Application of fixed-frequency ultrasound in the cultivation of Saccharomyces cerevisiae for rice wine fermentation.

BACKGROUND: Rice wine (RW) fermentation is limited by its long fermentation time, weak taste and unpleasant flavors such as oil and odor. In this study, a novel ultrasound technology of Saccharomyces cerevisiae was used with the aim of improving fermentation efficiency and volatile flavor quality of RW. RESULTS: The results showed that fixed-frequency ultrasonic treatment (28 kHz, 45 W L-1, 20 min) of S. cerevisiae seed culture at its logarithmic metaphase significantly increased the biomass and alcohol yield by 31.58% and 26.45%, respectively, and reduced fermentation time by nearly 2 days. Flavor analysis indicated that the flavor compounds in RW, specifically the esters and alcohols, were also increased in quantity after the ultrasonic treatment of S. cerevisiae seed liquid. Isobutyl acetate, ethyl butyrate, ethyl hexanoate and phenethyl acetate contents were increased by 78.92%, 129.19%, 7.79% and 97.84%, respectively, as compared to the control. CONCLUSION: Ultrasonic treatment of S. cerevisiae reduced fermentation time and enhanced the flavor profile of RW. This study could provide a theoretical and/or technological basis for the research and development of RW. © 2024 Society of Chemical Industry.

Effect of carbonyl-amino condensation, non-covalent cross-linking and conformational changes induced by ultrasound-assisted Maillard reaction on lysinoalanine formation in silkworm pupa protein.

The inhibition of cross-linked lysinoalanine (LAL) formation in silkworm pupa protein isolates (SPPI) by Maillard reaction (using varying xylose concentration) and ultrasound treatment was studied. Results showed that sonicated SPPI was effectively grafted with high concentration of xylose (5 %), resulting in the lowest LAL content, which was 48.75 % and 30.64 % lower than the control and ultrasound-treated samples, respectively. Chemical bond analysis showed that the combined treatment destroyed the ionic bonds, intrachain (g-g-t), and interchain (g-g-g) disulfide bonds, but stimulated the polymerization of hydrogen and hydrophobic bonds between SPPI and xylose, and as well enhanced the net negative charge between SPPI/Xylose complexes. The particles of the complexes were more loose, dispersed and rough, and had a stronger hydrophilic microenvironment, accompanied by alterations in microscopic, secondary and tertiary structures. Ultrasound treatment induced the breakdown of the oxidative cross-linking in SPPI, and promoted the sulfhydryl group-dehydroalanine binding and the carbonyl-amino condensation of the protein and xylose, and thus inhibited the formation of cross-linked LAL. Furthermore, the physicochemical and structural parameters were highly interrelated with cross-linked LAL content (|r| > 0.9). The outcomes provided a novel avenue and theoretical basis for minimizing LAL formation in SPPI and improving the nutrition and safety of SPPI.

Comparative Transcriptome Analysis of Candidate Genes Associated with Mycelia Growth from a He-Ne Laser with Pulsed Light Mutant of Phellinus igniarius (Agaricomycetes).

A mutant Phellinus igniarius JQ9 with higher mycelial production was screened out by He-Ne laser with pulsed light irradiation, the mechanism underlying the higher mycelial production is still unknown. This study aims to obtain a comprehensive transcriptome assembly during the Ph. igniarius liquid fermentation and characterize the key genes associated with the mycelial growth and metabolism in Ph. igniarius JQ9. Our transcriptome data of Ph. iniarius JQ9 and the wild strain were obtained with the Illumina platform comparative transcriptome sequencing technology. The results showed that among all the 346 differentially expressed genes (DEGs), 245 were upregulated and 101 were downregulated. Candidate genes encoding endoglucanase, beta-glucosidase, cellulose 1,4-beta-cellobiosidase, glycoside hydrolase family 61 protein, were proposed to participate in the carbohydrate utilization from KEGG enrichment of the starch and sucrose metabolism pathways were upregulated in Ph. igniarius JQ9. In addition, three candidate genes encoding the laccase and another two candidate genes related with the cell growth were higher expressed in Ph. igniarius JQ9 than in the wild type of strain (CK). Analysis of these data revealed that increased these related carbohydrate metabolism candidate genes underlying one crucial way may cause the higher mycelia production.

Inhibition of cross-linked lysinoalanine formation in pH12.5-shifted silkworm pupa protein, and functionality thereof: Effect of ultrasonication and glycation.

