Effects of ultrasound pretreatment on functional property,antioxidant activity,and digestibility of soy protein isolate nanofibrils

Nanofibrils, an effective method to modulate the functional properties of proteins, can be promoted by ultrasound pretreatment. This study investigated the effect of ultrasound pretreatment on the structure, functional property, antioxidant activity and digestibility of soy protein isolate (SPI) nanofibrils. The results showed that high amplitude ultrasound had a significant effect on structure of SPI nanofibrils. SPI nanofibrils pretreated by 80% amplitude ultrasound showed a blueshift of the amide II band in Fourier transform infrared spectroscopy (FTIR), resulted in more tryptophan residues being buried and increased the crystallinity. Low amplitude ultrasound (20%) pretreatment significantly improved the solubility, emulsifying activity index (EAI) and water absorption capacity (WAC) of SPI nanofibrils, but 80% amplitude ultrasound pretreatment of SPI nanofibrils reduced emulsifying stability index (ESI). High amplitude ultrasound (60% and 80%) pretreatment of SPI nanofibrils improved the foaming capacity and foaming stability and decreased denaturation temperature. DPPH radical scavenging activity of SPI nanofibrils were significantly improved by ultrasound pretreatment. 20% amplitude ultrasound pretreatment improved DPPH, ABTS radical scavenging activity and ferric reducing antioxidant power of SPI nanofibrils. The digestion rate of 80% amplitude ultrasound-pretreated nanofibrils were consistently higher, and SPI nanofibrils pretreated by ultrasound were more fragmented and shorter after simulating gastrointestinal digestion. This study would expand the application of food-grade protein nanofibrils in the food industry.     

Effect mechanism of ultrasound pretreatment on fibrillation Kinetics,physicochemical properties and structure characteristics of soy protein isolate nanofibrils

Self-assembly of soy proteins into nanofibrils is gradually considered as an effective method to improve their technical and functional properties. Ultrasound is a non-thermal, non-toxic and environmentally friendly technology that can modulate the formation of protein nanofibrils through controlled structural modification. In this research, the effect of ultrasound pretreatment on soy protein isolate nanofibrils (SPIN) was evaluated by fibrillation kinetics, physicochemical properties and structure characteristics. The results showed that the optimum ultrasound condition (20% amplitude, 15 min, 5 s on-time and 5 s off-time) could increase the formation rate of SPIN by 38.66%. Ultrasound reduced the average particle size of SPIN from 191.90 ± 5.40 nm to 151.83 ± 3.27 nm. Ultrasound could increase the surface hydrophobicity to 1547.67 in the initial stage of nanofibrils formation, and extend the duration of surface hydrophobicity increased, indicating ultrasound could expose more binding sites, creating more beneficial conditions for nanofibrils formation. Ultrasound could change the secondary and tertiary structure of SPIN. The reduction of α-helix content of ultrasound-pretreated soy protein isolate nanofibrils (USPIN) was 12.1% (versus 5.3% for SPIN) and the increase of β-sheet content was 5.9% (versus 3.5% for SPIN) during fibrillation. Ultrasound could accelerate the formation of SPIN by promoting the unfolding of SPI, exposure of hydrophobic groups and formation of β-sheets. Microscopic images revealed that USPIN generated a curlier and looser shape. And ultrasound reduced the zeta potential, free sulfhydryl groups content and viscosity of SPIN. SDS-PAGE results showed that ultrasound could promote the conversion of SPI into low molecular weight peptides, providing building blocks for the nanofibrils formation. The results indicated that ultrasound pretreatment could be a promising technology to accelerate SPIN formation and promote its application in food industry, but further research is needed for the improvement of the functional properties of SPIN.     

