Understanding Stabilization of Oil-in-Water Emulsions with Pea Protein─Studies of Structure and Properties.

This study investigates the stability and structure of oil-in-water emulsions stabilized by pea protein. Of the wide range of emulsion compositions explored, a region of stability at a minimum of 5% w/v pea protein and 30-50% v/v oil was determined. This pea protein concentration is more than what is needed to form a layer covering the interface. X-ray scattering revealed a thick, dense protein layer at the interface as well as hydrated protein dispersed in the continuous phase. Shear-thinning behavior was observed, and the high viscosity in combination with the thick protein layer at the interface creates a good stability against creaming and coalescence. Emulsions in a pH range from acidic to neutral were studied, and the overall stability was observed to be broadly similar independently of pH. Size measurements revealed polydisperse protein particles. The emulsion droplets are also very polydisperse. Apart from understanding pea protein-stabilized emulsions in particular, insights are gained about protein stabilization in general. Knowledge of the location and the role of the different components in the pea protein material suggests that properties such as viscosity and stability can be tailored for various applications, including food and nutraceutical products.

Effects of Sequential Induction Combining Thermal Treatment with Ultrasound or High Hydrostatic Pressure on the Physicochemical and Mechanical Properties of Pea Protein-Psyllium Hydrogels as Elderberry Extract Carriers.

Entrapping bioactive ingredients like elderberry extract in hydrogels improves their stability and functionality in food matrices. This study assessed the effect of sequential thermal treatment with ultrasound (US) or high hydrostatic pressure (HHP) and treatment duration on pea protein-psyllium hydrogels as elderberry extract carriers. Measurements included color parameters, extract entrapment efficiency, physical stability, textural properties, microrheology, FT-IR, thermal degradation (TGA), SEM images, total polyphenols content, antioxidant activity, and reducing power. The control hydrogel was obtained using only thermal induction. Both treatments impacted physical stability by affecting biopolymer aggregate structures. Thermal and US combined induction resulted in hydrogels with noticeable color changes and reduced entrapment efficiency. Conversely, thermal and HHP-combined induction, especially with extended secondary treatment (10 min), enhanced hydrogel strength, uniformity, and extract entrapment efficiency (EE = 33% for P10). FT-IR and TGA indicated no chemical structural alterations post-treatment. Sequential thermal and HHP induction preserved polyphenol content, antioxidant activity (ABTS = 5.8 mg TE/g d.m.; DPPH = 11.1 mg TE/g d.m.), and reducing power (RP = 1.08 mg TE/g d.m.) due to the dense hydrogel structure effectively enclosing the elderberry extract. Sequential thermal and HHP induction was more effective in developing pea protein-psyllium hydrogels for elderberry extract entrapment.

Role of pea protein isolate in modulating pea starch digestibility: insights from physicochemical and microstructural analysis.

BACKGROUND: Understanding the interactions between protein and starch is crucial in revealing the mechanisms by which protein influences starch digestibility. The present study investigated the impact of different contents of pea protein isolate (PPI) on the physicochemical properties and digestibility of pea starch (PS). RESULTS: The results demonstrated that as the content of PPI increased from 0% to 12%, and the digestion of PS decreased by 12.3%. Rheological analysis indicated that PPI primarily interacted with molecular chains of PS through hydrogen bonds. Increasing the content of PPI resulted in a 30.6% decrease in the hardness of the composite gels, accompanied by a 10% reduction in the short-ordered structure of PS. This hindered the formation of molecular aggregation and resulted in a loose and disordered gel network structure. The microstructure confirmed that the attachment of PPI to PS served as a physical barrier, impeding starch digestibility. CONCLUSION: In summary, the primary mechanism by which PPI inhibited PS digestion involved steric hindrance exerted by PPI and its interaction with PS via hydrogen bonds. These findings contribute to a better understanding of the interaction mechanisms between PS and PPI and offer insights for the optimal utilization of pea resources. © 2024 Society of Chemical Industry.

Pea protein isolate-soluble soybean polysaccharides electrostatic assembly: effect of pH, biopolymer mass ratio and heat treatment.

