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Preparation and application of natural protein polymer-based Pickering emulsions

  • Qianqian Ma , Sensen Ma , Jie Liu , Ying Pei , Keyong Tang , Jianhua Qiu , Jiqiang Wan , Xuejing Zheng EMAIL logo and Jun Zhang EMAIL logo
Published/Copyright: April 25, 2023
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e-Polymers
From the journal e-Polymers Volume 23 Issue 1

Abstract

Emulsification is the effect of a liquid being uniformly dispersed as tiny droplets in another liquid that is immiscible. Traditional emulsification requires the addition of suitable surfactant to stabilize the emulsion. When the surfactant molecules are replaced by solid particles, the emulsion is known as Pickering emulsions (PEs). PEs with dispersed phase volume fraction above 74% are also named high internal phase Pickering emulsions (HIPPEs). The use of solid particles of natural origin allows PEs to be highly physically stable, environmentally compatible, and biodegradable compared to traditional emulsions. Among them, protein-based solid particles are well suited to stabilize PEs for their great emulsification properties and nutritional value. In this article, we reviewed the preparation of different forms of proteins and their emulsion stabilization properties and summarized the applications of protein-based PEs in various fields, including food, biomedicine, porous materials, biodegradable packaging films, sewage treatment, 3D printing, etc.

Graphical abstract

Particle morphology and applications of protein-based PEs.

Keywords: Pickering emulsion; natural polymer; morphology; synthesis; application

1 Introduction

Emulsions are created by dispersing one liquid (dispersed phase) into another liquid (continuous phase) by means of tiny droplets (typically 0.1–10 µm) (1). Traditional emulsions stabilized by amphiphilic polymers or surfactants are thermodynamically unstable and delaminated by various physical or chemical forces (2). Pickering (3) discovered in the early 1900s that solid particles could help stabilize the emulsion polymerization process. Such solid particles were named Pickering particles, and emulsions stabilized by solid particles were called Pickering emulsions (PEs). PEs with dispersed phase volume higher than 74% were called high internal phase Pickering emulsions (HIPPEs) (4). In contrast to small molecule surfactants, solid particles in PEs can act as a physical barrier with size exclusion that prevents droplet contact and interfacial interactions. PEs have a higher surface loading and thickness than traditional emulsions, and its higher stability has potential applications for encapsulation and controlled release of active substances (5). Therefore, in-depth research on Pickering particles, commercial production, and practical applications is required.

Not all polymers are suitable as Pickering particles since the particles must maintain their insolubility and integrity in both phases of the emulsion over the life of the emulsion system (6,7). Studies have shown that considerable deformability in terms of shape, aspect ratio, and morphology can be found in naturally sourced or biodegradable nanoparticles, which can reduce interfacial tension and thus stabilize emulsions (8). Proteins can be easily prepared as particles for PEs stabilization due to their amphiphilic nature and simple conformational adjustment (9). The use of proteins and their derivatives from different sources can facilitate the production of emulsions with different properties. In addition, proteins can also be physically, chemically, or enzymatically modified by changing temperature (10,11), pH (12), particle strength (13), and shear stress (14) to enhance their solubility and functional properties. Emulsions can also be stabilized more effectively by forming protein–polysaccharide (5), protein–polyphenol (15), and protein–protein (16) complex particles.

This review explored the effects and applications of protein polymer stabilized PEs. The current preparation and emulsion stabilization capabilities of different forms of protein particles are discussed. Application areas for protein-based PEs are also presented, including food, biomedicine, packaging materials, 3D printing, etc. This review could provide comprehensive information for the understanding and development of protein-based PEs.

2 Types of protein particles for PE stabilization

2.1 Protein nanoparticles

Plant- and animal protein-based nanoparticles, which can be prepared by solvent evaporation (17), pH-driven (12), or antisolvent precipitation (18) methods, have been widely used to stabilize PEs. Some proteins that are inherently amphiphilic can be used directly as emulsion stabilizers and can have high storage stability for bioactive compounds without further treatment. The easily cultured and high yielding spirulina proteins have inherent emulsification ability (19). By forming spirulina protein into a gel through thermal coagulation followed by controlled shearing and high-pressure homogenization, the obtained spirulina protein nanoparticles showed good emulsion stability, which opened a new way to design novel plant-based products in the future (20). HIPPEs stabilized by gelatin were reported by Tan et al. (21), which were able to improve β-carotene bioavailability. The droplet size distribution, morphology, and digestive properties of HIPPEs can be readily adjusted by the concentration of gelatin particles. Due to the self-assembly behavior of the molecular chains, the regenerated silk fibroin leads to the predominance of folded β-sheet secondary structures after regeneration. The use of regenerated nanosilk as stabilizers for PEs could avoid unstable structural transformations during practical use. PEs insensitive to environmental stresses such as heating and high ionic strength but sensitive to pH changes can be obtained (22).

