Effect of ultrasonic pretreatment on the rheology and structure of grass pea (Lathyrus sativus L.) protein emulsion gels induced by transglutaminase

In this study, emulsion gels were prepared by sonicated grass pea protein isolates (GPPI) at different ultrasonic amplitudes (25, 50 and 75 %) and times (5, 10 and 20 min). Formation of emulsion gels was induced by transglutaminase. Enzymatic gelation of emulsions stabilized by sonicated GPPI occurred in two stages. A relatively fast stage led to the formation of a weak gel which was followed by a slow stage that strongly reinforced the gel structure. Emulsion gels fabricated by sonicated GPPIs showed a homogeneous and uniform droplet distribution with higher elastic modulus compared to the native protein. A stiffer emulsion gel with a higher G' was formed after the protein was treated at 75 % amplitude for 10 min. After sonication of GPPI, the water holding capacity (WHC) of emulsion gels increased in accordance with the mechanical properties. Higher intermolecular cross-linking within the gel network increased the thermal stability of emulsion gels fabricated by sonicated GPPI. Although sonicated-GPPI emulsion gels clearly displayed homogenous microstructure in comparison to that made with native GPPI, the microstructures of these gels were nearly identical for all sonication amplitudes and times.     

Improvement in enzymolysis efficiency and changes in conformational attributes of corn gluten meal by dual-frequency slit ultrasonication action

The influences of dual-frequency slit ultrasound (DFSU) pretreatment with various working parameters on the enzymolysis efficiency and conformational characteristics of corn gluten meal (CGM) were studied. Results indicated that under the conditions of ultrasonic power density of 80 W/L, time of 30 min, ultrasonic intermittent ratio of 5:2 s/s, temperature of 30 °C, and substrate concentration of 50 g/L, the relative enzymolysis efficiency (REE) of CGM reached a maximum of 21.05%, and the protein dissolution rate was 68.50%. In addition, ultrasonication had considerable impact on the conformation of CGM and consequently improved the susceptibility to alcalase proteolysis. Changes in free sulfhydryl (SH_F) and disulfide bonds (SS) groups indicated spatial conformation of CGM was altered following sonication (sonochemical) treatment. Fourier Transform Infrared Spectrum (FITR) analysis showed a reduction in α-helix and β-turn content; and an increase in β-sheet and random coil content of CGM. Alterations in the particle size, particle size distribution, microstructure and surface roughness (R_a, R_q) indicated generation of smaller and more uniform protein fragments of CGM by sonochemical pretreatment. The proposed mechanism of sonicated CGM was elaborated. Our findings suggest that using DFSU in pretreating CGM may be an efficacious approach to enhance proteolysis.     

Effects of sonication on the in vitro digestibility and structural properties of buckwheat protein isolates

The present work investigated the effects of sonication at different amplitudes and durations on the in vitro digestibility of buckwheat protein isolates (BPIs). The conformation, particle size and microstructures of the BPIs were also studied to explicate the possible mechanisms of the sonication-induced changes. The results showed that sonication conditions of 20 kHz, pulsed on-time 10 s, off-time 5 s, amplitude of 60% and duration of 10 min (SA6T10) improved the digestibility of BPIs from 41.4% (control) to 58.2%. The tertiary structure analysis showed that sonication exposed the hydrophobic core buried inside the protein molecules and broke the intramolecular crosslinks, based on the increase in the surface hydrophobicity and intrinsic fluorescence and the decrease in the disulphide content. The secondary structure analysis showed that SA6T10 decreased the content of β-turn and β-sheet by 40.9% and 22.4%, respectively, and increased the content of anti-parallel β-sheet, random coil, and α-helix by 40.9%, 30.6%, and 25.5%, respectively. The particle size of the control BPIs (427.7 ± 76.7 nm) increased to 2130.8 ± 356.2 nm in the SA6T10 sonicated sample with a corresponding decrease in the polydispersity index from 0.97 ± 0.04 to 0.51 ± 0.13. Moreover, scanning electron microscopy indicated that sonication broke the macroparticles into smaller fragments and changed the surface state of the proteins. Taken together, sonication has proven to be a promising approach for improving the digestibility of buckwheat proteins, which can be explored as a source of plant-based alternative protein for food applications.     