BACKGROUND: In past years, thousands of protein-polysaccharide complexes have been investigated to modify protein characteristics and functionality in food systems. However, the interaction between pea protein isolate (PPI) and soluble soybean polysaccharide (SSPS) has not been thoroughly characterized yet. RESULTS: In the present study, the phase behavior of PPI and SSPS mixtures was analyzed as a function of PPI:SSPS mixing ratio (1:1 to 1:0.10) and pH (7.0 to 2.0), showing that these biopolymers could be electrostatically assembled at 1:1 to 1:0.25 mixing ratios and 4.0 to 3.0 pH values. Then, the characteristics of the PPI-SSPS complexes were studied before and after heating (90 °C and 30 min) by ζ-potential, surface hydrophobicity, protein solubility, particle size distribution and physical stability for 56 days. By lowering the pH and PPI:SSPS mixing ratio, the complexes showed increased solubility, changed 𝜁-potential and higher physical stability. By heating, the complexes presented increased hydrophobicity and physical stability. CONCLUSION: Overall, PPI-SSPS complexes increased the protein solubility, reduced the particle size, and changed both the ζ-potential and the surface hydrophobicity with respect to PPI control, allowing stabilization of the colloidal system and broadening the possible applications of these high-quality proteins in acidic food systems. © 2024 Society of Chemical Industry.

In vitro gastrointestinal simulated digestion of three plant proteins: determination of digestion rate, free amino acids and peptide contents.

Cassava protein (CP), barley protein (BP) and yellow pea protein (YPP) are important nutrient and integral constituent of staple in pet foods. It is known that the digestion of proteins directly influences their absorption and utilisation. In the present work, we performed in vitro simulated gastrointestinal digestion of three plant proteins as a staple for dog and cat food. The digestion rate of CP, BP and YPP in dog food was 56.33 ± 0.90%, 48.53 ± 0.91%, and 66.96 ± 0.37%, respectively, whereas the digestion rate of CP, BP, and YPP in cat food was 66.25 ± 0.72%, 43.42 ± 0.83%, and 58.05 ± 0.85%, respectively. Using SDS-polyacrylamide gel electrophoresis to determine the molecular weight (MW) of each protein and the products of their digestion, it was revealed that MW of digestion samples decreased, and MW during the small intestine phase was lower than that during the gastric phase. Peptide sequences of digested products were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and it was found that the total number of peptides in the small intestine digestion samples was higher than that in the gastric phase samples. The MW of peptides obtained from CP was within the range of 1000-1500 Da, while MW of peptides derived from BP and YPP was within the range of 400-2000 Da. In addition, free amino acids were mainly produced in the small intestine phase. Furthermore, the percentage of essential amino acids in the small intestine phase (63 ~ 82%) was higher than that in the gastric phase (37 ~ 63%). Taken together, these findings contribute to the current understanding of the utilisation of plant proteins in dog and cat foods and provide important insights into the selection and application of plant proteins as a staple in dog and cat foods.

Characterization, Bioaccesibility and Antioxidant Activities of Phenolic Compounds Recovered from Yellow pea (Pisum sativum) Flour and Protein Isolate.

This study focused on studying the bioaccesible phenolic compounds (PCs) from yellow pea flour (F) and protein isolate (I). Total phenolic contents (TPC), PCs composition and antioxidant activities were analysed in ethanol 60% extracts obtained by applying ultrasound assisted extraction (UAE, 15 min/40% amplitude). The preparation of I under alkaline conditions and the elimination of some soluble components at lower pH produced a change of PCs profile and antioxidant activity. After simulated gastrointestinal digestion (SGID) of both ingredients to obtain the digests FD and ID, notable changes in the PCs concentration and profiles could be demonstrated. FD presented a higher ORAC activity than ID (IC50 = 0.022 and 0.039 mg GAE/g dm, respectively), but lower ABTS•+ activity (IC50 = 0.8 and 0.3 mg GAE/g dm, respectively). After treatment with cholestyramine of extracts from FD and ID in order to eliminate bile salts and obtain the bioaccesible fractions FDb and IDb, ROS scavenging in H2O2-induced Caco2-TC7 cells was evaluated, registering a greater activity for ID respect to FD (IC50 = 0.042 and 0.017 mg GAE/mL, respectively). These activities could be attributed to the major bioaccesible PCs: OH-tyrosol, polydatin, trans-resveratrol, rutin, (-)-epicatechin and (-)-gallocatechin gallate for FD; syringic (the most concentrated) and ellagic acids, trans-resveratrol, and (-)-gallocatechin gallate for ID, but probably other compounds such as peptides or amino acids can also contribute.

Fabrication and Characterization of Chitosan-Pea Protein Isolate Nanoparticles.

