Abstract Fibrous mats from an α-amino acid based poly(ester amide) have been prepared due to the potential applications of this kind of polymers in the biomedical field thanks to the expected non-toxicity of their degradation products. Specifically, the electrospinning technique has been applied and both solution properties and operational parameters have been optimized to get continuous fibers in the micrometer range. Furthermore, fibrous mats have been loaded with biguanide compounds differing on molecular size but having a well-proved bactericide activity (i.e., chlorhexidine, CHX and polyhexamethylenebiguanide, PHMB). The high solubility of the poly(ester amide) constituted by l-phenylalanine, adipic acid and 1,4-butanediol also allowed getting appropriate electrospinning conditions to incorporate degrading enzymes like α-chymotrypsin without significant denaturation. Degradability of fibrous mats has been evaluated in distinct enzymatic media (lipase, proteinase K and α-chymotrypsin) being found a similar behavior that contrasts with the significant differences detected when film samples were employed. An accelerated degradation was clearly found for fibrous mats loaded with α-chymotrypsin even when they were exposed to a non-enzymatic aqueous medium. Release of bactericide agents was evaluated and a specific delay was determined when the polymeric biguanide was employed. Nevertheless, PHMB showed a clearly enhanced activity. Biocompatibility of the new fibrous mats was verified being also determined an increase on the cell adhesion with respect to film samples as a consequence of the increased porosity.