PANGMA nanofibers and nanomats with fiber diameters of 200–300 nanometers were fabricated by electrospinning. Cal-B was covalently immobilized onto the PANGMA nanomats via three different immobilization routes. The properties of the Cal-B-immobilized PANGMA nanomats were assayed and compared with the free Cal-B. The observed Cal-B loading on these nanomats is up to ≈50mg · g-1, and their hydrolytic activity is up to ≈2500 nmol · min-1 · mg-1, much higher than free enzyme powder and also slightly higher than Novozyme 435. Cal-B immobilized PANGMA nanomats have better reusability, thermal stability, and storage ability than free Cal-B. They retain over 50% of their initial activity after 15 cycles, over 65% after 10 h heat incubation, and over 75% after 30 d storage.