Chronic wound infections, especially due to the emergence of multidrug resistance in bacteria, require the urgent development of alternative antibacterial therapies. Here, we developed a new class of hydrogel nanofibrous membranes that show visible light-induced disinfection. The presented photocatalytic disinfection is based on the generation of reactive singlet oxygen from a conjugated microporous polymer upon visible light irradiation. Therefore, sustained protection of the wound area can be provided in the presence of visible light. Fabrication of the photoactive wound dressing consists of first synthesizing photoactive conjugated microporous polymer nanoparticles by miniemulsion polymerization and subsequently embedding the nanoparticles in polyvinyl alcohol hydrogel nanofibers by colloid-electrospinning. The fibers were then crosslinked in glutaraldehyde/HCl vapor to be water-insoluble. This nanoparticle-in-nanofiber structure allows for a flexible combination of the properties of the nanoparticles and supporting nanofibers. The disinfecting properties of the membranes were evaluated with the inactivation of Escherichia coli K-12 and Bacillus subtilis as model systems of Gram-negative and Gram-positive bacteria, as well as the inhibition of biofilm growth under irradiation of visible light. Cytotoxicity tests on fibroblast cells revealed a high cytocompatibility of the membranes. Furthermore, the good mechanical properties of the membranes allow for their facile removal after use and prevent the leakage of the embedded nanoparticles into the wound, making the photoactive hydrogel membranes a promising candidate for active wound dressing materials.