiber mats were composed by non-woven nanofibers, with diameters ranging from 200nm to 1.3μm. The solvent (TFA) as well as the chitosan addition influenced morphology, hydrophobicity and mechanical properties of PLA nanofibers. It was observed that even for lower concentration of lipase (5U) higher enzyme activity retention was detected in the PLA/chitosan blends. In addition, a remarkable improvement of lipase activity on pure PLA fiber mat was verified, indicating that most of the enzymes were probably in their active form.