Controlling Solvent Polarity to Regulate Protein Self-Assembly Morphology and Its Universal Insight for Fibrillation Mechanism.

The mechanism of ethanol-induced fibrillation of ß-lactoglobulin (ß-lg) in the acidic aqueous solution upon heating was investigated using various techniques, mainly thioflavin T fluorescence, atomic force microscopy, nonreducing electrophoresis, mass spectrometry, Fourier transform infrared spectroscopy, and circular dichroism spectroscopy. The results showed that fibrillation occurred with a heating time increase, but high ethanol content slowed down the process. At a low ethanol volume fraction, peptides existed after heating for 2 h, with long and straight fibrils formed after 4-6 h, while at a high ethanol volume fraction, the proteins aggregated with very few peptides appeared at the early stage of heating, and short and curved fibrils formed after heating for 8 h. Ethanol weakened the hydrophobic interactions between proteins in the aqueous solution; therefore the latter could not completely balance the electrostatic repulsion, and thus suppressing the fibrillation process. It is believed that the fibrillation of ß-lg in the acidic solution upon heating is mainly dominated by the polypeptide model; however, ethanol inhibited the hydrolysis of proteins, and the self-assembly mechanism changed to the monomer model.

Effect of emulsifiers on the properties of corn starch films incorporated with Zanthoxylum bungeanum essential oil.

The use of natural and safe ingredients in green food packaging material is a hot research topic. This study investigated the effect of different emulsifiers on starch film properties. Three types of emulsifiers, including Tween 80 as a small-molecule surfactant, sodium caseinate (CAS), whey protein isolate (WPI), and gelatin (GE) as macromolecule emulsifiers, whey protein isolate fibril (WPIF) as a particle emulsifier, were utilized to prepare Zanthoxylum bungeanum essential oil (ZBO) emulsions. The mechanical, physical, thermal, antibacterial properties, microstructure and essential oil release of starch films were investigated. CAS-ZBO nanoemulsion exhibited the smallest particle size of 198.6 ± 2.2 nm. The film properties changed with different emulsifiers. CAS-ZBO film showed the highest tensile strength value. CAS-ZBO and WPIF-ZBO films exhibited lower water vapor permeability than Tween-ZBO. CAS-ZBO film showed good dispersion of essential oil, the slowest release rate of essential oils in all food simulants, and the best antibacterial effect against Staphylococcus aureus and Listeria monocytogenes. The films composed of CAS-ZBO nanoemulsion, corn starch, and glycerol are considered more suitable for food packaging. This work indicated that natural macromolecule emulsifiers of CAS and WPIF are expected to be used in green food packaging material to offer better film properties.

Influence of interfacial properties/structure on oxygen diffusion in oil-in-water emulsions.

Oxygen diffusion played an important role in the lipid oxidation of food emulsions. In this study, a simple method was developed to quantitatively observe the oxygen diffusion in the oil-water biphasic system, and it was further applied to investigate the relationship between the oxygen diffusion and lipid oxidation in O/W emulsions. Various factors that related to the emulsion oxidation were considered, from their influence on the oxygen diffusion and lipid oxidation in the emulsions. Results showed that there was obvious correlation between the oxygen diffusion and lipid oxidation in O/W emulsions, which reveals the inhibition of oxygen diffusion could apparently slow down the lipid oxidation. Moreover, the changes of oil phase, water phase and interfacial layer of the emulsions, which were related to the oxygen diffusion, could improve the oxidative stability of the emulsions effectively. Our findings are helpful for deep understanding the mechanisms of the lipid oxidation in food emulsions.

Formation and application of edible oleogels prepared by dispersing soy fiber particles in oil phase.