Effect of high-intensity ultrasound on the structural,rheological, emulsifying and gelling properties of insoluble potato protein isolates

The denaturation and lower solubility of commercial potato proteins generally limited their industrial application. Effects of high-intensity ultrasound (HIU) (200, 400, and 600 W) and treatment time (10, 20, and 30 min) on the physicochemical and functional properties of insoluble potato protein isolates (ISPP) were investigated. The results revealed that HIU treatment induced the unfolding and breakdown of macromolecular aggregates of ISPP, resulting in the exposure of hydrophobic and R–SH groups, and reduction of the particle size. These active groups contributed to the formation of a dense and uniform gel network of ISPP gel and insoluble potato proteins/egg white protein (ISPP/EWP) hybrid gel. Furthermore, the increase of solubility and surface hydrophobicity and the decrease of particle size improved the emulsifying property of ISPP. However, excessive HIU treatment reduced the emulsification and gelling properties of the ISPP. Meanwhile, HIU treatment changes the secondary structure of ISPP. It could be speculated that the formation of a stable secondary structure of ISPP initiated by cavitation and shearing effect might play a dominant role on gel strengthens and firmness. Meanwhile, the decrease in relative content of β-turn had a positive effect on the formation of small particle to improve emulsifying property of ISPP.     

Ultrasound driven conformational and physicochemical changes of soy protein hydrolysates

The effect of ultrasound on the conformational and physicochemical properties of soy protein isolate hydrolysates (SPHs) was investigated. SPHs were prepared at hydrolysis times of 20 min, 60 min, and 180 min, then treated with ultrasound for 10 min, 20 min, and 30 min at a frequency of 20 kHz and output powers of 150 W and 450 W. The structural properties and antioxidant capacities of the aqueous layer of SPHs (ASPHs) after sonication were evaluated by Fourier-transform infrared spectroscopy (FTIR), intrinsic fluorescence, DPPH radical scavenging activity assays, and microscopy observations. Results obtained showed that ultrasound treatment significantly disrupted the peptide aggregates formed during protein hydrolysis. The protein solubility was significantly increased after sonication (by up to 18.33%), as did the percentage of proteins with MW < 1 kDa in ASPHs. The antioxidant capacity of ASPHs also increased, as measured by DPPH assay. FTIR analysis of ASPHs indicated that the protein secondary structures were different, with an increase in β-sheet and a decrease in α-helix and β-turn. Furthermore, the changes in fluorescence spectra of ASPHs showed the transition of protein tertiary structure with a greater exposure of Trp residues in the side chains. Scanning electron microscope (SEM) and atomic force microscope (AFM) observations of the morphological structure of ASPHs further confirmed the significant effect of sonication on disrupting peptide aggregates. In conclusion, ultrasound can be used as an efficient treatment to promote the solubility of protein hydrolysates.     

Influence of ultrasound-assisted ionic liquid pretreatments on the functional properties of soy protein hydrolysates

In this work, the effect of dual-frequency ultrasound-assisted ionic liquids (ILs) pretreatment on the functional properties of soy protein isolate (SPI) hydrolysates was investigated. The degree of hydrolysis (DH) of SPI pretreated by ultrasound and [BMIM][PF₆] increased by 12.53% as compared to control (P < 0.05). More peptides with low molecular weight were obtained, providing support for the changes in DH. The trichloroacetic acid-nitrogen soluble index presented an increase, suggesting a better protein hydrolysate property. The increase in the calcium-binding activity showed the ultrasound-assisted ILs pretreatment could potentially improve bone health. The foaming capacity and stability of SPI hydrolysates pretreated by ultrasound-assisted [BMIM][PF₆] always increased remarkably as compared to ultrasound-assisted [BDMIM][Cl] pretreatment. However, the synergistic effect of ultrasound-assisted [BMIM][PF₆] on the emulsifying activity and antioxidant activities (DPPH and hydroxyl radical scavenging activity) was not as ideal as ultrasound-assisted [BDMIM][Cl] pretreatment, which may be affected by the structure of peptide. In conclusion, these results indicated the combination of dual-frequency ultrasound and ionic liquids would be a promising method to improve the functional properties of SPI hydrolysates and broaden the application scope of compound modification in proteolysis industry.     