We added three different carbohydrates (Xylose/Xyl, Maltose/Mal, and Sodium alginate/Sal) to pH12.5-shifted silkworm pupa protein isolates (SPPI), and examined the influence of multi-frequency ultrasound (US) on them, with reference to lysinoalanine (LAL) formation, changes in conformational characteristics and functionality. Results showed that, the LAL content of the glycoconjugates - SPPI-Xyl, SPPI-Mal, and SPPI-Sal decreased by 1.47, 1.39, and 1.54 times, respectively, compared with the control. Notably, ultrasonication further reduced the LAL content by 45.85 % and brought SPPI-Xyl highest graft degree (57.14 %). SPPI-Xyl and SPPI-Mal were polymerized by different non-covalent bonds, and SPPI-Sal were polymerized through ionic, hydrogen, and disulfide (covalent/non-covalent) bonds. Significant increase in turbidity, Maillard reaction products and the formation of new hydroxyl groups was detected in grafted SPPI (p < 0.05). US and glycation altered the structure and surface topography of SPPI, in which sugars with high molecular weight were more likely to aggregate with SPPI into enormous nanoparticles with high steric hindrance. Compared to control, the solubility at pH 7.0, emulsifying capacity and stability, and foaming capacity of SPPI-US-Xyl were respectively increased by 244.33 %, 86.5 %, 414.67 %, and 31.58 %. Thus, combined US and xylose-glycation could be an effective approach for minimizing LAL content and optimizing functionality of SPPI.

Effect of pectin concentration on emulsifying properties of black soldier fly (Hermetia illucens) larvae albumin modified by pH-shifting and ultrasonication.

The effect of pectin concentration on the structural and emulsifying properties of black soldier fly larvae albumin (BSFLA) modified by pH-shifting (pH12) and ultrasound (US) was studied. The results (intrinsic fluorescence, surface hydrophobicity, Fourier transform infrared spectrum, and disulfide bonds) showed that modified BSFLA samples, especially pH12-US, were more likely to bind to pectin through hydrogen bonding, electrostatic interactions, and hydrophobic interactions due to the unfolding of BSFLA, the collapse of disulfide bonds and exposure of hydrophobic groups. Thus, a BSFLA-pectin complex with smaller particle size, more negative charges, and a relatively loose structure was formed. The emulsifying activity (EAI) and stability index (ESI) of pH12-US modified BSFLA were significantly enhanced by the addition of pectin, reaching the highest values (associated with 174.41 % and 643.22 % increase, respectively) at pectin concentration of 1.0 %. Furthermore, the interface modulus of the emulsion prepared by the modified BSFLA was mainly viscous, and had higher apparent viscosity, smaller particle size and droplet size, contributing to higher EAI and ESI. The study findings suggest the addition of pectin to pH12-US treated BSFLA could be used in industry to prepare BSFLA-pectin emulsion with exceptional/desirable properties.

Preparation of housefly (Musca domestica) larvae protein hydrolysates: Influence of dual-sweeping-frequency ultrasound-assisted enzymatic hydrolysis on yield, antioxidative activity, functional and structural attributes.

Dual-sweeping-frequency ultrasound (DSFU) was utilized in the preparation of polypeptides from housefly (Musca domestica) larvae protein (HLP). Results indicated that ultrasonication (20 ± 2/28 ± 2 kHz, 42 W/L, 25 min) significantly increased peptide yield and DPPH scavenging capacity by 8.25 % and 14.83 %, respectively. Solubility, foaming and emulsification properties of polypeptides were improved by 19.89 %, 33.33 % and 38.74 % over the control; along with notable reduction in particle size and increase in zeta potential. Tertiary structural changes of the sonicated hydrolysates were illustrated by UV and fluorescence spectra. FTIR showed that ultrasonication increased α-helix, ß-turn, and random coil by 38.23 %, 46.35 % and 16.36 %, respectively, but decreased ß-sheet by 48.03 %, indicating partial unfolding in HLP hydrolysate conformation and reduction in intermolecular interactions. The research results demonstrated that dual-sweeping-frequency ultrasonication has a great prospect in industry application for the purpose of improving enzymolysis efficiency and product quality for housefly larvae protein hydrolysates production.

Hexagonal plate ultrasound pretreatment on the correlation between soy protein isolate structure and cholesterol-lowering activity of peptides, and protein's enzymolysis kinetics, thermodynamics.

