Ultrasound combined with slightly acidic electrolyzed water thawing of mutton: Effects on physicochemical properties,oxidation and structure of myofibrillar protein

The effects of air thawing (AT), water immersion thawing (WT), microwave thawing (MT) and ultrasound combined with slightly acidic electrolyzed water thawing (UST) on the myofibrillar protein (MP) properties (surface hydrophobicity, solubility, turbidity, particle size and zeta potential), protein oxidation (carbonyl content and sulfhydryl content) and structure (primary, secondary and tertiary) of frozen mutton were investigated in comparison with fresh mutton (FM). The solubility and turbidity results showed that the MP properties were significantly improved in the UST treatment. UST treatment could effectively reduce the MP aggregation and enhance the stability, which was similar to the FM. In addition, UST treatment could effectively inhibit protein oxidation during thawing as well. The primary structure of MP was not damaged by the thawing methods. UST treatment could reduce the damage to MP secondary and tertiary structure during the thawing process compared to other thawing methods. Overall, the UST treatment had a positive influence in maintaining the MP properties by inhibiting protein oxidation and protecting protein structure.     

Effect of ultrasound treatment on interactions of whey protein isolate with rutin

Rutin is a biologically active polyphenol, but its poor water solubility and low bioavailability limit its application to the food industry. We investigated the effect of ultrasound treatment on the properties of rutin (R) and whey protein isolate (WPI) using spectral and physicochemical analysis. The results revealed that there was covalent interaction between whey protein isolate with rutin, and the binding degree of whey isolate protein with rutin increased with ultrasound treatment. Additionally, solubility and surface hydrophobicity of WPI-R complex improved with ultrasonic treatment, and a maximum solubility of 81.9 % at 300 W ultrasonic power. The ultrasound treatment caused the complex to develop a more ordered secondary structure, resulting in a three-dimensional network structure with small and uniform pore sizes. This research could provide a theoretical reference for studying protein–polyphenol interactions and their applications in food delivery systems.     

Molecular aggregation and property changes of egg yolk low-density lipoprotein induced by ethanol and high-density ultrasound

Solvent and physical treatment are widely used in egg yolk processing, but the detailed changes in the molecular structure of egg yolk proteins during processing are unclear. The aim of this study was to investigate the effects of ethanol and ultrasonic treatments on chicken egg yolk low-density lipoprotein (LDL). The solubility, emulsifying activity and emulsifying stability decreased by 74.75%, 46.91%, and 81.58% after ethanol treatment, respectively. The average particle size of ethanol-treated LDL increased 13.3-fold to 937.85 nm. These results suggested that ethanol treatment induced wide-ranging aggregation of LDL. In contrast to ethanol treatment, ultrasonic treatment promoted the solubility and emulsifying stability of LDL and enhanced its zeta-potential (119.56%) and surface hydrophobicity (10.81%). Based on particle size analysis and transmission electron microscopy, approximately 34.65% of LDL had undergone aggregation and the molecular interface became more flexible after ultrasonic treatment. These results revealed the detailed changes in egg yolk LDL structure and properties during solvent (ethanol) and physical (ultrasound) processing.     

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.     

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.     

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.     

Modifying the physicochemical properties,solubility and foaming capacity of milk proteins by ultrasound-assisted alkaline pH-shifting treatment

This study investigated the effects of different treatment of alkaline pH-shifting on milk protein concentrate (MPC), micellar casein concentrate (MCC) and whey protein isolate (WPI) assisted by the same ultrasound conditions, including changes in the physicochemical properties, solubility and foaming capacity. The solubility of milk proteins had a significant increase with gradual enhancement of ultrasound-assisted alkaline pH-shifting (p < 0.05), especially for MCC up to 99.50 %. Also, treatment made a significant decline in the particle size of MPC and MCC, as well as the turbidity of the proteins (p < 0.05). The foaming capacity of MPC, MCC, and WPI was all improved, especially at pH 11, and at this pH, the milk protein also showed the highest surface hydrophobicity. The best foaming capacity at pH 11 was the result of the combined effect of particle size, potential, protein conformation, solubility, and surface hydrophobicity. In conclusion, ultrasound-assisted pH-shifting treatment was found to be effective in improving the physicochemical properties and solubility and foaming capacity of milk proteins, especially MCC, with promising application prospect in food industry.     