However, some natural proteins cannot provide sufficient PEs stability due to insufficient hydrophilicity or hydrophobicity, requiring physical treatment (23), cross-linking (24), thermal denaturation (25), and other methods to improve their PEs stabilizing ability. Gelatin was cross-linked with sodium alginate using calcium ions (Ca2+) and transglutaminase (TG) to form gelatin–sodium alginate cohesive particles (Figure 1a). The PEs stabilized at pH = 3.5 had surface loading protein capacity (97.4 ± 5.0%), high oil sealing efficiency (91.8 ± 2.0%), and viscoelasticity (26). PEs were prepared by cross-linking potato protein dissolved in the water phase and zein dissolved in the oil phase by TyrBm (tyrosinase from Bacillus megaterium). The enzymatically cross-linked emulsions were stable for more than a month without significant separation compared to the uncross-linked emulsions, probably due to covalent forces between the low-molecular-weight potato protein and the α-zein fraction (27). In addition, proteins can be combined with various polyphenols or polysaccharides to better stabilize PEs. The electrostatic complexes of chitosan (COS) and glycosylated whey protein isolate (gWPI) were used as emulsifiers to stabilize PEs, which can be used as effective camellia oil carrier systems (Figure 1b). The emulsions showed good oxidative stability, thermal stability, and shear resistance (28). Macadamia protein isolates (MPI) have lower oil holding capacity and dispersion in the aqueous phase compared with other oilseed proteins, such as peanut and almond (29). As a positively charged chitosan derivative, chitosan hydrochloride can act as a co-stabilizer, enhancing the physicochemical stability of emulsions and modifying interfacial, rheological, or gel properties. PEs stabilized by MPI with chitosan hydrochloride complex polymer (MCCP) through a percolating network structure has higher stability and viscosity (30), which facilitates the creation of gel-like emulsion systems (Figure 1c). Nanoparticles prepared by non-covalent bonding of whey protein isolate (WPI) and phytosterols exhibited good interfacial wettability and antioxidant activity. PEs stabilized by the composite particles exhibited good storage and oxidative stability, and promoted the process of reducing lipid digestion (31) (Figure 1d).

Figure 1 (a) Diagram representation of the gliadin–SA complex formation cross-linked by TG and Ca2+ (26). Copyright © 2020; Elsevier. (b) Diagram of the electrostatic complex of gWPI with COS used to stabilize the PEs (28). Copyright © 2021; Elsevier. (c) Illustration of the preparation of MPI and MCCP stabilized emulsions, respectively (30). Copyright © 2020; Elsevier. (d) Diagram of the preparation of WPI and PS composite nanoparticle stabilized emulsion and can be used for lipid digestion (31). Copyright © 2022; Elsevier.
Figure 1

(a) Diagram representation of the gliadin–SA complex formation cross-linked by TG and Ca2+ (26). Copyright © 2020; Elsevier. (b) Diagram of the electrostatic complex of gWPI with COS used to stabilize the PEs (28). Copyright © 2021; Elsevier. (c) Illustration of the preparation of MPI and MCCP stabilized emulsions, respectively (30). Copyright © 2020; Elsevier. (d) Diagram of the preparation of WPI and PS composite nanoparticle stabilized emulsion and can be used for lipid digestion (31). Copyright © 2022; Elsevier.

2.2 Protein microgels

Protein microgels, assembled by protein interactions, are highly hydrated porous surface-active particles with high stability and can be used as stabilizers for PEs preparation. Protein microgels can be cross-linked to form structured networks to trap emulsion droplets (32). Commonly used animal proteins for preparing gel particles include whey protein (33), gelatin (34), casein (35), and collagen (36); whereas, plant proteins include soy protein (37) and zein (38). PEs stabilized with casein gel particles exhibited higher stability and resistance to agglomeration than emulsions stabilized with casein only, primarily due to the greater density and higher spatial potential resistance of casein gel particles at the interface (35). Whey protein microgels (WPM) of various stiffnesses could be prepared from 5%, 10%, and 20% (w/w) WPI. The soft microgel particles (5%) adsorbed at the water–oil interface exhibited good deformability compared to the stiffer particles (10% and 20%). In addition, the soft gel particles exhibited higher interfacial coverage, faster interfacial adsorption rate, and excellent ability to reduce interfacial tension. PEs stabilized by soft WPM exhibited high stability and delayed lipid digestibility (39).