Effects of ultrasound on the structural and emulsifying properties and interfacial properties of oxidized soybean protein aggregates

Oxidative attack leads to the oxidative aggregation and structural and functional feature weakening of soybean protein. We aimed to investigate the impact of ultrasonic treatment (UT) with different intensities on the structure, emulsifying features and interfacial features of oxidized soybean protein aggregates (OSPI). The results showed that oxidative treatment could disrupt the native soy protein (SPI) structure by promoting the formation of oxidized aggregates with β1-sheet structures through hydrophobic interactions. These changes led to a decrease in the solubility, emulsification ability and interfacial activity of soybean protein. After low-power ultrasound (100 W, 200 W) treatment, the relative contents of β1-sheets, β2-sheets, random coils, and disulfide bonds of the OSPI increased while the surface hydrophobicity, absolute ζ-potential value and free sulfhydryl content decreased. Moreover, protein aggregates with larger particle sizes and poor solubility were formed. The emulsions prepared using the OSPI showed bridging flocculation and decreased protein adsorption and interfacial tension. After applying medium-power ultrasound (300 W, 400 W, and 500 W) treatments, the OSPI solubility increased and particle size decreased. The α-helix and β-turn contents, surface hydrophobicity and absolute ζ-potential value increased, the structure unfolded, and the disulfide bond content decreased. These changes improved the emulsification activity and emulsion state of the OSPI and increased the protein adsorption capacity and interfacial tension of the emulsion. However, after a high-power ultrasound (600 W) treatment, the OSPI showed a tendency to reaggregate, which had a certain negative effect on the emulsification activity and interfacial activity. The results showed that UT at an appropriate power could depolymerize OSPI and improve the emulsification and interfacial activity of soybean protein.     

Influence of high-intensity ultrasound on physicochemical and functional properties of a guamuchil Pithecellobium dulce (Roxb.) seed protein isolate

In this study, the influence of ultrasound on the physicochemical and functional properties of guamuchil seed protein isolate (GSPI) was investigated. The GSPI was prepared by alkaline extraction and isoelectric precipitation method followed by treating with ethanol (95%), from defatted guamuchil seed flour. GSPI suspensions (10%) were sonicated with a probe (20 kHz) at 3 power levels (200 W, 400 W, 600 W) for 15 and 30 min, in addition, to control treatment without ultrasound. Moisture content, water activity, bulk and compact densities and the L*, a* and b* color parameters of the GSPI decreased due to the ultrasound. Glutelin (61.1%) was the main protein fraction in GSPI. Results through Fourier transform infrared and fluorescence spectroscopy showed that ultrasound modified the secondary and tertiary protein structures of GSPI, which increased the surface hydrophobicity, molecular flexibility and in vitro digestibility of GSPI proteins by up to 114.8%, 57.3% and 12.5%, respectively. In addition, maximum reductions of 11.9% in particle size and 55.2% in turbidity of GSPI suspensions, as well as larger and more porous aggregates in GSPI lyophilized powders were observed by ultrasound impact. These structural and physicochemical changes had an improvement of up to 115.5% in solubility, 39.8% in oil absorption capacity, while the increases for emulsifying, foaming, gelling, flow and cohesion properties of GSPI were 87.4%, 74.2%, 40.0%, 44.4%, and 8.9%, respectively. The amelioration of the functional properties of GSPI by ultrasound could represent an alternative for its possible use as a food ingredient in industry.     