Ultrasonic pretreatment of corn slurry for saccharification: A comparison of batch and continuous systems

The effects of ultrasound on corn slurry saccharification yield and particle size distribution was studied in both batch and continuous-flow ultrasonic systems operating at a frequency of 20 kHz. Ground corn slurry (28% w/v) was prepared and sonicated in batches at various amplitudes (192–320 μm_{peak-to-peak (p–p)}) for 20 or 40 s using a catenoidal horn. Continuous flow experiments were conducted by pumping corn slurry at various flow rates (10–28 l/min) through an ultrasonic reactor at constant amplitude of 12 μm_p–p. The reactor was equipped with a donut shaped horn. After ultrasonic treatment, commercial alpha- and gluco-amylases (STARGEN^TM 001) were added to the samples, and liquefaction and saccharification proceeded for 3 h. The sonicated samples were found to yield 2–3 times more reducing sugars than unsonicated controls. Although the continuous flow treatments released less reducing sugar compared to the batch systems, the continuous flow process was more energy efficient. The reduction of particle size due to sonication was approximately proportional to the dissipated ultrasonic energy regardless of the type of system used. Scanning electron microscopy (SEM) images were also used to observe the disruption of corn particles after sonication. Overall, the study suggests that both batch and continuous ultrasonication enhanced saccharification yields and reduced the particle size of corn slurry. However, due to the large volume involve in full scale processes, an ultrasonic continuous system is recommended.     

Functional properties of ultrasonically generated flaxseed oil-dairy emulsions

This study reports on the functional properties of 7% flaxseed oil/milk emulsion obtained by sonication (OM) using 20 kHz ultrasound (US) at 176 W for 1–8 min in two different delivery formulae, viz., ready-to-drink (RTD) and lactic acid gel. The RTD emulsions showed no change in viscosity after sonication for up to 8 min followed by storage up to a minimum of 9 days at 4 ± 2 °C. Similarly, the oxidative stability of the RTD emulsion was studied by measuring the conjugated diene hydroperoxides (CD). The CD was unaffected after 8 min of ultrasonic processing. The safety aspect of US processing was evaluated by measuring the formation of CD at different power levels. The functional properties of OM gels were evaluated by small and large scale deformation studies. The sonication process improved the gelation characteristics, viz., decreased gelation time, increased elastic nature, decreased syneresis and increased gel strength. The presence of finer sono-emulsified oil globules, stabilized by partially denatured whey proteins, contributed to the improvements in the gel structure in comparison to sonicated and unsonicated pasteurized homogenized skim milk (PHSM) gels. A sono-emulsification process of 5 min followed by gelation for about 11 min can produce gels of highest textural attibutes.     

Sonochemical synthesis of aluminium and aluminium hybrids for remediation of toxic metals

Spherical shaped nano-size aluminium oxide and its hybrids with indole and indole derivatives have been synthesized using sol–gel and post grafting methods coupled with sonication (Branson Digital SonifierS-250D; 20 kHz; 40%) for the remediation of toxic metals (lead and mercury). Different spectroscopic techniques (FTIR, SEM, BET, XRD, and XPS) have been applied to assess the properties of synthesized aluminium oxide and its hybrids. FTIR spectra showed the absorption bands of aluminium oxide (Al-O-Al) and aluminium hybrids (Al-O-C) at 800–400 cm⁻¹ and 1650–1100 cm⁻¹ region, respectively. SEM showed spherical shaped clusters of aluminium oxide which changed into the net-shape structure after the hybrid synthesis. It is worth noting that sonication energy increases the total surface area of aluminium oxide when it gets hybridized with indole and its derivatives from 82 m²/g to 167 m²/g; it also improved the product yield from 68% to 78%. Simultaneously, FTIR, SEM and BET analysis of non-sonicated aluminium oxide and its hybrids were also recorded for comparison. While XRD and XPS analysis were only conducted for sonicated aluminium oxide and its hybrids to manifest the structural and compositional properties. XRD patterns indexed as the cubic crystal system with an average 41 nm crystallite size of sonicated aluminium oxide which remains unaffected after hybrid synthesis. A survey scan under XPS confirmed the presence of all expected elements (aluminium, oxygen, carbon, nitrogen) and deconvolution of each recorded peak showed binding of element with its neighboring elements. The performance of aluminium oxide and its hybrids synthesize with and without sonication are also evaluated using a time-dependent batch adsorption protocol optimize for one hour. The maximum adsorption of lead (37%) and mercury (40%) are found onto sonicated aluminium oxide. The sonicated aluminium hybrids showed 43–63% of lead and 55–67% of mercury at pH 7. The fitness of experimental data using adsorption kinetics and isotherms revealed that adsorption follows Pseudo-second-order kinetic, Langmuir, and Freundlich isotherms.     