Gel particles can be prepared by separating and breaking the gel or colloidal particle solution using mechanical methods, such as extrusion (40), spray drying (41), shearing (42), and high-pressure homogenization (1), to obtain solutions containing micron or nano-sized gel particles (43). Soy protein isolate (SPI)-based gel particles can be prepared by a two-step method. Soy protein was first gelated with TG, and then microgel particles were obtained in a homogenizer (Figure 2). Compared with SPI, microgel particles with neutral wettability showed strong adsorption on the surface of oil droplets, and emulsions stabilized by microgels had higher storage stability (44). Based on this method, peanut protein isolate (PPI) (45) and bamboo fungus proteins (46) were also used to prepare microgel particles. The TG cross-linked PPI microgel particles could also be combined with xanthan gum to stabilize the PEs through electrostatic interactions. The presence of xanthan gum could increase the apparent viscosity of the emulsion, giving it good storage stability (47).

Figure 2 (a) Flow chart for the preparation of SPI microgel particles. (b) Zeta-potential, hydrodynamic diameter, and surface morphology of microgel particles at various pH conditions. (c) SEM of microgel particles captured from the water–oil interface. (d) Protein surface load (Γ s) of PEs at various pH conditions (44). Copyright © 2016; Elsevier.
Figure 2

(a) Flow chart for the preparation of SPI microgel particles. (b) Zeta-potential, hydrodynamic diameter, and surface morphology of microgel particles at various pH conditions. (c) SEM of microgel particles captured from the water–oil interface. (d) Protein surface load (Γ s) of PEs at various pH conditions (44). Copyright © 2016; Elsevier.

2.3 Protein fibrils

Protein fibrils are anisotropic particles that can be used as stabilizers in PEs. Proteins can be hydrolyzed into specific peptides, which form protofibrils through self-assembly (48). Due to the large number of polar groups of the amino acid residue on the protein molecular chains, proteins are usually electrically charged. The electrostatic repulsion between peptides causes the protofibril chains to grow laterally rather than longitudinally (49). The properties of protein protofibrils are controlled by the balance of electrostatic interactions, hydrogen bonding, and hydrophobic bonding. Anisotropic protofibrils have a specific interfacial behavior due to the capillary forces between the fibrils (50,51). Whey protein nanogenic fibrils (WPNFs) with large aspect ratio and flexibility could bend over the droplet surface for better encapsulation of oil droplets, as low as 5 wt% of WPNFs exhibited good stability to HIPPEs with internal phase fractions up to 90% (52). Complexes of WPI protofibrils with cellulose nanocrystals could also be prepared by electrostatic complexation. The introduction of cellulose nanocrystals could form a thick charged coating around the oil droplets. Emulsions stabilized by the complexes showed greater resistance to creaming than those stabilized by WPI alone. The self-supporting gel emulsions formed at 70% oil concentration could protect curcumin from photodegradation and thermal degradation, potentially serving as a curcumin delivery system (53).

The structure and morphology of protein protofibrils are controlled by the concentration and type of protein, temperature, ionic strength, and pH (54,55). Ovotransferrin fibrils with high aspect ratio stabilized PEs were shown to be highly stable at different ionic strengths (0–1,000 mM) and pH values (2–7) (56). WPI fibrils could be effectively adsorbed at the oil–water interface to stabilize the oil droplets. The cross-sectional radius of the highly pH-dependent WPI fibrils increased from 1.87 ± 0.12 to 7.75 ± 0.33 nm when the pH was increased from 2.0 to 5.0. As the pH increased to 7.0, the fibrils decreased to a single strand (57). Emulsions stabilized by β-lactoglobulin protofibrils exhibited excellent physical stability at concentrations from 5 to 20 mg·mL−1 and at pHs far from the isoelectric point (pI) of β-lactoglobulin. While at pHs close to the pI of β-lactoglobulin (pH = 5.2) or high protofibril concentrations (≥25 mg·mL−1), protofibrils tended to aggregate into emulsions with large droplet sizes (58) (Figure 3).

