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Phospholipid and nucleotide syntheses are fundamental metabolic processes in eukaryotic organisms, with their dysregulation implicated in various disease states. Despite their importance, the interplay between these pathways remains poorly understood. Using genetic and metabolic analyses in Saccharomyces cerevisiae, we elucidate how cytidine triphosphate usage in the Kennedy pathway for phospholipid synthesis influences nucleotide metabolism and redox balance. We find that deficiencies in the Kennedy pathway limit nucleotide salvage, prompting compensatory activation of de novo nucleotide synthesis and the pentose phosphate pathway. This metabolic shift enhances the production of antioxidants such as NADPH and glutathione. Moreover, we observe that the Kennedy pathway for phospholipid synthesis is inhibited during replicative aging, indicating its role in antioxidative defense as an adaptive mechanism in aged cells. Our findings highlight the critical role of phospholipid synthesis pathway choice in the integrative regulation of nucleotide metabolism, redox balance and membrane properties for cellular defense.
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Cytochrome P450s are one of the most versatile oxidases that catalyze significant and unique chemical transformations for the construction of complex structural frameworks during natural product biosynthesis. Here, we discovered a set of P450s, including SdnB, SdnH, SdnF, and SdnE, that cooperatively catalyzes the reshaping of the inert cycloaraneosene framework to form a highly oxidized and rearranged sordarinane architecture. Among them, SdnB is confirmed to be the first P450 (or oxidase) that cleaves the C-C bond of the epoxy residue to yield formyl groups in pairs. SdnF selectively oxidizes one generated formyl group to a carboxyl group and accelerates the final Diels-Alder cyclization to furnish the sordarinane architecture. Our work greatly enriches the enzyme functions of the P450 superfamily, supplies the missing skills of the P450 synthetic toolbox, and supports them as biocatalysts in further applications toward the synthesis of new chemical entities.
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Sistema Enzimático del Citocromo P-450 , Diterpenos , Catálisis , Sistema Enzimático del Citocromo P-450/metabolismo , Oxidación-Reducción , Metabolismo SecundarioRESUMEN
Eight new polyketides, including three dimeric benzophenones, named dipleosporones A-C (1-3), three benzophenones (4-6), one xanthone (7), and one phenylbenzoate (8), along with seven known polyketides (9-15) were isolated from the fungus Pleosporales sp. YY-4. The structures of the new compounds were established on the basis of spectroscopic methods, including high-resolution electrospray ionization mass spectrometry and one- and two-dimensional nuclear magnetic resonance. This is the first report of a benzophenone dimer connection via a C bridge from natural sources. An anti-inflammatory assay indicated that the dimeric benzophenones (1-3) inhibited lipopolysaccharide-induced NO production in RAW 264.7 cells, with half-maximal inhibitory concentration (IC50) values ranging from 8.8 to 18.1 µM, being more potent than the positive control, dexamethasone (IC50 = 22.2 µM).
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Antiinflamatorios/farmacología , Ascomicetos/química , Benzofenonas/aislamiento & purificación , Benzofenonas/farmacología , Animales , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Espectroscopía de Resonancia Magnética con Carbono-13 , Dimerización , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Estructura Molecular , Óxido Nítrico/biosíntesis , Espectroscopía de Protones por Resonancia Magnética , Células RAW 264.7RESUMEN
Adding essential oil into the gelatin-based film can enhance the antibacterial activity of the film, but excessive amounts of addition will bring the film an unpleasant flavor and reduce its mechanical performance. Hence, we prepared functional gelatin-based films by incorporating low content of ginger essential oil (GEO). The flavor of GEO was not detected from the films containing less than 1% GEO. The antimicrobial activity of films was found to be proportional to GEO content. As GEO content increased from 0 to 1%, the value of water vapor permeability (WVP) and elongation at break (EAB) increased, whereas the value of tensile strength (TS) of film decreased. The Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy analysis revealed the vibration of gelatin film was affected by adding low content of essential oil. Surface morphologies demonstrated oil droplets and a discontinuous structure, and cross-section morphologies proved the formation of a loose structure as GEO was incorporated in the film through SEM. Sensory evaluation revealed that composite films incorporated with 0.5% GEO exhibited the best performance. The resulting films can be used as antimicrobial packaging materials with good physical properties and sensory performance.
