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Functionalized iron oxide nanoparticles (IONPs) have unique physical and chemical properties, which make them potential candidates for biomedical applications. In this study, a facile one-pot method is reported for the preparation of poly(acrylic acid) (PAA) functionalized IONPsthrough in situ free radical solution polymerization of AA and subsequent coprecipitation of Fe ^3+ and Fe ^2+ ions. The FTIR spectroscopic and TGA results indicated the successful formation and surface functionalization of IONPs with PAA. Electron micrographs showed that the prepared particles were of nano-sized and their shape is dependent on the concentration of PAA. pH-dependent variation of average hydrodynamic diameter confirmed the pH-responsivity of PAA-functionalized IONPs. Magnetic measurement suggested that the PAA functionalized IONPs were strongly paramagnetic (53.0 emug ^−1 ). Fenton-like catalytic generation is carried out to measure toxicity associated with the nanoparticles. The suppression ability for reactive oxygen species (ROS) generation associated with PAA-functionalized IONPs was studied via methylene blue degradation assay to address their toxicity profile. PAA-functionalized IONPs exhibited better suppression ability than that of the bare IONPs. The adsorption behavior of trypsin was also studied at different pH levels and a maximum adsorption is occurred on PAA-functionalized IONPs at pH 5.0. Catalytic behavior study confirmed higher activity of trypsin immobilized on PAA-functionalized IONPs than that of the reference IONPs. Therefore, the functionalized IONPs can be of high interest for magnetically recyclable biocatalyst carrier.