Figure 3 (a) Schematic illustration of the influence of β-lactoglobulin fibril concentration on the structure of PEs. Storage stability of PEs stabilized by β-lactoglobulin fibrils at different pH conditions (b) or at different fibril concentrations at pH 7.0 (c) (58). Copyright © 2016; Elsevier.
Figure 3

(a) Schematic illustration of the influence of β-lactoglobulin fibril concentration on the structure of PEs. Storage stability of PEs stabilized by β-lactoglobulin fibrils at different pH conditions (b) or at different fibril concentrations at pH 7.0 (c) (58). Copyright © 2016; Elsevier.

2.4 Protein nanogels

Protein nanogels can be prepared by covalent cross-linking of self-assemblies or by simple thermal denaturation methods (59). As shown in Figure 4, the acylated rapeseed isolated protein nanogels were prepared using a simple thermal denaturation self-assembly method. The nanogels were effective in stabilizing droplets at pH = 5.5–8.5 and salt concentrations up to 0.2 M. PEs stabilized by acylated rapeseed isolate protein nanogels at concentrations higher than 0.5% showed storage stability up to 30 days (60). Casein nanogels prepared by cross-linking could be used as effective stabilizers for HIPPEs without any other additives. Casein nanogel concentration, ionic strength, and pH affected the droplet size distribution of the emulsion, and the gel state rheological properties of the emulsion could be used as a carrier for the hydrophobic drug indomethacin for controlled release purposes (61). Araiza-Calahorra and Sarkar (62) found that 1.0 wt% of nanogel particles were enough to form monolayers at the oil–water interface. PEs stabilized by whey protein nanogels not only resisted aggregation for more than a year, but also served as delivery carriers for curcumin. The desalted duck egg white nanogels prepared by top-down method could be used as Pickering emulsifiers (63). The introduction of tannic acid (64), sodium alginate (65), and κ-carrageenan (66) made the desalted duck egg white nanogel particles close to neutral wettability and had great potential as a high-quality food grade emulsifier.

Figure 4 (a) Diagram of the preparation of PEs stabilized by acylated rapeseed isolate protein nanogels. Storage stability of PEs stabilized by acylated rapeseed isolate protein nanogels at different oil fractions (b) or different nanogel concentrations (c) (60). Copyright © 2020; American Chemical Society.
Figure 4

(a) Diagram of the preparation of PEs stabilized by acylated rapeseed isolate protein nanogels. Storage stability of PEs stabilized by acylated rapeseed isolate protein nanogels at different oil fractions (b) or different nanogel concentrations (c) (60). Copyright © 2020; American Chemical Society.

2.5 Other types of protein-based particles

In addition to the protein types mentioned above, other structural types of protein particles have been developed to stabilize PEs (67). Compared to solid zein protein nanoparticles, hollow zein protein nanoparticles with diameters less than 100 nm have also been developed to stabilize emulsions. More sustained and controlled release properties during the release of metformin were reported (68). Self-assembling tobacco mosaic virus (TMV) capsid protein (TMVCP) into TMV-like nanorods could be used for long-term stabilization of PEs. As shown in Figure 5a, two steps were involved: the adsorption of amphiphilic TMVCP at the water–oil interface and the self-assembly of TMVCP into nanorods with enlarged size. Combining the amphiphilic nature of TMVCP with the advantages of in situ self-assembly for size growth, PEs stabilized with virus-like nanorods had considerable potential for virus recognition and drug delivery (69). E2 protein is part of the pyruvate dehydrogenase multi-enzyme complex found in thermophilic bacterium Geobacillus stearothermophilus. Sarker et al. (70) proposed dodecahedral hollow protein nanocages self-assembled by 60 E2 protein subunits as pH-switchable Pickering emulsifiers (Figure 5b). The nanocage could effectively stabilize the emulsion at pH ≥ 7.0, while the emulsion delaminated significantly at pH < 7.0. PEs can also be prepared from plant-based nano-assemblies by high shear homogenization. Nasrabadi et al. (71) prepared a composite nanocomponent stabilized emulsion using flaxseed protein and flaxseed soluble mucilage. The addition of flaxseed soluble mucilage reduced the electrostatic repulsion between protein molecules, so the composite nanocomponent formed a protective layer on the surface of the oil droplets to prevent flocculation and agglomeration. This plant-based composite nanocomponent has the potential to be a natural food-grade Pickering stabilizer. Hu et al. (72) used 3D brush-like silk nanostructures as stabilizers to prepare PEs. The lowest interfacial tension value of the silk nanobrush was 21.27 mN·m−1, and the concentration of 0.3 wt% was able to form a complex interconnected network in the oil–water interface, which remained stable at an optimal ultrasonic intensity of 40%. The biocompatible silk nanobrush has the potential to replace all-natural green emulsions in the food and biomedical fields.

