Multicompartmented nanostructures with tunable interior complexities co-assembled by two plant proteins

Multicompartmented nanostructures, which mirror the complexity of living cells, are promising for bio-related applications. However, controlling the spatial arrangement within natural biopolymer assemblies, like protein nanoparticles, is challenging. In our study, we reveal a facile method for compartmentalizing the interiors of protein nanoparticles by co-assembling rice proteins (RPs) and soy protein isolates (SPIs) through simultaneous folding. The assembly of these two protein types results in nanoparticles with a hydrophobic core to accommodate hydrophobic substances, such as eugenol, leading to its internalization. The protein–ligand (SPI-eugenol) interaction then triggers SPI gelation within the nanoparticle cores, forming distinct physical barriers hindering the complete merging of eugenol droplets. Subsequent dialysis of the nanoparticles containing eugenol against water facilitated the removal of eugenol. This step was crucial for forming multicompartmented nanoparticles (MCNPs) with adjustable interior complexities, including varying numbers of compartments and different hollowing rates related to various SPI:RP mass ratios. This allows us to endow MCNPs with adjustable barrier effects tailoring controllable responses of the encapsulated payload, vitamin D 3 (VD 3 ), under different physicochemical conditions. Our study demonstrates a straightforward method for constructing biomacromolecular architectures with precise control over their compartmentalization.