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The ongoing COVID-19 global pandemic caused by SARS-CoV-2 inspires the development of effective inhibitors to block the SARS-CoV-2 spike-ACE2 interaction. A chemical investigation on the fruiting bodies of Phellinus pini led to the isolation of five aromatic cadinane sesquiterpenoids including four new ones, named piniterpenoids A-D (1-4), as well as three known lignans. Their structures were determined by extensive spectroscopic analysis including HRMS and 1D and 2D NMR. All of the aromatic cadinane sesquiterpenoids inhibited the SARS-CoV-2 spike-ACE2 interaction, with IC50 values ranging from 64.5 to 99.1 µM. A molecular docking study showed the disruption of the interaction of compound 1 via hydrogen interactions with Arg403, Asp405, and Arg408 of SARS-CoV-2 RBD and Arg393 and His34 residues of ACE2. These results suggested that aromatic cadinane sesquiterpenoids might be useful in developing agents for COVID-19.
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Enzima Convertidora de Angiotensina 2/antagonistas & inhibidores , Antivirales/química , Antivirales/farmacología , Cuerpos Fructíferos de los Hongos/química , Phellinus/química , Sesquiterpenos Policíclicos/química , Sesquiterpenos Policíclicos/farmacología , SARS-CoV-2/efectos de los fármacos , Sesquiterpenos/química , Sesquiterpenos/farmacología , Glicoproteína de la Espiga del Coronavirus/antagonistas & inhibidores , Humanos , Enlace de Hidrógeno/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Simulación del Acoplamiento MolecularRESUMEN
Bovine ß-lactoglobulin (ß-LG) is the major allergen in milk powder. The IgG/IgE binding capacity and structural characteristics of ß-LG after spray drying in the presence or absence of α-lactose at 120 and 180°C were investigated by ELISA and mass spectrometry. At a drying temperature of 120°C, no change was found in the IgG/IgE binding capacity of ß-LG and no change was observed in free amino group content, fluorescence intensity, or detectable glycation. At a drying temperature of 180°C, aggregation of ß-LG occurred, leading to a decrease in the IgG/IgE binding capacity. When α-lactose was also present, 7 lysine side-chains in ß-LG were modified by glycation and the IgG/IgE binding capacity was further decreased. Therefore, the glycation and structural changes in ß-LG were responsible for the reduction in the IgG/IgE binding capacity during high temperature (180°C) spray drying.
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Inmunoglobulina E/metabolismo , Inmunoglobulina G/metabolismo , Lactoglobulinas/metabolismo , Alérgenos/metabolismo , Animales , Bovinos , Desecación , Ensayo de Inmunoadsorción Enzimática , Femenino , Glicosilación , Calor , Inmunoglobulina G/química , Lactosa/metabolismo , Lisina/metabolismo , Masculino , Espectrometría de Masas , Unión Proteica , Relación Estructura-ActividadRESUMEN
Cadmium (Cd) is known to be a potentially toxic heavy metals to the fish health and growth. Carassius auratus gibelio (C. a. gibelio) specimens were exposed to waterborne Cd (0, 0.05, 0.10, 0.15, and 0.20 mg/L CdCl2) for 14 days. Cd accumulation, liver and intestine histopathology, and intestinal microorganism were investigated in the present study. The results indicated that Cd accumulation in the gill, liver, intestine, and muscle gradually decreased as Cd concentration increased. The gill accumulated higher amounts of Cd than other tissues. The histopathology of liver and intestine underwent changes with different Cd concentrations, including hepatocyte hypertrophy, aggregation of blood cells, sinusoids, lipidosis, necrosis of hepatic tissues, the erosion of villi, necrosis in the mucosal layer, the appearance of vacuoles in the lamina propria, hyperplasia, and swelling of goblet cells. Moreover, the core gut microbiota existed in the intestinal microorganism and did not change as Cd concentration increased. However, the diversity of intestinal microorganism was significantly reduced compared with that of the control sample. The present results indicated that C. a. gibelio exposed to Cd suffered toxicity, and Cd could affect the biodiversity of the intestinal microbiota of C. a. gibelio.
