Impact of Lactiplantibacillus plantarum and casein fortification on angiotensin converting enzyme inhibitory peptides in yogurt: identification and in silico analysis.

In this study, the effects of Lactiplantibacillus plantarum M11 (Lb. plantarum M11) in conjunction with sodium caseinate on the characteristics and angiotensin converting enzyme (ACE) inhibitory activity of yogurt were investigated. ACE inhibitory peptides (ACEIPs) in yogurt were identified by nano-LC-MS/MS and potential ACEIPs were predicted by in silico and molecular docking methods. The results showed that the ACE-inhibitory activity of yogurt was significantly enhanced (p < 0.05), while maintaining the quality characteristics of the yogurt. Thirteen ACEIPs in the improved yogurt (883 + M11-CS group) were identified, which were more abundant than the other yogurt groups (control 883 group, 883 + M11 group and 883-CS group). Two novel peptides with potential ACE inhibitory activity, YPFPGPIH and NILRFF, were screened. The two peptides showed PeptideRanker scores above 0.8, small molecular weight and strong hydrophobicity, and were non-toxic after prediction. Molecular docking results showed that binding energies with ACE were -9.4 kcal mol-1 and -10.7 kcal mol-1, respectively, and could bind to the active site of ACE. These results indicated that yogurt with Lb. plantarum M11 and sodium caseinate has the potential to be utilized as a functional food with antihypertensive properties. The combination of ACEIP-producing strains and casein fortification could be an effective method to promote the release of ACEIPs from yogurt.

Investigation into potential allergenicity of DBD plasma-treated casein digestion products based on immunoglobulin E linear epitopes and the sensitized-cell model.

The study aimed to investigate the allergenicity change in casein treated with dielectric barrier discharge (DBD) plasma during in vitro simulated digestion, focusing on the immunoglobulin E (IgE) linear epitopes and utilizing a sensitized-cell model. Results indicated that prior treatment with DBD plasma treatment (4 min) before simulated digestion led to a 10.5% reduction in the IgE-binding capacity of casein digestion products. Moreover, the release of biologically active substances induced from KU812 cells, including ß-HEX release rate, human histamine, IL-4, IL-6, and TNF-α, decreased by 2.1, 28.1, 20.6, 11.6, and 17.3%, respectively. Through a combined analysis of LC-MS/MS and immunoinformatics tools, it was revealed that DBD plasma treatment promoted the degradation of the IgE linear epitopes of casein during digestion, particularly those located in the α-helix region of αs1-CN and αs2-CN. These findings suggest that DBD plasma treatment prior to digestion may alleviate casein allergic reactions.

Characteristics of casein phosphopeptides in Chinese human milk and its correlation with infant growth: A cross-sectional study.

This research aimed to investigate the characteristics of casein phosphopeptides in Chinese human milk, and their potential relationship to infant growth. Using the liquid chromatography-Orbitrap-mass spectrometry technique, a total of 15 casein phosphopeptides were identified from 200 human milk samples. Also, our results indicate that casein phosphopeptides were phosphorylated with only one phosphate. The relative concentrations of casein phosphopeptides at 6 months postpartum were increased compared with milk at 2 months (FDR < 0.05). Significantly positive correlations were observed between casein phosphopeptides and infant growth, as shown by four casein phosphopeptides were positively correlated with the infants' weight-for-age Z-scores (rs range from 0.20 to 0.29), and three casein phosphopeptides were positively correlated with the infants' length-for-age Z-scores (rs range from 0.19 to 0.27). This study is the first to reveal the phosphorylated level and composition of casein phosphopeptides in Chinese human milk, and their potential relationship with infant growth.

Structural Analysis of Breast-Milk αS1-Casein: An α-Helical Conformation Is Required for TLR4-Stimulation.

Breast-milk αS1-casein is a Toll-like receptor 4 (TLR4) agonist, whereas phosphorylated αS1-casein does not bind TLR4. The objective of this study was to analyse the structural requirements for these effects. In silico analysis of αS1-casein indicated high α-helical content with coiled-coil characteristics. This was confirmed by CD-spectroscopy, showing the α-helical conformation to be stable between pH 2 and 7.4. After in vitro phosphorylation, the α-helical content was significantly reduced, similar to what it was after incubation at 80 °C. This conformation showed no in vitro induction of IL-8 secretion via TLR4. A synthetic peptide corresponding to V77-E92 of αS1-casein induced an IL-8 secretion of 0.95 ng/mL via TLR4. Our results indicate that αS1-casein appears in two distinct conformations, an α-helical TLR4-agonistic and a less α-helical TLR4 non-agonistic conformation induced by phosphorylation. This is to indicate that the immunomodulatory role of αS1-casein, as described before, could be regulated by conformational changes induced by phosphorylation.

