DNA-loaded porous polysulfone (PSf) microspheres were prepared using a liquid-liquid phase separation technique. The microspheres were then used in environmental application. The DNA-loaded microspheres were stable in water. They could effectively accumulate the DNA-binding harmful pollutants and endocrine disruptors, such as ethidium bromide (EB), acridine orange (AO), biphenyl (BP), dibenzofuran (DBF), and dibenzo-p-dioxin (DBD), in their aqueous solutions. On the other hand, the microspheres were found to selectively remove heavy metal ions from their aqueous solutions. The DNA-loaded PSf microspheres have a potential ability to serve as functional materials in environmental application.