Electrospun nanofibers, renowned for their high specific surface area, robust mechanical properties, and versatile chemical functionalities, offer a promising platform for enzyme immobilization. Over the past decade, significant strides have been made in developing enzyme-induced electrospun nanofibers (EIEN). This review systematically summarizes the advanced applications of EIEN which are fabricated using both non-specific immobilization methods including interfacial adsorption (direct adsorption, cross-linking, and covalent binding) and encapsulation, and specific immobilization techniques (coordination and affinity immobilization). Future research should prioritize optimizing immobilization techniques to achieve a balance between enzyme activity, stability, and cost-effectiveness, thereby facilitating the industrialization of EIEN. We elucidate the rationale behind various immobilization methods and their applications, such as wastewater treatment, biosensors, and biomedicine. We aim to provide guidelines for developing suitable EIEN immobilization techniques tailored to specific future applications.