Improvement of structural,physicochemical, and rheological properties of porcine myofibrillar proteins by high-intensity ultrasound treatment for application as Pickering stabilizers

This study aimed to evaluate the potential of time-dependent (0, 15, 30, 60, 120 min) treatment of porcine-derived myofibrillar proteins (MPs) with high-intensity ultrasound (HIU) for utilizing them as a Pickering stabilizer and decipher the underlying mechanism by which HIU treatment increases the emulsification and dispersion stability of MPs. To accomplish this, we analyzed the structural, physicochemical, and rheological properties of the HIU-treated MPs. Myosin heavy chain and actin were observed to be denatured, and the particle size of MPs decreased from 3,342.7 nm for the control group to 153.9 nm for 120 min HIU-treated MPs. Fourier-transformed infrared spectroscopy and circular dichroism spectroscopy confirmed that as the HIU treatment time increased, α-helical content increased, and β-sheet decreased, indicating that the protein secondary/tertiary structure was modified. In addition, the turbidity, apparent viscosity, and viscoelastic properties of the HIU-treated MP solution were decreased compared to the control, while the surface hydrophobicity was significantly increased. Analyses of the emulsification properties of the Pickering emulsions prepared using time-dependent HIU-treated MPs revealed that the emulsion activity index and emulsion stability index of HIU-treated MP were improved. Confocal laser scanning microscopy images indicated that small spherical droplets adsorbed with MPs were formed by HIU treatment and that dispersion stabilities were improved because the Turbiscan stability index of the HIU-treated group was lower than that of the control group. These findings could be used as supporting data for the utilizing porcine-derived MPs, which have been treated with HIU for appropriate time periods, as Pickering stabilizers.     

Structural and physicochemical properties of the different ultrasound frequency modified Qingke protein

There is a burgeoning demand for modified plant-based proteins with desirable physicochemical and functional properties. The cereal Qingke is a promising alternative protein source, but its use has been limited by its imperfect functional characteristics. To investigate the effect of ultrasound treatment on Qingke protein, we applied single- (40 kHz), dual- (28/40 kHz), and tri- (28/40/50 kHz) frequency ultrasound on the isolated protein and measured subsequent physicochemical and structural changes. The results showed that the physicochemical properties of proteins were modified following ultrasound treatment, and many of these changes significantly increased with increasing frequency. Compared with the native Qingke protein (control), the solubility, foaming activity, stability, and water or oil holding capacity of tri-frequency ultrasound modified Qingke protein increased by 43.54%, 20.83%, 20.51%, 28.9%, and 45.2%, respectively. Furthermore, ultrasound treatment altered the secondary and tertiary structures of the protein resulting in more exposed chromophoric groups and inner hydrophobic groups, as well as reduced β-sheets and increased random coils, relative to the control. Rheological and texture characterization indicated that the values of G' and G'', hardness, gumminess, and chewiness decreased after ultrasound treatment. This study could provide a theoretical basis for the application of multi-frequency ultrasonic technology for modification of Qingke protein to expand its potential use as an alternative protein source.     

High intensity ultrasound treatment of protein isolate extracted from dephenolized sunflower meal: Effect on physicochemical and functional properties

The influence of high intensity ultrasound (HIUS) on physicochemical and functional properties of sunflower protein isolates was investigated. Protein solutions (10% w/v) were treated with ultrasound probe (20 kHz) and ultrasound bath (40 kHz) for 5, 10, 20 and 30 min. Thermal stability of protein isolates was reduced as indicated by differential scanning calorimetry. Minimum thermal stability was observed at 20 min of sonication and increased further with increase in treatment time indicating aggregation at prolonged sonication. SDS-PAGE profile of proteins showed a significant reduction in molecular weight. Further, surface hydrophobicity and sulfhydryl content increased after HIUS treatment indicating partial unfolding of proteins and reduction in the intermolecular interactions. The particle size analysis showed that HIUS treatment reduced the particle size. Less turbid solution were observed largely due to reduction in particle size. HIUS decreased the available lysine content in protein isolates. Solubility, emulsifying capacity, emulsion stability, foaming capacity, foam stability and oil binding capacity were improved significantly, while as, water binding capacity was decreased. The effect of HIUS on physicochemical and functional properties of sunflower protein isolates was more pronounced in probe sonication rather than bath sonication. Protein isolates with improved functional properties can be obtained using high intensity ultrasound technology.     