Chitosan (CS) and pea protein isolate (PPI) were used as raw materials to prepare nanoparticles. The structures and functional properties of the nanoparticles with three ratios (1:1, 1:2 1:3, CS:PPI) were evaluated. The particle sizes of chitosan-pea protein isolate (CS-PPI) nanoparticles with the ratios of 1:1, 1:2, and 1:3 were 802.95 ± 71.94, 807.10 ± 86.22, and 767.75 ± 110.10 nm, respectively, and there were no significant differences. Through the analysis of turbidity, endogenous fluorescence spectroscopy and Fourier transform infrared spectroscopy, the interaction between CS and PPI was mainly caused by electrostatic mutual attraction and hydrogen bonding. In terms of interface properties, the contact angles of nanoparticles with the ratio of 1:1, 1:2, and 1:3 were 119.2°, 112.3°, and 107.0°, respectively. The emulsifying activity (EAI) of the nanoparticles was related to the proportion of protein. The nanoparticle with the ratio of 1:1 had the highest potential and the best thermal stability. From the observation of their morphology by transmission electron microscopy, it could be seen that the nanoparticles with a ratio of 1:3 were the closest to spherical. This study provides a theoretical basis for the design of CS-PPI nanoparticles and their applications in promoting emulsion stabilization and the delivery of active substances using emulsions.

Impact of Ageing on Pea Protein Volatile Compounds and Correlation with Odor.

Vegetal proteins are of high interest for their many positive aspects, but their 'beany' off-flavor is still limiting the consumer's acceptance. The aim of this work was to investigate the conservation of pea protein isolate (PPI) during time and especially the evolution of their organoleptic quality under two storage conditions. The evolution of the volatile compounds, the odor and the color of a PPI has been investigated during one year of storage. PPI was exposed to two treatments mimicking a lack of control of storage conditions: treatment A with light exposition at ambient temperature (A-Light 20 °C) and treatment B in the dark but with a higher temperature (B-Dark 30 °C). For each sampling time (0, 3, 6, 9, 12 months), the volatile compounds were determined using HS-SPME-GC-MS, the odor using direct sniffing, and the color using the measurement of L*, a*, b* parameters. Treatment A was the most deteriorating and led to a strong increase in the total volatile compounds amount, an odor deterioration, and a color change. Furthermore, a tentative correlation between instrumental data on volatile compounds and the perceived odor was proposed. By the representation of volatile compounds sorted by their sensory descriptor, it could be possible to predict an odor change with analytical data.

Influence of protein extraction and texturization on odor-active compounds of pea proteins.

BACKGROUND: The use of plant proteins as food ingredients might be limited due to the presence of foreign or 'off' flavors, which may evolve during extraction and subsequent processing. In this study, the influence of dry (TVP) and wet (WTP) texturization on characteristic volatile compounds of two different pea protein isolates was assessed using gas chromatography-mass spectrometry-olfactometry (GC-MS-O) after direct immersion stir bar sorptive extraction (DI-SBSE). RESULTS: Twenty-four odor-active compounds were found, with a prevalence of carbonyls from fat oxidation. Nine of these compounds which are also known as major (off-) flavor contributors in peas were distinctively impacted in all texturates: hexanal, nonanal, 2-undecanone, (E)-2-octenal, (E, Z)-3,5-octadiene-2-one, (E, E)-2,4-decadienal, 2-pentyl-furan, 2-pentyl-pyridine, and γ-nonalactone. For example, hexanal, a characteristic green odorant, was reduced by up to sixfold by wet texturization, from 3.29 ± 1.05% (Pea Protein I) to 0.52 ± 0.02% (Pea WTP I). Furthermore, (E,Z)-3,5-Octadiene-2-one and (E,E)-2,4-decadienal were decreased by 1.5- and 1.8-fold when Pea Protein I and Pea TVP I were compared. CONCLUSION: An overall reduction in fat oxidation products and of green and fatty odor-active compounds was observed. The results represent a first insight into the process-related modulation of pea protein (off-) flavors to broaden the applicability of pea proteins as food ingredients.

The structure and stability analysis of the pea seed legumin glycosylated by oligochitosan.

