Effects of multi-frequency power ultrasound on the enzymolysis and structural characteristics of corn gluten meal

The aim of this study was to investigate the effect of multi-frequency power ultrasound (sweeping frequency and pulsed ultrasound (SFPU) and sequential dual frequency ultrasound (SDFU)) on the enzymolysis of corn gluten meal (CGM) and on the structures of the major protein fractions (zein, glutelin) of CGM. The results showed that multi-frequency power ultrasound pretreatments improved significantly (P < 0.05) the degree of hydrolysis and conversion rate of CGM. The changes in UV–Vis spectra, fluorescence emission spectra, surface hydrophobicity (H₀), and the content of SH and SS groups indicated unfolding of zein and glutelin by ultrasound. The circular dichroism analysis showed that both pretreatments decreased α-helix and increased β-sheet of glutelin. The SFPU pretreatment had little impact on the secondary structure of zein, while the SDFU increased the α-helix and decreased the β-sheet remarkably. Scanning electron microscope indicated that both pretreatments destroyed the microstructures of glutelin and CGM, reduced the particle size of zein despite that the SDFU induced aggregation was observed. In conclusion, multi-frequency power ultrasound pretreatment is an efficient method in protein proteolysis due to its sonochemistry effect on the molecular conformation as well as on the microstructure of protein.     

Effects of multi-frequency ultrasound on the freezing rates,quality properties and structural characteristics of cultured large yellow croaker (Larimichthys crocea)

This research evaluated the effects of multi-frequency ultrasound assisted freezing (UAF) on the freezing rate, structural characteristics, and quality properties of cultured large yellow croaker. The freezing effects with triple ultrasound-assisted freezing (TUF) at 20, 28 and 40 kHz under 175 W was more obvious than that of single ultrasound-assisted freezing (SUF) at 20 kHz and dual ultrasound-assisted freezing (DUF) at 20 and 28 kHz. The results showed that UAF significantly increased the freezing rate and better preserved the quality of frozen large yellow croaker samples. Specifically, the quality parameters of the TUF-treated samples were closer to those of the fresh samples, with greater texture characteristics, a larger water holding capacity (lower thawing loss and cooking loss), lower K values and lower thiobarbituric acid reactive substances values. Light microscopy observation images revealed that the ice crystals formed by TUF were fine and evenly distributed, resulting in less damage to the frozen large yellow croaker samples. Therefore, multi-frequency UAF could improve the quality properties of the large yellow croaker samples.     

Effects of ultrasound and ultrasound assisted alkaline pretreatments on the enzymolysis and structural characteristics of rice protein

The objectives of this study were to investigate the effects of multi-frequency energy-gathered ultrasound (MFEGU) and MFEGU assisted alkaline pretreatments on the enzymolysis and the mechanism of two pretreatments accelerating the rice protein (RP) proteolysis process. The results showed that MFEGU and MFEGU assisted alkaline pretreatments improved significantly (P < 0.05) the degree of hydrolysis (DH) and the protein elution amount of RP. Furthermore under the same DH conditions, ultrasound and ultrasound assisted alkaline pretreatments were more save the enzymolysis time than the unpretreatment. The changes in UV–vis spectra, fluorescence emission spectra indicated unfolding and destruction of RP by MFEGU and MFEGU assisted alkaline pretreatments. The circular dichroism analysis showed that both pretreatments decreased α-helix but increased β-sheet and random coil of RP. Amino acid composition revealed that MFEGU and MFEGU assisted alkaline pretreatments could increase the protein elution amount and the ratio of hydrophobic amino acids. Atomic force microscopy (AFM) indicated that both pretreatments destroyed the microstructures and reduced the particle size of RP. Therefore, MFEGU and MFEGU assisted alkaline pretreatments are beneficial to improving the degree of hydrolysis due to its sonochemistry effect on the molecular conformation as well as on the microstructure of protein.     

