Pretreatment of defatted wheat germ proteins (by-products of flour mill industry) using ultrasonic horn and bath reactors: Effect on structure and preparation of ACE-inhibitory peptides

The ultrasonic horn and bath reactors were compared based on production of angiotensin-converting-enzyme (ACE) inhibitory peptides from defatted wheat germ proteins (DWGP). The DWGP was sonicated before hydrolysis by Alcalase. Degree of hydrolysis, ACE-inhibitory activity, surface hydrophobicity, fluorescence intensity, free sulfhydryl (SH), and disulfide bond (SS) were determined. The highest ACE-inhibitory activity of DWGP hydrolysate was obtained at power intensity of 191.1 W/cm² for 10 min in the ultrasonic horn reactor. The fixed frequency of 33 kHz and the sweep frequency of 40 ± 2 kHz resulted in the maximum ACE-inhibitory activity. The combined irradiation of dual fixed frequency (24/68 kHz) produced significant increase in ACE-inhibitory activity compared with single frequency (33 kHz). The ultrasonic probe resulted in significant higher ACE-inhibitory activity compared with ultrasonic bath operating at single or dual fixed and sweep frequencies. The changes in conformation of the DWGP due to sonication were confirmed by the changes in fluorescence intensity, surface hydrophobicity, SH_f and SS contents and they were found in conformity with the ACE-inhibitory activity in case of the ultrasonic horn reactor but not in bath reactor.     

Effects of ultrasound pretreatment with different frequencies and working modes on the enzymolysis and the structure characterization of rice protein

The effects of ultrasound pretreatment with different frequencies and working modes, including mono-frequency ultrasound (MFU), dual-frequency ultrasound (DFU) and tri-frequency ultrasound (TFU), on the degree of hydrolysis (DH) of rice protein (RP) and angiotensin-I-converting enzyme (ACE) inhibitory activity of RP hydrolysate were investigated. Ultraviolet–visible (UV) spectroscopy, fourier transform infrared (FTIR) spectroscopy, surface hydrophobicity and scanning electron microscopy (SEM) of RP pretreated with ultrasound were measured. The results showed that ultrasound pretreatment did not increase DH of RP significantly (p > 0.05). However, all the ultrasound pretreatment increased the ACE inhibitory activity of RP hydrolysate significantly (p < 0.05). The MFU of 20 kHz showed higher ACE inhibitory activity compared to that of other MFU. The ACE inhibitory activity of sequential DFU was higher than that of simultaneous with the same frequency combination. Sequential TFU of 20/35/50 kHz produced the highest increase in ACE inhibitory activity in contrast with other ultrasound frequencies and working modes. All the results under ultrasound pretreatment showed that ultrasound frequencies and working modes were of great effect on the ACE inhibitory activity of RP. The changes in UV–Vis spectra and surface hydrophobicity indicated the unfolding of protein and exposure of hydrophobic groups by ultrasound. The FTIR analysis showed that all the ultrasound pretreatment with different frequencies and working modes decreased α-helix, β-turn content and increased β-sheet, random coil content of RP. The SEM results indicated that ultrasound pretreatment resulted in the deformation of RP. In conclusion, the frequency selection of ultrasound pretreatment of RP is essential for the preparation of ACE inhibitory peptide.     

Enzymolysis reaction kinetics and thermodynamics of defatted wheat germ protein with ultrasonic pretreatment

This research explores the mechanism of ultrasonic pretreatment on enzymolysis of defatted wheat germ protein (DWGP). The enzymolysis reaction kinetics and thermodynamics were studied after ultrasonic pretreatments using a probe-type sonicator and an ultrasonic cleaning bath, and the results were compared with traditional enzymolysis. The results showed that both the traditional and ultrasonic pretreated enzymolysis fit well to first-order kinetics. Both the temperature and ultrasound had a positive effect on the enzymolysis of DWGP, with temperature playing a dominant role. Under the optimized conditions of DWGP concentration of 1% (w/v), Alcalase concentration of 2000 U/g, time of 10 min and temperature of 50 °C, both the probe and cleaning bath ultrasonic pretreated enzymolysis showed high polypeptide concentrations (231.019 and 231.320 μg/mL) and low energy requirements. In comparison with traditional enzymolysis, these methods significantly increased the reaction rate constant (k) by 166.7% and 144.4%, 92.9% and 85.7%, 28.0% and 28.0%, 16.1% and 12.9% at 20, 30, 40 and 50 °C, and decreased the activation energy (E_a), enthalpy of activation (ΔH), Gibbs free energy of activation (ΔG) and entropy of activation (ΔS) by 68.6% and 62.4%, 74.1% and 67.5%, 34.3% and 31.2%, 1.4% and 1.3%. It can be concluded that ultrasonic pretreatment of DWGP can remarkably improve the enzymolysis efficiency and consequently leads to the production of higher polypeptide yield.     