In this study, a hexagonal plate ultrasound (HPU) pretreatment technology was employed to modify soy protein isolate (SPI) and enhance the hypocholesterolemic activity of enzymatic digests from SPI. Results demonstrated that under the condition of ultrasound power density of 40 W/L, the hypocholesterolemic activity of enzymatic digests from HPU-pretreated SPI (HPU-SPI) increased by 88.40 % compared to control group after gastrointestinal digestion. The sulfhydryl content of HPU-SPI increased by a maximum of 45.32 % compared to control group. Fourier transform infrared and scanning electron microscopy revealed that HPU pretreatment partially unfolded the SPI conformation, reduced the intermolecular interactions, and exposed the internal hydrophobic regions. Pearson correlation analysis showed that sulfhydryl groups (r = 0.860), disulfide bonds (r = -0.875) and random coil (r = 0.917) were strongly correlated with the cholesterol-lowering activity of soy protein hydrolysate (SPH), following a simulated gastrointestinal digestion. Finally, the effects of HPU pretreatment on enzymolysis kinetics and thermodynamics of the SPI enzymatic process showed that HPU pretreatment significantly reduced the Mie's constant, activation energy, activation enthalpy, activation entropy and Gibbs free energy. Overall, the study outcome suggested that HPU pretreatment could positively influence the hypocholesterolemic peptide activity, and thus, may be beneficial to the pharmaceutical/food industry.

Improving the Flavour of Enzymatically Hydrolysed Beef Liquid by Sonication.

Beef potentiator is an important flavour enhancer in the food industry, while it is prone to generating insufficient compounds with umami and sweet tastes and compounds with a fishy odour during enzymatic hydrolysis of beef, resulting in poor flavour of beef potentiator. It has been extensively reported that sonication is capable of improving food flavour. However, the effect of sonication on the flavour of enzymatically hydrolysed beef liquid (EHBL) was scarcely reported. Herein, we investigated the effect of sonication on the flavour of EHBL using quantitative descriptive analysis (QDA), physicochemical analysis and SPME-GC-olfactometry/MS. QDA showed that sonication had a significant effect on taste improvement and off-odour removal of EHBL. Compared with the control, sonication (40 kHz, 80 W/L) increased the contents of total nitrogen, formaldehyde nitrogen, total sugars, reducing sugars, free amino acids (FAAs) and hydrolysis degree of EHBL by 19.25%, 19.80%, 11.83%, 9.52%, 14.37% and 20.45%. Notably, sonication markedly enhanced the contents of sweet FAAs, umami FAAs and bitter FAAs of EHBL by 19.66%, 14.04% and 9.18%, respectively, which contributed to the taste improvement of EHBL. SPME-GC-olfactometry/MS analysis showed that aldehydes and alcohols were the main contributors to aroma compounds of EHBL, and sonication significantly increased the contents of key aroma compounds and alcohols (115.88%) in EHBL. Notably, sonication decreased the contents of fishy odorants, hexanoic acid and nonanal markedly by 35.29% and 26.03%, which was responsible for the aroma improvement of EHBL. Therefore, sonication could become a new potential tool to improve the flavour of EHBL.

Ultrasound Intensify the Flavonoid Production of the Willow Bracket Mushroom, Phellinus igniarius (Agaricomycetes), Fermentation Mycelia.

This research aimed to use a novel and effective ultrasound (US) approach for obtaining high bio-compound production, hence proposing strategies for boosting active ingredient biosynthesis. Furthermore, the US promotes several physiological effects on the relevant organelles in the cell, morphological effects on the structure of Phellinus igniarius mycelium, and increases the transfer of nutrients and metabolites. One suitable US condition for flavonoid fermentation was determined as once per day for 7-9 days at a frequency 22 + 40 kHz, power density 120 W/L, treated 10 min, treatment off time 7 s. The flavonoid content and production increased about 47.51% and 101.81%, respectively, compared with the untreated fermentation (P < 0.05). SEM showed that sonication changes the morphology and structure of Ph. igniarius mycelium; TEM reveals the ultrasonic treatment causes organelle aggregation. The ultrasound could affect the metabolism of the biosynthesis of the active ingredients.

The latest advances on soy sauce research in the past decade: Emphasis on the advances in China.

