Whey Proteins Isolate-Based Biopolymeric Combinations to Microencapsulate Supercritical Fluid Extracted Oleoresins from Sea Buckthorn Pomace

In this study, high-value, carotenoid-rich oleoresin obtained by supercritical carbon dioxide (SFE-CO2) extraction was used to develop five variants of microencapsulated delivery system, based on whey proteins isolate (WPI), in combination with inulin (I), pectin (P) or lactose (L). The WPI:I and WPI:L variants were also obtained by conjugation via Maillard reaction. The microencapsulation of the SFE-CO2 sea buckthorn pomace oleoresin was performed by emulsion, complex coacervation and freeze-drying, which allowed for the obtaining of five powders, with different phytochemicals profile. The WPI:I conjugate showed the highest level of total carotenoids, whereas the counterpart WPI:L showed the highest content in linoleic acid (46 ± 1 mg/g) and palmitoleic acid (20.0 ± 0.5 mg/g). The β-tocopherol and β-sitosterol were identified in all variants, with the highest content in the conjugated WPI:L variant. Both WPI:L and WPI:I conjugate samples presented similar IC50 value for inhibitory activity against pancreatic lipase and α-amylase; the highest activity was observed for the conjugated WPI:I. The WPI:P combination allowed the highest release of carotenoids in the gastro-intestinal environment. All the powders exhibited poor flowing properties, whereas water activity (aw) ranged from 0.084 ± 0.03 to 0.241 ± 0.003, suggesting that all variants are stable during storage. In case of solubility, significant differences were noticed between non-heated and glycated samples, with the highest value for the WPI:I and the lowest for glycated WPI:I. The structural analysis revealed the presence of finer spherosomes in WPI:I and WPI:L, with a reduced clustering capacity, whereas the particles in the conjugated samples were more uniform and aggregated into a three-dimensional network.

RNA length has a non-trivial effect in the stability of biomolecular condensates formed by RNA-binding proteins

Biomolecular condensates formed via liquid-liquid phase separation (LLPS) play a crucial role in the spatiotemporal organization of the cell material. Nucleic acids can act as critical modulators in the stability of these protein condensates. Here, we present a multiscale computational strategy, exploiting the advantages of both a sequence-dependent coarse-grained representation of proteins and a minimal coarse-grained model that represent proteins as patchy colloids, to unveil the role of RNA length in regulating the stability of RNA-binding protein (RBP) condensates. We find that for a constant RNA/protein ratio in which phase separation is enhanced, the protein fused in sarcoma (FUS), which can phase separate on its own-i.e., via homotypic interactions-only exhibits a mild dependency on the RNA strand length, whereas, the 25-repeat proline-arginine peptide (PR25), which does not undergo LLPS on its own at physiological conditions but instead exhibits complex coavervation with RNA-i.e., via heterotypic interactions-shows a strong dependence on the length of the added RNA chains. Our minimal patchy particle simulations, where we recapitulate the modulation of homotypic protein LLPS and complex coacervation by RNA length, suggest that the strikingly different effect of RNA length on homotypic LLPS versus complex coacervation is general. Phase separation is RNA-length dependent as long as the relative contribution of heterotypic interactions sustaining LLPS is comparable or higher than that committed by protein homotypic interactions. Taken together, our results contribute to illuminate the intricate physicochemical mechanisms that influence the stability of RBP condensates through RNA inclusion.