This work presents the synthesis of polyvinylpyrrolidone (PVP) and chitosan (CS) coated magnetite (Fe₃O₄) nanoparticles (NPs) by co-precipitation technique. The objective of this research was to investigate the catalytic, antibacterial efficacy with evidential molecular docking analysis of CS/PVP- Fe₃O₄ and PVP/CS- Fe₃O₄. The coating of PVP and CS reduced the particle size of Fe₃O₄, providing a large surface area that increased catalytic and antibacterial potency. These binary coated magnetite NPs used as ideal catalytic agent for the reduction of rhodamine B dye as well as antibacterial agent against Escherichia coli. The findings of a molecular docking investigation were in reasonable accord with in vitro microbicidal activity and revealed that CS-Fe₃O₄, Fe₃O₄-doped PVP and CS are promising inhibitors of dihydrofolate reductase_{E. Coli} (DHFR) and DNA gyrase_{E. Coli}.