Gastric coagulation and postprandial amino acid absorption of milk is affected by mineral composition: a randomized crossover trial.

Background: In vitro studies suggest that casein coagulation of milk is influenced by its mineral composition, and may therefore affect the dynamics of protein digestion, gastric emptying and appearance of amino acids (AA) in the blood, but this remains to be confirmed in vivo. Objective: This study aimed to compare gastrointestinal digestion between two milks with the same total calcium content but different casein mineralization (CM). Design: Fifteen males (age 30.9 ± 13.8 years, BMI 22.5 ± 2.2 kg m-2) participated in this randomized cross-over study with two treatments. Participants underwent gastric magnetic resonance imaging (MRI) scans at the baseline and every 10 min up to 90 min after consumption of 600 ml milk with low or high CM. Blood samples were taken at the baseline and up to 5 hours postprandially. Primary outcomes were postprandial plasma AA concentrations and gastric emptying rate. Secondary outcomes were postprandial glucose and insulin levels, gastric coagulation as estimated by image texture metrics, and appetite ratings. Results: Gastric content volume over time was similar for both treatments. However, gastric content image analysis suggested that the liquid fraction emptied quicker in the high CM milk, while the coagulum emptied slower. Relative to high CM, low CM showed earlier appearance of AAs that are more dominant in casein, such as proline (MD 4.18 µmol L-1, 95% CI [2.38-5.98], p < 0.001), while there was no difference in appearance of AAs that are more dominant in whey protein, such as leucine. The image texture metrics homogeneity and busyness differed significantly between treatments (MD 0.007, 95% CI [0.001, 0.012], p = 0.022; MD 0.005, 95% CI [0.001, 0.010], p = 0.012) likely because of a reduced coagulation in the low CM milk. Conclusions: Mineral composition of milk can influence postprandial serum AA kinetics, likely due to differences in coagulation dynamics. The clinical trial registry number is NL8959 (https://clinicaltrials.gov).

Effects of protein genetic variants on their phosphorylation levels, milk composition, milk proteome, and milk coagulation ability in Chinese Holstein bovine milk.

Milk samples were collected from 3625 Chinese Holstein cows to assess the effects of κ-casein (κ-CN) and ß-lactoglobulin (ß-LG) genetic variants on its milk coagulation properties. The results show that Chinese Holstein cows have a higher frequency of the κ-CN AA and AB variants, and ß-LG of the AB and AA variants. Of these, κ-CN B variants, the ß-LG AA and BB variants were more frequent in milk showing good coagulation. The effects of the genetic variants on milk composition, milk proteome, and protein phosphorylation sites were studied. The results showed that higher concentrations of protein and dry matter were found in κ-CN BE variant. Moreover, large variations in milk proteome among different κ-CN and ß-LG variants were observed. Highly phosphorylated for κ-CN, especially Ser97, was observed in cows with the κ-CN BE variant, but no effect of ß-LG variants on phosphorylation site was found. Of the various factors examined, variation of κ-CN phosphorylation sites Ser97 may be the most important in affecting casein structure and milk coagulation ability. Some milk protein contents were found to be negative factors for milk coagulation. In summary, this study showed that κ-CN genetic variants contained different milk compositions and phosphorylation site Ser97 influenced milk coagulation.

Digestive properties and peptide profiles exhibited significant differences between skim camel milk and bovine milk powder after static in vitro simulated infant gastrointestinal digestion.

This study aims to analyze the differences in digestion properties and peptide profiles between the skim camel and bovine milk powder after static in vitro simulated infant gastrointestinal digestion. The hydrolysis degree of camel milk proteins exceeded by 13.18% that of bovine milk. The concentration and release rate of free amino groups in the camel milk digesta was higher than that of bovine milk powder, which was likely due to the higher ß-/αs-casein ratio and larger casein micelle size in camel milk. Camel milk powder presented higher ß-CN coverage and comparatively shorter bioactive peptides compared to bovine milk powder. The anti-inflammatory peptide KVLPVPQ displayed the highest abundance in camel milk powder. Outcomes of this study showed that camel milk proteins possessed superior digestibility and unique peptides, which outlined the potential nutritional implications of camel milk for infants.