Effect of high intensity ultrasound on transglutaminase-catalyzed soy protein isolate cold set gel

The effects of high intensity ultrasound (HIU, 105–110 W/cm² for 5 or 40 min) pre-treatment of soy protein isolate (SPI) on the physicochemical properties of ensuing transglutaminase-catalyzed soy protein isolate cold set gel (TSCG) were investigated in this study. The gel strength of TSCG increased remarkably from 34.5 to 207.1 g for TSCG produced from SPI with 40 min HIU pre-treatment. Moreover, gel yield and water holding capacity also increased after HIU pre-treatments. Scanning electron microscopy showed that HIU of SPI resulted in a more uniform and denser microstructure of TSCG. The content of free sulfhydryl (SH) groups was higher in HIU TSCG than non-HIU TSG, even though greater decrease of the SH groups present in HIU treated SPI was observed when the TSCG was formed, suggesting the involvement of disulfide bonds in gel formation. Protein solubility of TSCG in both denaturing and non-denaturing solvents was higher after HIU pretreatment, and changes in hydrophobic amino acid residues as well as in polypeptide backbone conformation and secondary structure of TSCG were demonstrated by Raman spectroscopy. These results suggest that increased inter-molecular ε-(γ-glutamyl) lysine isopeptide bonds, disulfide bonds and hydrophobic interactions might have contributed to the HIU TSCG gel network. In conclusion, HIU changed physicochemical and structural properties of SPI, producing better substrates for TGase. The resulting TSCG network structure was formed with greater involvement of covalent and non-covalent interactions between SPI molecules and aggregates than in the TSCG from non-HIU SPI.     

Improving the gel properties of salted egg white/cooked soybean protein isolate composite gels by ultrasound treatment: Study on the gelling properties and structure

In this study, the effects of ultrasound treatment on the texture, physicochemical properties and protein structure of composite gels prepared by salted egg white (SEW) and cooked soybean protein isolate (CSPI) at different ratios were investigated. With the increased SEW addition, the ζ-potential absolute values, soluble protein content, surface hydrophobicity and swelling ratio of composite gels showed overall declining trends (P < 0.05), while the free sulfhydryl (SH) contents and hardness of exhibited overall increasing trends (P < 0.05). Microstructural results revealed that composite gels exhibited denser structure with the increased SEW addition. After ultrasound treatment, the particle size of composite protein solutions significantly decreased (P < 0.05), and the free SH contents of ultrasound-treated composite gels were lower than that of untreated composite gels. Moreover, ultrasound treatment enhanced the hardness of composite gels, and promoted the conversion of free water into non-flowable water. However, when ultrasonic power exceeded 150 W, the hardness of composite gels could not be further enhanced. FTIR results indicated that ultrasound treatment facilitated the composite protein aggregates to form a more stable gel structure. The improvement of ultrasound treatment on the properties of composite gels was mainly by promoting the dissociation of protein aggregates, and the dissociated protein particles further interacted to form denser aggregates through disulfide bond, thus facilitating the crosslinking and reaggregation of protein aggregates to form denser gel structure. Overall, ultrasound treatment is an effective approach to improve the properties of SEW-CSPI composite gels, which can improve the potential utilization of SEW and SPI in food processing.     

Ultrasonic pretreatment improves the gelation properties of low-salt Penaeus vannamei (Litopenaeus vannamei) surimi

