Abstract
This study investigates the potential of an undervalued cephalopod species, Eledone moschata, for producing a freeze-dried protein concentrate via acid solubilisation and isoelectric precipitation. Although nutritionally rich, the processing route significantly affected the aggregation state of the recovered proteins, as demonstrated by differential scanning calorimetry (DSC) and SDS-PAGE electrophoresis. We systematically examined pretreatments of the lyophilised protein concentrate (PC) by dispersing it across a pH range (2-10) and applying ultrasonication (US), characterising the resulting aggregates in terms of protein solubility, surface hydrophobicity, dynamic light scattering (DLS), and ζ-potential. Subsequently, ultrasound-treated protein dispersions at different pH values were used to produce protein-quercetin nanoparticles (PQ), which were analysed for particle size (DLS), yield, and quercetin entrapment efficiency. PQ dispersions at pH 2, 4, and 7 were evaluated as stabilising agents in US-treated sunflower oil emulsions containing 10% oil and were characterised by rheological properties, microstructure, and DLS particle sizing. Confocal laser scanning microscopy (CLSM) revealed that nanoparticles at pH 2 produced small, uniformly distributed fat droplets with a particle diameter of 1.5 μm. This study provides insights into how processing conditions modulate the structural and interfacial behaviour of cephalopod proteins and highlights their potential application in designing low-fat, fluid emulsions for innovative food formulations.