Formation of bovine serum albumin-galangin nanoparticles and their potential to inhibit ROS-induced inflammation: Enthanol desolvation vs. pH-shiftng method.

pH-shifting method, as an eco-friendly approach, is a promising alternative to desolvation method, yet systematic comparison of their property is still lacking. In this study, bovine serum albumin-galangin nanoparticles (BSA-GA NPs) were designed for alleviating ROS-mediated macrophage inflammation by the 2 separate methods. Compared with the desolvation method, BSA exhibited a higher loading capacity for GA under the pH-shifting method, which was attributed to the exposure of the binding site leading to enhanced affinity for GA and a more compact particle structure. Further analyses evidenced that the electron arrangement and crystal structure of GA changed with different methods. The content of random coil of BSA was elevated after pH-shifting method. Besides, the smaller size rendered the pH-shifting treated BSA-GA NPs easier to be taken up by macrophages, while the enhanced specific surface area conferred excellent ROS scavenging and anti-inflammatory performances. This study may provide new insights into the choice of loading methods.

Co-assembled whey protein and proanthocyanidins as a promising biocarrier for hydrophobic pterostilbene: Fabrication, characterization, and cellular antioxidant potential.

The usage of food-derived polyphenols with different polarities has been limited by their instability and incompatibility. Therefore, a biocarrier was developed by co-assembly of whey protein isolate (WPI) and hydrophilic proanthocyanidin (PC) for loading hydrophobic pterostilbene (PTE). Such biocarrier has superior affinity for PTE than WPI alone, as determined by encapsulation efficiency and loading capacity assay, fluorescence quenching analysis, and molecular docking, whereas the assembly process was characterized by particle size and zeta potential, 3-dimensional fluorescence, and scanning electron microscopy. Circular dichroism and Fourier transform infrared spectroscopy spectra confirmed the α-helix to ß-sheet and random coil transition of proteins during the formation of nanocomplexes. Whey protein isolate acted as a mediator through altering the binding mode of PC and PTE, allowing them to perform significant synergistic effects in enhancing 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radical scavenging and reducing H2O2-induced cell damage. This research may serve to develop new protein/polyphenol co-loading systems and offer a reliable nutritional fortification.

Polymerized Whey Protein Concentrate-Based Glutathione Delivery System: Physicochemical Characterization, Bioavailability and Sub-Chronic Toxicity Evaluation.

Glutathione (GSH) is a powerful antioxidant, but its application is limited due to poor storage stability and low bioavailability. A novel nutrient encapsulation and delivery system, consisting of polymerized whey protein concentrate and GSH, was prepared and in vivo bioavailability, antioxidant capacity and toxicity were evaluated. Polymerized whey protein concentrate encapsulated GSH (PWPC-GSH) showed a diameter of roughly 1115 ± 7.07 nm (D50) and zeta potential of 30.37 ± 0.75 mV. Differential scanning calorimetry (DSC) confirmed that GSH was successfully dispersed in PWPC particles. In vivo pharmacokinetics study suggested that PWPC-GSH displayed 2.5-times and 2.6-fold enhancement in maximum concentration (Cmax) and area under the concentration-time curve (AUC) as compared to free GSH. Additionally, compared with plasma of mice gavage with free GSH, significantly increased antioxidant capacity of plasma in mice with PWPC-GSH was observed (p < 0.05). Sub-chronic toxicity evaluation indicated that no adverse toxicological reactions related to oral administration of PWPC-GSH were observed on male and female rats with a diet containing PWPC-GSH up to 4% (w/w). Data indicated that PWPC may be an effective carrier for GSH to improve bioavailability and antioxidant capacity.

Formation, stability and in vitro digestion of curcumin loaded whey protein/ hyaluronic acid nanoparticles: Ethanol desolvation vs. pH-shifting method.

Curcumin (CUR) was encapsulated in whey protein isolate/hyaluronic acid (WPI/HA) electrostatic nanoparticles at pH 5.4, 4.4, 3.4 and 2.4 using ethanol desolvation (DNP) or pH-shifting (PSNP) method. The prepared nanoparticles were characterized and compared for physiochemical properties, structure, stability, and in vitro digestion. PSNPs had smaller particle size, more uniform distribution, and higher encapsulation efficiency than DNPs. Main driving forces involved for fabricating the nanoparticles were electrostatic forces, hydrophobic forces, and hydrogen bonds. PSNP exhibited better resistance towards salt, thermal treatment, and long-term storage while DNPs showed stronger protection for CUR against thermal degradation and photodegradation. Stability of nanoparticles increased with decreasing pH values. In vitro simulated digestion exhibited that DNPs had lower release rate of CUR in SGF and higher antioxidant activity of its digestion products. Data may provide a comprehensive reference for selection of loading approach when constructing nanoparticles based on proteins/polysaccharides electrostatic complexes.

Binding of Licochalcone A to Whey Protein Enhancing Its Antioxidant Activity and Maintaining Its Antibacterial Activity.

Incorporating LA into whey protein by forming whey protein isolate-LA (WPI-LA) and polymerized whey protein-LA (PWP-LA) complexes is a good way to maintain its bioactivity and improve its functional performance within food matrices. Herein, we found that WPI and PWP were able to interact with LA as suggested by multi-spectroscopy, molecular docking, and molecular dynamics simulations. The interaction between whey protein and LA was a spontaneous non-covalent binding process, while PWP had a higher affinity for LA than WPI, resulting from its more negatively binding free energy with LA. Hydrogen bonds, van der Waals forces, and electrostatic interactions were responsible for WPI-LA interactions. Hydrophobic forces, van der Waals, and hydrogen bonds positively accounted for PWP-LA interactions. The antioxidant activity of LA was improved by complexation with whey proteins as identified by DPPH and ABTS. The antimicrobial efficiency of LA was partially screened by complexation with whey protein with MIC values increased by seven-fold compared to free LA but successfully recovered to its original efficiency upon isolating it from the complex. This work demonstrates the promising antioxidant and antibacterial activities of the whey protein-LA complex and provides a good candidate for developing a new class of natural functional ingredients for food systems.

Characterization of interactions between whey protein isolate and hyaluronic acid in aqueous solution: Effects of pH and mixing ratio.

Interactions between whey protein isolate (WPI) and hyaluronic acid (HA) were characterized as functions of pH (6.0-1.0) and protein to polysaccharide ratio (R, 1:4-10:1). Intramolecular soluble complexes formed at pHc of 5.6-5.8, followed by intermolecular insoluble complexes formed at pHΦ1 of 4.4-4.6. Complexes at ratios below 4:1 reached maximum optical value at pH 2.4 while samples above 4:1 peaked at pH 3-3.4 then precipitated. WPI/HA coacervates completely dissociated into soluble complex at pH 1.6-1.8 (pHΦ2). WPI/HA mixtures showed shear thinning behavior and elastic property. Whey protein underwent significant α-helix structure change when interacting with HA in range of pHΦ1>pH > pHΦ2 and at low R values (1:4 and 1:2). Scanning electronic microscope (SEM) pictures showed pH and mixing ratio dependent microstructural changes corresponding with phase transition. Data may provide helpful information for further application of WPI/HA complexes in medical, food and cosmetic fields.