Transglutaminase-induced gelation properties of soy protein isolate and wheat gluten mixtures with high intensity ultrasonic pretreatment

Soy protein isolate (SPI) and wheat gluten (WG) are widely used in commercial food applications in Asia for their nutritional value and functional properties. However, individually each exhibits poor gelation. In this study, we examined the microbial transglutaminase (MTGase)-induced gelation properties of SPI and WG mixtures with high intensity ultrasonic pretreatment. Ultrasonic treatment reduced the particle size of SPI/WG molecules, which led to improvements in surface hydrophobicity (H_o) and free sulfhydryl (SH) group content. However, MTGase crosslinking facilitated the formation of disulfide bonds, markedly decreasing the content of free SH groups. Ultrasonic treatment improved the gel strength, water holding capacity, and storage modulus and resulted in denser and more homogeneous networks of MTGase-induced SPI/WG gels. In addition, ultrasonic treatment changed the secondary structure of the gel samples as determined by Fourier transform infrared spectroscopic analysis, with a reduction in α-helices and β-turns and an increase in β-sheets and random coils. Thus, ultrasound is useful in facilitating the gelation properties of MTGase-induced SPI/WG gels and might expand their utilization in the food protein gelation industry.     

Multi-frequency ultrasound:A potential method to improve the effects of surface decontamination and structural characteristics on large yellow croaker (Pseudosciaena crocea) during refrigerated storage

The effects of multi-frequency ultrasound on surface decontamination and structural characteristics of large yellow croaker (Pseudosciaena crocea) during refrigerated storage were evaluated. The results of total viable counts (TVCs) and psychrophilic bacteria counts (PBCs) demonstrated that multi-frequency ultrasound retarded the growth of microorganisms. The bacteriostatic effect was positively correlated with the increase of ultrasound frequencies. However, compared with triple-frequency ultrasound (TUS, 20/28/40 kHz) treatment, dual-frequency ultrasound (DUS, 20/28 kHz) treatment had higher water-holding capacity (WHC) and immobilized water content, better texture characteristics, lower pH and total volatile basic nitrogen (TVB-N). Through the results of myofibrillar fragmentation index (MFI), intrinsic fluorescence intensity (IFI) and atomic force microscope (AFM), multi-frequency ultrasound could effectively stabilize the myofibrillar protein structure of refrigerated large yellow croaker, which could maintain better texture characteristics. The effects of DUS were the most significant. Therefore, multi-frequency ultrasound treatment could inhibit the growth of microorganisms and improve the structural characteristics of large yellow croaker during refrigerated storage.     

Effect of multi-frequency ultrasound thawing on the structure and rheological properties of myofibrillar proteins from small yellow croaker

The influence of multi-frequency combined ultrasound thawing on primary, secondary, and tertiary structures, electrophoresis pattern, particle size distribution, zeta potential values, thermal stability, rheological behavior, and microstructure of small yellow croaker myofibrillar proteins (MPs) were studied. Four treatments were used for thawing small yellow croakers: flow water thawing (FWT), mono-frequency ultrasonic thawing (MUT), dual-frequency ultrasonic thawing (DUT), and tri-frequency ultrasonic thawing (TUT). Compared with fresh samples (FS), the MPs of the sample pretreated by DUT had non-significant effect on protein primary (including free amino groups and surface hydrophobicity), secondary, tertiary structures, electrophoresis pattern, and microstructure. MPs pretreated by DUT had less aggregation and degradation. Besides, DUT treatment increased the thermal stability of MPs. The ultrasound had significant effects on the rheological properties of MPs. Overall, DUT effectively minimized the changes in MPs structure and protected the protein thermal stability and rheological behavior during the thawing process.     

Effects of multi-frequency ultrasound on freezing rates and quality attributes of potatoes

The effects of multi-frequency ultrasound assisted freezing on the freezing rate, microstructure, quality properties (drip loss, firmness, total calcium content, l-ascorbic acid content and total phenol content) of potatoes were studied. The results indicated that the freezing effects of multi-frequency ultrasound was better than those of single-frequency ultrasound. Multi-frequency ultrasound could significantly increase the freezing rate and preserve the quality of frozen samples better. With increase in the number of ultrasonic frequencies, the freezing effect was more obvious. In addition, scan electron microscopy (SEM) images showed that the ice crystals formed by the multi-frequency ultrasonic treatment were fine and uniformly distributed, which caused less damage to the frozen potato samples. From the analysis of the quality attributes, the nutritional values of the samples after multi-frequency ultrasonic treatment was higher, but attention should be paid to the negative influence of the hydroxyl radical generated by the multi-frequency ultrasound.     

