In this study, 2-hydroxyethyl methacrylate (HEMA) was used as the monomers for surface grafting on electrospun PU/RC nanofiber membrane via atom transfer radical polymerization (ATRP) method, and the PU/RC-poly(HEMA) nanofiber membrane was investigated as a carrier for LAC. Free and immobilized LAC was characterized, and efficiency of bisphenol A (BPA) removal was determined. The results indicated that the PU/RC-poly(HEMA)-LAC showed relatively higher pH stability, temperature stability, and storage stability than free and PU/RC-LAC; moreover, more than 60% of the PU/RC-poly(HEMA)-LAC activity was retained after 10 cycles of ABTS treatment. Notably, the BPA removal efficiency of PU/RC-poly(HEMA)-LAC membrane generally ranged from 87.3 to 75.4% for the five cycles. Therefore, the PU/RC-poly(HEMA) nanofiber membrane has great potential as a carrier for the LAC immobilization for various industrial applications and bioremediation.