Sonication enhances polyphenolic compounds,sugars, carotenoids and mineral elements of apple juice

A study was initiated with the objective of evaluating the effects of sonication treatment on quality characteristics of apple juice such as polyphenolic compounds (chlorogenic acid, caffeic acid, catechin, epicatechin and phloridzin), sugars (fructose, glucose and sucrose), mineral elements (Na, K, Ca, P, Mg, Cu and Zn), total carotenoids, total anthocyanins, viscosity and electrical conductivity. The fresh apple juice samples were sonicated for 0, 30 and 60 min at 20 °C (frequency 25 kHz and amplitude 70%), respectively. As results, the contents of polyphenolic compounds and sugars significantly increased (P < 0.05) but the increases were more pronounced in juice samples sonicated for 30 min whereas, total carotenoids, mineral elements (Na, K and Ca) and viscosity significantly increased (P < 0.05) in samples treated for 60 min sonication. Losses of some mineral elements (P, Mg and Cu) also occurred. Total anthocyanins, Zn and electrical conductivity did not undergo any change in the sonicated samples. Findings of the present study suggest that sonication technique may be applied to improve phytonutrients present naturally in apple juice.     

Ultrasonic processing of dairy systems in large scale reactors

High intensity low frequency ultrasound was used to process dairy ingredients to improve functional properties. Based on a number of lab-scale experiments, several experimental parameters were optimised for processing large volumes of whey and casein-based dairy systems in pilot scale ultrasonic reactors. A continuous sonication process at 20 kHz capable of delivering up to 4 kW of power with a flow-through reactor design was used to treat dairy ingredients at flow rates ranging from 200 to 6000 mL/min. Dairy ingredients treated by ultrasound included reconstituted whey protein concentrate (WPC), whey protein and milk protein retentates and calcium caseinate. The sonication of solutions with a contact time of less than 1 min and up to 2.4 min led to a significant reduction in the viscosity of materials containing 18% to 54% (w/w) solids. The viscosity of aqueous dairy ingredients treated with ultrasound was reduced by between 6% and 50% depending greatly on the composition, processing history, acoustic power and contact time. A notable improvement in the gel strength of sonicated and heat coagulated dairy systems was also observed. When sonication was combined with a pre-heat treatment of 80 °C for 1 min or 85 °C for 30 s, the heat stability of the dairy ingredients containing whey proteins was significantly improved. The effect of sonication was attributed mainly to physical forces generated through acoustic cavitation as supported by particle size reduction in response to sonication. As a result, the gelling properties and heat stability aspects of sonicated dairy ingredients were maintained after spray drying and reconstitution. Overall, the sonication procedure for processing dairy systems may be used to improve process efficiency, improve throughput and develop value added ingredients with the potential to deliver economical benefits to the dairy industry.     

