Unveiling the applications of membrane proteins from oil bodies: leading the way in artificial oil body technology and other biotechnological advancements.

Oil bodies (OBs) function as organelles that store lipids in plant seeds. An oil body (OB) is encased by a membrane composed of proteins (e.g., oleosins, caleosins, and steroleosins) and a phospholipid monolayer. The distinctive protein-phospholipid membrane architecture of OBs imparts exceptional stability even in extreme environments, thereby sparking increasing interest in their structure and properties. However, a comprehensive understanding of the structure-activity relationships determining the stability and properties of oil bodies requires a more profound exploration of the associated membrane proteins, an aspect that remains relatively unexplored. In this review, we aim to summarize and discuss the structural attributes, biological functions, and properties of OB membrane proteins. From a commercial perspective, an in-depth understanding of the structural and functional properties of OBs is important for the expansion of their applications by producing artificial oil bodies (AOB). Besides exploring their structural intricacies, we describe various methods that are used for purifying and isolating OB membrane proteins. These insights may provide a foundational framework for the practical utilization of OB membrane proteins in diverse applications within the realm of AOB technology, including biological and probiotic delivery, protein purification, enzyme immobilization, astringency detection, and antibody production.

Advance in dietary polyphenols as dipeptidyl peptidase-IV inhibitors to alleviate type 2 diabetes mellitus: aspects from structure-activity relationship and characterization methods.

Dietary polyphenols with great antidiabetic effects are the most abundant components in edible products. Dietary polyphenols have attracted attention as dipeptidyl peptidase-IV (DPP-IV) inhibitors and indirectly improve insulin secretion. The DPP-IV inhibitory activities of dietary polyphenols depend on their structural diversity. Screening methods that can be used to rapidly and accurately identify potential polyphenol DPP-IV inhibitors are urgently needed. This review focuses on the relationship between the structures of dietary polyphenols and their DPP-IV inhibitory effects. Different characterization methods used for polyphenols as DPP-IV inhibitors have been summarized and compared. We conclude that the position and number of hydroxyl groups, methoxy groups, glycosylated groups, and the extent of conjugation influence the efficiency of inhibition of DPP-IV. Various combinations of methods, such as in-vitro enzymatic inhibition, ex-vivo/in-vivo enzymatic inhibition, cell-based in situ, and in-silico virtual screening, are used to evaluate the DPP-IV inhibitory effects of dietary polyphenols. Further investigations of polyphenol DPP-IV inhibitors will improve the bioaccessibility and bioavailability of these bioactive compounds. Exploration of (i) dietary polyphenols derived from multiple targets, that can prevent diabetes, and (ii) actual binding interactions via multispectral analysis, to understand the binding interactions in the complexes, is required.

Construction of protein-, polysaccharide- and polyphenol-based conjugates as delivery systems.

Natural polymers, such as polysaccharides and proteins, have been used to prepare several delivery systems owing to their abundance, bioactivity, and biodegradability. They are usually modified or combined with small molecules to form the delivery systems needed to meet different needs in food systems. This paper reviews the interactions of proteins, polysaccharides, and polyphenols in the bulk phase and discusses the design strategies, coupling techniques, and their applications as conjugates in emulsion delivery systems, including traditional, Pickering, multilayer, and high internal-phase emulsions. Furthermore, it explores the prospects of the application of conjugates in food preservation, food development, and nanocarrier development. Currently, there are seven methods for composite delivery systems including the Maillard reaction, carbodiimide cross-linking, alkali treatment, enzymatic cross-linking, free radical induction, genipin cross-linking, and Schiff base chemical cross-linking to prepare binary and ternary conjugates of proteins, polysaccharides, and polyphenols. To design an effective target complex and its delivery system, it is helpful to understand the physicochemical properties of these biomolecules and their interactions in the bulk phase. This review summarizes the knowledge on the interaction of biological complexes in the bulk phase, preparation methods, and the preparation of stable emulsion delivery system.

Encapsulation of cinnamaldehyde: an insight on delivery systems and food applications.