Oleogels containing less saturated and trans-fats were considered as an ideal option to replace the solid fats in foods. In this research, oleogel was fabricated by dispersing soy fiber particles (SFP) in soy oil, and further it was used in bread preparation. Effect of the particle size, particle content and the second fluid content on the formation of oleogels were evaluated, based on the appearance and rheological properties. Results showed that the suspension of SFP in soy oil (24%, w/w) could be transformed into gel-like state, upon the addition of the second fluid. The SFP based networks were dominated by the capillary force which was originated from the second fluid. The rheological properties and yield stress of the oleogels could be modulated by particle size and particle content of SFP in oil phase, as well as the second fluid content in the system. When the oleogels were applicated in bread preparation, a layered structure could be formed in the bread, indicating the possibility of replacing the solid fats in bakery products by our oleogels. Our results offered a feasibility approach for oil structuring with natural raw materials, and developed a new approach to replace the solid fats in foods.

Advances in Bioactivity of MicroRNAs of Plant-Derived Exosome-Like Nanoparticles and Milk-Derived Extracellular Vesicles.

MicroRNA (miRNA) is a class of small noncoding RNA involved in physiological and pathological processes via the regulation of gene expression. Naked miRNAs are unstable and liable to degradation by RNases. Exosome-like nanoparticles (ELNs) secreted by plants and extracellular vesicles (EVs) found in milk are abundant in miRNAs, which can be carried by ELNs and EVs to target cells to exert their bioactivities. In this review, we describe the current understanding of miRNAs in plant ELNs and milk EVs, summarize their important roles in regulation of inflammation, intestinal barrier, tumors, and infantile immunological functions, and also discuss the adverse effect of EV miRNAs on human health. Additionally, we prospect recent challenges centered around ELN and EV miRNAs for interventional applications and provide insights of grain-derived ELNs and miRNAs interventional use in human health. Overall, plant ELNs and milk EVs can transfer miRNAs to mitigate the pathological status of recipient cells by mediating the expression of target genes but may also exert some side effects. More studies are required to elucidate the in-depth understanding of potential interventional effects of ELN and EV miRNAs on human health.

Nonlinear dilatational rheology of different protein aggregates at the oil-water interface.

Proteins tend to self-assemble into different morphological aggregates such as nanoparticles or fibrils during heat treatment depending on the processing conditions. The protein aggregates exhibit excellent interfacial activity and even better ability to stabilize emulsions than native proteins. The interfacial rheological properties at the oil-water interface play a very important role in emulsion stability, among which the interfacial nonlinear rheology is closely related to their ability to resist large perturbation. However, there are very few studies reporting the nonlinear interfacial rheological behavior of protein aggregates at the oil-water interface. In this study, ß-lactoglobulin fibrous aggregates (F) and nanoparticle aggregates (NP) were prepared, and the adsorption kinetics and dilatational nonlinear rheological behavior of ß-lactoglobulin aggregates at the oil-water interface under large amplitude deformation were studied using a pendant drop tensiometer, and compared with those of native proteins. From the adsorption experiments, the adsorption of protein aggregates, especially fibrils, was faster than that of native proteins in the early stage, while in the late stage, the native proteins displayed a significantly higher degree of rearrangement than the fibrils. The surface hydrophobicity and the short fibrils present mainly determine the properties of the fibril interface, while the behavior of the nanoparticle interface was significantly influenced by the size and charge properties of the nanoparticles. From the dilatational experiment, the Lissajous plots revealed that the F interface at all pHs evaluated and the ßlg interface at pH 5.8 displayed strain softening in both expansion and compression processes, while the NP interface at all pHs and ßlg interface at pH 2 and pH 7 displayed strain softening in expansion and strain hardening in compression processes. The nonlinear response of the protein aggregates at the oil-water interface was more obvious at pH 5.8. The modulus change from frequency sweeps revealed that the fibril interface was strong but not very structured in contrast to that formed by the native proteins which displays high structuration although weak in strength, whereas the strength of the interface formed by protein nanoparticles is in between, but more sensitive to the surface charge.

Interfacial behaviour of ß-lactoglobulin aggregates at the oil-water interface studied using particle tracking and dilatational rheology.