The effects of different ultrasonic pretreatments (120–600 W, 20 min; 360 W, 10–30 min) on the gel properties of shrimp surimi were investigated. Gel properties and protein functional properties were analysed to clarify the mechanism of action of ultrasound. The gel strength, water holding capacity and surface hydrophobicity of shrimp surimi gel increased initially and then decreased with the increase in ultrasound power or time, but the change in total sulfhydryl content showed the opposite trend, which indicated that proper ultrasound pretreatment could improve the gel properties of shrimp surimi, expand the protein to a greater extent and expose more SH groups and hydrophobic groups. According to scanning electron microscopy observation, ultrasound made shrimp surimi gel form a denser gel network. Fourier transform infrared analysis indicated that the α-helix content in shrimp surimi gel decreased initially and then increased with the increase of in ultrasound power or time, whereas the change in β-sheet content showed the opposite trend. And the protein was the most stable in 360 W/20 min pretreatment. SDS-PAGE patterns showed that proper ultrasound inhibited the degradation of actin and troponin C. In addition, dynamic rheology illustrated that the G′ values of the ultrasonic pretreatment group were higher than that of the control group, indicating that ultrasound could improve the elasticity and stability of shrimp surimi gel. The results suggested that the shrimp surimi gel pretreated by 360 W/20 min ultrasound showed the best gel properties. Furthermore, the correlation between the indexes affecting the properties of the gel was analyzed. This study provides a new technical means to improve the gel properties of shrimp surimi.     

Effects of ultrasonic pretreatment of soybean protein isolate on the binding efficiency,structural changes,and bioavailability of a protein-luteolin nanodelivery system

The combination of protein and flavonoids can ameliorate the problems of poor solubility and stability of flavonoids in utilization. In this study, soybean protein isolate pretreated by ultrasonication was selected as the embedding wall material, which was combined with luteolin to form a soybean protein isolate-luteolin nanodelivery system. The complexation effect and structural changes of soybean protein isolate (SPI) and ultrasonic pretreatment (100 W, 200 W, 300 W, 400 W and 500 W) of soybean protein isolate with luteolin (LUT) were compared, as well as the changes in digestion characteristics and antioxidant activity in vitro. The results showed that proper ultrasonic pretreatment increased the encapsulation efficacy, loading amount and solubility to 89.72%, 2.51 μg/mg and 90.56%. Appropriate ultrasonic pretreatment could make the particle size and the absolute value of ζ-potential of SPI-LUT nanodelivery system decrease and increase respectively. The FTIR and fluorescence results show that appropriate ultrasonic pretreatment could reduce α-helix, β-sheet and random coil, increase β-turn, and enhance fluorescence quenching. The thermodynamic evaluation results indicate that the ΔG < 0, ΔH > 0 and ΔS > 0, so the interaction of LUT with the protein was spontaneous and mostly governed by hydrophobic interactions. The XRD results show that the LUT was amorphous and completely wrapped by SPI. The DSC results showed that ultrasonic pretreatment could improve the thermal stability of SPI-LUT nanodelivery system to 112.66 ± 1.69 °C. Digestion and antioxidant analysis showed that appropriate ultrasonic pretreatment increased the LUT release rate and DPPH clearance rate of SPI-LUT nanodelivery system to 89.40 % and 55.63 % respectively. This study is a preliminary source for the construction of an SPI nanodelivery system with ultrasound pretreatment and the deep processing and utilization of fat-soluble active substances.     

Effects of high intensity unltrasound on structural and physicochemical properties of myosin from silver carp

Myosin from silver carp was sonicated with varying power output (100, 150, 200 and 250 W) for 3, 6, 9, and 12 min. The changes in the structure and physicochemical properties of myosin were evaluated by dynamic light scattering, SDS-PAGE and some physicochemical indexes. The ultrasound treatments induced a significant conversion of myosin aggregates to smaller ones with a more uniform distribution, and obvious enhancement in solubility. The structure of myosin was also notably changed by sonication, with a decrease in Ca²⁺-ATPase activity and SH content, and an increase in surface hydrophobicity. Furthermore, SH groups were oxidized, leading to a decrease in reactive SH and total SH contents. SDS-PAGE analysis revealed that ultrasound could induce the degradation of myosin heavy chain and change the protein fraction of myosin. Collectively, the ultrasonic treatment of 100 W for 3 min showed slight influence on the SH content, S₀-ANS, and electrophoretic patterns, and the extent of changes in myosin structure and physicochemical properties tended to increase with ultrasonic power and time. The integrated data indicate that ultrasonic treatment can facilitate the improvement of the solubility and dispersion of myosin, but the choice of a suitable ultrasonic condition to avoid oxidation and degradation of myosin is very important.     