Lysinoalanine formation and conformational characteristics of rice dreg protein isolates by multi-frequency countercurrent S-type sonochemical action

The influences of multi-frequency countercurrent S-type ultrasound (MFSU), with various frequency modes, on lysinoalanine (LAL) formation and conformational characteristics of rice dreg protein isolates (RDPI) were investigated. The ultrasonic operating mode with dual-frequency combination (20/40 kHz) indicated lower LAL content and higher protein dissolution rate of RDPI compared with that of other ultrasound operating modes. Under the dual-frequency ultrasound mode of 20/40 kHz, acoustic power density of 60 W/L, time of 20 min, and temperature of 35 °C, the relative reduction rate of LAL of RDPI reached the highest with its value of 26.95%, and the protein dissolution rate was 71.87%. The changes in chemical interactions between protein molecules indicated that hydrophobic interactions and disulfide bonds played a considerable role in the formation of LAL of RDPI, especially the reduction of g-g-g and g-g-t disulfide bond. Alterations in microstructure showed that ultrasonication loosened the protein structure and created more uniform protein fragments of RDPI. In conclusion, using MFSU in treating RDPI was an efficacious avenue for minimizing LAL content and modifying the conformational characteristics of RDPI.     

Effect of ultrasound treatments on functional properties and structure of millet protein concentrate

In this study, the effect of high power ultrasound (US) probe in varying intensities and times (18.4, 29.58, and 73.95 W/cm² for 5, 12.5 and 20 min respectively) on functional properties of millet protein concentrate (MPC) was investigated, and also the structural properties of best modified treatment were evaluated by FTIR, DSC, Zeta potential and SDS-PAGE techniques. The results showed the solubility in all US treated MPC was significantly (p < .05) higher than those of the native MPC. Foaming capacity of native MPC (271.03 ± 4.51 ml) was reduced after US treatments at low intensities (82.37 ± 5.51 ml), but increased upon US treatments at high intensities (749.7 ± 2 ml). In addition, EAI and ES increased after US treatments. One of the best US treatments that can improve the functional properties of MPC was 73.95 W/cm² for 12.5 min that resulted in reduction of molecular weight and increase nearly 36% in the negative surface charge that was confirmed by SDS-page and Zeta potential results, respectively.     

High-intensity ultrasound improved the physicochemical and gelling properties of Litopenaeus vannamei myofibrillar protein

The effects of high-intensity ultrasound on the physicochemical and gelling properties of Litopenaeus vannamei (L. vannamei) myofibrillar protein (MP) were investigated. MP solutions were subjected to ultrasound treatment (power 100 W, 300 W, and 500 W). It was found that the carbonyl and free amino contents of MP increased significantly with increasing ultrasound power, accompanied by enhanced emulsification properties. The increase of free radical and carbonyl content indicated that ultrasound induced the oxidation of MP. With the increase of ultrasound power, it was found that the total sulfhydryl content of the shrimp MP decreased, but the surface hydrophobicity increased significantly, which might be closely related to the conformational changes of MP. Meanwhile, a significant increase of β-sheet but a decrease of α-helix in the secondary structure of MP was observed with increasing ultrasound power, indicating that ultrasound treatment induced the stretching and flexibility of MP molecules. SDS-PAGE showed that L. vannamei MP consisted of myosin heavy chain, actin, myosin light chain, paramyosin and tropomyosin. Ultrasound treatment could lead to some degree of oxidative aggregation of MP. The results of rheological properties indicated that ultrasound treatment enhanced the viscoelasticity of MP and further improved the gel strength of MP gel. This study can provide a theoretical basis for the functional modification of shrimp MP and the processing of its surimi products.     

Effects of multi-frequency power ultrasound on the enzymolysis of corn gluten meal: Kinetics and thermodynamics study

The effects of multi-frequency power ultrasound (MPU) pretreatment on the kinetics and thermodynamics of corn gluten meal (CGM) were investigated in this research. The apparent constant (K_M), apparent break-down rate constant (k_A), reaction rate constants (k), energy of activation (E_a), enthalpy of activation (ΔH), entropy of activation (ΔS) and Gibbs free energy of activation (ΔG) were determined by means of the Michaelis–Menten equation, first-order kinetics model, Arrhenius equation and transition state theory, respectively. The results showed that MPU pretreatment can accelerate the enzymolysis of CGM under different enzymolysis conditions, viz. substrate concentration, enzyme concentration, pH, and temperature. Kinetics analysis revealed that MPU pretreatment decreased the K_M value by 26.1% and increased the k_A value by 7.3%, indicating ultrasound pretreatment increased the affinity between enzyme and substrate. In addition, the values of k for ultrasound pretreatment were increased by 84.8%, 41.9%, 28.9%, and 18.8% at the temperature of 293, 303, 313 and 323 K, respectively. For the thermodynamic parameters, ultrasound decreased E_a, ΔH and ΔS by 23.0%, 24.3% and 25.3%, respectively, but ultrasound had little change in ΔG value in the temperature range of 293–323 K. In conclusion, MPU pretreatment could remarkably enhance the enzymolysis of CGM, and this method can be applied to protein proteolysis industry to produce peptides.     