Interaction between curcumin and ultrafiltered casein micelles or whey protein, and characteristics of their complexes.

This work evaluated the interaction between micellar casein (MC) or whey protein (WP) in ultrafiltration retentate with curcumin (Cur), as well as the physicochemical and functional properties of Cur-MC and Cur-WP complexes. The MC had a higher affinity for Cur than WP, shown by higher binding constants of Cur-MC at various temperatures. Thermodynamic analysis of the binding process indicated that the interaction between Cur and MC or WP was hydrophobic in nature. Cur promoted the size and polydispersity index of MC and WP at 4 mM but did not alter the morphology of spray-dried MC and WP. The Cur-MC complexes showed better aqueous solubility at pH 2-3 and 6-10 compared to free MC. Combination with MC or WP improved the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical scavenging activity of Cur. In addition, combination with MC and WP promoted cumulative release of Cur during simulated gastrointestinal digestion, especially for WP. Thus, MC and WP in retentates can be good alternative protein-based carriers for Cur delivery, whereas their complexes in powder form have good functional properties that could be used as active food ingredients in several food formulations. PRACTICAL APPLICATION: Microfiltration is a cheap and convenient approach that can be used to easily produce micellar casein (MC), with whey protein (WP) as one byproduct. In this study, we proved that MC and WP in retentates have strong interaction with curcumin (Cur), whereas their complexes have good functional properties. Thus, spray-dried MC-Cur or WP-Cur complexes could be used as active food ingredients in several food formulations.

High-pressure microfluidization enhanced the stability of sodium caseinate-EGCG complex-stabilized fish oil emulsion.

Improving the emulsion-stabilizing effect of protein by chemical or physical modification has been paid much attention recently. Here, sodium caseinate (CS) was treated by high-pressure-microfluidization (HPM) under 0-100 MPa, and was further complexed with (-)-epigallocatechin-3-gallate (EGCG) to form an excellent emulsifier that stabilized fish oil emulsions. Results showed that HPM treatment (especially 80 MPa) significantly changed the secondary structure of CS, and 80 MPa-PCS-EGCG had the best emulsifying and antioxidant activities. In addition, after HPM treatment and EGCG bonding, CS formed a thicker interface layer on the surface of oil droplets, which could better protect the fish oil from the influence by oxygen, temperature and ion concentration. Moreover, the fish oil emulsion stabilized by PCS-EGCG complex significantly delayed the release of free fatty acids subjected to in vitro digestion. Conclusively, HPM-treated CS-EGCG complex could be a potential emulsifier to improve the stability of fish oil emulsions.

Casein-based conjugates and graft copolymers. Synthesis, properties, and applications.

Biobased natural polymers, including polymers of natural origin such as casein, are growing rapidly in the light of the environmental pollution caused by many mass-produced commercial synthetic polymers. Although casein has interesting intrinsic properties, especially for the food industry, numerous chemical reactions have been carried out to broaden the range of its properties, most of them preserving casein's nontoxicity and biodegradability. New conjugates and graft copolymers have been developed especially by Maillard reaction of the amine functions of the casein backbone with the aldehyde functions of sugars, polysaccharides, or other molecules. Carried out with dialdehydes, these reactions lead to the cross-linking of casein giving three-dimensional polymers. Acylation and polymerization of various monomers initiated by amine functions are also described. Other reactions, far less numerous, involve alcohol and carboxylic acid functions in casein. This review provides an overview of casein-based conjugates and graft copolymers, their properties, and potential applications.

Effect of composition, casein genetic variants and glycosylation degree on bovine milk whipping properties.

This study investigated the individual and combined effects of ĸ-Casein (ĸ-CN; AA, AB, BB), ß-Casein (ß-CN; A1A1, A1A2, A2A2) and high and low ratios of glycosylated ĸ-CN to total ĸ-CN, referred to as the glycosylation degree (GD), on bovine cream whipping properties. The genetic variants of individual cows were identified using reversed-phase high-performance liquid chromatography (RP-HPLC) and verified through liquid chromatography-mass spectrometry (LC-MS). A previously discovered relationship between days-in-milk and GD was validated and used to obtain high and low GD milk. Whipped creams were created through the mechanical agitation of fat standardised cream from milk of different ĸ-CN, ß-CN, and GD combinations, and whipping properties (the ability to whip, overrun, whipping time and firmness) were evaluated. No significant correlation was measured in whipping properties for cream samples from milks with different ĸ-CN and ß-CN genetic variants. However, 80 % of samples exhibiting good whipping properties (i.e., the production of a stiffened peak) were from milk with low GD suggesting a correlation between whipping properties and levels of glycosylation. Moreover, cream separated from skim milk of larger casein micelle size showed superior whipping properties with shorter whipping times (<5 min), and higher firmness and overrun. Milk fat globule (MFG) size, on the other hand, did not affect whipping properties. Results indicate that the GD of κ-CN and casein micelle size may play a role in MFG adsorption at the protein and air interface of air bubbles formed during whipping; hence, they govern the dynamics of fat network formation and influencing whipping properties.