During processing, proteins are easily self-assembled into different aggregates, such as nanoparticles and fibrils. Protein aggregates exhibit a strong interfacial activity due to their morphologies and functional groups on the surface. Their interfacial structure and rheological properties at the oil-water interface have a significant effect on the stability and fat digestion of emulsions in food. In this study, ß-lactoglobulin (ß-lg) aggregates including ß-lg nanoparticles (ß-lg NP) and ß-lg fibrils (ß-lg F) were prepared in solution by controlling the heating temperature and pH, and their surface properties including the electric potential, hydrophobicity, and density of free thiol groups were characterized. The adsorption kinetics, interfacial rheology, and displacement by bile salts (BSs) of native ß-lg and its aggregates at the oil (decane)/water interfaces were studied using particle tracking microrheology and dilatational rheology. From the movement of tracer particles at the interface, ß-lg NP and ß-lg F were found to adsorb faster than native ß-lg, and they were found to form interfacial films with a marginally higher elasticity. During the process of protein adsorption, the films of ß-lg and its aggregates are not uniform. In the process of protein displacement, ß-lg NP has the strongest ability while native ß-lg has the weakest ability to resist BS substitution, which is consistent with the results from in vitro digestion experiments. The present study reveals the microrheological behaviour of protein aggregates at the oil-water interface and demonstrates that ß-lg thermal aggregates exhibit an excellent emulsification ability and can be used to control fat digestion. The study also illustrates the applicability of microrheological methods to the study of interfacial rheology and its complementarity with dilatational rheological methods.

Effect of simulated saliva components on the in vitro digestion of peanut oil body emulsion.

The digestion properties of natural oil bodies (OBs) are very important to their potential applications such as traditional fat replacement or bioactive delivery systems. However, study on the complete digestion behaviours of OBs has not been reported yet. In this paper, peanut OBs were extracted by an aqueous medium method, and their digestion behaviour was studied using completed in vitro oral-gastric-intestinal digestion simulation. In particular, the effects of saliva components, mainly α-amylase and mucin, on the digestion of the peanut OBs were systematically investigated. The OB emulsion microstructure, average particle size d 4,3, ζ-potential, and surface protein compositions during oral, gastric and intestinal digestion, and the free fatty acid (FFA) release rate of the peanut OBs during intestinal digestion were determined. Interestingly, it was revealed from both the periodic acid-Schiff staining technique and the confocal laser microscopy characterization that glycosidic bonds exist on the surface of the peanut OBs, though how they were produced was unknown. The results from the digestion measurements showed that α-amylase in saliva can break the glycosidic bonds in oral digestion, promoting the digestion of the OBs in the gastric and intestinal environments. Saliva mucin caused bridging flocculation of OBs by electrostatic attraction in the gastric tract, and depletion flocculation of OBs in the intestinal tract. The former hindered the fusion of oil droplets, and the latter promoted FFA release rate by increasing the contacting surface area of OBs with bile salts.

Molar mass effect in food and health.

It is demanded to supply foods with good quality for all the humans. With the advent of aging society, palatable and healthy foods are required to improve the quality of life and reduce the burden of finance for medical expenditure. Food hydrocolloids can contribute to this demand by versatile functions such as thickening, gelling, stabilising, and emulsifying, controlling texture and flavour release in food processing. Molar mass effects on viscosity and diffusion in liquid foods, and on mechanical and other physical properties of solid and semi-solid foods and films are overviewed. In these functions, the molar mass is one of the key factors, and therefore, the effects of molar mass on various health problems related to noncommunicable diseases or symptoms such as cancer, hyperlipidemia, hyperglycemia, constipation, high blood pressure, knee pain, osteoporosis, cystic fibrosis and dysphagia are described. Understanding these problems only from the viewpoint of molar mass is limited since other structural characteristics, conformation, branching, blockiness in copolymers such as pectin and alginate, degree of substitution as well as the position of the substituents are sometimes the determining factor rather than the molar mass. Nevertheless, comparison of different behaviours and functions in different polymers from the viewpoint of molar mass is expected to be useful to find a common characteristics, which may be helpful to understand the mechanism in other problems.