Cinnamaldehyde is an essential oil extracted from the leaves, bark, roots and flowers of cinnamon plants (genus Cinnamomum). Cinnamaldehyde has shown biological functions such as antioxidants, antimicrobials, anti-diabetic, anti-obesity and anti-cancer. However, poor solubility in water as well as molecular sensitivity to oxygen, light, and high temperature limit the direct application of cinnamaldehyde. Researchers are using different encapsulation techniques to maximize the potential biological functions of cinnamaldehyde. Different delivery systems such as liposomes, emulsions, biopolymer nanoparticles, complex coacervation, molecular inclusion, and spray drying have been developed for this purpose. The particle size and morphology, composition and physicochemical properties influence the performance of each delivery system. Consequently, the individual delivery system has its advantages and limitations for specific applications. Given the essential role of cinnamaldehyde in functional food and food preservation, appropriate approaches should be applied in the encapsulation and application of encapsulated cinnamaldehyde. This review systematically analyzes available encapsulation techniques for cinnamaldehyde in terms of their design, properties, advantages and limitations, and food application status. The information provided in this manuscript will assist in the development and widespread use of cinnamaldehyde-loaded particles in the food and beverage industries.

Stability and digestibility of encapsulated lycopene in different emulsion systems stabilized by acid-modified soybean lipophilic protein.

BACKGROUND: Owing to the harsh acidic environment of the stomach, acid-resistant emulsion products have wide-ranging applications in the food industry. Herein, natural soybean lipophilic protein (LP) was used to establish coarse emulsions, nanoemulsions, emulsion gels, and high internal phase Pickering emulsions (HIPPE) under acidic conditions. Furthermore, the carrying characteristics of the acid-resistant emulsion system with lycopene were explored. RESULTS: Comparisons of particle sizes, potentials, microstructures, and rheology of the four carrier systems revealed that HIPPE has a single particle-size distribution, the largest zeta potential, and an elastic gel-like network structure. Comparison of encapsulation rates indicated that HIPPE had the best effect on encapsulating lycopene, reaching approximately 90%. The pH stability, storage stability, and simulated in vitro digestion experiments showed that the four emulsions that were stable under acidic conditions had good acid resistance. Among them, the acid-induced LP-stabilized HIPPE had the best storage stability and superior compatibility with the harsh acidic environment of the stomach, which not only achieved the purpose of delaying the release of lipids but also conferred better protection to lycopene in the gastric tract; moreover, it achieved the best bioavailability. CONCLUSION: LP-stabilized HIPPE has the best stability and can yield better absorption and utilization of lycopene by the body. The results of this study are helpful for the development of acid-resistant functional emulsion foods that are conducive to the absorption of lycopene. © 2022 Society of Chemical Industry.

Development of high-internal-phase emulsions stabilized by soy protein isolate-dextran complex for the delivery of quercetin.

BACKGROUND: Protein-polysaccharide complexes have been widely used to stabilize high-internal-phase emulsion (HIPEs). However, it is still unknown whether soy protein isolate-dextran (SPI-Dex) complexes can stabilize HIPEs or what is the effect of Dex concentration on the HIPEs. Furthermore, the non-covalent interaction mechanism between SPI and Dex is also unclear. Therefore, we fabricated SPI-Dex complexes and used them to stabilize HIPEs-loaded quercetin and explore the interaction mechanism between SPI and Dex, as well as the effect of Dex concentration on the particle size, ζ-potential, microstructure, rheology, quercetin encapsulation efficiency, and gastrointestinal fate of the HIPEs. RESULTS: Spectral analysis (fourier transform infrared spectroscopy, ultraviolet spectroscopy, and fluorescence spectroscopy) results identified the formation of SPI-Dex complexes, and indicated that the addition of Dex changed the spatial structure of SPI, whereas thermodynamic analysis (ΔH > 0, ΔS > 0) showed that hydrophobic interactions were the main driving forces in the formation of SPI-Dex complexes. Compared with HIPEs stabilized by SPI, the SPI-Dex complex-stabilized HIPEs had smaller particles (3000.33 ± 201.22 nm), as well as higher ζ-potential (-21.73 ± 1.10 mV), apparent viscosities, modulus, and quercetin encapsulation efficiency (98.19 ± 0.14%). In addition, in vitro digestion revealed that SPI-Dex complex-stabilized HIPEs significantly reduced the release of free fatty acid and improved quercetin bioaccessibility. CONCLUSION: HIPEs stabilized by SPI-Dex complexes delayed the release of free fat acid and improved the bioaccessibility of quercetin, and may be help in designing delivery systems for bioactive substances with specific properties. © 2022 Society of Chemical Industry.