Impact of pulsed electric field intensity on the cream separation efficiency from bovine milk and physico-chemical properties of the cream.

Low-temperature (9-12 °C) pulsed electric field (PEF) was investigated in milk before cream separation at different intensities (9-27 kV/cm, 66 µs, 16-28 kJ/L) regarding its potential to render processing more sustainable, retain a high physico-chemical quality, enhance functional properties, and gently modify the structure of the milk fat globule membrane (MFGM). Cream volume per L milk were most efficiently increased by 31 % at the lowest PEF intensity in comparison to untreated milk and cream (P < 0.05). Untreated and PEF-treated milk and obtained cream were assessed with compositional (fat, protein, casein, lactose, and total solids content) and particle size distribution analyses, showing no significant differences (P ≥ 0.05) and, thus, indicating retention of 'native-like' product quality. Overrun and stability of cream, whipped for 20 and 60 s at 15000 rpm using a high-shear mixer, were improved most notably by the lowest and the highest PEF intensities, achieving up to 69 % enlarged overrun and up to 22 % higher stability, respectively (P < 0.05), than in untreated whipped cream. Protein component analyses for milk and cream were carried out by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Noticeable differences between untreated and PEF-treated milk were not observed, but the SDS-PAGE results for cream showed noticeably different bands for some of the protein components, indicating structural changes in MFGM-, whey-, and phospho-proteins due to PEF and/or separator processing effects. More intense bands of xanthine oxidase, xanthine dehydrogenase, butyrophilin, bovine serum albumine, adipophilin (ADPH), and glycoproteins PAS6/7 were observed specifically at 21 kV/cm. Gentle electroporation of both MFGM layers by PEF was determined based on the changes in MFGM monolayer components, such as ADPH and PAS 6/7, exhibiting intensified bands. PEF intensity-dependent impact on the structure of MFGM and casein, leading to a reconfiguration of the cream matrix due to different structuring interactions among proteins, among milk fat globules, and between fat and protein components, was suggested. Overall, low-temperature PEF applied at different intensities showed great potential for gentle, efficient, and functional properties-tailored dairy processing and may also enable effective extraction of highly bioactive ingredients from dairy sources.

A SAXS and USAXS study of the influence of pH on the casein micelle structure.

Changes in milk pH significantly influence the behavior and physical properties of casein micelles; however, the effects of these changes on casein micelle structures are still unclear. The aim of this study was to elucidate the effect of changes in pH range from 5.9 to 7.1 on the structure of casein micelles in milk using small-angle X-ray scattering (SAXS) and ultra small-angle X-ray scattering (USAXS). The casein micelles formed one-dimensional aggregates. The micelle radius decreased with decreasing pH, whereas the size of the water domain increased. The distance between colloidal calcium phosphates (CCP) remained unchanged, whereas the CCP radius decreased with decreasing pH. Voluminosity, which was calculated from scattering intensities, increased at increased pH. In conclusion, the micelle structure changed significantly in response to changes in pH. Our findings help to understand the changes in the physical properties of milk at various pH levels in terms of the microscopic structure.

Preparation and characterization of sodium caseinate-apricot tree gum/gum Arabic nanocomplex for encapsulation of conjugated linoleic acid (CLA).