As an indispensable soybean-fermented condiment, soy sauce is extensively utilized in catering, daily cooking and food industry in East Asia and Southeast Asia and is becoming popular in the whole world. In the past decade, researchers began to pay great importance to the scientific research of soy sauce, which remarkably promoted the advances on fermentation strains, quality, safety, function and other aspects of soy sauce. Of them, the screening and reconstruction of Aspergillus oryzae with high-yield of salt and acid-tolerant proteases, mechanism of soy sauce flavor formation, improvement of soy sauce quality through the combination of novel physical processing technique and microbial/enzyme, separation and identification of soy sauce functional components are attracting more attention of researchers, and related achievements have been reported continually. Meanwhile, we pointed out the drawbacks of the above research and the future research directions based on published literature and our knowledge. We believe that this review can provide an insightful reference for international related researchers to understand the advances on soy sauce research.

Enhancing the growth of thermophilic Bacillus licheniformis YYC4 by low-intensity fixed-frequency continuous ultrasound.

The effect of low-intensity fixed-frequency continuous ultrasound (LIFFCU) on the growth of Bacillus licheniformis YYC4 was investigated. The changes in morphology and activity of the organism, contributing to the growth were also explored. Compared with the control, a significant increase (48.95%) in the biomass of B. licheniformis YYC4 (at the logarithmic metaphase) was observed following the LIFFCU (28 kHz, 1.5 h and 120 W (equivalent to power density of 40 W/L)) treatment. SEM images showed that ultrasonication caused sonoporation, resulting in increased membrane permeability, evidenced by increase in cellular membrane potential, electrical conductivity of the culture, extracellular protein and nucleic acid, and intracellular Ca2+ content. Furthermore, LIFFCU action remarkably increased the extracellular protease activity, volatile components of the culture medium, microbial metabolic activity, and spore germination of the strain. Therefore, LIFFCU could be used to efficiently promote the growth of targeted microorganisms.

Influence of Multi-Frequency Ultrasound Treatment on Conformational Characteristics of Beef Myofibrillar Proteins with Different Degrees of Doneness.

This study evaluated the effect of multi-frequency sonication (20 kHz, 25 kHz, 28 kHz, 40 kHz, 50 kHz) on structural characteristics of beef myofibrillar proteins (MPs) with different degrees of doneness (Rare 52~55 °C, Medium Rare 55~60 °C, Medium 60~65 °C, Medium Well 65~69 °C, Well Down 70~80 °C, and Overcooked 90 °C). The results showed that surface hydrophobicity and sulfhydryl content increased with the increase in degree of doneness. At the same degree of doneness, the sulfhydryl group contents reached the maximum at a frequency of 28 kHz. In addition, the absolute value of ζ-potential was significantly decreased after ultrasonic treatment (p < 0.05). SDS gel electrophoresis showed that the bands of beef MPs were not significantly affected by various ultrasonic frequencies, but the bands became thinner when the degree of doneness reached overcooked. Fourier transform infrared spectrum showed that with the increase of ultrasonic frequency, α-helix content decreased, and random coil content significantly increased (p < 0.05). The results of atomic force microscopy indicated that the surface structure of beef MPs was damaged, and the roughness decreased by sonication, while the roughness significantly increased when the degree of doneness changed from medium to overripe (p < 0.05). In conclusion, multi-ultrasound combined with degree of doneness treatment alters the structural characteristics of beef MPs.

Efficient degradation of soybean protein B3 subunit in soy sauce by ultrasound-assisted prolyl endopeptidase and its primary mechanism.

Prolyl endopeptidase can partially degrade soybean protein B3 subunit and alleviate soy sauce secondary precipitate. In this study, the influences of ultrasound-assisted prolyl endopeptidase on the degradation of soybean protein B3 subunit of soy sauce and primary mechanism were investigated using SDS-PAGE, MALDI-TOF-MS, circular dichromatic spectrometer, fluorescence spectra, etc. Results showed that ultrasound-assisted prolyl endopeptidase enhanced 72% degradation rate of B3 subunit and reduced soy sauce secondary precipitate remarkably, meanwhile significantly increased content of organic taste compounds of soy sauce compared with control (p < 0.05). Sonication markedly reduced percentage of α-helix and increased percentage of random coil, made hydrophobic amino acids inside prolyl endopeptidase exposed to its surface and enhanced its flexibility, which facilitated the binding of prolyl endopeptidase active center with B3 subunit and finally enhanced the latter's degradation rate and appearance quality of soy sauce. This work laid a foundation for solving soy sauce secondary precipitate.