超声波处理对脱脂麦胚分离蛋白结构的变化研究

应用傅里叶红外光谱(FTIR)、荧光光谱研究超声处理对脱脂麦胚分离蛋白(DWGP)结构的变化,建立DWGP高效酶解与其结构变化之间的关系.研究发现,超声处理均可提高DWGP的酶解效果,特别是经超声功率600 W、时间10 min处理后,酶解液抑制活性与对照组相比提高了23.96%.通过荧光光谱发现,超声作用改变DWGP溶液荧光强度,适当超声处理可使蛋白分子伸展、生色基团外露,有助于蛋白酶解后获得活性更高的抑制肽.采用对FTIR谱酰胺I带进行曲线拟合的方法,定量分析了不同超声功率、时间对DWGP二级结构的影响,发现超声作用使β-折叠相对含量下降而β-转角含量增加,这可能是DWGP酶解后获得高效抑制肽的主要原因.     

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 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.     

Preparation of corn ACE inhibitory peptide-ferrous chelate by dual-frequency ultrasound and its structure and stability analyses

In order to improve iron chelating ability and retain the activity of functional peptide, corn peptide was chelated with iron to form corn ACE inhibitory peptide-ferrous chelate (CP-Fe) treated by dual-frequency ultrasound. Furthermore, the chelating mechanism was revealed by analyzing various structural changes, and the stability was further evaluated. Under this study condition, the iron-binding capacity of corn ACE inhibitory peptide (CP) and chelate yield reached 66.39% and 82.87%, respectively. Ultrasound-treated CP exhibited a high iron chelating ability, meanwhile, chelation reaction had no significant effect on the ACE inhibition activity (82.21%) of the peptide. CP-Fe was formed by binding the peptides amino, carbonyl and carboxyl groups with Fe²⁺ demonstrated by Ultra-violet spectroscopy, Fourier transform infrared characterization, X-ray diffraction, energy dispersion spectrum, zeta potential, amino acid composition and other multi-angle analyses. Moreover, ultrasound-treated CP-Fe chelate exhibited porous surface and uniform nanoparticle shape. Furthermore, ultrasound-treated CP-Fe chelate exhibited an excellent stability towards various pH (retention rate ≥ 95.47% at pH 6–10), temperatures (retention rate ≥ 85.10% at 25–70 °C), and gastrointestinal digestion (retention rate 79.18%). Overall, ultrasound-treated CP-Fe chelate possessed high iron-chelating ability, ACE inhibition activity and stability. This study provides a novel synthesis method of the iron-chelating corn ACE inhibitory peptide, which is promising to be applied as iron supplements with high efficiency, bioactivity, and stability.     

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.     

Surface topography,nano-mechanics and secondary structure of wheat gluten pretreated by alternate dual-frequency ultrasound and the correlation to enzymolysis

The effects of alternate dual-frequency ultrasound (ADFU) pretreatment on the degree of hydrolysis (DH) of wheat gluten (WG) and angiotensin I-converting enzyme (ACE) inhibitory activity were investigated in this research. The surface topography, nano-mechanics and secondary structure of WG were also determined using atomic force microscope (AFM) and circular dichroism (CD). The correlations of ACE inhibitory activity and DH with surface topography, nano-mechanics and secondary structure of WG were determined using Pearson’s correlation analysis. The results showed that with an increase in either pretreatment duration or power, the ACE inhibitory activity of the hydrolysate also increases, reaching maximum at 10 min and 150 W/L, respectively, and then decreases thereafter. Similarly, AFM analysis showed that as the pretreatment duration or power increases, the surface roughness also increase and again a decrease occurs thereafter. As the pretreatment duration or power increased, the Young’s modulus and adhesion of WG also increased and then declined. Young’s modulus and adhesions average values were compared with ACE inhibitory activity reversely. The result of the CD spectra analysis exhibited losses in the relative percentage of α-helix of WG. Pearson’s correlation analysis showed that the average values of Young’s modulus and the relative percentage of α-helix correlated with ACE inhibitory activity of the hydrolysates linearly and significantly (P < 0.05); the relative percentage of β-sheet correlated linearly with DH of WG significantly (P < 0.05). In conclusion, ADFU pretreatment is an efficient method in proteolysis due to its physical and chemical effect on the Young’s modulus, α-helix and β-sheet of WG.     