Figure 5 (a) Schematic diagram of TMVCP self-assembled into nanorod-like capsules at the water–oil interface (69). Copyright © 2017; American Chemical Society. (b) Schematic diagram of the structure of E2 protein nanocage (left) with its stable PEs (right) (70). Copyright © 2017; American Chemical Society.
Figure 5

(a) Schematic diagram of TMVCP self-assembled into nanorod-like capsules at the water–oil interface (69). Copyright © 2017; American Chemical Society. (b) Schematic diagram of the structure of E2 protein nanocage (left) with its stable PEs (right) (70). Copyright © 2017; American Chemical Society.

3 Applications of protein-based PE

3.1 Food science

3.1.1 Promising food substitute

Partially hydrogenated oils (PHOs) containing trans-fatty acids are used as a substitute for animal fats. However, excessive intake of PHOs increases the risk of diabetes and cardiovascular disease. HIPPEs in solid-like form can be a good substitute for PHOs. HIPPEs stabilized by chitosan/gliadin composite particles could construct liquid oils directly into gel-like soft solids, which may be a potential substitute for PHOs (73). Plant protein wheat gluten obtained by low-cost emulsification-evaporation could also be used to stabilize HIPPEs, which provided a similar creaminess, smoothness, and viscosity to mayonnaise and were more thermally stable than mayonnaise (17). Li et al. (74) developed emulsions with similar viscosity to commercial mayonnaise by studying the effects of the three main components of commercial mayonnaise (sucrose [0–10 wt%], NaCl [0–650 mmol], and acetic acid [pH from 2.5 to 6.5]) on the quality of PEs. Oxidation of oils and fats in food emulsions not only destroys the flavor and nutritional value of the food, but also produces toxic substances. Pickering particles can enhance the antimicrobial properties of emulsions by using their loading properties to carry antimicrobial agents. Cinnamon oil-enriched zein nanoparticle-stabilized PEs could be used to replace butter in cakes. The use of cinnamon oil not only reduced calorie intake, but also inhibited the growth of mold, thus extending the shelf lifespan of the product (75). Cinnamaldehyde can also be added to the oil phase of emulsions stabilized by WPI, which is very effective in inhibiting salt-induced droplet aggregation. This strategy helps to develop salt- and gastric-resistant emulsions, which have potential applications in high salt-sauces and diet foods (76).

3.1.2 Delivery of nutraceuticals

The application of some nutraceuticals, such as β-carotene, hesperidin, and curcumin, is limited due to their poor stability. PEs have good physical stability and oxidative stability, and are compatibility with food matrices. Solid particles can act as physical shields to retard nutrient degradation; therefore, PEs can be an ideal delivery system for protecting unstable biological active compounds. Ovotransferrin–gallic acid conjugates and carboxymethyl dextran-based particles were constructed using electrostatic assembly techniques. PEs stabilized by particles can improve lipolysis and enhance the bioaccessibility of curcumin, which is important for the design of PEs with good protective properties for nutraceutical loading (77). The PEs stabilized by an electrostatic complex composed of xanthan gum and protein nanoparticles could be effectively loaded with β-carotene, which improved the chemical degradation stability of β-carotene during storage and its bioaccessibility in the simulated gastrointestinal digestive tract (78). PEs can also be applied for the controlled delivery and release of nutrients at target sites. The release of β-carotene during gastrointestinal digestion could be regulated by altering the oil fraction of PEs, and PEs stabilized by PPI could be used as intestinal targeting and slow-release delivery mechanism for β-carotene (79).

The application of lutein in the food industry is usually restricted by its poor chemical stability and low water solubility. The β-lactoglobulin-gum Arabic composite particles could be used to stabilize lutein-rich emulsions. Lutein-rich emulsion gels stabilized with β-lactoglobulin-gum Arabic complex particles exhibited high flocculation resistance and significant chemical stability, with lutein retention up to 91.1% in 70% oil-phase emulsion gels after 12 weeks of storage (80). PEs stabilized by tea seed cake protein nanoparticles were superior to traditional emulsions in terms of lutein encapsulation (96.6 ± 1.0% vs 82.1 ± 1.4%) and bioaccessibility (56.0 ± 1.1% vs 35.2 ± 1.2%), with potential as an effective carrier for lutein (81).