Novel strategy for enhancing the color intensity of ß-Carotene: Enriching onto the oil-water interface.

A novel strategy to enhance the color intensity of ß-carotene (BC), namely, "interfacial enriching", was developed in this work. As the sole emulsifier in W/O emulsion, BC particles were enriched onto the droplet surface through emulsifying process. By increasing the concentration of BC in oil phase from 1 mg/g to 5 mg/g, the average droplet size of the emulsion decreased from 92.2 ± 5.1 µm to 34.0 ± 5.4 µm. Too low (e.g. ≤ 1 mg/g) or too high (e.g. ≥25 mg/g) concentration of BC in the oil phase yielded an insufficient coverage or flocculation of the droplets. By enriching onto the interface, the color intensity of BC were enhanced apparently at the reflectance wavelength ranging from 500 nm to 700 nm, compared with that of the BC encapsulated within the emulsion droplets. This enhancement was due to the higher availability of incident light for the BC particles on the interface than that of the BC particles buried inside the droplets.

Iron encapsulated microstructured gel beads using an emulsification-gelation technique for an alginate-caseinate matrix.

Iron-deficiency anemia is an important health problem in global public issues, and development of iron fortifiers in diets is essential for the decrease of iron deficiency. However, there are problems for iron fortification in food because the common bioavailable iron compounds would contribute to iron-promoted lipid oxidation and unpleasant iron odor, presenting an adverse food quality. Ferrous fumarate loaded microstructured gel beads were prepared by an emulsification-gelation method using an alginate-caseinate matrix, and the gel network was formed by crosslinking of Ca2+ or Fe2+. Internal gelated beads showed relatively symmetrical and homogeneous spheres with no adhesion due to the simultaneous release of Fe2+ to initiate gelation in situ. External gelated beads displayed an irregular and adhesive structure, probably because the random contact between Na-ALG and Ca2+ occurred on the droplet surface, and the immediately gelated hardening layer provided a delay for further Ca2+ diffusion. The gel beads exhibited a lag phase in the promotion of lipid oxidation of the emulsion and restrained the iron odor release from ferrous fumarate. Ferrous ion release from microstructured gel beads in the simulated gastric juice was obviously delayed before a more progressive high release in the simulated intestinal juice, beneficial for iron absorption in the duodenum. The iron encapsulated microstructured gel beads might be developed as a promising safe iron fortifier by relieving lipid oxidation and iron odor.

In situ nanomechanical properties of natural oil bodies studied using atomic force microscopy.

Natural oil bodies (OBs) from plant organs represent an important category of functional ingredients and materials in a variety of industrial sectors. Their applications are closely related to the membrane mechanical properties on a single droplet level, which remain difficult to determine. In this research, the mechanical properties of the membranes of OBs from soybean, sesame, and peanut were investigated in-situ by atomic force microscopy (AFM). Different regions of the force-deformation curves obtained during compression were analyzed to extract the stiffness Kb or Young's modulus of the OB membranes using Hooke's law, Reissner theory, and the elastic membrane theory. At higher strains (ε = 0.15-0.20), the elastic membrane theory breaks down. We propose an extension of the theory that includes a contribution to the force from interfacial tension based on the Gibbs energy, allowing effective determination of Young's modulus and interfacial tension of the OB membranes in the water environment simultaneously. The mechanical properties of the OBs of different sizes and species, as well as a comparison with other phospholipid membrane materials, are discussed and related to their membrane compositions and structures. It was found that the natural OBs are soft droplets but do not rupture and can fully recover following compressive strains as large as 0.3. The OBs with higher protein/oil ratio, have smaller size and stronger mechanical properties, and thus are more stable. The low interfacial tension due to the existence of phospholipid-protein membrane also contributes to the stability of the OBs. This is the first report measuring the mechanical properties of OB membranes in-situ directly.

