Mechanism of ultrasound-induced soybean/egg white composite gelation: Gel properties, morphological structure and co-aggregation kinetics.

This study aims to develop ultrasound-assisted acid-induced egg white protein (EWP)-soy protein isolate (SPI) composite gels and to investigate the mechanistic relationship between the co-aggregation behavior of composite proteins and gel properties through aggregation kinetics monitored continuously by turbidity. The results showed that the inclusion of EWP caused the attenuation of gel properties and maximum aggregation (Amax) because EWP could aggregate with SPI at a higher rate (Kapp), which impeded the interaction between SPI and the formation of a continuous gelling network. In the EWP-dominated system, SPI with higher molecular weights also increased the fractal dimension of gels. Ultrasound improved properties of composite gels, especially the SPI-dominated system. After ultrasound treatment, the small, uniform size of co-aggregates and the decrease in potential led to an increase in the aggregation rate and formation of dense particles, consistent with an increase in gelation rate and texture properties. Excessively fast aggregation generated coarse chains and more pores. Still, the exposure of free sulfhydryl groups assisted the gel structure units to form a compact network through disulfide bonding. On the whole, the study could provide theoretical support for a deeper understanding on the interaction mechanism and gelation of composite proteins.

Insight into the improvement mechanism of gel properties of pea protein isolate based on the synergistic effect of cellulose nanocrystals and calcium ions.

Here, the influence and potential mechanism by which cellulose nanocrystals (CNC) collaborated with Ca2+ enhancing the heat-induced gelation of pea protein isolate (PPI) were investigated. It was found that the combination of 0.45% CNC and 15 mM Ca2+ synergistically increased the gel strength (from 14.18 to 65.42 g) and viscoelasticity of PPI while decreased the water holding capacity. The improved particle size, turbidity, and thermostability as well as the reduced solubility, crystallinity, and gel porosity were observed in CNC/CaCl2 composite system. CNC fragments bind to specific amino acids in 11S legumin and 7S vicilin mainly through hydrogen bonding and van der Waals forces. Moreover, changes in the protein secondary structure and enhancement of the molecular interaction induced by CNC and Ca2+ could favor the robust gel network. The results will provide a new perspective on the functional regulation of pea protein and the creation of pea protein gel-based food.

Viscoelastic behavior, gelation properties and structural characterization of Deccan hemp seed (Hibiscus cannabinus) protein: Influence of protein and ionic concentrations, pH, and temperature.

This study investigates the viscoelastic behavior, gelling properties, and structural characteristics of Deccan hemp seed protein (DHSP) to overcome limitations in its application in food formulations. Small amplitude oscillatory shear measurements were employed to investigate the impact of protein concentration, pH, ionic concentration, and temperature on DHSP's rheological features. The study revealed that the 20 % protein dispersion had the highest storage modulus (G') and yield stress at 63.96 ± 0.23 Pa and 0.61 Pa, respectively. DHSP dispersion exhibited pseudo-plastic behavior across various conditions. The gelling performance was higher at pH 4 and 8 and at ionic concentration in the range of 0.1 M - 0.5 M. Gelation time and temperature were observed from the temperature ramp test. Structural characterizations, including fluorescence spectroscopy, circular dichroism spectra, FTIR spectra, SEM, AFM images, zeta potential analysis, and DSC, provided insights into DHSP's tertiary and secondary conformation, surface characteristics, and thermal properties. Notably, the study highlighted DHSP's exceptional rheological properties, making it a promising gelling material for the food and nutraceutical industries. The findings also offer new insights into DHSP's structural characteristics, suggesting potential applications in food packaging and product development within the food industry.

Insight into the Mechanism of Porcine Myofibrillar Protein Gel Properties Modulated by κ-Carrageenan.

The purpose of this study is to explain the mechanism of porcine myofibrillar protein gel properties modulated by κ-carrageenan. The textural properties results showed that the stress at fracture of the composite gel with 0.4% κ-carrageenan had the highest value (91.33 g), which suggested that the 0.4% κ-carrageenan addition was the limitation. The strain at fracture was significantly reduced with κ-carrageenan addition. The composite gel with 0.4% κ-carrageenan had the lowest proportion of T22 (7.85%) and the shortest T21 relaxation time (252.81 ms). The paraffin section showed that the phase separation behavior of the composite gel transformed from single-phase behavior to dispersed phase behavior to bi-continuous phase behavior, and the ratio of CG/MP phase area significantly increased from 0.06 to 1.73. The SEM showed that the three-dimensional network of myofibrillar protein transformed from a loose structure to a compact structure to an unaggregated structure with κ-carrageenan addition. The myofibrillar protein network of the treatment with 0.4% κ-carrageenan had the highest DF value (1.7858) and lowest lacunary value (0.452). The principal component analysis was performed on the data of microstructure and textural properties, and the results showed that the dispersed phase behavior and moisture stabilization promoted the aggregation of myofibrillar protein and the composite gel had better water holding capacity and textural properties, while bi-continuous phase behavior hindered the aggregation of myofibrillar protein and the composite gel had worse water holding capacity and textural properties.