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.     

Study of ultrasonic treatment on the structural characteristics of gluten protein and the quality of steamed bread with potato pulp

Physicochemical properties and microstructure of gluten protein, and the structural characteristics of steamed bread with 30 % potato pulp (SBPP) were investigated by ultrasonic treatments. Results showed that 400 W ultrasonic treatment significantly (P < 0.05) increased the combination of water and substrate in the dough with 30 % potato pulp (DPP). The contents of wet gluten, free sulfhydryl (SH), and disulfide bond (SS) were influenced by ultrasonic treatment. Moreover, UV-visible and fluorescence spectroscopy demonstrated that the conformation of gluten protein was changed by ultrasonic treatment (400 W). Fourier transform infrared (FT-IR) illustrated that the β-sheet content was significantly (P < 0.05) increased (42 %) after 400 W ultrasonic treatment, and the surface hydrophobicity of gluten protein in SBPP increased from 1225.37 (0 W ultrasonic treatment) to 4588.74 (400 W ultrasonic treatment). Ultrasonic treatment facilitated the generation of a continuous gluten network and stabilized crumb structure, further increased the specific volume and springiness of SBPP to 18.9 % and 6.9 %, respectively. Those findings suggested that ultrasonic treatment would be an efficient method to modify gluten protein and improve the quality of SBPP.     

Effect of ultrasound pretreatment on structural,physicochemical, rheological and gelation properties of transglutaminase cross-linked whey protein soluble aggregates

A solution (10%, w/v) of whey protein soluble aggregates (WPISA) was pretreated with high-intensity ultrasound (HUS, 20 kHz) for different durations (10–40 min) before incubation with transglutaminase (TGase) to investigate the effect of HUS on the structural, physicochemical, rheological, and gelation properties of TGase cross-linked WPISA. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results showed that HUS increased the amounts of high-molecular-weight polymers/aggregates in WPISA after incubation with TGase. HUS significantly increased (P < 0.05) the degree of TGase-mediated cross-linking in WPISA, as demonstrated by a reduction in free amino group contents. HUS significantly increased (P < 0.05) the particle size, intrinsic fluorescence intensity, and surface hydrophobicity of TGase cross-linked WPISA, but had no significant impact (P > 0.05) on the zeta-potential or total free sulfhydryl group content of TGase cross-linked WPISA. The apparent viscosity and the consistency index of TGase cross-linked WPISA were significantly increased by HUS (P < 0.05), which indicated that HUS facilitated the formation of more high-molecular-weight polymers. HUS significantly increased (P < 0.05) the water holding capacity and gel strength of glucono-δ-lactone (GDL)-induced TGase cross-linked WPISA gels. The results indicated that HUS could be an efficient tool for modifying WPISA to improve its degree of TGase-mediated cross-linking, which would lead to improved rheological and gelation properties.     

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.     

Effect of high-intensity ultrasound on the physicochemical properties,microstructure, and stability of soy protein isolate-pectin emulsion

In this study, an emulsion stabilized by soy protein isolate (SPI)-pectin (PC) complexes was prepared to investigate the effects of high-intensity ultrasound (HIU) treatment (150–600 W) on the physicochemical properties, microstructure, and stability of emulsions. The results found that the emulsion treated at 450 W showed the best emulsion stability index (ESI) (25.18 ± 1.24 min), the lowest particle size (559.82 ± 3.17 nm), the largest ζ-potential absolute value (16.39 ± 0.18 mV), and the highest adsorbed protein content (27.31%). Confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) revealed that the emulsion aggregation was significantly improved by ultrasound treatment, and the average roughness value (Rq) was the smallest (10.3 nm) at 450 W. Additionally, HIU treatment reduced the interfacial tension and apparent viscosity of the emulsion. Thermal stability was best when the emulsion was treated at 450 W, D₄₃ was minimal (907.95 ± 31.72 nm), and emulsion separation also improved. Consequently, the creaming index (CI) was significantly decreased compared to the untreated sample, indicating that the storage stability of the emulsion was enhanced.     