Mono-frequency ultrasonic-assisted thawing of frozen goose meat: Influence on thawing efficiency, product quality and microstructure.

This study aimed to investigate the influences of mono-ultrasound assisted thawing on the thawing efficiency, product quality and conformational characteristics of frozen goose meat. The thawing time, thawing loss, muscle quality, and microstructure of frozen goose meat were studied. The results displayed that ultrasonic-assisted thawing effectively reduced the thawing time by 45.37-57.58% compared with non-sonicated group, and significantly decreased the thawing loss. For the quality properties of goose meat tissue, ultrasound-assisted thawing with single-frequency of 50 kHz indicated a lower protein turbidity; meanwhile, hardness values were also significantly increased, and displayed a higher springiness, gumminess and chewiness of goose meat tissue. The microstructure analysis exhibited that the conformation of goose myofibrillar protein (MP) was modified following ultrasonic-assisted thawing, and became closer and more irregular. Therefore, ultrasound-assisted thawing treatments at 50 kHz mono-frequency (temperature 25℃) have a high potential application value in the thawing research of frozen goose meat, and lay a theoretical foundation for use in the meat process industries.

Characterization of the interaction between allicin and soy protein isolate and functional properties of the adducts.

BACKGROUND: Soybean meal, a by-product of the soybean oil production industry, has a high protein content but the compact globular structure of the protein from soybean meal limits its wide application in food processing. Allicin has been found to have numerous functional properties. In this study, allicin was interacted with soy protein isolate (SPI). The functional properties of the adducts were investigated. RESULTS: Binding with allicin significantly quenched the fluorescence intensity of SPI. Static quenching was the main quenching mechanism. The stability of adducts decreased with increasing temperature. The greatest extent of binding between allicin and sulfhydryl groups (SH) of SPI was obtained at an allicin/SH molar ratio of 1:2. The amino groups of SPI did not bind with allicin covalently. Soy protein isolate was modified by allicin through covalent and non-covalent interactions. Compared with SPI, the emulsifying activity index and foaming capacity of adducts with a ratio of 3:1 were improved by 39.91% and 64.29%, respectively. Soy protein isolate-allicin adducts also exhibited obvious antibacterial effects. The minimum inhibitory concentrations (MICs) of SPI-allicin adducts on Escherichia coli and Staphylococcus aureus were 200 and 160 µg mL-1 , respectively. CONCLUSION: The interaction of allicin with SPI is beneficial for the functional properties of SPI. These adducts can be used in different food formulations as emulsifiers, foamers, and transport carriers. © 2023 Society of Chemical Industry.

Synergistic effects of pH shift and heat treatment on solubility, physicochemical and structural properties, and lysinoalanine formation in silkworm pupa protein isolates.

The application of silkworm pupa protein isolates (SPPI) in food industry was limited because SPPI's solubility is poor and it contains a potential harmful component of lysinoalanine (LAL) which formed during protein extraction. In this study, combined treatments of pH shift and heating were performed to improve the solubility of SPPI and to reduce the content of LAL. The experimental results showed that the promoting effect on SPPI's solubility by alkaline pH shift + heat treatment was greater than that by acidic pH shift + heat. And an 8.62 times increase of solubility was observed after pH 12.5 + 80 â„ƒ treatment compared to the control SPPI sample which was extracted at pH 9.0 without pH shift treatment. Very strong positive correlation was found between alkali dosage and SPPI solubility (Pearson's correlation coefficient r = 0.938). SPPI with pH 12.5 shift treatment showed the highest thermal stability. Alkaline pH shift combined with heat treatment altered the micromorphology of SPPI and destroyed the disulfide bonds between macromolecular subunits (72 and 95 kDa), resulting in reduced particle size and increased zeta potential and free sulfhydryl content of the isolates. The fluorescence spectra analysis showed red shifts phenomena with pH increasing and fluorescence intensity increase with temperature increasing, implying the alterations in the tertiary structure of protein. Compared to the control SPPI sample, the amount of LAL reduced by 47.40 %, 50.36 % and 52.39 % using pH 12.5 + 70 â„ƒ, pH 12.5 + 80 â„ƒ and pH 12.5 + 90 â„ƒ treatment, respectively. These findings provide fundamental information for the development and application of SPPI in food industry.