Trivalent iron induced gelation in Artemisia sphaerocephala Krasch. polysaccharide.

Artemisia sphaerocephala Krasch. polysaccharide (ASKP) has attracted growing attention in the field of food and medical engineering due to its biological activity and colloidal property. In this study, the binding between ASKP and ferric ions was found and the binding mechanism was explored. The results showed that ASKP could form a hydrogel with three-dimensional network structure in the presence of ferric ions. Ferric ions could specifically bind with the carboxyl and hydroxyl groups of the high molecular weight fraction of 60P with the binding stoichiometry of [M3+]/[repeating unit] = 2.5. The possible mechanism of the formation of ASKP-Fe3+ complex was proposed as two binding modes of monodentate and bridging binding. ASKP-Fe3+ complex exhibited higher thermal stability than ASKP revealed with DSC thermograms. The study indicated that ASKP would be a novel gelation biopolymer and the ASKP-Fe3+ complex hydrogel could be exploited as a new iron fortifier.

Improving the Stability of Oil Body Emulsions from Diverse Plant Seeds Using Sodium Alginate.

In this study, peanut, sesame, and rapeseed oil bodies (OBs) were extracted by the aqueous medium method. The surface protein composition, microstructure, average particle size d 4 ,   3 , ζ-potential of the extracted OBs in aqueous emulsion were characterized. The stability of the OB emulsions was investigated. It was found that different OB emulsions contained different types and contents of endogenous and exogenous proteins. Aggregation at low pHs (<6) and creaming at high pHs (7 and 8) both occurred for all of three OB emulsions. Sodium alginate (ALG) was used to solve the instability of OB emulsions under different conditions-low concentration of ALG improved the stability of OB emulsions below and near the isoelectric point of the OBs, through electrostatic interaction. While a high concentration of ALG improved the OB emulsion stability through the viscosity effect at pH 7. The OB emulsions stabilized by ALG were salt-tolerant and freeze-thaw resistant.

Preparation and stability of nano-scaled gel beads of λ-carrageenan bound with ferric ions.

Iron-deficiency anemia (IDA) is a major global public health problem, and the iron fortifiers in diet are clearly needed in the prevention and improvement of IDA for humans. A novel nano-scaled gel beads of λ-carrageenan (λ-car) specifically binding with ferric ions was developed to be a promising iron fortifier with no adverse organoleptic changes on food. Turbidity measurement, thermogravimetric analysis and Fourier transform infrared spectroscopy confirmed the successful chelating. The gel beads of λ-car-Fe3+ complex showed good dispersibility and solvent stability. The in vitro cell viability of HepG2 cells treated with λ-car-Fe3+ was over 75% at 5 mg/mL of ferric ions, indicating a significant cytotoxicity reduction of ferric ions. The stability of λ-car-Fe3+ complex powder was obviously increased against browning during 60 d storage with zein coating, which was attributed to the prevention of moisture permeation. Zein coated gel beads also performed a slow release of ferric ions in simulated gastrointestinal juices, resulting from the compact and hydrophobic zein surface delaying the dissociation of λ-car-Fe3+ in acidic environment. This λ-car-Fe3+ complex would have a great potential as a safe iron fortifier and facilitate iron supplementary with the advantage to relieve the side effects of iron ions.

Effect of sodium alginate on the stability of natural soybean oil body emulsions.