Effect of multi-frequency power ultrasound (MFPU) treatment on enzyme hydrolysis of casein

Effect of multi-frequency power ultrasound (MFPU) pretreatments on the degree of hydrolysis (DH) and mechanism of casein during alcalase enzymolysis was investigated. Results showed that MFPU pretreatment in tri-frequency 20/40/60 kHz mode significantly (p < 0.05) improved the DH value of casein. Variation of intrinsic fluorescence spectrum indicated the unfolding and degradation of casein occurred after MFPU pretreatment. Fourier transform infrared spectra showed that α-helix and β-sheet content of MFPU pretreated casein decreased, while β-turn and random coil content increased. Surface topography and nanostructures of caseins were found modified after MFPU pretreatments by the analysis of scanning electron microscopy (SEM) and atomic force microscopy (AFM). The SEM analysis also indicated that the enzymolysis residues of casein pretreated by MFPU were smaller than untreated samples. In conclusion, the MFPU can be used as an efficient pretreatment method to promote the enzymolysis of casein.     

High-intensity ultrasound pretreatment influence on whey protein isolate and its use on complex coacervation with kappa carrageenan: Evaluation of selected functional properties

The aim of this work was to evaluate the influence of high-intensity ultrasound (HIUS) treatment on whey protein isolate (WPI) molecular structure as a previous step for complex coacervation (CC) with kappa-carrageenan (KC) and its influence on CC functional properties. Protein suspension of WPI (1% w/w) was treated with an ultrasound probe (24 kHz, 2 and 4 min, at 50 and 100% amplitude), non HIUS pretreated WPI was used as a control. Coacervation was achieved by mixing WPI and KC dispersions (10 min). Time and amplitude of the sonication treatment had a direct effect on the molecular structure of the protein, FTIR-ATR analysis detected changes on pretreated WPI secondary structure (1600–1700 cm⁻¹) after sonication. CC electrostatic interactions were detected between WPI positive regions, KC sulfate group (1200–1260 cm⁻¹), and the anhydrous oxygen of the 3,6 anhydro-D-galactose (940–1066 cm⁻¹) with a partial negative charge. After ultrasound treatment, a progressive decrease in WPI particle size (nm) was detected. Rheology results showed pseudoplastic behavior for both, KC and CC, with a significant change on the viscosity level. Further, volume increment, stability, and expansion percentages of CC foams were improved using WPI sonicated. Besides, HIUS treatment had a positive effect on the emulsifying properties of the CC, increasing the time emulsion stability percentage. HIUS proved to be an efficient tool to improve functional properties in WPI-KC CC.     

Adsorption of methylene blue onto sonicated sepiolite from aqueous solutions

The aim of the present study is to enhance the methylene blue (MB) adsorption of sepiolite by ultrasonic treatment. The natural sepiolite was pretreated by sonication to improve the surface characteristics and enhance the dye uptake capacity. Sonication process resulted in a significant increase in the specific surface area (SSA) of sepiolite. The FTIR spectrum of the sonicated sepiolite indicates that the tetrahedral sheet is probably distorted after sonication process. The effect of various parameters such as sonication, pH, initial dye concentration and temperature on dye adsorption has been investigated. The adsorbed amount of MB on sepiolite increased after sonication as well as with increasing pH and temperature. The experimental data were evaluated by applying the pseudo-first- and second-order, and the intraparticle diffusion adsorption kinetic models. Adsorption process of MB onto sepiolite followed the pseudo-second-order rate expression. The experimental data were analyzed by Langmuir and Freundlich isotherms, and found that the isotherm data were reasonably well correlated by Langmuir isotherm. Maximum monolayer adsorption capacity of sepiolite for MB increased from 79.37 to 128.21 mg/g after the sonication. Various thermodynamic parameters, such as ΔG⁰, ΔH⁰ and ΔS⁰ were calculated. The thermodynamics of MB/sepiolite system indicated spontaneous and endothermic nature of the process. Adsorption measurements showed that the process was very fast and physical in nature.     