BACKGROUND: The functionality of pea proteins is relatively weak relative to that of soybean proteins, which limits the application of pea proteins in food and nutritional applications. Glycosylation is a promising approach to influence the protein structure and in turn change the functional properties of pea proteins. RESULTS: In this study, the effect of transglutaminase-induced oligochitosan glycosylation on the structural and functional properties of pea seed legumin was studied. Different oligochitosan-modified legumin complexes (OLCs) were prepared by applying different molar ratios of legumin to oligochitosan (1:1 to 1:4) induced by transglutaminase (10 U g-1 protein). Results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), glucosamine, and free amino analysis showed that the legumin could be covalently bonded with the oligochitosan and were influenced by the applying dose of the oligochitosan. Infrared spectroscopy, fluorescence, and scanning electron microscopy analysis indicated that the structure of the different OLC samples could be changed to different extents. Moreover, although the emulsifying activity decreased, the emulsification stability, thermal stability, and in vitro digestive stability of the OLCs were remarkably improved relative to that of the untreated legumin. CONCLUSION: Oligochitosan glycosylation could change the structure of the legumin and consequently improve its emulsification stability, thermal stability, and in vitro digestive stability. This study will facilitate the legumin functionalization by the glycosylation approach to fabricate protein-oligochitosan complex for potential food and nutritional applications. © 2020 Society of Chemical Industry.

Composition, physicochemical properties of pea protein and its application in functional foods.

Field pea is one of the most important leguminous crops over the world. Pea protein is a relatively new type of plant proteins and has been used as a functional ingredient in global food industry. Pea protein includes four major classes (globulin, albumin, prolamin, and glutelin), in which globulin and albumin are major storage proteins in pea seeds. Globulin is soluble in salt solutions and can be further classified into legumin and vicilin. Albumin is soluble in water and regarded as metabolic and enzymatic proteins with cytosolic functions. Pea protein has a well-balanced amino acid profile with high level of lysine. The composition and structure of pea protein, as well as the processing conditions, significantly affect its physical and chemical properties, such as hydration, rheological characteristics, and surface characteristics. With its availability, low cost, nutritional values and health benefits, pea protein can be used as a novel and effective alternative to substitute for soybean or animal proteins in functional food applications.

Application of Maillard reaction product of xylose-pea protein enzymatic hydrolysate in 3D printing.

BACKGROUND: This paper aims to provide a basis for the application of Maillard reaction products and ideas for 3D printing of functional food. RESULTS: We evaluated the effects of pea protein enzymatic hydrolysate-xylose Maillard reaction products (xMRPs) on the printability of 3D printing materials and the structure using various techniques. It was found that compared with the addition of enzymatic hydrolysate, the materials with xMRPs and 3D printing could significantly improve the structure, physical properties, formability of 3D printing materials and 3D print products, among which the xMRPs with 6 g of xylose had the most significant effect. CONCLUSIONS: This study has important scientific value and practical significance for the high-valued application of Millard reaction products and the promotion of the practical application of 3D printing technology in the food industry. © 2020 Society of Chemical Industry.

Pea Protein for Hempseed Oil Nanoemulsion Stabilization.

In this paper, we present the possibility of using pea protein isolates as a stabilizer for hempseed oil (HSO)-based water/oil emulsions in conjunction with lecithin as a co-surfactant. A Box-Behnken design was employed to build polynomial models for optimization of the ultrasonication process to prepare the emulsions. The stability of the system was verified by droplet size measurements using dynamic light scattering (DLS) as well as centrifugation and thermal challenge tests. The z-ave droplet diameters of optimized emulsion were 209 and 207 nm after preparation and 1 week storage, respectively. The concentration of free Linoleic acid (C18:2; n-6) was used for calculation of entrapment efficiency in prepared nanoemulsions. At optimum conditions of the process, up to 98.63% ± 1.95 of entrapment was achieved. FTIR analysis and rheological tests were also performed to evaluate the quality of oil and emulsion, and to verify the close-to-water like behavior of the prepared samples compared to the viscous nature of the original oil. Obtained results confirmed the high impact of lecithin and pea protein concentrations on the emulsion droplet size and homogeneity confirmed by microscopic imaging. The presented results are the first steps towards using hempseed oil-based emulsions as a potential food additive carrier, such as flavor. Furthermore, the good stability of the prepared nanoemulsion gives opportunities for potential use in biomedical and cosmetic applications.

Chemical acylation of pea protein isolate hydrolysate with fatty acid N-hydroxysuccinimide esters: Effect on structure and functional properties.