In this work, the influence of an anionic polysaccharide, sodium alginate (ALG), on the stability of soybean oil body (OB) emulsions under different environmental conditions, including NaCl, pH and freeze-thaw cycling, was studied by analyzing the particle electric charge, particle size and distribution, and using optical and fluorescence microscopy. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the proteins on the surface of oil bodies were mainly oleosins. It was found that ALG can be adsorbed to the surface of oil bodies by strong electrostatic interactions at pH 4.5 and the optimal concentration of ALG was 0.35 wt% for 1 wt% OB emulsions. At pH 4-8, ALG-coated OB emulsions were more stable than uncoated OB emulsions with smaller particle size and more uniform size distribution due to the interaction between OB and ALG. The ALG-coated OB emulsions were also all stable against NaCl within the concentration range of 0-250 mM at pH 7 while uncoated OB emulsions aggregated gradually with the increase of NaCl concentration. For OB emulsions with higher concentration of 10-40 wt% which are frequently used in the food industry, the minimal concentration of ALG required to make stable emulsions was found to be 0.5-1 wt%, correspondingly. Coating oil bodies with ALG also significantly improved the stability of natural oil body emulsions against freeze-thaw cycling, which is of great significance to the further development of natural oil body-based products in food industry.

Stability and Oil Migration of Oil-in-Water Emulsions Emulsified by Phase-Separating Biopolymer Mixtures.

Oil-in-water (O/W) emulsions with varying concentration of oil phase, medium-chain triglyceride (MCT), were prepared using phase-separating gum arabic (GA)/sugar beet pectin (SBP) mixture as an emulsifier. Stability of the emulsions including emulsion phase separation, droplet size change, and oil migration were investigated by means of visual observation, droplet size analysis, oil partition analysis, backscattering of light, and interfacial tension measurement. It was found that in the emulsions prepared with 4.0% GA/1.0% SBP, when the concentration of MCT was greater than 2.0%, emulsion phase separation was not observed and the emulsions were stable with droplet size unchanged during storage. This result proves the emulsification ability of phase-separating biopolymer mixtures and their potential usage as emulsifiers to prepare O/W emulsion. However, when the concentration of MCT was equal or less than 2.0%, emulsion phase separation occurred after preparation resulting in an upper SBP-rich phase and a lower GA-rich phase. The droplet size increased in the upper phase whereas decreased slightly in the lower phase with time, compared to the freshly prepared emulsions. During storage, the oil droplets exhibited a complex migration process: first moving to the SBP-rich phase, then to the GA-rich phase and finally gathering at the interface between the two phases. The mechanisms of the emulsion stability and oil migration in the phase-separated emulsions were discussed.

Protein/Polysaccharide Electrostatic Complexes and Their Applications in Stabilizing Oil-in-Water Emulsions.

Consumers are becoming increasingly fastidious in demanding food products with improved quality and functionality. This largely relies on rational design of food structures. As the two key food ingredients, protein and polysaccharides play important roles in food structuring. The combination of protein and polysaccharide provides rich opportunities for food structure and function designs through molecular interaction and assembly. This paper provides a brief review on the formation and characterization of protein/polysaccharide electrostatic complexes and their applications in stabilizing oil-in-water emulsions, particularly those containing polyunsaturated fatty acids.

Aggregation behaviour and stability of maize germ oil body suspension.

To utilize maize germ oil bodies as ingredients in the food industry, zeta potential determination and particle diameter analysis were used in this study as indicators of the stability of maize germ oil body suspensions. The stability and aggregation properties of maize germ oil body suspensions were studied at different pH and ion strength conditions, and different thermal treatments. Zeta potential measurement, mean particle size determination, and confocal laser scanning microscopy were also performed. The zeta potential and mean particle diameter of the oil bodies were easily affected by salt (7.05 mV and d32=1.43 µm at 100mM NaCl) and pH (23.30, 15.00, -16.43 mV and d32=1.02, 2.55, and 0.95 µm at pH 3, pH 4, and pH 7, respectively). Results demonstrated that aggregation and instability of the oil bodies were promoted by high salt concentrations and acidic pH but not by heating. The association between oil bodies and surfactant properties resulted in the disruption of hydrophobic interactions among oil body surface proteins because of the smaller mean particle sizes and the reduced negative charges (-75.73 mV and d32=0.46 µm at pH 3). Thus, the oil body aggregation behavior and stability of proteins are based on hydrophobic interactions present on the surface of the oil bodies.