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Cadmio/metabolismo , Cadmio/toxicidad , Carpa Dorada , Intestinos/microbiología , Contaminantes Químicos del Agua/toxicidad , Animales , Branquias , Intestinos/efectos de los fármacosRESUMEN
BACKGROUND: Protein-polysaccharide complex coacervations have been considered extensively for the development of functional foods. The main problem of the complex coacervates is that they are highly unstable under different conditions and that cross-linking is necessary to stabilize them. In this study, the effects of pectin at different concentrations on the gel and structural properties of fish scale gelatin (FSG)-high methoxyl citrus pectin (HMP) coacervate enhanced by microbial transglutaminase (MTGase) were studied. RESULTS: The gelation rates and gel strength of the MTGase-enhanced FSG-HMP coacervate gels decreased with increasing HMP concentration. However, the enhanced coacervate gels exhibited better thermal behavior and mechanical properties compared with the original gels. Also, TG-P8 exhibited the highest melting point (27.15 ± 0.12 °C), gelation point (15.65 ± 0.01 °C) and stress (15.36 ± 0.48 kPa) as HMP was 8 g kg-1 . Particle size distribution, fluorescence emission and UV absorbance spectra indicated that MTGase and HMP could make FSG form large aggregates. Moreover, confocal laser scanning microscopy of treated coacervate gels showed a continuous protein phase at low HMP concentrations. CONCLUSION: FSG and HMP could form soluble coacervate, and MTGase could improve the thermal and mechanical properties of coacervate gels. © 2017 Society of Chemical Industry.
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Escamas de Animales/química , Proteínas Bacterianas/química , Proteínas de Peces/química , Gelatina/química , Pectinas/química , Transglutaminasas/química , Animales , Biocatálisis , Peces , Geles/química , CinéticaRESUMEN
BACKGROUND: Hen eggs are widely used in food industry as a result of their excellent nutritional and processing properties and ovalbumin (OVA) is the major allergen of hen egg whites. Ultrasound has been widely used in the food processing industry, although the effect of high-intensity ultrasound on the immunoglobulin (Ig)G binding, IgE binding and conformational structure of OVA remains unclear. RESULTS: Ultrasound treatment at 600-800 W dramatically increased the IgG and IgE binding of OVA, with the highest values observed at 600 W. The free sulfhydryl content, surface hydrophobicity and ultraviolet absorption were gradually increased when the power was increased from 200 to 600 W, implying the occurrence of molecule unfolding. When the power was 800 W, the binding ability was slightly decreased and was associated with the aggregation of OVA molecules. Polyacrylamide gel electrophoresis analysis implied that ultrasound treatment at 800 W could induce the aggregation of OVA molecules via disulfide bonds and/or hydrophobic interactions. CONCLUSION: High-intensity ultrasound can increase the potential allergenicity of OVA. Therefore, high-intensity ultrasound processing of some egg products alone may improve the risk of an allergenic reaction in egg allergy patients to some extent. © 2016 Society of Chemical Industry.
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Hipersensibilidad al Huevo/inmunología , Inmunoglobulina E/inmunología , Inmunoglobulina G/inmunología , Ovalbúmina/química , Ovalbúmina/inmunología , Electroforesis en Gel de Poliacrilamida , Manipulación de Alimentos , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Conformación Proteica , Pliegue de Proteína , UltrasonidoRESUMEN
In this study, microbial transglutaminase (MTGase) and pectin were compared to modify bighead carp (Hypophthalmichthys nobilis) scale gelatin. The functional properties of modified fish scales gelatin (FSG) were largely improved, including melting temperature and rheological behavior. While, modification methods decreased the triple-helix content and destroyed the single left-hand helix chain of modified FSG as investigated by X-ray diffraction. MTGase could induce the denser and finer gels network, but had none significant effect on nanostructural properties of fish gelatin. Pectin inserted itself into the fish gelatin gels network and caused aggregations, forming crystalline peaks and various nanostructures. In particular, compared with pectin modified FSG, MTGase produced FSG with lower storage modulus and apparent viscosity, but higher gel points and melting points.