Soy/whey protein isolates: interfacial properties and effects on the stability of oil-in-water emulsions.

BACKGROUND: The adsorption of proteins at oil/water interfaces can reduce interfacial tension and increase emulsion stability. However, emulsions stabilized by soy protein isolate (SPI) are not sufficiently stable. Using SPI as a control, a theoretical basis for the adsorption behavior of mixed SPI and whey protein isolate (WPI) at the oil/water interface was established and the effects of the protein ratio and content on the emulsion stability were studied. RESULTS: Compared to SPI solution, SPI-WPI mixed solutions were found to reduce the size distribution of emulsion droplets and significantly improve the emulsion stability. Among the studied protein contents and ratios, the protein content of 0.2 g kg-1 and SPI/WPI mass ratio of 1:9 offered the lowest creaming stability index (15%), the smallest droplet size (278 nm), and the largest absolute value ζ-potential (35 mV), i.e. the emulsion stability was excellent. The largest dilatational modulus (10.08 mN m-1 ), dilatational elasticity (10.01 mN m-1 ), and dilatational viscosity (1.18 mN m-1 ), were observed with a protein content of 0.15 g kg-1 (SPI/WPI ratio of 1:9), along with a high interfacial protein adsorption capacity (47.33%). SPI-WPI complexes form a thick adsorption layer around oil droplets, resulting in an increase of the expansion modulus of the interfacial layer. CONCLUSION: SPI-WPI complexes can form a thick adsorption layer around oil droplets, resulting in increased expansion modulus of the interfacial layer, which improves emulsion stability. © 2020 Society of Chemical Industry.

Characterizing the Structural and Functional Properties of Soybean Protein Extracted from Full-Fat Soybean Flakes after Low-Temperature Dry Extrusion.

The soy protein isolates (SPI) extracted from different extruded full-fat soybean flakes (FFSF), and their conformational and functional properties were characterized. Overall, the free thiol (SH) content of SPI increased when the extrusion temperature was below 80 °C and decreased at higher temperatures. Soy glycinin (11S) showed higher stability than ß-conglycinin (7S) during extrusion. Results also indicated that the increase in some hydrophobic groups was due to the movement of hydrophobic groups from the interior to the surface of the SPI molecules at extrusion temperatures from 60 to 80 °C. However, the aggregation of SPI molecules occurred at extrusion temperatures of 90 and 100 °C, with decreasing levels of hydrophobic groups. The extrusion temperature negatively affected the emulsifying activity index (EAI); on the other side, it positively affected the emulsifying stability index (ESI), compared to unextruded SPI.

Effect of ultrasound treatment on the wet heating Maillard reaction between mung bean [Vigna radiate (L.)] protein isolates and glucose and on structural and physico-chemical properties of conjugates.

BACKGROUND: The objective of this study was to determine the effect of ultrasound treatment on the wet heating Maillard reaction between mung bean protein isolates (MBPIs) and glucose, and on structural and physico-chemical properties of the conjugates. RESULTS: The degree of glycosylation of MBPI-glucose conjugates treated by ultrasound treatment and wet heating (MBPI-GUH) was higher than that of MBPI-glucose conjugates only treated by wet heating (MBPI-GH). Solubility, emulsification activity, emulsification stability and surface hydrophobicity of MBPI-GUH were higher than that of MBPI-GH. Grafted MBPIs had a lower content of α-helix and unordered coil, but a higher content of ß-sheet and ß-turn structure than MBPIs. No significant structural changes were observed in ß-turn and random coil structure of MBPI-GUH, while α-helix content increased with ultrasonic time, and decreased at 300 W ultrasonic power with the increase of ß-sheet. MBPI-GUH had a less compact tertiary structure compared to MBPI-GH and MBPI. Grafting MBPIs with glucose formed conjugates of higher molecular weight, while no significant changes were observed in electrophoresis profiles of MBPI-GUH. CONCLUSION: Ultrasound-assisted wet heating Maillard reaction between MBPIs and glucose could be a promising way to improve functional properties of MBPIs.

Raman Spectroscopy Study Structural Changes in Black Bean Protein Isolate upon Ultrasonic-Treatment.