Nanocomplexes (NCs) were formed through electrostatic complexation theory using Na-caseinate (NaCa), gum Arabic (GA), and Prunus armeniaca L. gum exudates (PAGE), aimed to encapsulate Conjugated linoleic acid (CLA). Encapsulation was optimized using NaCa (0.1 %-0.5 %), GA/PAGE (0.1 %-0.9 %) and CLA (1 %-5 %), and central composite design (CCD) was employed for numerical optimization. The optimum conditions for NC containing GA (NCGA) were 0.336 %, 0.437 %, and 3.10 % and for NC containing PAGE (NCPAGE) were 0.403 %, 0.730 %, and 4.177 %, of NaCa, GA/PAGE, and CLA, respectively. EE and particle size were 92.46 % and 52.89 nm for NCGA while 88.23 % and 54.76 nm for NCPAGE, respectively. Fourier transform infrared spectroscopy (FTIR) indicated that CLA was physically entrapped. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed the electrostatic complex formation. The elastic modulus was predominant for NCGA and NCPAGE dispersions while the complex viscosity of NCPAGE suspension was slightly higher than that of NCGA. The CLA in NCGA-CLA and NCPAGE-CLA exhibited higher oxidative stability than free CLA during 30 days of storage without a significant difference between the results of CLA oxidative stability tests obtained for NCs. Consequently, NCPAGE and NCGA could be applied for the entrapment and protection of nutraceuticals in the food industry.

Effect of casein phosphopeptide-amorphous calcium phosphate, Riva Star, and Moringa oleifera-based nano-silver fluoride on caries affected dentin remineralization bonded to composite resin.

OBJECTIVE: This study aims to assess the effects of the most recent remineralizing agents, casein phosphopeptide-amorphous calcium phosphate (CPP-ACP), CO2 laser irradiation + topical fluoride (CO2 laser + TF), and Nanosilver fluoride - M. oleifera (NSF-MOLE), on the shear bond strength (SBS) and bond failure between resin composite and remineralized caries affected dentin (CAD). MATERIALS AND METHODS: Fifty human molars with occlusal caries reaching approximately halfway through the dentin were immersed in a 4% thymol solution. The infected dentin was removed using an excavator and the CAD surface was exposed. The sample was allocated into five groups (n=10) based on the remineralizing agent applied. Group 1: no remineralizing agent, group 2: CPP-ACP, group 3: Riva Star, group 4: NSF:MOLE, and group 5: (CO2 laser + TF). The shear bond testing procedure was conducted utilizing a universal testing machine and a stereo-microscope was used to study the failure pattern. The researchers utilized a one-way analysis of variance. The Tukey post hoc test was conducted for multiple comparison tests. RESULTS: Group 4 (NSF-MOLE) (13.77±1.94 MPa) treated testers recognized the highest bond values of tooth color restoration to the CAD surface. Nonetheless, group 1 test samples with no mineralization unveiled the minimum outcome of bond integrity (9.12±1.14 MPa). Intergroup comparison exploration showed that group 2 (CPP-ACP), group 4 (NSF-MOLE) (13.77±1.94 MPa), and group 5 (CO2 laser + TF) established comparable values of SBS. Furthermore, group 3 (Riva Star) displayed better SBS than group 1 but lower than group 2, group 4, and group 5. CONCLUSIONS: Remineralization of CAD using modern regimes (CPP-ACP, NSF-MOLE, and CO2 laser + TF has the potential to be used to enhance the bond strength of CAD to composite restoration.

The aggregation of casein micelles induced by Ca2+ during in vitro digestion: effects on the release of loaded anthocyanins.

Colloidal calcium phosphate (CCP) confers a modifiable structure to micellar casein (MC), which endows it with potential advantages as a delivery carrier. However, it is difficult to achieve multipattern release of the core material in the intestine with MC as a single wall. In this study, we prepared an anthocyanin-casein-based delivery system utilizing MC with different freezing degrees as the wall material with the objective of achieving the controlled release of anthocyanin as the model core in the intestine. The results showed that freezing could significantly reduce the CCP level up to 50%. Static in vitro simulated digestion with the addition of exogenous Ca2+ showed that the designed delivery system exhibited low anthocyanin release (15%-35%) in the gastric tract. The pattern of release in the intestine depended on the CCP dissociation degree. High and low dissociation degrees corresponded to slow release (from 15% to 65% within 2 h) and burst release (from 35% to 90% within 5 min), respectively. WAXS/SAXS analysis revealed that exogenous serum Ca2+ inherent in simulated gastric fluid and endogenous serum Ca2+ induced by CCP dissociation was synergistically involved in the reconstitution of CCP-mediated nanoclusters and large aggregates. The freezing degree of MC determined the endogenous serum Ca2+ level, which influenced the gastric aggregation behavior of wall MC and ultimately led to a fairly different gastrointestinal release behavior of anthocyanins.

Exploring the effect of high pressure in the denaturation of casein micelles by in-situ SERS.

