Comparison of lipidome profiles in human milk from Chinese Han and Korean ethnic groups based on high-throughput lipidomic techniques.

The composition of milk lipids varies across different ethnic sources. The lipidome profiles of Chinese Han human milk (HHM) and Chinese Korean human milk (KHM) were investigated in this study. A total of 741 lipids were identified in HHM and KHM. Twenty-eight differentially expressed lipids (DEL) were screened between the 2 milk groups; among these, 6 triacylglycerols (TG), 13 diacylglycerols (DG), 7 free fatty acids (FFA), and 1 monoglyceride (MG) were upregulated in KHM. Carnitine (CAR) was upregulated in HHM. Most DEL showed a single peak distribution in both groups. The correlations, related pathways and diseases of these DEL were further analyzed. The results demonstrated that DG, MG, and FFA showed highly positive correlations with each other (r > 0.8). The most enriched Kyoto Encyclopedia of Genes and Genomes (https://www.kegg.jp/kegg/) and Human Metabolome Database (http://www.hmdb.ca) pathways were inositol phosphate metabolism, and α-linolenic acid and linolenic acid metabolism, respectively. Major depressive disorder-related FFA (20:5) and FFA (22:6) were more abundant in KHM, whereas HHM showed more obesity-related CAR. These data potentially provide lipidome information regarding human milk from different ethnicities in China.

A review on polyphenols and their potential application to reduce food allergenicity.

This review summarized recent studies about the effects of polyphenols on the allergenicity of allergenic proteins, involving epigallocatechin gallate (EGCG), caffeic acid, chlorogenic acid, proanthocyanidins, quercetin, ferulic acid and rosmarinic acid, etc. Besides, the mechanism of polyphenols for reducing allergenicity was discussed and concluded. It was found that polyphenols could noncovalently (mainly hydrophobic interactions and hydrogen bonding) and covalently (mainly alkaline, free-radical grafting, and enzymatic method) react with allergens to induce the structural changes, resulting in the masking or/and destruction of epitopes and the reduction of allergenicity. Oral administration in murine models showed that the allergic reaction might be suppressed by regulating immune cell function, changing the levels of cytokines, suppressing of MAPK, NF-κb and allergens-presentation pathway and improving intestine function, etc. The outcome of reduced allergenicity and suppressed allergic reaction was affected by many factors such as polyphenol types, polyphenol concentration, allergen types, pH, oral timing and dosage. Moreover, the physicochemical and functional properties of allergenic proteins were improved after treatment with polyphenols. Therefore, polyphenols have the potential to produce hypoallergenic food. Further studies should focus on active concentrations and bioavailability of polyphenols, confirming optimal intake and hypoallergenic of polyphenols based on clinical trials.

Proteomic characterization and comparison of milk fat globule membrane proteins of Saanen goat milk from 3 habitats in China using SWATH-MS technique.

Saanen goats are among the major dairy goats in China. In present study, variation of milk fat globule membrane proteins profile of Saanen goat milk caused by geographic location was investigated using sequential window acquisition of all theoretical fragment ions data-independent acquisition mass spectrometry based proteomic approach. A total of 1,001 proteins were quantified in goat milk collected from 3 habitats of China [Guangdong (GD); Inner Mongolia (IM); Shannxi (SX)]. Most of the proteins were found to act cellular process of biological process, cell of cellular component, binding of molecular function after Gene Ontology annotation and metabolic of pathway indicated by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Differentially expressed proteins (DEP) for GD versus IM, GD versus SX, IM versus SX were identified to be 81, 91, and 44, respectively. Gene Ontology enrichment analysis showed that the greatest DEP for 3 groups (GD vs. IM, GD vs. SX, IM vs. SX) were cellular process, cellular process and organonitrogen compound biosynthetic process/immune system process for biological process. For cellular component, the largest number of DEP for 3 comparison groups were organelle, organelle and organelle/intracellular. For molecular function, DEP of the 3 comparison groups were expressed most in structural molecule activity, binding and anion binding, respectively. Pathways with the majority of DEP were ribosome, systemic lupus erythematosus and primary immunodeficiency/systemic lupus erythematosus/amoebiasis/PI3K-Akt signaling pathway for GD versus IM, GD versus SX and IM versus SX, severally. Protein-protein interaction network analysis showed that DEP interacted most were 40S ribosomal protein S5, fibronectin and Cytochrome b-c1 complex subunit 2, mitochondrial for GD versus IM, GD versus SX and IM versus SX, separately. Data may give useful information for goat milk selection and milk authenticity in China.