This article focused on the assessment of the potential of Raman spectroscopy for the determination of structural changes in black-bean protein isolate (BBPI) dispersions with low-frequency (20 kHz) ultrasonication applied at various powers (150, 300 or 450 W) and for different durations (12 or 24 min). It also reported on differential scanning calorimetry analyses. A decrease in TD at low- and medium-power ultrasonication confirmed these ultrasonication treatment disrupted internal hydrophobic interactions of protein molecules and broke up unstable aggregates to smaller soluble protein aggregates, while an increase in TD at high-power was attributed to repolymerization of aggregates. Raman spectroscopy analysis revealed a decrease in the α-helix proportion and an increase in ß-sheets after ultrasonic treatment except Sample E (300 W, 24 min). Transformation of aggregation results in a reconstruction in secondary structure of BBPI, especially in ß-sheet structure. Ultrasonic-treatment induced a decrease in the normalized intensity of the Raman band near 760 cm-1 which indicated that Tryptophan residues tended to expose and also indicated protein partially unfolding. No significant difference was found in Tyr doublet ratios between unheated and ultrasound-treated BBPI indicated that ultrasound did not change the microenvironment around tyrosyl residues. While the intensity of 1 450 cm-1 band increased with increasing ultrasonic intensity and treatment time, and then decreased with further increase in power and treatment time. In general, the formation of aggregation transferred g-g-t conformation to t-g-t conformation. Though some mechanism of aggregation-repolymerization of BBPI remains to be clearly defined, Raman spectroscopy provide a feasible tool to study the structural changes of BBPI prepared under different ultrasonic conditions, give a new perspective to elucidation of protein structure.

Effect of extruding full-fat soy flakes on trans fat content.

To evaluate the effects of extrusion process on the trans fatty acids (TFAs) formation in soybean crude oils, three different extrusion parameters, namely, extrusion temperature (80-160 °C), feed moisture (10-26%), and screw speed (100-500 rpm), were carried out. It was found that only five different types of TFAs were detected out using gas chromatography-mass spectrometry. Before the extrusion started, the initial amount of total TFAs was 3.04 g/100 g. However, after extruding under every level of any variable, the total amounts of TFAs were significantly higher than those in the control sample (P < 0.05). For example, taking the effect of extrusion temperature into account, we can find that the highest amount of total of trans fatty acid (TTFA) was 1.62 times the amount of that in the control sample, whereas the lowest amount of TTFA was 1.54 times the amount of that in the control sample. Importantly, it was observed that the amounts of every type of trans fatty acid were not continuously increasing with the increase of the level of any extrusion variable. This phenomenon demonstrated that the formation and diversification were intricate during extruding process and need to be further studied.

Comparison of pH-induced protein-polyphenol self-assembly methods: Binding mechanism, structure, and functional characteristics.

Herein, we used pH-shifted and pH-driven methods to assemble kidney-bean protein isolate (KPI) and luteolin (Lut) into a nanocomplex and subsequently investigated their binding mechanism, structure, and functional properties. Results showed that the nanocomplex prepared by the pH-driven method exhibited a better encapsulation effect and controlled release of Lut. Fluorescence spectroscopy and molecular docking analysis showed that the binding affinities under alkaline conditions were higher than those under acidic and neutral conditions. Various spectral techniques were used to determine the structural changes in the KPI-Lut nanocomplex, including the transformation of α-helices and ß-sheets and alteration of specific amino acid microenvironments, which were more pronounced in the pH-driven nanocomplex. The structural changes in the nanocomplex further affected their surface hydrophobicity and thermal stability. Additionally, the combination of KPI and Lut significantly improved the antioxidant activity and α-glucosidase inhibitory ability of the resultant nanocomplexes, particularly the one prepared by the pH-driven method.

Ultrasound-assisted modification of soybean protein isolate with L-histidine: Relationship between structure and function.