Effects of ultrasound-assisted basic electrolyzed water (BEW) extraction on structural and functional properties of Antarctic krill (Euphausia superba) proteins

A novel protein extraction method of ultrasound-assisted basic electrolyzed water (BEW) was proposed, and its effects on the structural and functional properties of Antarctic krill proteins were investigated. Results showed that BEW reduced 30.9% (w/w) NaOH consumption for the extraction of krill proteins, and its negative redox potential (−800 ~ −900 mV) protected the active groups (carbonyl, free sulfhydryl, etc.) of the proteins from oxidation compared to deionized water (DW). Moreover, the ultrasound-assisted BEW increased the extraction yield (9.4%), improved the solubility (8.5%), reduced the particle size (57 nm), favored the transition of α-helix and β-turn to β-sheet, promoted the surface hydrophobicity and disulfide bonds formation of krill proteins when compared to BEW without ultrasound. These changes contributed to the enhanced foam capacity, foam stability and emulsifying capacity of the krill proteins. Notably, all the physicochemical, structural and functional properties of the krill proteins were comparable to those extracted by the traditional ultrasound-assisted DW. This study suggests that the ultrasound-assisted BEW can be a potential candidate to extract proteins, especially offering an alternative way to produce marine proteins with high nutritional quality.     

Impacts of sonication and high hydrostatic pressure on the structural and physicochemical properties of quinoa protein isolate dispersions at acidic,neutral and alkaline pHs

Herein, 1 wt% quinoa protein isolate (QPI) was exposed to sonication using a 20 kHz ultrasonicator equipped with a 6 mm horn (14.4 W, 10 mL, up to 15 min) or high hydrostatic pressure (HHP, up to 600 MPa, 15 min) treatments at pH 5, pH 7, and pH 9. The changes to physicochemical properties were probed by SDS-PAGE, FTIR, free sulfhydryl group (SH), surface hydrophobicity (H₀), particle size and solubility. As revealed by SDS-PAGE, substantial amounts of 11S globulin participated in the formations of aggregates via SS bond under HHP, particularly at pH 7 and pH 9. However, protein profiles of QPI were not significantly affected by the sonication. Free SH groups and surface hydrophobicity were increased after the sonication treatment indicating protein unfolding and exposure of the embedded SH and/or hydrophobic groups. An opposite trend was observed in HHP treated samples, implying aggregation and reassociation of structures under HHP. HHP and sonication treatments induced a decrease in ordered secondary structures (random coil and β-turn) accompanied with an increase in disordered secondary structures (α-helix and β-sheet) as probed by FTIR. Finally, the sonication treatment induced a significant improvement in the solubility (up to ∼3 folds at pH 7 and ∼2.6 folds at pH 9) and a reduction in particle sizes (up to ∼3 folds at pH 7 and ∼4.4 folds at pH 9). However, HHP treatment (600 MPa) only slightly increased the solubility (∼1.6 folds at pH 7 and ∼1.2 folds at pH 9) and decreased the particle size (∼1.3 folds at pH 7 and ∼1.2 folds at pH 9). This study provides a direct comparison of the impacts of sonication and HHP treatment on QPI, which will enable to choose the appropriate processing methods to achieve tailored properties of QPI.     

Effects of homogeneous and ultrasonic treatment on casein/phosphatidylcholine complex-emulsions: Stability and bioactivity insights

Casein (CAS), a typical protein emulsifier, has functional properties limited by its chemical structure in practical production applications. This study aimed to combine phosphatidylcholine (PC) and casein to form a stable complex (CAS/PC) and improve its functional properties through physical modification (homogeneous and ultrasonic treatment). To date, few studies have explored the effects of physical modification on the stability and biological activity of CAS/PC. Interface behavior analysis showed that compared to homogeneous treatment, PC addition and ultrasonic treatment could decrease the mean particle size (130.20 ± 3.96 nm) and increase the zeta potential (−40.13 ± 1.12 mV), indicating the emulsion is more stable. The chemical structural analysis of CAS showed that PC addition and ultrasonic treatment promoted changes in its sulfhydryl content and surface hydrophobicity, exposing more free sulfhydryl groups and hydrophobic binding sites, thereby enhancing its solubility and improving the stability of the emulsion. Additionally, storage stability analysis revealed that the incorporation of PC with ultrasonic treatment could improve the root mean square deviation value and radius of gyration value of CAS. These modifications resulted in an increase the binding free energy between CAS and PC (−238.786 kJ/mol) at 50 °C, leading to an improvement in the thermal stability of the system. Furthermore, digestive behavior analysis indicated that PC addition and ultrasonic treatment could increase the total FFA release from 667.44 ± 22.33 μmol to 1250.33 ± 21.56 μmol. In conclusion, the study underscores the effectiveness of PC addition and ultrasonic treatment in enhancing the stability and bioactivity of CAS, offering novel ideas for designing stable and healthy emulsifiers.     