3.2 Applications in biomedicine

3.2.1 Enhanced oral bioavailability

Improving the dissolution and absorption of drugs is an important goal of oral administration since poor solubility or permeability affects the efficacy of orally administered drugs. Owing to the unique structures, PEs possess characteristics including high stability and biocompatibility, giving PEs the potential in oral drug administration (82). Similar to traditional emulsions, PEs can dissolve poorly soluble drugs and improve their ability to penetrate gastrointestinal biofilms. HIPPEs stabilized by chitosan–caseinophosphopeptides nanocomplexes could increase the bioaccessibility of curcumin from 20.49% (bulk oil) to 49.21% (HIPPEs) (83). Zein–pectin–proanthocyanidins composites stabilized PEs with an elastic gel-like structure could increase the bioavailability of curcumin up to 39.7% (84). Due to the unique structure of the particle-loading interface, PEs exhibit resistance to coalescence, giving it a better physical stability than surfactant-stabilized emulsions. The droplets encapsulated by gliadin/chitosan colloidal particles could build a permeable 3D network that converts DHA-rich liquid algal oil into soft solid to protect it from oxidation. In addition, the bioaccessibility of curcumin by this method increased from 2.13% (bulk algal oil) to 53.61% (core curcumin), and the bio-accessibility of shell curcumin reached 76.82%, showing potential applications in oral administration of nutritional products (85).

3.2.2 Enhanced drug stability in the gastrointestinal tract

Particles loaded at the water–oil interface can effectively retard or inhibit the lipolysis of the oil, thereby protecting the drug in the droplet (86). However, protein-based stabilized PEs is sensitive to degradation by gastrointestinal enzymes and offers limited protection against carrying drugs (87). The composite granule layer of zein and tannin has a gel-like structure that inhibits the hydrolysis process of proteins during digestion and weakens the interaction between gastric digestive enzymes and oil droplets, thus resisting the harsh gastric environment and maintaining curcumin release (88). PEs stabilized by ovotransferrinogen fibrils showed a protofibril layer at the interface. The ionic strength and pH of the gastrointestinal tract attenuated the electrostatic repulsion between ovotransferrinogen fibrils, and the protofibril layer at the interface became thicker and denser, providing better protection for the carried curcumin. Such pH-responsive PEs facilitate the protection of acid-unstable drugs (56,89). Antarctic krill oil is a functional marine oil with low solubility and oxidative stability. HIPPEs stabilized by bamboo protein gel particles exhibited excellent krill oil loading efficiency and good functional activity, which helped to inhibit the overexpression of pro-inflammatory cytokines and protect the intestinal barrier function (90).

In addition, protein-stabilized PEs can inhibit pepsin degradation of proteins by reducing their sensitivity to environmental factors. Protein glycosylation can change its spatial conformation and improve functional properties of protein, such as acid stability and thermal stability. The half-life of curcumin encapsulated in the WPI–chitosan oligosaccharides complex was 40 h longer than that of the free oil phase, indicating that the glycosylated protein had good stability to curcumin (91).