Herein, the effects of ultrasound-assisted L-histidine (L-His) on the physicochemical properties and conformation of soybean protein isolate (SPI) were investigated. Particle size, zeta potential, turbidity, and solubility were used to evaluate protein aggregation, and the relationship between structural and functional changes of the proteins was characterized using spectral analysis, surface hydrophobicity, emulsification, and antioxidant properties. After ultrasound-assisted L-His treatment, SPI exhibited a smaller particle size, higher solubility, and more homogeneous micromorphology owing to the decrease in alpha-helix content and subsequent increases in zeta potential and active sulfhydryl content. In addition, spectral analysis showed that L-His and SPI could form a complex, which changed the microenvironment of the amino acid residues in SPI, thus improving its emulsification and antioxidant properties. At the concentration of L-His was 0.3 % w/w, the nanocomplex had a smaller particle size (140.03 nm), higher ζ-potential (-23.63 mV), and higher emulsification stability (22.48 min).

Casein-phosphatidylcholine emulsifier remodels LPS-induced intestinal barrier disfunction via regulating ferroptosis and lipid metabolism.

Recently, the biosafety of synthetic emulsifier in intestinal barrier has raised significant concerns. Casein- phosphatidylcholine (CP), which is a natural emulsifier, has better emulsification and stability. However, the effect of CP on intestinal barrier remains unknow. Intestinal permeability and lipomics analysis showed that carboxymethyl cellulose (CMC) and CP have no significant effect on intestinal barrier in normal intestinal barrier model, whereas CP increased transmembrane resistance value and remodeled lipid homeostasis in LPS induced intestinal barrier dysfunction model, indicating its superior biosafety. To explore the underlying molecular mechanism of emulsifier on intestinal barrier dysfunction, the bioinformatics analysis of six original microarray datasets including 168 cases in NCBI-Gene Expression Omnibus database showed ferroptosis-related genes showed a significant differential expression. The quantitative polymerase chain reaction analysis demonstrated that CP can repair the imbalance of lipid homeostasis induced by LPS and restore normal intestinal permeability by regulating the expression of ferroptosis-related genes, while CMC could can enhance intestinal permeability by inducing ferroptosis of intestinal epithelial cells through lipid peroxidation. In conclusion, this study highlighted CP could remodel LPS-induced intestinal barrier disfunction via regulating ferroptosis and lipid metabolism. These findings can be used as a new insight for the design of new healthy emulsifier.

Fibrillation of soy protein isolate in the presence of metal ions: Structure and gelation behavior.

The structure and functional properties of protein fibrils are closely related to environmental factors in fibrillation. Herein, soy protein isolate fibrils (SPIFs, 22 mg/mL) were prepared under acid-heating conditions in the presence of 100 mM metal ions (K+, Na+, Ca2+, Mg2+, and Fe3+). Except for Fe3+, fibrillation and subsequent larger fibril aggregates were promoted, ultimately leading to gel formation. Compared with K+ or Na+, the addition of Ca2+ or Mg2+ resulted in more organized SPIF structures with increased ß-sheet contents and higher ThT fluorescence intensities. Furthermore, both of them resulted in longer fibrils with an average contour length of 700-800 nm, which significantly enhanced the storage modulus. However, the presence of Fe3+ accelerated protein hydrolysis and inhibited SPIF formation, resulting in samples consistently exhibited liquid behavior. These findings provide a foundation for understanding the influence of metal ions on regulating the fibrillation and gelling properties of SPIFs.

Comparison of the physical stability, microstructure and protein-lipid co-oxidation of O/W emulsions stabilized by l-arginine/l-lysine-modified soy protein hydrolysate.

This work investigated the physical stability, microstructure, and oxidative stability of the emulsions prepared by soy protein hydrolysate (SPH) after modification with different concentrations of l-arginine and l-lysine. l-Arginine and l-lysine significantly increased the absolute zeta potential values, and decreased droplet sizes of the emulsions, thereby improving the physical stability of the emulsions. Meanwhile, l-arginine and l-lysine markedly decreased the apparent viscosity of the emulsions. The measurement of interfacial protein adsorption percentage showed that l-arginine (≤0.5 %) promoted the adsorption of SPH at the oil-water interface, whereas l-lysine (≤1%) reduced the adsorption of SPH at the oil-water interface. In addition, l-arginine and l-lysine (≤0.5 %) could retard lipid and protein oxidation. Correlation analysis indicated that the improvement in the physical stability of the emulsions by l-arginine and l-lysine also enhanced the oxidative stability of the emulsions. In summary, l-arginine and l-lysine could be effective modifiers for the protein-based emulsion systems.

