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We report, for the first time, the synthesis of superparamagnetic and fluorescent nickel (Ni) nanoparticles (consisting of a single material) directly conjugated to an enzyme, bovine pancreatic α-chymotrypsin (CHT), by chemical reduction in aqueous solution. The structural characterization of Ni–CHT nanobioconjugates was carried out using UV-VIS absorption/photoluminescence spectroscopy and high-resolution transmission electron microscopy. The temperature dependence of the magnetization M(T) taken in zero field cooling and field cooling conditions, exhibits the main features of superparamagnetism. Circular dichroism studies were performed to monitor the structural perturbation to the structure of the enzyme after conjugation with the nickel nanoparticles (Ni NPs). The functional integrity of the enzyme conjugated to the Ni NPs was investigated by monitoring the enzymatic activity of the Ni–CHT conjugates …