Impact of ultrasound pretreatment on hydrolysate and digestion products of grape seed protein

The effects of ultrasound pretreatment with different working modes, including mono frequency ultrasound (MFU), simultaneous dual frequency ultrasound (SDFU) and alternate dual frequency ultrasound (ADFU) using energy-gather counter flow ultrasound equipment, on the degree of hydrolysis (DH) of grape seed protein (GSP) hydrolysate and IC₅₀ of GSP digestion products were studied. Amino acid composition analysis (AACA), ultraviolet–visible (UV) spectroscopy and atomic force microscopy (AFM) of GSP with different ultrasound pretreatments were measured. The results showed that MFU, SDFU and ADFU pretreatments improved the DH and reduced the IC₅₀ of GSP significantly (P < .05). The MFU of 20 kHz and SDFU of 20/40 kHz showed higher ACE inhibitory activity within the MFU and SDFU groups, respectively. ADFU of 20/35 kHz produced the highest ACE inhibitory activity among the three working modes (MFU, SDFU and ADFU). AACA showed that all the working modes of the ultrasound pretreatment could increase the amount of hydrophobic amino acids and the total amino acids. The changes in UV spectra and amino acid analysis indicated the unfolding of protein structure and exposure of more hydrophobic groups by SDFU and ADFU pretreatments. AFM analysis of the GSP indicated that the microstructures were destroyed and the particle size reduced after dual-frequency ultrasound pretreatments. Therefore, energy-gather counter flow ultrasound pretreatment is an effective method to improve the DH and reducing the IC₅₀ due to the changes of molecular conformation and effects on the microstructure by sonochemistry of GSP. In conclusion, it is necessary to select the frequency and working modes of ultrasound pretreatment for the preparation of ACE inhibitory peptide of GSP.     

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.     

Effect of ultrasonic treatment on the polyphenol content and antioxidant capacity of extract from defatted hemp,flax and canola seed cakes

The effectiveness of ultrasonic extraction of phenolics and flavonoids from defatted hemp, flax and canola seed cakes was compared to the conventional extraction method. Ultrasonic treatment at room temperature showed increased polyphenol extraction yield and antioxidant capacity by two-fold over the conventional extraction method. Different combinations of ultrasonic treatment parameters consisting of solvent volume (25, 50, 75 and 100 mL), extraction time (20, 30 and 40 min) and temperature (40, 50, 60 and 70 °C) were selected for polyphenol extractions from the seed cakes. The chosen parameters had a significant effect (p < 0.05) on the polyphenol extraction yield and subsequent antioxidant capacity from the seed cakes. Application of heat during ultrasonic extraction yielded higher polyphenol content in extracts compared to the non-heated extraction. From an orthogonal design test, the best combination of parameters was 50 mL of solvent volume, 20 min of extraction time and 70 °C of ultrasonic temperature.     

Improving the enzymolysis efficiency of potato protein by simultaneous dual-frequency energy-gathered ultrasound pretreatment: Thermodynamics and kinetics

The thermodynamics and kinetics of traditional and simultaneous dual frequency energy-gathered ultrasound (SDFU) assisted enzymolysis of potato protein were investigated to get the knowledge of the mechanisms on the SDFU’s promoting efficiency during enzymolysis. The concentration of potato protein hydrolysate and parameters of thermodynamic and kinetic during traditional and SDFU assisted enzymolysis were determined. The results showed that potato protein hydrolysate concentration of SDFU assisted enzymolysis was higher than traditional enzymolysis at the hydrolysis time of 60 min (p < 0.05) whereas not significantly different at 120 min (p > 0.05). In some cases, SDFU assisted enzymolysis took less hydrolysis time than traditional enzymolysis when the similar conversion rates of potato protein were obtained. The thermodynamic papameters including the energy of activation (E_a), enthalpy of activation (△H), entropy of activation (△S) were reduced by ultrasound pretreatment while Gibbs free energy of activation (△G) increased little (1.6%). Also, kinetic papameters including Michaelis constant (K_M) and catalytic rate constant (k_cat) decreased by ultrasound pretreatment. On the contrary, reaction rate constants (k) of SDFU assisted enzymolysis were higher than that of traditional enzymolysis (p < 0.05). It was indicated that the efficiency of SDFU assisted enzymolysis was higher than traditional enzymolysis in a limited time. The higher efficiency of SDFU assisted enzymolysis was related with the decrease of E_a and K_M by lowering the energy barrier between ground and active state and increasing affinity between substrate and enzyme.     