Effect of multiple-frequency ultrasound-assisted transglutaminase dual modification on the structural,functional characteristics and application of Qingke protein

Qingke protein rich in restricted amino acids such as lysine, while the uncoordination of ratio of glutenin and gliadin in Qingke protein has a negative impact on its processing properties. In this study, the effect of multiple-frequency ultrasound combined with transglutaminase treatment on the functional and structural properties of Qingke protein and its application in noodle manufacture were investigated. The results showed that compared with the control, ultrasound-assisted transglutaminase dual modification significantly increased the water and oil holding capacity, apparent viscosity, foaming ability, and emulsifying activity index of Qingke protein, which exhibited a higher storage modulus G' (P < 0.05). Meanwhile, ultrasound combined with transglutaminase treatment enhanced the cross-linking degree of Qingke protein (P < 0.05), as shown by decreased free amino group and free sulfhydryl group contents, and increased disulfide bond content. Moreover, after the ultrasound-assisted transglutaminase dual modification treatment, the fluorescence intensity, the contents of α-helix and random coil in the secondary structure of Qingke protein significantly decreased, while the β-sheet content increased (P < 0.05) compared with control. SDS-PAGE results showed that the bands of Qingke protein treated by ultrasound combined with transglutaminase became unclear. Furthermore, the quality of Qingke noodles made with Qingke powder (140 g/kg dual modified Qingke protein mixed with 860 g/kg extracted Qingke starch) and wheat gluten 60–70 g/kg was similar to that of wheat noodles. In summary, multiple-frequency ultrasound combined with transglutaminase dual modification can significantly improve the physicochemical properties of Qingke protein and the modified Qingke proteins can be used as novel ingredients for Qingke noodles.     

Influence of ultrasound treatment on the physicochemical and antioxidant properties of mung bean protein hydrolysate

This study aimed to investigate influence of ultrasonic treatment on physicochemical and antioxidant properties of mung bean protein hydrolysate (MPH). Physicochemical properties of MPH were evaluated by Tricine-SDS-PAGE, particle size distribution, fourier transform infrared spectroscopy (FTIR) and fluorescence spectroscopy, among others. Radicals scavenging activities of ABTS, hydroxyl, superoxide anion, Fe²⁺ chelating ability and reducing power characterized antioxidant activities of MPH. MPH contained four bands of 25.6, 12.8, 10.6 and 4.9 kDa, in which 4.9 kDa was the most abundant. Ultrasonic treatment increased the contents of aromatic and hydrophobic amino acids in MPH. Ultrasonic treatment decreased the content of α-helix of MPH and increased β-sheet and β-turn compared to MPH. MPH-546 W (ultrasonic treatment 546 W, 20 min) had the lowest average particle size (290.13 nm), zeta potential (-36.37 mV) and surface hydrophobicity (367.95 A.U.). Antioxidant activities of ultrasonicated-MPH increased with the ultrasonic power, achieving the lowest IC₅₀ (mg/mL) of 0.1087 (ABTS), 1.796 (hydroxyl), 1.003 (superoxide anion) and 0.185 (Fe²⁺ chelating ability) in 546 W power. These results indicated ultrasonic treatment would be a promising method to improve the antioxidant properties of MPH, which would broaden the application scope of MPH as bioactive components in the food industry.     

