It is of considerable significance to develop efficient and robust water purification technologies because of the increasing population and the serious water pollution. Herein, we proposed a facile strategy to synthesize a multifunctional water treatment agent, via one-pot grafting of antibacterial polyhexamethylene biguanide (PHMB) onto porous microspheres (PMSs) constructed by poly(N-isopropylacrylamide-co-methacrylic acid) and Fe 3 O 4 nanoparticles. The resulting PHMB-g-PMS exhibited prominent disinfection performance and a high adsorption capacity for dye pollutants. The minimum inhibitory concentrations (MICs) of the microspheres against Staphylococcus aureus and Escherichia coli were 2.0 and 3.9 μg/mL, respectively. The adsorption capacities for Congo red (CR) and acid fuchsin (AF) were 2422 and 885 mg/g, respectively, and the adsorption processes of the microspheres toward CR and AF match with the Langmuir and pseudo-second-order models. Moreover, pollutant-loaded microspheres were readily collected using a magnet and regenerated effectively using NaOH solution. After seven adsorption− desorption cycles, PHMB-g-PMS still showed excellent reusability with 100% inactivation of Escherichia coli and around 80% removal efficiency for AF. In summary, the prepared magnetic porous microspheres exhibit tremendous potential for removing multiple contaminants by an all-in-one process.