Fabrication and characterization of a cannabidiol-loaded emulsion stabilized by a whey protein-maltodextrin conjugate and rosmarinic acid complex.

A cannabidiol (CBD)-loaded oil-in-water emulsion stabilized by a whey protein (WP)-maltodextrin (MD) conjugate and rosmarinic acid (RA) complex was fabricated, and its stability characteristics were investigated under various environmental conditions. The WP-MD conjugates were formed via dry-heating. The interaction between WP and MD was assessed by browning intensity, reduced amount of free amino groups, the formation of high molecular weight components in sodium dodecyl sulfate-PAGE, and changes in secondary structure of whey proteins. The WP-MD-RA noncovalent complex was prepared and confirmed by fluorescence quenching and Fourier-transform infrared spectroscopy spectra. Emulsions stabilized by WP, WP-MD, and WP-RA were used as references to evaluate the effect of WP-MD-RA as a novel emulsifier. Results showed that WP-MD-RA was an effective emulsifier to produce fine droplets for a CBD-loaded emulsion and remarkably improved the pH and salt stabilities of emulsions in comparison with WP. An emulsion prepared with WP-MD-RA showed the highest protection of CBD against UV and heat-induced degradation among all emulsions. The ternary complex kept emulsions in small particle size during storage at 4°C. Data from the current study may offer useful information for designing emulsion-based delivery systems which can protect active substance against environmental stresses.

Effects of polymerized goat milk whey protein on physicochemical properties and microstructure of recombined goat milk yogurt.

Goat milk whey protein concentrates were manufactured by microfiltration (MF) and ultrafiltration (UF). When MF retentate blended with cream, which could be used as a starting material in yogurt making. The objective of this study was to prepare goat milk whey protein concentrates by membrane separation technology and to investigate the effects of polymerized goat milk whey protein (PGWP) on the physicochemical properties and microstructure of recombined goat milk yogurt. A 3-stage MF study was conducted to separate whey protein from casein in skim milk with 0.1-µm ceramic membrane. The MF permeate was ultrafiltered using a 10 kDa cut-off membrane to 10-fold, followed by 3 step diafiltration. The ultrafiltration-diafiltration-treated whey was electrodialyzed to remove 85% of salt, and to obtain goat milk whey protein concentrates with 80.99% protein content (wt/wt, dry basis). Recombined goat milk yogurt was prepared by mixing cream and MF retentate, and PGWP was used as main thickening agent. Compared with the recombined goat milk yogurt without PGWP, the yogurt with 0.50% PGWP had desirable viscosity and low level of syneresis. There was no significant difference in chemical composition and pH between the recombined goat milk yogurt with PGWP and control (without PGWP). Viscosity of all the yogurt samples decreased during the study. There was a slight but not significant decrease in pH during storage. Bifidobacterium and Lactobacillus acidophilus in yogurt samples remained above 106 cfu/g during 8-wk storage. Scanning electron microscopy of the recombined goat milk yogurt with PGWP displayed a compact protein network. Results indicated that PGWP prepared directly from raw milk may be a novel protein-based thickening agent for authentic goat milk yogurt making.

Comparative analysis of caseins in Saanen goat milk from 3 different regions of China using quantitative proteomics.

A quantitative proteomic technique based on data-independent acquisition (DIA) was used to analyze differentially expressed caseins of Saanen goat milk samples collected from 3 regions in China (Guangdong, GD; Inner Mongolia, IM; Shaanxi, SX). A total of 345 proteins were quantified in each sample. Gene Ontology (GO) analysis showed that proteins were mainly involved in cellular process and cell and binding functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that proteins were mainly involved in metabolic pathways. Differentially expressed proteins (DEP) between goat milk from 3 comparison groups composed of paired regions were compared and analyzed. The number of DEP was 114, 69, and 79 for GD versus IM, GD versus SX, and IM versus SX, respectively. The GO enrichment analysis of the 3 comparison groups showed that differences were mainly related to the regulation of biological quality, biological regulation, and response to stimulus in terms of biological process; extracellular region for cellular component; and binding function for molecular function. Pathways in which DEP of GD versus IM, GD versus SX, and IM versus SX were mostly protein processing in endoplasmic reticulum for the first 2 groups and metabolic pathways for the last. Protein-protein interaction network analysis demonstrated that the most prominent DEP was heat shock protein 90 ß family member 1 for both the GD versus IM and the GD versus SX groups, and haptoglobin for the IM versus SX group. Data from this study may offer useful information for further investigation of the protein composition of Saanen goat milk and its application in the dairy industry.