Applications of pea protein in the food industry have been greatly restricted by its poor functional properties. In order to solve this problem, a novel technique combining enzymatic hydrolysis and fatty acid acylation has been applied in this work to construct a pea protein-fatty acid covalent complex that aims to improve its functional properties. The processed pea protein with increased water solubility tends to decrease the chance of self-aggregation. Additionally, emulsifying and antioxidant properties have also been found after this process. On top of that, the modified pea protein has been characterized by Fourier transform infrared and circular dichroism spectroscopy. These results demonstrate that these properties were mainly caused by the acylation of the amino group from hydrolyzed pea protein and the carboxyl group from the fatty acid. The enzymatic hydrolysis/fatty acid acylation research provides insights into manufacturing high-quality functional lipoproteins from inexpensive pea protein for the food industry.

Structural evolution of pea-derived albumins during pH and heat treatment studied with light and X-ray scattering.

Pea albumins are found in the side stream during the isolation of pea proteins. They are soluble at acidic pH and have functional properties which differ from their globulin counterparts. In this study, we have investigated the aggregation and structural changes occurring to pea albumins under different environmental conditions, using a combination of size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALS) and small-angle X-ray scattering (SAXS). Albumins were extracted from a dry fractionated pea protein concentrate by precipitating the globulin fraction at acidic pH. The albumins were then studied at different pH (3, 4, 4.5, 7, 7.5, and 8) values. The effect of heating at 90 °C for 1, 3, and 5 min on their structural changes was investigated using SAXS. In addition, size exclusion of the albumins showed 4 distinct populations, depending on pH and heating conditions, with two large aggregates peaks (∼250 kDa): a dimer peak (∼24 kDa) containing predominantly pea albumin 2 (PA2), and a monomer peak of a molar mass of about 12 kDa (PA1). X-ray scattering intensities as a function of q were modeled as polydisperse spheres, and their aggregation was followed as a function of heating time. Albumins was most stable at pH 3, showing no aggregation during heat treatment. While albumins at pH 7.5 and 8 showed aggregation after heating, solutions at pH 4, 4.5, and 7 already contained aggregates even before heating. This work provides new knowledge on the overall structural development of albumins under different environmental conditions, improving our ability to employ these as future ingredients in foods.

Plant and animal protein mixed systems as wall material for microencapsulation of Manuka essential Oil: Characterization and in vitro release kinetics.

Combination of plant and animal protein diet is becoming a valuable source of nutrition in the modern diet due to the synergistic functional properties inherent in these protein complexes. Moreover, the synergy between animal and plant proteins can contribute to the high stability and improved solubility of the encapsulated bioactive ingredients (e.g., essential oils). Therefore, the study was designed to evaluate the plant (pea protein (PP) and lupine protein (LP)) and animal protein (whey protein, WP) mixed systems as a wall material for microencapsulation of manuka essential oil, as an example of bioactive compound. Moreover, physicochemical properties and in vitro release profile of encapsulated manuka essential oil were studied. Manuka essential oil microcapsules exhibited low moisture content (5.3-7.1 %) and low water activity (0.33-0.37) with a solubility of 53.7-68.1 %. Change in wall material ratio significantly affected the color of microcapsules, while microcapsules prepared with 1:1 protein/oil ratio demonstrated a high encapsulation efficiency (90.4 % and 89.4 %) for protein mixed systems (PP + WP and LP + WP), respectively. Microcapsules further showed low values for lipid oxidation with a high oxidative stability and antioxidant activity (62.1-87.0 %). The zero order and Korsmeyer-Peppas models clearly explained the release mechanism of encapsulated oil, which was dependent on the type and concentration of the protein mixed used. The findings demonstrated that the protein mixed systems successfully encapsulated the manuka essential oil with controlled release and high oxidative stability, indicating the suitability of the protein mixed systems as a carrier in encapsulation and application potential in development of encapsulated functional foods.

Solubility, (micro)structure, and in vitro digestion of pea protein dispersions as affected by high pressure homogenization and environmental conditions.