Comparison of soy protein isolate-(–)-epigallocatechin gallate complexes prepared by mixing,chemical polymerization,and ultrasound treatment

The effects of the preparation method (mixing, chemical polymerization, or ultrasound treatment) on the structure and functional properties of soy protein isolate-(–)-epigallocatechin-3-gallate (SPI-EGCG) complexes were examined. The mixing treated SPI-EGCG samples (M−SE) were non-covalently linked, while the chemical polymerization and ultrasound treated SPI-EGCG samples (C-SE and U-SE, respectively) were bound covalently. The covalent binding of EGCG with protein improved the molecular weight and changed the structures of the SPI by decreasing the α-helix content. Moreover, U-SE samples had the lowest particle size (188.70 ± 33.40 nm), the highest zeta potential (−27.82 ± 0.53 mV), and the highest polyphenol binding rate (59.84 ± 2.34 %) compared with mixing and chemical polymerization-treated samples. Furthermore, adding EGCG enhanced the antioxidant activity of SPI and U-SE revealed the highest DPPH (84.84 ± 1.34 %) and ABTS (88.89 ± 1.23 %) values. In conclusion, the SPI-EGCG complexes prepared by ultrasound formed a more stable composite system with stronger antioxidant capacity, indicating that ultrasound technology may have potential applications in the preparation of protein-polyphenol complexes.     

Analysis in protein profile,antioxidant activity and structure-activity relationship based on ultrasound-assisted liquid-state fermentation of soybean meal with Bacillus subtilis

This study investigated effects of ultrasound on the contents of peptide and soluble protein, antioxidant activity, functionalities and structural characteristics of fermented soybean meal (FSBM) with Bacillus subtilis systematically. The results showed that there were significant effects of ultrasound treatments (frequency, treatment time and power density) on the contents of peptide and soluble protein (p < 0.05). Under the optimum ultrasound conditions (power density of 0.08 W/mL, frequency of 33 kHz and treatment time of 1 h) by single factor experiment, the contents of peptide and soluble protein increased by 31.27% and 18.79% compared to those of the control, respectively. Additionally, the in vitro antioxidant activity (•OH scavenging rate, Fe²⁺ chelating capacity and DPPH radical scavenging rate) and functional properties (emulsifying activity and emulsifying stability) of FSBM were found to be noticeably improved by ultrasound (p < 0.05). The fourier transform infrared (FTIR) spectroscopy revealed that ultrasound caused protein molecules to unfold with a decrease content of α-helix and β-turn and an increase in the proportion of β-sheet and random coil. Besides, atomic force microscope (AFM) results indicated that ultrasonication generally increased the surface roughness of protein and the protein sonicated with higher frequency (≥33 kHz) exhibited a greater height compared to lower frequency ultrasonication. Structure-activity relationship analysis illustrated that there was a good linear relationship between •OH scavenging rate and β-sheet/β-turn with Pearson’s correlation coefficient r of −0.86/0.90. Collectively, the selection of ultrasonic parameters is essential for the preparation of functional protein and bioactive peptide by enhancing fermentation of agroindustrial by-products.     

Covalent binding of ultrasound-treated japonica rice bran protein to catechin: Structural and functional properties of the complex

Due to the existence of many disulfide bonds in japonica rice bran protein (JRBP) molecules, their solubility is poor, which seriously affects other functional properties. To improve the functional characteristics of JRBP molecules, they were processed by ultrasound technology, and JRBP-catechin (CC) covalent complex was prepared. The structural and functional properties of indica and japonica rice bran proteins and their complexes were compared; furthermore, the changes in the structural and functional properties of JRBP-CC under different ultrasound conditions were investigated. The results showed that compared with indica rice bran protein (IRBP), the secondary structure of JRBP-CC was very different, the water holding capacity (WHC) was higher, and the emulsification performance was better. Different ultrasound conditions had different effects on the functional properties of JRBP-CC. When the ultrasound power was 200 W, the λ_max redshift of the JRBP-CC complex was the most significant, the particle size was the smallest, the absolute value of the zeta potential was the largest, and the hydrophobicity and microstructure of the JRBP-CC complex were the best. Concurrently, the maximum WHC and oil holding capacity (OHC) of JRBP-CC under these conditions were 7.54 g/g and 6.87 g/g, respectively. Moreover, the emulsifying activity index (EAI) and emulsifying stability index (ESI) were 210 m²/g and 47.8 min, respectively, and the scavenging activities of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ABTS⁺ were 71.96 % and 80.07 %, respectively.