Effect of synbiotic yogurt fortified with monk fruit extract on hepatic lipid biomarkers and metabolism in rats with type 2 diabetes.

Monk fruit extract (MFE) is widely used as a sweetener in foods. In this study, the effects of the consumption of MFE-sweetened synbiotic yogurt on the lipid biomarkers and metabolism in the livers of type 2 diabetic rats were evaluated. The results revealed that the MFE-sweetened symbiotic yogurt affected the phosphatidylcholines, phosphatidylethanolamines, phosphatidylglycerol, lysophosphatidic acids, lysophosphatidylcholines, lysophosphatidylethanolamines, lysophosphatidylglycerols, lysophosphatidylinositols, lysophosphatidylserines, and fatty acid-hydroxy fatty acids biomarkers in the livers of type 2 diabetic rats. In addition, the consumption of the MFE-sweetened synbiotic yogurt significantly altered 12 hepatic metabolites, which are involved in phenylalanine metabolism, sphingolipid metabolism, bile secretion, and glyoxylate and dicarboxylate metabolism in the liver. Furthermore, a multiomics (metabolomic and transcriptomic) association study revealed that there was a significant correlation between the MFE-sweetened synbiotic yogurt and the metabolites and genes involved in fatty acid biosynthesis, bile secretion, and glyoxylate and dicarboxylate metabolism. The findings of this study will provide new insights on exploring the function of sweeteners for improving type 2 diabetes mellitus liver lipid biomarkers.

Peptide profiles and antioxidant capacity of extensive hydrolysates of milk protein concentrate.

Milk protein concentrate was hydrolyzed using one-step enzymatic hydrolysis. Both the peptide profiles and antioxidant activities of the resulting extensive hydrolysates of milk protein concentrate (EMPH) were analyzed using a peptidomics approach based on liquid chromatography-tandem mass spectrometry. The results demonstrated that the degrees of hydrolysis of the 4 EMPH by Alcalase-Protamex, Alcalase-Protease A 2SD, Alcalase-Flavorzyme, and Alcalase-ProteAXH were 12.02%, 16.85%, 15.87%, and 15.77%, respectively. Using size exclusion chromatography, 99.85% of the peptides in the Alcalase-Protease A 2SD hydrolysate were shown to have a molecular weight of <3 kDa. A total of 33 common peptides were identified in the EMPH by liquid chromatography-tandem mass spectrometry, 16 of which were identified as bioactive peptides using bioinformatics. The peptide profiles and the coverage of master proteins of the 4 EMPH were different. The EMPH also exhibited strong free radical scavenging capacity, as indicated by the results of the 1,1-diphenyl-2-picrylhydrazyl radical, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), hydroxyl radical, and reducing power assays. The results of this study provided useful information on the peptide profiles and antioxidant activity of EMPH.

Safety assessment of two strains and anti-obese effects on mice fed a high-cholesterol diet.

Previous study documented that Lactobacillus paracasei S0940 and Streptococcus thermophilus ldbm1 have obvious cholesterol-lowering abilities in vitro. In this study, the safety of two strains were evaluated by nitroreductase test, hemolysis test and antibiotic sensitivity test and to evaluate the cholesterol-reducing abilities in vivo. The results indicated that two strains did not exhibit nitroreductase activities and were ɤ-hemolytic on blood agar plates. Further, both strains did not represent a health risk by antibiotic sensitivity test, and significantly reduced serum and liver cholesterol and triglyceride levels of high fat-fed mice. Compared with the high-fat model group, administration of the strains to mice fed a high-cholesterol diet increased fecal water content and fecal cholesterol and significantly improved the intestinal microbiota, which indicating that Lactobacillus paracasei S0940 and Streptococcus thermophilus ldbm1 have a positive effect on reducing cholesterol levels and may be used in functional food.

Proteomic analysis of differentially expressed whey proteins in Saanen goat milk from different provinces in China using a data-independent acquisition technique.