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Insoluble dietary fiber from soybean residue (SIDF) was treated with dynamic high-pressure microfluidization (DHPM) and used as adsorbent for Pb(II) ion. The effects of pressure on the Pb(II) adsorption capacity, primary cilia structure and surface topography of SIDF were determined using a gastrointestinal simulated model in vitro. SIDF (at pH 7.0) showed maximum binding capacity (261.42 ± 2.77 µmol/g), which was about 1.13 times higher than that of untreated sample (233.47 ± 1.84 µmol/g), when pressure reached 80 MPa. However, the net adsorption value of SIDF in a simulated small intestine (~ 9 µmol/g) was significantly lower than that in the stomach (~ 48 µmol/g), because of the competitive adsorption of Pb(2+) by pancreatin, cholate and several enzymes in the small intestine. In addition, the adsorption capacity of SIDF exhibited good linear relationship with the physicochemical properties of total negative charges, and the adsorption behavior presumably occurred on the surface area of granules fiber.
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This study aimed at evaluating the performance of peony seed oil microencapsulated by spray drying during encapsulation and storage. Four different combinations of gum arabic (GA), corn syrup (CS), whey protein concentrate (WPC) and sodium caseinate (CAS) were used to encapsulate peony seed oil. The best encapsulation efficiency was obtained for CAS/CS followed by the CAS/GA/CS combination with the encapsulation ratio of 93.71 and 92.80 %, respectively, while the lowest encapsulation efficiency was obtained for WPC/GA/CS (85.96 %). Scanning electron microscopy and confocal laser scanning microscopy revealed that the particles were spherical in shape and did not exhibit apparent cracks or fissures, and gum arabic was uniformly distributed across the wall of the microcapsules. Oxidative stability study indicated that the CAS/GA/CS combination presented the best protection against lipid oxidation and the smallest loss of polyunsaturated fatty acid content among all of the formulas as measured by gas chromatography. Therefore, CAS/GA/CS could be promising materials encapsulate peony seed oil with high encapsulation efficiency and minimal lipid oxidation.
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Ginger and garlic have long been used in Asian countries to enhance the flavor and to neutralize any unpleasant odors present in fish soup. The purpose of this study was to evaluate the change in the amount of volatile components present in fish soup compared to boiled water solutions of ginger and garlic. The fish soup was prepared by boiling oil-fried grass carp (Ctenopharyngodon idella) with or without ginger and/or garlic. Generally, boiling garlic and ginger in water led to a decrease in the amount of the principal volatile constituents of these spices, together with the formation of some new volatiles such as pentanal, hexanal, and nonanal. The results showed that 16 terpenes present in raw ginger, predominantly camphene, ß-phellandrene, ß-citral, α-zingiberene, and (E)-neral, were detected in fish soup with added ginger and thus remained in the solution even after boiling. Similarly, 2-propen-1-ol and three sulfur compounds (allyl sulfide, diallyl disulfide, and diallyl trisulfide) present in raw garlic, were present in trace amounts in the boiled garlic solution, but were present in considerably larger amounts in the boiled fish solution with garlic or garlic plus ginger. In conclusion, the effect of adding spices on the volatile profile of grass carp soup can be attributed to the dissolution of flavor volatiles mainly derived from raw spices into the solution, with few additional volatiles being formed during boiling. In addition, boiling previously fried grass carp with spices led to enhanced volatile levels compared to boiled spice solutions.
