Synthesis of gold nanoclusters-loaded lysozyme nanoparticles for ratiometric fluorescent detection of cyanide in tap water, cyanogenic glycoside-containing plants, and soils

The modification of protein-stabilized gold nanoclusters with fluorophores has been intensively applied for the ratiometric detection of biomolecules, metal ions, and anions. This study developed a straightforward strategy to prepare lysozyme nanoparticle-encapsulated gold nanoclusters (LysNP-AuNCs) as a dual-emission probe for the ratiometric sensing of cyanide through fluorescence resonance energy transfer (FRET) without the conjugation of additional fluorophores. The reduction of gold ion precursors with lysozyme generated lysozyme-stabilized AuNCs under an alkaline pH, which were demonstrated to self-assemble into nanoaggregates during the formation of AuNCs. The aggregated lysozyme molecules on the AuNCs were treated with glutaraldehyde, triggering the conversion of the aggregated lysozymes into blue-emitting lysozyme nanoparticles. As a result, the AuNCs were well distributed inside a single lysozyme nanoparticle, as demonstrated by transmission electron microscopy. The presence of cyanide triggered the etching of the AuNCs in the LysNP-AuNCs, leading to the suppression of FRET from lysozyme nanoparticle to AuNCs. The LysNP-AuNC probe was implemented for FRET detection of cyanide with a linear range of 3–100 μM. Additionally, the selectivity of the LysNP-AuNC probe for cyanide toward other anions was remarkably high. The practicality of the proposed probe was evaluated by quantifying cyanide in tap water and soils and monitoring the liberation of hydrogen cyanide from cyanogenic glycoside-containing foods.