Whey proteins of Saanen goat milk samples from 3 provinces in China (Guangdong, GD; Inner Mongolia, IM; Shaanxi, SX) were characterized and compared using data-independent acquisition quantitative proteomics technique. A total of 550 proteins were quantified in all 3 samples. There were 44, 44, and 33 differentially expressed proteins (DEP) for GD versus IM, GD versus SX, and IM versus SX, respectively. Gene ontology annotation analysis showed that the largest number of DEP for the 3 comparisons were as follows: for biological processes: response to progesterone, glyceraldehyde-3-phosphate metabolic process, and negative regulation of megakaryocyte differentiation; for molecular functions: antioxidant activity, binding, and peroxiredoxin activity; and for cellular components: the same category of extracellular regions for the 3 comparisons, respectively. Pathways for the DEP of 3 comparisons were (1) disease; (2) synthesis and metabolism; and (3) synthesis, degradation, and metabolism. Protein-protein interaction network analysis showed that DEP for GD versus SX had the most interactions.

Polymerized Whey Protein Concentrate-Based Glutathione Delivery System: Physicochemical Characterization, Bioavailability and Sub-Chronic Toxicity Evaluation.

Glutathione (GSH) is a powerful antioxidant, but its application is limited due to poor storage stability and low bioavailability. A novel nutrient encapsulation and delivery system, consisting of polymerized whey protein concentrate and GSH, was prepared and in vivo bioavailability, antioxidant capacity and toxicity were evaluated. Polymerized whey protein concentrate encapsulated GSH (PWPC-GSH) showed a diameter of roughly 1115 ± 7.07 nm (D50) and zeta potential of 30.37 ± 0.75 mV. Differential scanning calorimetry (DSC) confirmed that GSH was successfully dispersed in PWPC particles. In vivo pharmacokinetics study suggested that PWPC-GSH displayed 2.5-times and 2.6-fold enhancement in maximum concentration (Cmax) and area under the concentration-time curve (AUC) as compared to free GSH. Additionally, compared with plasma of mice gavage with free GSH, significantly increased antioxidant capacity of plasma in mice with PWPC-GSH was observed (p < 0.05). Sub-chronic toxicity evaluation indicated that no adverse toxicological reactions related to oral administration of PWPC-GSH were observed on male and female rats with a diet containing PWPC-GSH up to 4% (w/w). Data indicated that PWPC may be an effective carrier for GSH to improve bioavailability and antioxidant capacity.

Proteomic analysis of whey proteins in the colostrum and mature milk of Xinong Saanen goats.

Xinong Saanen goats are among the major dairy goats in China, and their milk is one of the major milk supplies for the Chinese dairy industry. To explore the whey proteome of Xinong Saanen goat milk, we analyzed the whey proteins of goat colostrum and mature milk using proteomic techniques. We identified a total of 314 and 524 proteins in goat colostrum and mature milk, respectively. Our data showed the first 3 functional categories of signal, disulfide bond, and secreted in both milk types. The most abundant Gene Ontology annotations in both milks were the biological process of positive regulation of extracellular regulated protein kinases (ERK)1 and ERK2 cascade; the cellular component of extracellular exosome; and the molecular function of calcium ion binding. Goat colostrum whey proteins showed more disease-related pathways, and mature milk showed more pathways associated with metabolism. Moreover, we observed several pathways involved in intestinal mucosal immunity only in colostrum. Protein-protein interaction network and module analysis revealed that complement and coagulation cascades and Staphylococcus aureus infection were significant in the whey proteins of both milks, and carbon metabolism was more common for mature milk than for colostrum. These findings could provide useful information for the use of goat milk whey proteins in the Chinese dairy industry.

Characterization of the milk fat globule membrane proteome in colostrum and mature milk of Xinong Saanen goats.

Xinong Saanen goat milk is a major source of milk in the Chinese dairy industry. Milk fat globule membrane (MFGM) proteomes of goat colostrum and mature milk were analyzed and compared using proteomic technology. A total of 543 and 585 proteins were identified in goat colostrum and mature milk, respectively. Functional category analyses revealed that most of the MFGM proteins in both colostrum and mature milk were related to phosphoprotein and acetylation. The biological process of translation, cellular component of extracellular exosome, and molecular function of poly(A) RNA binding were the main gene ontology annotations of both colostrum and mature milk. Pathways associated with disease and genetic information processing involved large number of proteins in colostrum and mature milk, and more metabolism-related pathways were observed in mature milk. Protein-protein interaction network analyses showed that ribosome was abundant in both colostrum and mature milk. Colostrum showed more functions associated with protein processing in the endoplasmic reticulum, whereas mature milk had more oxidative phosphorylation functions. The results could provide further understanding of the unique biological properties of MFGM proteins of goat colostrum and mature milk.