Effects of atmospheric cold plasma and high-power sonication on rheological and gelling properties of mung bean protein dispersions.

The objective of this study was to evaluate the impact of high-power sonication (HPS) and atmospheric cold plasma (ACP) on gelling and rheological properties of mung bean protein dispersions. HPS at 250 J/mL for 2 min and ACP at 80 kV for 5 min were applied to different concentrations of mung bean protein isolate (MBPI). Control and HPS-treated MBPI dispersions showed a minimum gelling concentration (MGC) of 16% w/v, while ACP-treated dispersions started to gel at 14% w/v. Dynamic rheology of dispersions at 16 % concentrations showed that HPS and ACP treatments could reduce the initial gelling temperature to 52° and 65 °C, respectively, from 75 °C for no-treatment control. ACP-treated 16% protein dispersions showed a six-fold higher storage modulus (G') than the control. In addition, ACP treatment resulted in significantly more hydrophobic bonds (∼5.0 g/L) than control (∼1.4 g/L) and HPS-treated (∼1.1 g/L) MBPI gels; however, the net interaction of ionic, hydrogen, hydrophobic, and disulfide bonds was higher in HPS-treated MBPI gels. Thus, both ACP and HPS treatments altered the gelling characteristics of mung bean protein dispersions- ACP reduced MGC and improved firmness, whereas HPS improved the springiness, cohesiveness, gumminess, chewiness, and resilience of the gels.

Squid Suckerin-Spider Silk Fusion Protein Hydrogel for Delivery of Mesenchymal Stem Cell Secretome to Chronic Wounds.

Chronic wounds are non-healing wounds characterized by a prolonged inflammation phase. Excessive inflammation leads to elevated protease levels and consequently to a decrease in growth factors at wound sites. Stem cell secretome therapy has been identified as a treatment strategy to modulate the microenvironment of chronic wounds via supplementation with anti-inflammatory/growth factors. However, there is a need to develop better secretome delivery systems that are able to encapsulate the secretome without denaturation, in a sustained manner, and that are fully biocompatible. To address this gap, a recombinant squid suckerin-spider silk fusion protein is developed with cell-adhesion motifs capable of thermal gelation at physiological temperatures to form hydrogels for encapsulation and subsequent release of the stem cell secretome. Freeze-thaw treatment of the protein hydrogel results in a modified porous cryogel that maintains slow degradation and sustained secretome release. Chronic wounds of diabetic mice treated with the secretome-laden cryogel display increased wound closure, presence of endothelial cells, granulation wound tissue thickness, and reduced inflammation with no fibrotic scar formation. Overall, these in vivo indicators of wound healing demonstrate that the fusion protein hydrogel displays remarkable potential as a delivery system for secretome-assisted chronic wound healing.

Effect of temperature and pH on the gelation, rheology, texture, and structural properties of whey protein and sugar gels based on Maillard reaction.

This study aimed to determine the effect of initial pH and temperature on whey protein gel formation via the Maillard reaction, including changes in gel structure, rheological and texture properties. The color changes in the whey protein and glucose gels were not significant with increasing heat temperature. High temperature and alkaline conditions promoted exposure to hydrophobic groups such as -SH, which accelerated protein aggregation and gel formation. Moreover, the increased particle size and additional hydrophobic groups contributed to higher elastic modulus (G') in the whey protein gel. Fluorescence measurements revealed that more tryptophan on the protein surface decreased with increasing temperature, which indicated that exposure to tryptophan could increase the hydrophobicity of the protein gels. Whey proteins formed stronger, gummier, more elastic, and more cohesive gels at 70 ℃ under initial pH 9 conditions, which also increased with the addition of fructose.

Use of l-arginine-assisted ultrasonic treatment to change the molecular and interfacial characteristics of fish myosin and enhance the physical stability of the emulsion.

The effects of l-arginine (Arg)-assisted ultrasonic treatment on the molecular and interfacial characteristics of myosin and emulsifying properties of the emulsion were evaluated to ascertain the underlying mechanism in improving the emulsion stability. Ultrasonication induced the exposure of residues of native myosin, which was increased by the addition of Arg (40 mM). Furthermore, in terms of emulsions containing Arg, the higher the ultrasonication intensity was, the greater the increase in adsorbed protein (from 15.43 ± 0.28% to 50.49 ± 1.65%) and π value, and the decrease in droplet sizes (from 4098 nm to 2324 nm) (P < 0.05). Moreover, the increase in the ordered structures of interfacial myosin induced by Arg and ultrasonication favoured the formation of a protein gelation network. In summary, Arg-assisted ultrasonic treatment improved the stability of the emulsion by inducing the exposure of native myosin and facilitating the formation of ordered structures of interfacial myosin.

Influence of amino acids on thermal stability and heat-set gelation of bovine serum albumin.