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.     

Ultrasonic treatment on physicochemical properties of water-soluble protein from Moringa oleifera seed

Effect of ultrasonic power on the structure and functional properties of water-soluble protein extracted from defatted Moringa oleifera seed were explored. The results showed that ultrasonic treatment could reduce β-sheet and β-turn content of water-soluble protein from Moringa oleifera seed (MOWP) and increase the content of random coil and α-helix. Changes in intrinsic fluorescence spectra, surface hydrophobicity (H₀) and thermal behaviors indicated that ultrasonic had significant effect on the tertiary structure of MOWP. The results of SEM and SDS-PAGE showed that the MOWP was aggregated but not significantly degraded by ultrasound. The solubility, foaming properties and emulsifying properties of MOWP increased firstly and then decreased with the increase of ultrasonic power. Ultrasonic treatment altered the functional properties of MOWP, which might be attributed to the exposure of hydrophilic group and the change of and secondary and tertiary structure.     

Formation of soybean protein isolate-hawthorn flavonoids non-covalent complexes: Linking the physicochemical properties and emulsifying properties

In recent years, more and more attention had been paid to the combination of proteins and flavonoids, and several flavonoids had been reported to improve the physicochemical and emulsifying properties of proteins. This study investigated the effects of ultrasonic treatment (450 W for 10 min, 20 min, and 30 min) on the physicochemical properties, antioxidant activity, and emulsifying properties of soy protein isolate (SPI) -hawthorn flavonoids (HF) non-covalent complexes. The results showed that the addition of HF to SPI and 20 min of ultrasound could reduce α-helix and random coil, increase β-sheet and β-turn, and enhance fluorescence quenching. In addition, it decreased the particle size, zeta potential, surface hydrophobicity, and turbidity to 88.43 or 95.27 nm, −28.80 mV, 1250.42, and 0.23, respectively. The protein solubility, free sulfhydryl group, antioxidant activity, emulsifying activity index, and emulsifying stability index all increased to 73.93%, 15.07 μmol/g, 71.00 or 41.91%, 9.81 m²/g, and 67.71%, respectively. Moreover, high-density small and low-flocculation droplets were formed. Therefore, the combined ultrasound treatment and addition of HF to SPI is a more effective method for protein modification compared to ultrasound treatment alone. It provides a theoretical basis for protein processing and application in the future.     

Ultrasonic-assisted pH shift-induced interfacial remodeling for enhancing the emulsifying and foaming properties of perilla protein isolate

In order to expand the applications of plant protein in food formulations, enhancement of its functionalities is meaningful. Herein, the effects of ultrasonic (20 KHz, 400 W, 20 min)-assisted pH shift (pH 10 and 12) treatment on the structure, interfacial behaviors, as well as the emulsifying and foaming properties of perilla protein isolate (PPI) were investigated. Results showed that the solubility of PPI treated by ultrasonic-assisted pH shift (named UPPI-10/12) exceeded 90 %, which was at least 2 and 1.4 times that of untreated PPI and ultrasound-based PPI. Meanwhile, UPPI-10/12 possessed higher foamability (increasing by at least 1.2 times) and good emulsifying stability. Ultrasonic-assisted pH shift treatment decomposed large PPI aggregates into tiny particles, evident from the dynamic light scattering (DLS) and atomic force microscopy results. Besides, this approach induced a decrease in α-helix of PPI and an increase in β-sheet, which might result in the exposure of the hydrophobic group on the structural surface of PPI, thus leading to the increase of surface hydrophobicity. The smaller size and higher hydrophobicity endowed UPPI-10/12 faster adsorption rate, tighter interfacial structure, and higher elastic modulus at the air- and oil–water interfaces, evident from the cryo-SEM and interfacial dilatational rheological results. Thus, the emulsifying and foaming properties could evidently enhance. This study demonstrated that ultrasonic-assisted pH shift technique was a simple approach to effectively improve the functional performance of PPI.     

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.     

