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Cheap and efficient bioactive materials capable of producing potable water are in high demand. We synthesized bioactive prodigiosin-conjugated iron-oxide-activated carbon composite ([Ac]F@Fe3O4–PG) for the effective removal of microbial contaminants without biofilm formation. The disinfection effect of [Ac]F@Fe3O4–PG was investigated against antibiotic-resistant Escherichia coli and Bacillus cereus. The reactive oxygen species, glutathione concentration, and intracellular leakages assay, zeta potential, atomic force microscopy, scanning electron microscopy, and fluorescence microscopy were performed to understand the mechanistic route of bacterial disinfection. Results demonstrate the bacterial cell membrane damage due to the surface charge neutralization by cationic [Ac]F@Fe3O4–PG, thereby affecting the transport system of the cells. The generation of reactive oxygen species further triggered programmed bacterial cell death with apoptosis-like changes in cellular morphology. We demonstrate that the [Ac]F...