Biomolecule carrier systems based on metal-organic frameworks (MOFs) are highly promising owing to a high degree of structural tunability, high surface area, porosity, and higher chemical/thermal stability. UiO-66, one of the most stable Zr(IV)-based MOF, has a pore aperture of 0.6 nm and a theoretical pore volume of 0.77 cm3/g, ideal for high storage capacity of biomolecule cargo, however its potential to be implemented as an efficient carrier for fosfomycin has yet to be investigated. UiO-66 nanocrystal, synthesized at room temperature, as a fosfomycin carrier was reported. Drug modulated synthesis (in situ encapsulation) and post-synthesis (adsorption/impregnation) were applied to incorporate the fosfomycin in and/or on UiO-66 nanocrystals. We determined the effect of room temperature synthesis conditions on the structural properties of the UiO-66 nanocrystals as a functional carrier for fosfomycin. The incorporation of fosfomycin either by encapsulation or adsorption did not change the inherent crystal structure and UiO-66 nanocrystals kept their morphology. The addition of fosfomycin into the reaction medium led to an increase in particle size from 12745 nm to 20352 nm. Our FTIR results indicated the development of Zr-O-P connection due to the capture of drug molecules by adsorption. Antibacterial activity studies unveiled the drug concentration dependent bactericidal and bacteriostatic activities towards S.aureus and E.coli for fosfomycin loaded UiO-66 nanocrystals. Investigations with the help of density functional theory were performed to reveal the interaction mechanism between fosfomycin and UiO-66. Our theoretical findings indicated that fosfomycin strongly interacts with the metal center in the defected UiO-66 through its oxygen atom in the phosphite group with a Zr ̶ O distance of 2.087 Å. Additionally, the energy of interaction for the adsorption process was found to have a large negative value of -74.3 kcal/mol, which supports the strong interaction between the two systems.

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