Modifying the physicochemical properties of pea protein by pH-shifting and ultrasound combined treatments

The effect of a pH-shifting and ultrasound combined process on the functional properties and structure of pea protein isolate (PPI) was investigated. PPI dispersions were adjusted to pH 2, 4, 10, or 12, treated by power ultrasound for 5 min, and incubated for 1 h before the sample pH was brought back to neutral. After treatment, water solubility, protein aggregate size, solution turbidity, surface hydrophobicity (Ho), free sulfhydryl content (SH), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the soluble PPI were determined. pH-shifting at pH 12 and ultrasound combined treatment (pH12-US) significantly improved protein properties, while property modification of the samples treated under acidic conditions was less pronounced. The pH12-US treated PPI had a solubility seven times higher than the control, reaching an average particle size of 45.2 nm. In addition, the pH12-US treated PPI significantly improved Ho due to disulfide bonds disruption, and produced more protein sub-units than other treatments. The soluble PPI obtained through this process may be a promising emulsifier for the enrichment of fat-soluble nutrients in foods.     

High-intensity ultrasound modified the functional properties of Neosalanx taihuensis myofibrillar protein and improved its emulsion stability

This study aimed to investigate the effects of high-intensity ultrasound treatment on the functional properties and emulsion stability of Neosalanx taihuensis myofibrillar protein (MP). The results showed that the carbonyl groups, emulsification properties, intrinsic fluorescence intensity, and surface hydrophobicity of the ultrasound treated MP solution were increased compared to the MP without ultrasound treatment. The results of secondary structure showed that the ultrasound treatment could cause a huge increase of β-sheet and a decline of α-helix of MP, indicating that ultrasound induced molecular unfolding and stretching. Moreover, ultrasound reduced the content of total sulfhydryl and led to a certain degree of MP cross-linking. The microscopic morphology of MP emulsion indicated that the emulsion droplet decreased with the increase of ultrasound power. In addition, ultrasound could also increase the storage modulus of the MP emulsion. The results for the lipid oxidation products indicated that ultrasound significantly improved the oxidative stability of N. taihuensis MP emulsions. This study offers an important reference theoretically for the ultrasound modification of aquatic proteins and the future development of N. taihuensis deep-processed products represented by surimi.     

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.     

Ultrasound-assisted enzymatic hydrolysis of yeast β-glucan catalyzed by β-glucanase: Chemical and microstructural analysis

The purpose of this study was to investigate the effect of ultrasound-assisted enzymolysis on modified solubilization of yeast β-glucan and its related mechanism. The depolymerization effects of this system on the physicochemical properties and structural features of the degraded fragments were studied systematically. The structure and physicochemical properties of the samples showed that the solubility of yeast β-glucan achieved 75.35 % after modification; and ultrasonic enzymatic enhanced the degradation efficiency. The yeast β-glucan obtained after solubilization and modification owned better antioxidant activities. The yeast β-glucan particles become obviously smaller, sparsely dispersed in the aqueous solution and the stability was improved. In addition, the hydrogen bonds in yeast β-glucan native triple helix structure were partially broken. Moreover, the disruption of yeast β-glucan's original structure made it decreased thermostability and easier to dissolve in water. The atomic force microscope (AFM) imaging directly verified the branched-chain morphology of yeast β-glucan and the small-strand degradation fragments. Therefore, this research can provide a feasible and effective approach for improving solubility of water-insoluble yeast β-glucan to enlarge its food and biomedical applications.     

Functional and bioactive properties of Larimichthys polyactis protein hydrolysates as influenced by plasma functionalized water-ultrasound hybrid treatments and enzyme types

The effects of plasma functionalized water (PFW) and its combination with ultrasound (UPFW) on the functional and bioactive properties of small yellow croaker protein hydrolysates (SYPHs) produced from three enzymes were investigated. Fluorescence and UV–Vis spectroscopy indicated that SYPHs tended to unfold with increasing intensity and shift in wavelengths to more flexible conformations under PFW and UPFW treatments. Particle size distribution and microstructure analysis revealed that treatments could disrupt aggregation of protein molecules to increase the roughness, specific surface area, and decrease the particle size of peptides during hydrolysis. The partially denatured structure of SYPHs induced by treatments increased the susceptibility of the fish proteins to exogenous enzymes, thereby accelerating the hydrolytic process to yield peptides with improved solubility, decreased emulsifying and foaming properties, and improved enzyme-specific antioxidant properties. The results revealed that the functionality of SYPHs was influenced by the treatment method and the enzyme type employed.