To investigate the structural changes of casein in response to the pressurization process under varying pressure levels, this study carried out both ex-situ and in-situ high-pressure experiments. In the in-situ experiments, the surface-enhanced Raman scattering (SERS) technique was combined with a diamond anvil cell (DAC). The high-pressure experiments indicated that significant dissociation of casein occurred at 200 MPa. Over the range of 0-302 MPa, casein exhibited both dissociation and aggregation behaviors. However, casein tended towards aggregation at pressures of 302-486 MPa, with a further increase observed beyond 486 MPa.

Effect of electron beam irradiation on the structural characteristics and functional properties of goat's milk casein.

The effects of electron beam irradiation (EBI) at different doses (0, 2, 4, 6, 8, and 10 kGy) were investigated on the structural and functional properties of casein, including their interrelationship. A gradual reduction in the α-helix content of the secondary structure (as a stable structure) indicates that casein under EBI treatment mainly undergoes fragmentation and aggregation from a structural perspective. Furthermore, the hydrophobic group and tryptophan in the tertiary structure were exposed, which opened up the internal structure of the protein. In addition, a continuously increasing irradiation dose led to casein aggregation, as confirmed by electron microscopy. The structural changes affected its functional properties, such as solubility, emulsification, foaming, and rheological properties, all of which increased first and subsequently decreased. Finally, at irradiation doses of 4-6 kGy, casein was modified to exhibit optimal functional properties, which enhanced its food processing value and performance.

Fabrication of Citrus bergamia essential oil-loaded sodium caseinate/peach gum nanocomplexes: Physicochemical, spectral, and structural characterization.

`The objective of current research was to encapsulate citrus bergamia essential oil (CBEO) in nanocomplexes composed of sodium caseinate (SC) and peach gum polysaccharide (PG) in various ratios (SC/PG-1:0, 0:1, 1:1, 1:3, and 3:1). The nanocomplexes formed by the combination of SC and PG in a ratio of 1:3 exhibited a zeta potential of -21.36 mV and a PDI of 0.25. The CBEO-loaded SC/PG (1:3) nanocomplexes revealed the maximum encapsulation efficiency (82.47 %) and loading capacity (1.85 %). FTIR also confirmed the secondary structure variations in response to different ratios of CBEO-loaded SC/PG nanocomplexes. In addition, the XRD and fluorescence spectroscopy analysis also revealed structural changes among CBEO nanocomplexes. The thermal capability of CBEO-loaded SC/PG (1:3) nanocomplexes via TGA showed the minimum weight loss among other complexes. SEM and CLSM analysis demonstrated the uniform distribution and spherical morphology of CBEO-loaded SC/PG (1:3) nanocomplexes. The antioxidant activity of free CBEO was significantly improved in CBEO-loaded nanocomplexes. Likewise, the inhibitory activity of CBEO-loaded nanocomplexes exhibited significantly higher antibacterial action against S. aureus and E. coli. The aforementioned perspective suggests that SC/PG nanocomplexes have potent potential to serve as highly effective nanocarriers with a broad spectrum of uses in the pharmaceutical and food sectors.

Microencapsulation of green coffee oil by complex coacervation of soy protein isolate, sodium casinate and polysaccharides: Physicochemical properties, structural characterisation, and oxidation stability.

This study developed a combination method between protein-polysaccharide complex coacervation and freezing drying for the preparation of green coffee oil (GCO) encapsulated powders. Different combinations of soy protein isolate, sodium caseinate, sodium carboxymethylcellulose, and sodium alginate were utilised as wall materials. The occurrence of complexation between the biopolymers were compared to the final emulsion of the individual protein and confirmed by fourier transform infrared spectrometry and X-ray diffraction. The mean diameter and estimated PDI of GCO microcapsules were 72.57-295.00 µm and 1.47-2.02, respectively. Furthermore, the encapsulation efficiency of GCO microcapsules was between 61.47 and 90.01 %. Finally, oxidation kinetics models of GCO and its microcapsules demonstrated that the zero-order model of GCO microcapsules was found to have a higher fit, which could better reflect the quality changes of GCO microcapsules during storage. Different combinations of proteins and polysaccharides exhibited effective oxidative stability against single proteins because of polysaccharide addition. This research revealed that soy protein isolate, sodium caseinate combined with polysaccharides can be used as a promising microencapsulating agent for microencapsulation of GCO, especially with sodium carboxymethylcellulose and sodium alginate, and provided useful information for the potential use of GCO in the development of powder food.