Self-assembly and aggregation behavior of temperature-controlled modified glycinin and d-galactose colloidal particles.

The molecular structure and morphologies of complex colloidal particles with modified glycine (S-11S) and d-galactose were studied by multispectral, microscopic imaging and chromatographic techniques at different temperatures, and the self-assembly and aggregation mechanisms were determined. Overall, high-temperature-treated S-11S and d-galactose associate at cysteine and phenylalanine sites and self-assemble into colloidal particles of greater stability than glycinin and S-11S via ionic and disulfide bonds. The structure and subunit content of composite colloidal particles were changed. Assessing the sub-microstructure reveals that temperature can regulate the directional aggregation of complex colloidal particles. The elasticity of the complex colloidal particles is maximum enhanced at 95 ℃ as confirmed by the rheological. Thus, the heat-treated aggregation of the soy protein and its complex was evaluated to provide a new theoretical basis for the application of soy protein in gels and other areas and contribute to the design of new soy protein products.

Effect of sodium alginate block type on the physicochemical properties and curcumin release behavior of quaternized chitosan-oxidized sodium alginate Schiff base hydrogels.

Controlling bioactive ingredients release by modulating the 3D network structure of cross-linked hydrogels is important for functional food development. Hereby, oxidized sodium alginate (OSA) with varying aldehyde contents was formed by periodate oxidation of sodium alginate (SA) with different ß-d-mannuronic acid (M) and α-l-guluronic acid (G) ratios (M/G = 1:2, 1:1, and 2:1) and its structure was characterized. Moreover, hydrogels were prepared via Schiff base and electrostatic interactions between quaternized chitosan (QCS) and OSA. The properties of hydrogels such as microstructure, thermal stability, swelling and controlled release were investigated. The results showed that OSA with M/G = 1:2 had the highest content of aldehyde groups, and the hydrogel formed by it and QCS had higher thermal stability and a denser network structure with the lowest equilibrium swelling rate, which could better control the release of curcumin. Additionally, it had good self-healing and can recover rapidly after the rupture of its network structure.

Changes in the structure and functional properties of soybean isolate protein: Effects of different modification methods.

Soybean protein isolate (SPI) is an important plant protein in food processing; however, its spherical structure prevents the exposure of its hydrophobic residues and affects its functional properties. In this study, we elucidate the effects of deamidation, phosphorylation, and glycosylation on the structure (Fourier-transform infrared spectroscopy, circular dichroism, fluorescence, and scanning electron microscopy) and functional properties (solubility, emulsifying activity index (EAI), and emulsifying stability index (ESI)) of SPI. The zeta potentials of the deamidated, phosphorylated, and glycosylated (DSPI, PSPI, and MSPI, respectively) samples decreased significantly (p < 0.05) relative to those of SPI. The functional properties of the modified SPI samples were improved, with MSPI-2 showing the best solubility (86.73 ± 0.34%), EAI (118.89 ± 0.73 m2/g), and ESI (273.33 ± 0.59 min). Moreover, the effects of the three modifications on the SPI functional properties increase in the order MSPI > PSPI > DSPI. These results provide a theoretical understanding the relationship between the modifications and SPI structure.

Crosslinking of gelatin Schiff base hydrogels with different structural dialdehyde polysaccharides as novel crosslinkers: Characterization and performance comparison.

Few studies have been conducted on the relationship between the crosslinking ability of dialdehyde polysaccharides (DPs) with different structures and the structure and properties of hydrogels. Herein, the effects of dialdehyde sodium alginate (DSA), dialdehyde guar gum (DGG), and dialdehyde dextran (DDE) as crosslinking agents for gelatin (GE)-based hydrogels were comparatively studied. First, the structure and aldehyde content of DPs were evaluated. Subsequently, the structure, crosslinking degree, and physicochemical properties of GE/DP hydrogels were characterized. Compared with pure GE hydrogels, GE/DP hydrogels had higher thermal stability and mechanical properties. Moreover, the aldehyde content of DPs was ordered as follows: DSA < DGG < DDE. The higher crosslinking degree of the hydrogels formed by DPs with a higher aldehyde content resulted in smaller hydrogel pores, higher mechanical strength, and a lower equilibrium swelling rate. These observations provide a theoretical basis for selecting crosslinking candidates for hydrogel-specific applications.