Effects of ultrasound energy density on the non-thermal pasteurization of chocolate milk beverage

This study presents the emerging high-intensity ultrasound (HIUS) processing as a non-thermal alternative to high-temperature short-time pasteurization (HTST). Chocolate milk beverage (CMB) was subjected to different ultrasound energy densities (0.3–3.0 kJ/cm³), as compared to HTST pasteurization (72 °C/15 s) aimed to verify the effect of the HIUS processing on the microbiological and physicochemical characteristics of the beverage. The application of HIUS at an energy density of 3.0 kJ/cm³ was able to reduce 3.56 ± 0.02 logarithmic cycles in the total aerobic counts. In addition, the ultrasound energy density affected the physical properties of the beverage as the size distribution of fat globule and rheological behavior, as well as the chemical properties such as antioxidant activity, ACE inhibitory activity, fatty acid profile, and volatile profile. In general, the different energetic densities used as a non-thermal method of pasteurization of CMB were more effective when compared to the conventional pasteurization by HTST, since they improved the microbiological and physicochemical quality, besides preserving the bioactive compounds and the nutritional quality of the product.     

Effects of ultrasound pretreatment on the quality,nutrients and volatile compounds of dry-cured yak meat

The objective of the present study was to assess the effects of ultrasound pretreatment on the quality of dry-cured yak meat. The ultrasonic power with 0, 200, 300 and 400 W (ultrasonic frequency of 20 kHz) were used to assist processing of dry-cured yak meat. The meat quality, nutrient substances, sensory quality, electronic nose, electronic tongue and volatile compounds of dry-cured yak meat were determined. The results indicated that the moisture content and hardness value of ultrasonic treatment group was significantly lower compared to the control group (P < 0.05). Ultrasonic treatment increased the value of b*, and decreased the value of L*, a*, pH, chewiness, melting temperature and enthalpy. Springiness value significantly increased from control group to 300 W of ultrasonic power group. Shear force significantly decreased with the increase of ultrasonic power (P < 0.05). Ultrasonic treatment had no effect on the TVB-N content, but it could increase the TBARS content. Ultrasonic treatment could significantly increase the essential FAA (EFAA) and total FAA (P < 0.05). In addition, the saturated fatty acid (SFA) content significantly increased with the increase of ultrasonic power (P < 0.05). Ultrasound treatment negatively affected the meat’s color, smell, and taste but increased its tenderness and the overall acceptability. It also significantly increased alcohols and aldehydes contents (P < 0.05), which were consistent with the measurement of electronic nose and electronic tongue. The results demonstrated that the the appropriate ultrasonic power assisted in the processing improves quality of dry-cured yak meat, particularly for the power of 300 W.     

Effects of ultrasound on the degradation kinetics,physicochemical properties and prebiotic activity of Flammulina velutipes polysaccharide

The controllable ultrasonic modification was hindered due to the uncertainty of the relationship between ultrasonic parameters and polysaccharide quality. In this study, the ultrasonic degradation process was established with kinetics. The physicochemical properties and prebiotic activity of ultrasonic degraded Flammulina velutipes polysaccharides (U-FVPs) were investigated. The results showed that the ultrasonic degradation kinetic models were fitted to 1/M_t-1/M₀ = kt. When the ultrasonic intensity increased from 531 to 3185 W/cm², the degradation proceeded faster. The decrease of polysaccharide concentration contributed to the degradation of FVP, and the fastest degradation rate was at 60 °C. Ultrasound changed the solution conformation of FVP, and partially destroyed the stability of the triple helix structure of FVP. Additionally, the viscosity and gel strength of FVP decreased, but its thermal stability was improved by ultrasound. Higher ultrasonic intensity led to larger variations in physicochemical properties. Compared with FVP, U-FVPs could be more easily utilized by gut microbiota. U-FVPs displayed better prebiotic activity by promoting the growth of Bifidobacterium and Brautella and inhibiting the growth of harmful bacteria. Ultrasound could be effectively applied to the degradation of FVP to improve its physicochemical properties and bioactivities.     