Structural and physicochemical characterization of modified starch from arrowhead tuber (Sagittaria sagittifolia L.) using tri-frequency power ultrasound

Sagittaria sagittifolia L. is a well-known plant, belongs to the Alismataceae family. Sonication can improve the functional properties of starch; hence, the aim of this study was to develop ultrasonically modified arrowhead starch (UMAS) using a sophisticated and eco-friendly tri-frequency power ultrasound (20/40/60 kHz) method at 300, 600, and 900 W for 15 and 30 min. Significant (p < 0.05) increases in swelling power, solubility, and water and oil holding capacities were achieved. FTIR spectroscopy corroborated the ordered, amorphous, and hydrated crystals of the sonicated samples. Increases in sonication frequency and power led to significant (p < 0.05) increases in onset gelatinization temperatures. Scanning electron microscopic analysis of sonicated samples showed superficial cracks and roughness on starch granules appeared in a sonication power-dependent manner compared with that of untreated sample. Overall, the ultrasonically-treated samples showed improved physicochemical properties, which could be useful for industrial applications.     

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.     

Ultrasound assisted chitosan coated iron oxide nanoparticles: Influence of ultrasonic irradiation on the crystallinity,stability, toxicity and magnetization of the functionalized nanoparticles

Due to unique reaction conditions of the acoustic cavitation process, ultrasound-assisted synthesis of nanoparticles has attracted increased research attention. In this study, we demonstrate the effect of ultrasonic irradiation on the crystallinity, stability, biocompatibility, and magnetic properties of chitosan-coated superparamagnetic iron oxide nanoparticles (CS-SPIONs). CS solution and colloidal suspension of SPIONs were mixed and sonicated using an ultrasonic probe of 1.3 cm tip size horn, frequency (20 kHz), and power (750 W). Different samples were sonicated for 1.5, 5, and 10 min with corresponding acoustic powers of 67, 40 and 36 W, and the samples were denoted S_1.5, S_5, and S₁₀, respectively. The samples were characterized using X-ray diffractometer (XRD), Energy dispersive X-ray (EDX), Transmission electronic microscope (TEM), Fourier transform infrared spectroscopy (FTIR), Zeta sizer, and vibrating sample magnetometer (VSM). Cell cytotoxicity and cell uptake were investigated with human embryonic kidney 293 (HEK-293) cells through MTT assay and Prussian blue staining, respectively. The sharp peaks of the XRD pattern were disappearing with an increase in the sonication period but a decrease in acoustic power. EDX analysis also demonstrates that atomic and weight percentages of the various elements in the samples were decreasing with an increase in the sonication period. However, the Zeta potential (ζ) values increase with an increase in the sonication period.The saturation magnetization (M_s) of the S_1.5 before and after the coating is 62.95 and 86.93 emu/g, respectively. Cell cytotoxicity and uptake of the S_1.5 show that above 70% of cells were viable at the highest concentration and the longest incubation duration. Importantly, the CS-SPIONs synthesized by the sonochemical method are non-toxic and biocompatible.     

Molecular masking and unmasking of the paramagnetic effect of iron on the proton spin-lattice (T1) relaxation time in blood and blood clots

The contribution of hemolysis, proteolysis and the paramagnetic effect of iron on the proton spin-lattice (T1) relaxation time in blood was examined. Hemolysis induced by sonication resulted in a significant (10%) increase in the T1 relaxation time of whole blood. Proteolysis in both sonicated and unsonicated whole blood samples eventually yielded T1 values which correlated well with the relaxation times of free iron in plasma or water at concentrations comparable to the concentration of iron in whole blood. It is concluded that proteolysis allows the iron atom to express its paramagnetic effect on water relaxation by gradually destroying the hydrophobic nature of the pocket in which iron resides on the hemoglobin molecule. The contribution of various blood components to the T1 relaxation of whole blood was also studied. The T1 values for packed erythrocytes, intact whole blood, sonicated whole blood, plasma and serum proved to be significantly different from each other. Serum was found to have a significantly (12%) longer T1 relaxation time than plasma. Packed clotted blood in vitro showed no change in the T1 time for at least 13 days while packed erythrocytes showed a shortening of T1 time after 6-8 days.     