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The interaction between whey protein and acacia which were used as wall material was studied on the formation of the oils microcapsules by the FTIR Spectroscopy and Computer Aided Analysis. The results indicated that whey protein changed obviously in amide A and amide I by high pressured homogenization and spray-drying. The amide A moved from 3 406.5 cm(-1) to 3 425.4 cm(-1) which was possibly due to covalent cross-linking between whey protein and acacia. Furthermore the amide I moved from 1 648.6 cm(-1) to 1 654.7 cm(-1) for intramolecular hydrogen bonding of protein had been weaken. After Gaussian fitting on amide I , it was found that the content of secondary structure of α-helix content and ß-folding in whey protein reduced from 19.55% to 17.50% and from 30.59% to 25.63%, respectively. This suggests that protein intramolecular hydrogen bonding force was abated, resulting in abating the rigid structure of the protein molecules and enhancing of the toughness structure. The protein molecules showed some flexibility. The result of SDS-PAGE electrophoresis showed that whey protein--gum Arabic complexes produced covalent products in larger molecular weight. During the spray-drying process, covalent cross-linking produced between whey protein and gum Arabic which improved emulsifying activity of the complex whey protein and gum Arabic produced covalent cross-linking and improved the complex emulsifying activity. Observing the surface structure of the fish oil microcapsule by SEM, the compound of whey protein and acacia as wall material was proved better toughness, less micropore, and more compact structure.
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Acacia/química , Suplementos Dietéticos/análisis , Proteínas de la Leche/química , Amidas , Animales , Cápsulas , Desecación , Electroforesis en Gel de Poliacrilamida , Aceites de Pescado , Enlace de Hidrógeno , Peso Molecular , Presión , Estructura Secundaria de Proteína , Espectroscopía Infrarroja por Transformada de Fourier , Proteína de Suero de LecheRESUMEN
The aim of this study was to characterize the properties of ovalbumin (OVA) after glycated with glucose under microwave heating. For this purpose, microwave at 480 and 640 W power levels were used for heating the OVA-glucose system in solid-state for 0, 5, 10, 15, 20 and 25 min, respectively. The results indicated that the protein molecular weight was increased after glycated with glucose under microwave treatment, the pH of the system was decreased with the increase of microwave treatment power and time, while the UV absorbance, browning intensity, antioxidant activities as well as the emulsifying activity and emulsion stability of the Maillard reaction products (MRPs) were increased in according with the raise of microwave treatment power and time. The reaction time of microwave treatment is much shorter than those using traditional methods, suggesting that microwave irradiation is a novel and efficient approach to promote Maillard reaction (MR) in dry state and improve protein antioxidant and functional properties.
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Subcritical water is an emerging method in food industry. In this study, soybean protein isolate (SPI) was treated by subcritical water (SBW) at various temperatures (0, 120, 160, 200 °C) for 20 min. The changes in the appearances, physicochemical properties and structural changes were investigated. After SBW treatment, the color of SPI solution modified turned to be yellow. The mean particle size and turbidity of SPI had similar behaviors. The mean particle size was decreased from 263.7 nm to 116.8 nm at 120 °C and then reached the maximum at 160 °C (1446.1 nm) due to the aggregation of protein. Then it was decreased to 722.9 nm at 200 °C caused by the protein degradation. SBW treatment could significantly enhance the solubility, emulsifying and foaming properties of SPI. With increasing temperature, the crystalline structure of protein was gradually collapsed. The degradation of the protein advanced structure occurred, especially at 200 °C revealed by ultra-high resolution mass spectrometry. Better functional properties exhibited in hydrolysis products indicating that SBW treatment could be used as a good method to modify the properties of soy proteins isolate for specific purposes under appropriate treatment condition.
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In this study, gelatin was extracted from bighead carp (Hypophthalmichthys nobilis) scales by water bath (WB) and ultrasound bath (UB) at 60 °C for 1 h, 3 h and 5 h, named WB1, WB3, WB5, UB1, UB3 and UB5, respectively. The physicochemical properties of gelatin were investigated. The result indicated that gelatin extracted from bighead carp scales had a high protein content (84.15 ~ 91.85 %) with moisture (7.11 ~ 13.65 %), low ash content (0.31 ~ 0.97 %). All extracted gelatin contained α-and ß-chains as the predominant components. Gelatin extracted by UB obtained much higher yield (30.94-46.67 %) than that of WB (19.15-36.39 %). More voids and less sheets of gelatin structure were observed, when the gelatin was extracted by UB for longer time. Gelatin of UB-assisted extraction normally exhibited lower gel strength and melting points than that of WB, which may be resulted from the protein degradation reflected by the FTIR spectra and higher free amino group content. However, there was no significant difference between WB1 and UB1. Therefore, the ultrasound assisted extraction in a short time was a promising method to enhance the yield and obtain gelatin with high quality.