Proteomic analysis of differentially expressed whey proteins in Guanzhong goat milk and Holstein cow milk by iTRAQ coupled with liquid chromatography-tandem mass spectrometry.

Guanzhong goat and Holstein cow milk are the major milks supplied in China. Whey proteins play an important role in immune defense for newborn mammals. This study aimed to analyze the differentially expressed whey proteins of Guanzhong goat milk and Holstein cow milk by using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomics techniques. A total of 165 whey proteins were quantified, 114 of which differed significantly in abundance in goat and cow milks. According to the "up_keywords," in the online DAVID tool (https://david.ncifcrf.gov/home.jsp), 75% of these differentially expressed whey proteins were related to the category of "signal." Gene Ontology analyses classified these differentially expressed proteins into biological processes, cellular components, and molecular functions. The most common biological process was response to stress, the most common cellular component was related to extracellular region, and the most prevalent molecular function was binding. Kyoto Encyclopedia of Genes and Genomes pathway analyses showed that these proteins were mainly involved in the complement and coagulation cascade pathways. The results improve our understanding of the different biological properties of whey proteins in goat and cow milks.

Effects of a synbiotic yogurt using monk fruit extract as sweetener on glucose regulation and gut microbiota in rats with type 2 diabetes mellitus.

We developed a synbiotic yogurt using monk fruit extract as a sweetener and investigated the effects of feeding the yogurt to rats with type 2 diabetes induced by streptozotocin and a high-fat diet. The rats fed the synbiotic yogurt showed greater blood glucose regulation and a significant decrease in insulin resistance and glycosylated hemoglobin compared with rats fed yogurt sweetened with sucrose, and they showed a remarkable improvement in short-chain fatty acid levels and gut microbiota status. Liver and kidney damage was also ameliorated in the rats fed the synbiotic yogurt. Immunohistochemistry analysis showed that the synbiotic yogurt inhibited ß-cell loss compared with the control yogurt. Consuming the synbiotic yogurt helped to restore the islets of Langerhans. Our results indicated that monk fruit extract may be a good alternative to sucrose for synbiotic yogurt products in people with type 2 diabetes to delay the progression of diabetes and associated complications.

Physiochemical, rheological, microstructural, and antioxidant properties of yogurt using monk fruit extract as a sweetener.

A yogurt using monk fruit extract (MFE) as a sweetener was developed. The aim of the study was to investigate the viability of using MFE to develop sweetened yogurts without the calories of added sugar. The physiochemical, rheological, microstructural, and antioxidant properties of yogurt were studied. Rheological results showed that MFE affected the yogurt fermentation process and its rheological properties. Yogurt sweetened with MFE had similar microstructural properties to yogurt sweetened with sucrose. Yogurt with MFE showed higher levels of gly-pro-p-nitroanilide and dipeptidyl peptidase IV inhibitory activities, 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity, α-glucosidase inhibitory activities, and superoxide anion radical scavenging ability compared with other yogurt samples. Results indicated that MFE could be a novel sweetener and a food antioxidant for functional yogurt and related products.

Effects of High Intensity Ultrasound on Physiochemical and Structural Properties of Goat Milk ß-Lactoglobulin.

This study aimed to compare the effects of high intensity ultrasound (HIU) applied at various amplitudes (20~40%) and for different durations (1~10 min) on the physiochemical and structural properties of goat milk ß-lactoglobulin. No significant change was observed in the protein electrophoretic patterns by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Deconvolution and second derivative of the Fourier transform infrared spectra (FTIR) showed that the percentage of ß-sheet of goat milk ß-lactoglobulin was significantly decreased while those of α-helix and random coils increased after HIU treatment The surface hydrophobicity index and intrinsic fluorescence intensity of samples was enhanced and increased with increasing HIU amplitude or time. Differential scanning calorimetry (DSC) results exhibited that HIU treatments improved the thermal stability of goat milk ß-lactoglobulin. Transmission electron microscopy (TEM) of samples showed that the goat milk ß-lactoglobulin microstructure had changed and it contained larger aggregates when compared with the untreated goat milk ß-lactoglobulin sample. Data suggested that HIU treatments resulted in secondary and tertiary structural changes of goat milk ß-lactoglobulin and improved its thermal stability.