Effects of thermal treatment and sonication on quality attributes of Chokanan mango (Mangifera indica L.) juice

Ultrasonic treatment is an emerging food processing technology that has growing interest among health-conscious consumers. Freshly squeezed Chokanan mango juice was thermally treated (at 90 °C for 30 and 60 s) and sonicated (for 15, 30 and 60 min at 25 °C, 40 kHz frequency, 130 W) to compare the effect on microbial inactivation, physicochemical properties, antioxidant activities and other quality parameters. After sonication and thermal treatment, no significant changes occurred in pH, total soluble solids and titratable acidity. Sonication for 15 and 30 min showed significant improvement in selected quality parameters except color and ascorbic acid content, when compared to freshly squeezed juice (control). A significant increase in extractability of carotenoids (4–9%) and polyphenols (30–35%) was observed for juice subjected to ultrasonic treatment for 15 and 30 min, when compared to the control. In addition, enhancement of radical scavenging activity and reducing power was observed in all sonicated juice samples regardless of treatment time. Thermal and ultrasonic treatment exhibited significant reduction in microbial count of the juice. The results obtained support the use of sonication to improve the quality of Chokanan mango juice along with safety standard as an alternative to thermal treatment.     

Sono-thermal pre-treatment of waste activated sludge before anaerobic digestion

Sonication and thermalization can be applied successfully to disrupt the complex waste activated sludge (WAS) floc structure and to release extra and intra cellular polymeric substances into soluble phase along with solubilization of particulate organic matters, before sludge digestion. In this study, sonication has been combined with thermalization to improve its disintegration efficiency. It was aimed that rise in temperature occurring during the sonication of sludge was used to be as an advantage for the following thermalization in the combined pre-treatment. Thus, the effects of sonication, thermalization and sono-thermalization on physical and chemical properties of sludge were investigated separately under different pre-treatment conditions. The disintegration efficiencies of these methods were in the following descending order: sono-thermalization > sonication > thermalization. The optimum operating conditions for sono-thermalization were determined as the combination of 1-min sonication at 1.0 W/mL and thermalization at 80 °C for 1 h. The influences of sludge pre-treatment on biodegradability of WAS were experienced with biochemical methane potential assay in batch anaerobic reactors. Relative to the control reactor, total methane production in the sono-thermalized reactor increased by 13.6% and it was more than the sum of relative increases achieved in the sonicated and thermalized reactors. Besides, the volatile solids and total chemical oxygen demand reductions in the sono-thermalized reactor were enhanced as well. However, it was determined that sludge pre-treatment techniques applied in this study was not feasible due to their high energy requirements.     

Mechanism study of dual-frequency ultrasound assisted enzymolysis on rapeseed protein by immobilized Alcalase

The mechanism of ultrasound field promoting enzymolysis efficiency is difficult to study, because the reaction system mixes with enzymes, proteins and hydrolysates. Immobilized enzyme is a good option that can be used to investigate the mechanism by separating enzymes out from the system after enzymolysis. The objective of this study was by using immobilized Alcalase to investigate the effects and mechanisms of the promotion of dual-frequency ultrasound (DFU) assisted-enzymolysis on rapeseed protein. Based on single factor experiments, response surface methodology model with three factors – hydrolysis time, power density and solid–liquid ratio at three levels was utilized to optimize the degree of hydrolysis (DH). Circular dichroism (CD) was used to analyze the secondary structure change of the protein, scanning electron microscopy (SEM) was used to analyze the surface microstructure change of the enzyme. The results showed that with DFU assisted-enzymolysis, the DH increased by 74.38% at the optimal levels for power density 57 W/L, solid–liquid ratio 5.3 g/L and enzymolysis time 76 min. After DFU assisted-enzymolysis, the yield of soluble solids content, including protein, peptides and total sugar in hydrolysate increased by 64.61%, 40.88% and 23.60%, respectively. CD analysis showed that after DFU assisted-enzymolysis, the number of α-helix and random coil decreased by 10.7% and 4.5%, β-chain increased by 2.4%. SEM showed that the degree of surface roughness of immobilized Alcalase increased. The above results indicated that the improvement of hydrolysis by DFU assisted-enzymolysis was achieved by enhancing the solid solubility, changing the molecular structure of protein and increased the surface area of immobilized enzyme.     

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.