Insights into Ultrasonication Treatment on the Characteristics of Cereal Proteins: Functionality, Conformational and Physicochemical Characteristics.

BACKGROUND: It would be impossible to imagine a country where cereals and their byproducts were not at the peak of foodstuff systems as a source of food, fertilizer, or for fiber and fuel production. Moreover, the production of cereal proteins (CPs) has recently attracted the scientific community's interest due to the increasing demands for physical wellbeing and animal health. However, the nutritional and technological enhancements of CPs are needed to ameliorate their functional and structural properties. Ultrasonic technology is an emerging nonthermal method to change the functionality and conformational characteristics of CPs. Scope and approach: This article briefly discusses the effects of ultrasonication on the characteristics of CPs. The effects of ultrasonication on the solubility, emulsibility, foamability, surface-hydrophobicity, particle-size, conformational-structure, microstructural, enzymatic-hydrolysis, and digestive properties are summarized. CONCLUSIONS: The results demonstrate that ultrasonication could be used to enhance the characteristics of CPs. Proper ultrasonic treatment could improve functionalities such as solubility, emulsibility, and foamability, and is a good method for altering protein structures (including surface hydrophobicity, sulfhydryl and disulfide bonds, particle size, secondary and tertiary structures, and microstructure). In addition, ultrasonic treatment could effectively promote the enzymolytic efficiency of CPs. Furthermore, the in vitro digestibility was enhanced after suitable sonication treatment. Therefore, ultrasonication technology is a useful method to modify cereal protein functionality and structure for the food industry.

Effects of ultrasonic-assisted pH shift treatment on physicochemical properties of electrospinning nanofibers made from rapeseed protein isolates.

Electrospinning nanofibers (NFs) made from natural proteins have drawn increasing attention recently. Rapeseed meal is a by-product that rich in protein but not fully utilized due to poor properties. Therefore, modification of rapeseed protein isolates (RPI) is necessary to expand applications. In this study, pH shift alone or ultrasonic-assisted pH shift treatment was adopted, the solubility of RPI, along with the conductivity and viscosity of the electrospinning solution were detected. Moreover, the microstructure and functional characteristics of the electrospinning NFs, as well as the antibacterial activity of clove essential oil loaded-NFs were investigated. The tested parameters were remarkably improved after different treatments compared with the control, and synergistic effects were observed, especially under alkaline conditions. Hence, pH12.5 + US showed the maximum value of solubility, conductivity, and viscosity, which was more than 7-fold, 3-fold, and almost 1-fold higher than the control respectively. Additionally, SEM and AFM images showed a finer and smoother surface of NFs after treatments, and the finest diameter of 216.7 nm was obtained after pH12.5 + US treatment in comparison with 450.0 nm in control. FTIR spectroscopy of NFs demonstrated spatial structure changes of RPI, and improved thermal stability and mechanical strength of NFs were achieved after different treatments. Furthermore, an inhibition zone with a diameter of 22.8 mm was observed from the composite NFs. This study indicated the effectiveness of ultrasonic-assisted pH shift treatment on the physicochemical properties improvement and functional enhancement of NFs made from RPI, as well as the potential antibacterial application of the composite NFs in the future.

Application of simultaneous ultrasonic curing on pork (Longissimus dorsi): Mass transport of NaCl, physical characteristics, and microstructure.

This study aimed to investigate the effect of ultrasound curing with various working modes and frequency combinations, including mono-, dual- and tri-frequency, on the content of NaCl and tenderness of pork loins (Longissimus dorsi). The physical qualities, myoglobin, moisture migration, distribution, and microstructure of pork were also evaluated. The results displayed that the NaCl content of samples cured by simultaneous ultrasound (100 W/L) working mode with a frequency combination of 20, 40, and 60 kHz was higher than that of other ultrasound working modes. The effect of ultrasonic brining was significantly better than the static curing when the saline solution was >35 mL. In addition, the samples cured by simultaneous ultrasound had better physical qualities, including more pickling absorptivity, less cooking loss, and lower hardness, tenderness, and chewiness value. The intensity of lightness was reduced, although redness and yellowness remained unaltered compared to static curing. The myoglobin content decreased drastically without changing the oxygenation level, and the relaxation time of T2b and T21 was delayed. The microstructure indicated that the ultrasonic treatment could promote changes in meat texture. Overall, the simultaneous ultrasound at various frequencies could efficiently accelerate NaCl penetration and improve pork quality.