Physicochemical, texture properties, and microstructure of yogurt using polymerized whey protein directly prepared from cheese whey as a thickening agent.

The aim of this study was to investigate suitability of polymerized whey protein prepared directly from cheese whey on the physicochemical, texture properties, and microstructure of the yogurt. The results indicated that addition of polymerized whey protein obtained by heating the liquid whey protein concentrate at 75°C for 10 min had no significant differences in pH, titratable acidity, total solids, protein content, viscosity, texture, and syneresis between the yogurt with polymerized liquid whey protein (YWPS) and the yogurt with polymerized whey protein concentrate. However, the YWPS had significant differences in viscosity, texture, and syneresis compared with the control yogurt. Scanning electron micrographs of YWPS displayed a compact and homogeneous protein network for polymerized whey protein solution (PWPS) samples. The 4 yogurt samples were evaluated by the quantitative descriptive analysis method, and 14 sensory attributes were analyzed by principal component analysis. All 3 principal components had significant effects on the sensory profiles, accounting for 52.3, 24.32, and 10.8% of the variability in the results, respectively. Polymerized whey protein prepared directly from cheese whey may be a good protein base as a thickening agent for yogurt making.

Preparation and Characterization of Whey Protein Isolate-DIM Nanoparticles.

3,3'-Diindolylmethane (DIM) is a bioactive compound found in Cruciferous vegetables that possesses health benefits such as antioxidant, anticancer, and anti-inflammatory effects. However, hydrophobicity and photolabile limit its pharmaceutical applications. This study aims to prepare and characterize DIM-encapsulated whey protein isolate (WPI) nanoparticles mixed at different ratios of WPI and DIM using the combined heating-ultrasound method. Results showed that all the samples showed adequate physicochemical characteristics: The mean particle size of the nanoparticles could be controlled down to 96-157 nm depending on the DIM to WPI ratio used in the preparation with a low polydispersity index (<0.5), higher negative values of zeta potential (>-40 mV) as well as with greater encapsulation efficiency (>82%). Flow behavior indices showed the shear-thinning Non-Newtonian or pseudoplastic (n < 1) behavior of the nanoparticles. The thermal properties were characterized by differential scanning calorimetry (DSC), which showed that DIM was successfully entrapped in WPI nanoparticles. The secondary structure of WPI was changed after DIM incorporation; electrostatic interaction and hydrogen bonding were major facilitating forces for nanoparticles formation, confirmed by Fourier Transform Infrared Spectroscopy (FT-IR). Transmission electron microscopy (TEM) micrographs showed that all the samples had a smooth surface and spherical structure. The wall material (WPI) and encapsulation method provide effective protection to DIM against UV light and a broad range of physiologically relevant pH's (2.5, 3.5, 4.5, 5.5, and 7). In conclusion, whey protein isolate (WPI)-based nanoparticles are a promising approach to encapsulate DIM and overcome its physicochemical limitations with improved stability.

Heat-Induced Interactions between Whey Protein and Inulin and Changes in Physicochemical and Antioxidative Properties of the Complexes.

Whey protein and inulin at various weight ratios were dry heated at 60 °C for 5 days under relative humidity of 63%. The heated mixtures were found to have significant changes in browning intensity and zeta-potential compared to untreated mixture. Heated samples showed significantly lower surface hydrophobicity than untreated mixtures. Compared with untreated samples, dry-heated samples showed significantly higher 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) scavenging ability with whey protein to inulin mass ratios of 1:2 and 1:3 and significantly higher 2,2'-Azinobis(2-Ethylbenzothiazoline-6-Sulfonate) (ABTS) scavenging abilities and oxygen radical absorbance capacity (ORAC) at all weight ratios. Dry heat-induced interactions between whey protein and inulin was confirmed by changes in Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) protein profile, Fourier Transform Infrared Spectroscopy (FT-IR) and Far-ultraviolet Circular Dichroism (Far-UV CD) spectra. Dry heating caused physicochemical and structural changes of whey protein and therefore the complexes can be used to improve the antioxidative properties of the mixture under certain conditions.