Effects of multi-frequency ultrasound on physicochemical properties,structural characteristics of gluten protein and the quality of noodle

In this study, the influence of multi-frequency ultrasound irradiation on the functional properties and structural characteristics of gluten, as well as the textural and cooking characteristics of the noodles were investigated. Results showed that the textural and cooking characteristics of noodles that contain less gluten pretreated by multi-frequency ultrasonic were ultrasonic frequency dependent. Moreover, the noodles that contain a smaller amount of sonicated gluten could achieve the textural and cooking quality of commercial noodles. There was no significant difference in the cooking and texture characteristics between commercial noodles and noodles with 12%, 11%, and 10% gluten pretreated by single-frequency (40 kHz), dual-frequency (28/40 kHz), and triple-frequency sonication (28/40/80 kHz), respectively. Furthermore, the cavitation efficiency of triple-frequency ultrasound was greater than that of dual-frequency and single-frequency. As the number of ultrasonic frequencies increased, the solubility, water holding capacity and oil holding capacity of gluten increased significantly (p < 0.05), and the particle size was reduced from 197.93 ± 5.28 nm to 110.15 ± 2.61 nm. Furthermore, compared to the control group (untreated), the UV absorption and fluorescence intensity of the gluten treated by multi-frequency ultrasonication increased. The surface hydrophobicity of gluten increased from 8159.1 ± 195.87 (untreated) to 11621.5 ± 379.72 (28/40/80 kHz). Raman spectroscopy showed that the α-helix content of all sonicated gluten protein samples decreased after sonication, while the β-sheet and β-turn content increased, and tryptophan and tyrosine residues were exposed. Through scanning electron microscope (SEM) analysis, the gluten protein network structure after ultrasonic treatment was loose, and the pore size of the gluten protein network increased from about 10 μm (untreated) to about 26 μm (28/40/80 kHz). This work elucidated the effect of ultrasonic frequency on the performance of gluten, indicating that with increasing frequency combination increases, the ultrasound effect became more pronounced and protein unfolding increased, thereby impacting the functional properties and the quality of the final product. This study provided a theoretical basis for the application of multi-frequency ultrasound technology in the modification of gluten protein and noodle processing.     

Effect of multi-mode dual-frequency ultrasound pretreatment on the vacuum freeze-drying process and quality attributes of the strawberry slices

The effects of osmotic pretreatment assisted by ultrasound in different frequency modes before vacuum freeze-drying (VFD) on moisture migration and quality characteristics of strawberry slices were investigated. The frequency modes are single-frequency modes under 20, 40 kHz (SM-20, SM-40), and dual-frequency under 20/40 kHz including sequential mode (SeDM) and simultaneous mode (SiDM). The quality characteristics of dried strawberry products including rehydration, hardness, color, flavor, total anthocyanins, total phenols, vitamin C content, and active antioxidant components (DPPH and –OH) were determined. Results showed that drying time of the strawberry slices irradiated by ultrasound was reduced by 15.25%–50.00%, compared to the control samples. Besides, dual-frequency ultrasound shortened the drying time more than single-frequency ultrasound. The drying time of SeDM was the shortest. In addition to vitamin C content, the quality characteristics including rehydration, hardness, color, flavor, total anthocyanins, total phenols, and antioxidant activity of dried strawberry products pretreated by SeDM were significantly (p < 0.05) better than those of control and other pretreated samples. It can be concluded that the SeDM was an effective pretreatment method to produce high-quality vacuum freeze-dried strawberry products.     

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.     

Effects of ultrasound and additives on the function and structure of trypsin

Encapsulating proteins in polymeric microspheres is a useful mode of drug delivery, but the proteins are subjected to damage in the process of ultrasound emulsion microencapsulation. The objective of this study was to investigate the effects of ultrasound power and duration on the function and structure of trypsin, and the reason of protein denaturation when it was irradiated by 20 kHz ultrasound. The relatively stable enzyme, trypsin, was dissolved in aqueous solution in the presence and absence of additives to study the stability of trypsin during the ultrasound irradiation. The damage of the molecular structure of trypsin was detected via combined high performance liquid chromatogram and electrospray ionization mass spectrometry (HPLC–ESI-MS). The results showed that the activity of trypsin decreased with increasing ultrasound power from 100 to 500 W or extending the irradiation time from 1 to 20 min. This effect could be enhanced via aerating the solution for a duration 10 min at 300 W. Fragments of trypsin were detected in the treatment (300 W, 10 min) by HPLC–ESI-MS. The additives, Tween 80 and mannitol, could protect trypsin against the inactivation caused by ultrasound. The reason of inactivation was partly from the alteration of the molecular conformation and partly from the modification or damage of trypsin's molecular structure.