This study investigated the parallels in the influence of amino acid additives on thermal denaturation temperature (Td) and heat-set gelation of bovine serum albumin (BSA). Complete denaturation of BSA occurred only when the gelation temperature (TG) was 14 °C above Td. Under these conditions, the relative effects of various amino acid additives on elevation of Td and gel strength followed a particular order. Further, while zwitterionic amino acids increased both the strength of junction zones and participation of protein in the gel network, sucrose increased the gel strength primarily by strengthening the existing junction zone. The net increase in Td of BSA was linearly correlated with the net increase in gel strength (ΔGS), indicating that the underlying molecular mechanism in both cases might be same. The results suggest that the rheological properties of protein gels can be enhanced by using amino acids, instead of polyols and sugars, as additives.

Heat-induced whey protein isolate gels improved by cellulose nanocrystals: Gelling properties and microstructure.

Cellulose nanocrystals (CNC) were successfully prepared from wheat bran, and their effects on the gelling properties and microstructure of heat-induced whey protein isolate (WPI) gels were investigated. The results showed that the water holding capacity, gel strength, viscoelasticity, and thermal stability of the composite gels were improved by increasing the CNC concentration from 0 to 1.0 % (w/v). The incorporation of CNC restricted water mobility and facilitated conformation conversion of the secondary structure from α-helix to ß-sheet. CNC has good compatibility with the protein matrixes at relatively low concentrations. At higher CNC concentrations, the agglomerated CNC can serve as an active dehydrating agent to absorb moisture in the protein matrixes, which promotes unfolding and cross-linking of the protein molecules. Moreover, the active filling effects of CNC contributed to the formation of a compact and homogeneous gel structure. Therefore, naturally sourced CNC is suggested as a potential gel modifier in food industry.

Effect of κ-carrageenan on gelation and gel characteristics of Antarctic krill (Euphausia superba) protein isolated with isoelectric solubilization/precipitation.

Isoelectric solubilization/precipitation (ISP) was used to extract krill protein isolate (KPI). Due to krill endogenous proteases ISP-KPI barely forms gel which is a major hurdle in wide application of this tremendous protein source for direct human consumption. The objective was to improve gelation and elucidate interaction mechanism between κ-carrageenan and KPI. κ-Carrageenan was added to KPI at 0.00, 0.75, 1.50, 2.25, and 3.00 g/150 g and heated at 90 °C for 15 min. Added κ-carrageenan improved texture of KPI gels. However, the level of addition had minimal effect. SDS-PAGE revealed severe proteolysis of KPI even during heating. Total and free sulfhydryl were not different, while surface hydrophobicity and water-holding-capacity slightly decreased and increased, respectively with added κ-carrageenan. Fourier-transform-infrared-spectroscopy showed none-to-minimal shift of ß-sheet and α-helical protein structure; while CC and CO stretching as well as CH bending of κ-carrageenan showed the greatest shift, indicating that κ-carrageenan was mainly responsible for gel formation.

Surfactant Effects on Particle Generation in Antibody Formulations in Pre-filled Syringes.

Protein aggregation and particle formation have been observed when protein solutions contact hydrophobic interfaces, and it has been suggested that this undesirable phenomenon may be initiated by interfacial adsorption and subsequent gelation of the protein. The addition of surfactants, such as polysorbate 20, to protein formulations has been proposed as a way to reduce protein adsorption at silicone oil-water interfaces and mitigate the production of aggregates and particles. In an accelerated stability study, monoclonal antibody formulations containing varying concentrations of polysorbate 20 were incubated and agitated in pre-filled glass syringes (PFS), exposing the protein to silicone oil-water interfaces at the siliconized syringe walls, air-water interfaces, and agitation stress. Following agitation in siliconized syringes that contained an air bubble, lower particle concentrations were measured in the surfactant-containing antibody formulations than in surfactant-free formulations. Polysorbate 20 reduced particle formation when added at concentrations above or below the critical micelle concentration (CMC). The ability of polysorbate 20 to decrease particle generation in PFS corresponded with its ability to inhibit gelation of the adsorbed protein layer, which was assessed by measuring the interfacial diffusion of individual antibody molecules at the silicone oil-water interface using total internal reflectance fluorescence (TIRF) microscopy with single-molecule tracking.

Chlorogenic acid-mediated gel formation of oxidatively stressed myofibrillar protein.

The effect of chlorogenic acid (CA) at different concentration levels (0, 6, 30, and 150 µmol/g protein) on porcine myofibrillar protein (MP) gelling potential in relation to chemical and structural changes was investigated. The results showed that CA generally inhibited protein carbonyl formation but did not prevent sulphydryl and amine losses caused by oxidation. The presence of CA intensified oxidation-initiated loss of α-helix conformation as well as tertiary structure of MP. CA at 150 µmol/g produced the greatest increase in MP surface hydrophobicity and insolubility. The physicochemical changes with 6 and 30 µmol/g CA led to a remarkably enhanced gelling capacity of MP and augmented the positive effect of oxidation in building an elastic gel network. However, CA at 150 µmol/g was detrimental to the MP gelation. The result can explain why processed meats with phenolic-rich spices and herbs often exhibit variable texture-forming properties.