Bromo-substituted polyaniline (Br-PANI) is prepared by a novel strategy in which PANI is used as raw material, potassium bromate and potassium bromide as brominate regents. The obtained Br-PANI is characterized by scanning electron microscopy, X-ray photoelectron spectroscopy, elemental analysis, FTIR, UV-vis spectra, and TGA. The results clearly indicate that Br has successfully attached to the benzene ring of PANI by chemical bond. The behavior of Br-PANI toward bacteria is characterized by plate counting, minimal inhibitory concentration, and minimal bactericidal concentration methods. The experimental results indicate that both doped and dedoped Br-PANIs have long-term antibacterial activities as compared to doped/dedoped PANI. What's more, the antibacterial abilities increase with increasing the Br/N molar ratio of the Br-PANI. Thus Br-PANI should be a kind of efficient and environmentally friendly antibacterial agent.