In this work, dispersions were prepared with commercial pea protein isolate (PPI) and subjected to different (i) high pressure homogenization (HPH) intensities (0 - 200 MPa) (room temperature, pH 7) or (ii) environmental conditions (60 °C, pH 7 or pH 12) to generate dispersions with distinct protein molecular and microstructural characteristics, impacting protein solubility. Besides, protein digestion was analyzed following the static INFOGEST in vitro digestion protocol. Generally, increasing pressure of the homogenization treatment was linked with decreasing particle sizes and enhanced protein digestion. More specifically, the dispersion that did not undergo HPH (0 MPa) as well as the dispersion treated at 60 °C, pH 7, had highly similar microstructures, consisting of large irregular particles (10 - 500 µm) with shell-like structures, and exhibited low solubility (around 15 % and 28 %, respectively), which resulted in limited proteolysis (35 % and 42 %, respectively). In contrast, the dispersion subjected to HPH at 100 MPa and the dispersion treated at 60 °C, pH 12 also had similar microstructures with small and homogeneous particles (<1 µm), and exhibited relatively good solubility (54 % and 31 %, respectively), which led to enhanced protein digestion levels (87 % and 74 %, respectively). This study highlights the potential of food processing on macronutrient (micro)structure and further gastrointestinal stability and functionality.

Enhancing the usability of pea protein in emulsion applications through modification by various approaches: A comparative study.

The extensive utilization in food industry of pea protein is often impeded by its low water solubility, resulting in poor functional properties. Various methods, including pH-shifting (PS), ultrasonication (US), high-pressure micro-fluidization (MF), pH-shifting combined with ultrasonication (PS-US), and pH-shifting with micro-fluidization (PS-MF), were utilized to modify pea protein isolate (PPI) in order to enhance its functionality in emulsion formulation. The physicochemical properties and structural changes of the protein were investigated by assessing solubility, particle size, surface charge, protein profile, surface hydrophobicity, free sulfhydryl groups, and secondary structure content. The extent of modification induced by each treatment method on PPI-stabilized emulsions was compared based on parameters such as adsorbed interfacial protein concentration, particle size, zeta potential, and microstructure of the prepared emulsions. All modification increased the solubility of pea protein in the sequence of PS (4-fold) < MF (7-fold) < US (11-fold) < PS-US (13-fold) < PS-MF (14-fold). For single treatments, proteins dissolved more readily under US, resulting in the most uniform emulsions with small particle. The combined processes of PS-US and PS-MF further improved solubility, decreased emulsions particle size, promoted uniformity of emulsions. PS-US-stabilized emulsions displayed more smaller droplet size, narrower size distribution, and slightly higher stability than those prepared by PS-MF. The relatively higher emulsifying capacity of PPI treated by PS-US than those by PS-MF may be attributed to its higher surface hydrophobicity.

Acid-induced pea protein gels pretreated with media milling: Gelling properties and the formation mechanism.

This study explored the effect of stirred media mill (SMM) processing on the acid-induced gelling properties of pea protein. Results showed that SMM treatment enhanced the gel strength from 75.06 g to 183.89 g and increased the water holding capacity from 46.64 % to 73.50 %. The minimum gelation concentration achieved for SMM-treated pea protein was 4 %, significantly lower than that of heat-pretreated pea protein (9 %). SMM decreased protein aggregate size from 104 µm to 180 nm. Microscopy analysis revealed that the small aggregates facilitated the formation of uniform gel networks with tight connections. Linear rheology indicated that small protein aggregates resulted in slower gelation rates with a higher G' for the formed gels. The SMM-pretreated protein gel showed strain hardening, shear thinning behaviors, and satisfactory stability to withstand large-amplitude oscillatory shear. Overall, SMM emerges as a promising technology for producing protein gel products with strong mechanical attributes and customizable rheological properties.

Pea protein/carboxymethyl cellulose complexes prepared using a pH cycle strategy as stabilizers of high internal phase emulsions for 3D printing.

The development of food-grade high internal phase emulsions (HIPEs) for 3D printing and the replacement of animal fats have attracted considerable attention. In this study, in order to improve the rheological properties and stability of pea protein to prepare HIPE, pea protein/carboxymethyl cellulose (pH-PP/CMC) was prepared and subjected to pH cycle treatment to produce HIPEs. The results showed that pH cycle treatment and CMC significantly reduced the droplet size of HIPEs (from 143.33 to 12.10 µm). At higher CMC concentrations, the interfacial tension of the PP solution decreased from 12.84 to 11.71 mN/m without pH cycle treatment and to 10.79 mN/m with pH cycle treatment. The HIPEs with higher CMC concentrations subjected to pH cycle treatment showed shear thinning behavior and higher viscoelasticity and recovered their solid-like properties after being subjected to 50 % strain, indicating that they could be used for 3D printing. The 3D printing results showed that the pH-PP/CMC HIPE with 0.3 % CMC had the finest structure. Our work provides new insights into developing food-grade HIPEs and facilitating their use in 3D printing inks as nutrient delivery systems and animal fat substitutes.