Effects of high-intensity ultrasound on the structural,optical, mechanical and physicochemical properties of pea protein isolate-based edible film

Pea protein is a promising alternative to animal-based protein and the interest in its application in food industry has been rapidly growing. In this study, pea protein isolates (PPI) were used to form protein-based edible films and the effect of ultrasound treatment on the structure of PPI and the structural, optical, mechanical and physicochemical properties of PPI-films were investigated. Ultrasound induced unfolding of PPI and exposed interior hydrophobic groups to protein surface while both PPI dissociation and formation of large aggregates were observed, as confirmed by measuring intrinsic emission fluorescence, surface hydrophobicity, surface charge, and particle size distribution and polydispersity index, respectively. FE-SEM showed that ultrasound decreased the cracks and protein aggregates at the surface of PPI-film. The film structure was also investigated by FTIR, which showed peak shift in amide I and II region and noticeable difference of protein secondary structure as affected by ultrasound. As a result of such structural changes caused by ultrasound, the properties of PPI-films were improved. Results showed that ultrasound greatly improved the film transparency, significantly increased film tensile strength but not elongation at break, and decreased moisture content and water vapor permeability of the film. This study provided structural data as evidence for utilizing ultrasound technique to develop PPI-films with improved optical, mechanical and water barrier properties.     

Effects of pulsed ultrasound on rheological and structural properties of chicken myofibrillar protein

The effects of pulsed ultrasound (PUS) (power: 240 w) with varying time (0, 3, 6, 9, 12 and 15 min) on rheological and structural properties of chicken myofibrillar protein (CMP) were examined. PUS treatment significantly caused a decrease in the viscosity coefficients (k) but an increase in the flow index (n) value of CMP solutions within short time (0–6 min), while had no significant effect for longer time (9–15 min). Besides, at 6 min, the solubility and microstructure of CMP samples were optimum. The primary structure of CMP was not altered by PUS treatment. However, Raman spectroscopy revealed a decrease in the α-helix and β-sheets proportion and an increase in the β-turn of CMP following PUS treatment. Random coil reached a maximum at 6 min. The changes in tertiary and quaternary structure of CMP by PUS treatment also occurred. As PUS time extended, S₀-ANS for CMP increased measured by ANS fluorescence probe method. However, the normalized intensity of 760 cm⁻¹ increased from 0 min to 6 min, and then decreased to 15 min by Raman test. Moreover, the reactive sulphur (SH) contents and disulfide bonds (S-S) of samples increased while the total SH contents decreased within 0–6 min. At 9 min and above, the contents of reactive SH groups were almost equal to the contents of total SH groups. Differential scanning calorimetry (DSC) of CMP showed that peak temperature (T_d2) for myosin and peak temperature (T_d3) for actin were both reduced in the first 6 min, while T_d3 was not observed from 9 min following PUS treatment. Therefore, 6 min was the optimum PUS time to obtain better CMP rheological and structural properties.     

Effects and mechanism of dual-frequency power ultrasound on the molecular weight distribution of corn gluten meal hydrolysates

The impact of dual-frequency power ultrasound (DPU) on the molecular weight distribution (MWD) of corn gluten meal (CGM) hydrolysates and its mechanism were investigated in the present study. The mechanism was studied from aspects of structural and nano-mechanical characteristics of the major protein fractions of CGM, viz. zein and glutelin. The results of molecular weight distribution indicated that DPU pretreatment of CGM was beneficial to the preparation of peptides with molecular weights of 200–1000 Da. Moreover, FTIR spectral analysis and atomic force microscopy characterization showed that the DPU pretreatment changed the contents of secondary structure of proteins, decreased the particle height and surface roughness of glutelin, reduced the Young’s modulus and stiffness of zein while increased its adhesion force. In conclusion, DPU pretreatment of proteins before proteolysis is an efficient alternative method to produce short-chain peptides because of its positive effects originating from acoustic cavitation on the molecular conformation, nano-structures and nano-mechanical properties of proteins as well.     

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.