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This study found that, after microwave treatment at 560 W for 30 s, alkaline protease enzymolysis significantly reduced the allergenicity of ovalbumin (OVA). Furthermore, specific adsorption of allergenic anti-enzyme hydrolyzed peptides in the enzymatic products by immunoglobulin G (IgG) bound to magnetic bead further decreased the allergenicity of OVA. The results indicated that microwave treatment disrupts the structure of OVA, increasing the accessibility of OVA to the alkaline protease. A comparison between 17 IgG-binding epitopes identified through high-performance liquid chromatography-higher energy collisional dissociation-tandem mass spectrometry and previously reported immunoglobulin E (IgE)-binding epitopes revealed a complete overlap in binding epitopes at amino acids (AA)125-135, AA151-158, AA357-366, and AA373-381. Additionally, partial overlap was observed at positions AA41-59, AA243-252, and AA320-340. Consequently, these binding epitopes were likely pivotal in eliciting the allergic reaction to OVA, warranting specific attention in future studies. In conclusion, microwave-assisted enzymolysis synergized with magnetic bead adsorption provides an effective method to reduce the allergenicity of OVA.
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In this work, the effect of the addition of γ-polyglutamic acid (γ-PGA) on the rheology, physicochemical properties, and microstructure of fish gelatin (FG) emulsion gel was investigated. Samples of the emulsion gel were evaluated for rheological behavior and stability prior to gelation. The mechanical properties and water-holding capacity (WHC) of the emulsion were determined after gelation. The microstructure of the emulsion gel was further examined using confocal laser scanning microscopy (CLSM). The results indicated a gradual increase in the apparent viscosity and gelation temperature of the emulsion at a higher concentration of γ-PGA. Additionally, frequency scan results revealed that on the addition of γ-PGA, FG emulsion exhibited a stronger structure. The emulsion containing 0.1% γ-PGA exhibited higher stability than that of the control samples. The WHC and gel strength of the emulsion gel increased on increasing the γ-PGA concentration. CLSM images showed that the addition of γ-PGA modified the structure of the emulsion gel, and the droplets containing 0.1% γ-PGA were evenly distributed. Moreover, γ-PGA could regulate the droplet size of the FG emulsion and its size distribution. These findings suggest that the viscoelasticity and structure of FG emulsion gels could be regulated by adjusting the γ-PGA concentration. The γ-PGA-modified FG emulsion gel also exhibited improved rheology and physicochemical properties. The results showed that γ-PGA-modified FG emulsion gel may find potential applications in food, medicine, cosmetics, and other industries.
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The influence of ultrasonic pretreatment on the release and antioxidant activity of potential antioxidant peptides after in-vitro simulated gastrointestinal digestion of ß-lactoglobulin (BLG) were measured by HPLC-MS/MS, chemical and cellular-based assays. The gastrointestinal digest was fractionated into four fractions by Sephadex G-25 gel filtration column, and fractions showed a considerable ABTS·+ scavenging ability. The fraction with the strongest antioxidant activity was produced by ultrasonicated BLG after gastrointestinal digestion, which relies on ultrasonic-promoted proteolysis to produce many small-molecule antioxidant peptides. The best active fraction has better cellular antioxidant activity and protection of H2O2-induced oxidative HepG2 cell model, which significantly increases the activities of antioxidant enzyme, and is concentration-dependent. HPLC-MS/MS analysis showed that there were more potential antioxidant peptides in the best active fraction. This research will provide a basis for the further application of ultrasonic in dairy products, which can promote the release of more potential antioxidant peptides-derived from gastrointestinal digestion.