Different fluoroquinolon-type antibiotics were conjugated to gelatin with the aim to synthesize biomacromolecules with antimicrobial properties. The covalent linkage of the antibiotic was performed by a radical process involving the residues in the side chains of gelatin able to undergo oxidative modifications. The conjugation of antibiotic moieties onto the protein structure was confirmed by FT-IR, UV-Vis, fluorescence, and calorimetric analyses. Biocompatibility tests were performed on human bone marrow mesenchymal stromal cells and the antibacterial properties of bioactive polymers were investigated by appropriate tests against Klebsiella pneumoniae and Escherichia coli. With regard to the tests conducted in the presence of E. coli, a minimum inhibitory concentration (MIC) ranging from 0.05 to 0.40 μg mL(-1) was recorded, while in the presence of K. pneumoniae this concentration varies from 0.10 to 1.60 μg mL(-1). In all the conjugates, the drug moieties retain their biological activity and the MIC values are lower than the resistance parameters of fluoroquinolon-type antibiotics versus Enterobacteriacae. The collected data suggest a broad range of applications, from biomedical to pharmaceutical and food science for all conjugates.