3.3 Porous material

Recently, porous materials with large surface area and high porosity have been applied in a wide range of biomedical, adsorption, and catalytic fields. HIPPEs template methods enable the manufacture of well-defined porous materials. HIPPEs-derived porous materials have drawn the interest of researchers in various fields (92). HIPPEs stabilized by natural proteins and polysaccharides usually help to produce porous materials with high pore ratios, as well as with pore interconnectivity and superior biocompatibility (93). When preparing porous materials using the template method of HIPPEs, the polymerization reaction is usually carried out in the continuous phase. The protein particles dispersed in the continuous phase are cross-linked to form a gel-like network structure. The macroporous protein scaffolds are then formed directly by removing the volatile organic internal phase (94). All-natural gelatin nanoparticle-stabilized HIPPEs could construct hierarchical porous protein scaffolds. Proteins as low as 2.5 wt% in the dispersion medium could not only form scaffolds with connected pore morphology and high porosity, but also scaffolds with textured structures and smooth pore walls that could promote the adhesion and growth of L929 cells (95). Zhou et al. (96) developed a non-toxic porous material using HIPPEs template stabilized by gliadin–chitosan composite particles, which exhibited significant absorption of corn oil up to saturated absorption within 3 min. Through electrostatic interaction and hydrogen bonding, gliadin and propylene glycol alginate were combined to form colloidal particles. Porous materials were prepared from this particle-stabilized emulsion template. Heavy metal adsorption capacity of 202.71 mg·g−1 and 106.41 g·g−1 of oil absorption capacity were reported (97). Jiao et al. (45) used PPI microgel particles to stabilize HIPPEs as a substitute for PHOs with 87% edible oil as dispersed phase. When the dispersed phase was 88% hexane, such HIPPEs could be applied as template to prepare porous materials for promising applications in tissue engineering. Zein and hohenbuehelia serotina polysaccharides were also used to prepare nanoparticles to stabilize HIPPEs. Porous materials were prepared using HIPPEs as templates. The obtained porous materials showed not only good adsorption of water, oils, and pigments, but also superior adsorption of Pb2+ in water (98).

3.4 Biodegradable packaging film

The current environmental problems caused by non-biodegradable synthetic polymers are becoming increasingly serious, new natural polymer-based packaging materials need to be developed (99). The most promising biodegradable natural polymer-based food packaging is the active release packaging (100), where the preservatives, antioxidants, or antimicrobial agents are packed into the packaging rather than added directly to the food (101). Polysaccharides and proteins can be used to produce biodegradable packaging materials (102). PEs is a sustainable delivery system for bioactive compounds, and the encapsulation of essential oils (EO) in emulsions can improve the functional properties of EO by improving compatibility with different edible matrices (103). Liu et al. (104) encapsulated oregano essential oils (OEO) as an antimicrobial ingredient in PEs stabilized by soluble soy polysaccharide and soy protein, and subsequently incorporated the PEs into soluble soy polysaccharide solution to prepare the film. The curing effect of Pickering particles helped to retain more OEO within the film matrix, effectively prolonging the anti-bacterial properties of OEO. The incorporation of marjoram oil-loaded PEs stabilized by a mixture of WPI and inulin into the pectin film formulation allowed the pectin films to exhibit good mechanical and waterproof performance (105). The incorporation of EO-loaded PEs into film formulations enabled the film to have the sustained release properties of EO. The addition of clove oil-loaded PEs to the pullulan–gelatin film matrix resulted in blended films that exhibited not only water blocking properties and antioxidant properties, but also sustained release properties to clove oil (106). It is reported that by incorporating OEO loaded PEs into konjac glucomannan films, the OEO could be slowly released for more than 21 days and the films showed good preservation properties for fruits (107).

3.5 Sewage treatment

Recently, how to effectively remove harmful substances, such as heavy metal ions, from wastewater has received increasing research attention (108). Membrane technology is one of the most promising separation technologies for sewage treatment. The production of porous membranes with high permeability, anti-fouling properties, and good separation rates are essential for wastewater purification (109,110). Polymeric membranes with high efficiency and pollution control properties have attracted great interest in sewage treatment (111). The emulsion template allows for easy customization of the polymer pore size and size distribution, and therefore can be applied to membrane filtration technology. Nagarajan et al. (112) stabilized PEs with graphene oxide/gelatin. Membranes prepared using this emulsion had a uniform distribution of pores, were highly stable in water solutions, and provided purified water permeability values up to 5.8 ± 1.3 L·h−1·m−2·bar−1. Hexagonal boron nitride nanosheet/gelatin porous membranes could be prepared using the PEs strategy. By varying the curing and glutaraldehyde crosslinking time, the membrane pore size could be controlled between 0.39 and 1.60 μm, and the permeability of pure water between 150 and 506 L·h−1·m−2·bar−1, and an increase in free-iron(ii) ion removal from 14% (without acrylic acid) to 97% (113).

3.6 3D printing

3D printing is a technique for rapidly constructing objects by printing layer by layer. As the ink is continuously extruded, blueprints from specific design programs are fabricated into 3D structures (114,115). 3D printing technology has received a lot of attention in the food industry due to its convenience, customizable food design, and flexibility of arbitrary shapes. The composition and rheological behavior of ink will affect the effect of 3D printing. HIPPEs are considered an effective 3D printing ink because of their self-supporting structure and high stability. Emulsion templates have become a popular rapid casting technique due to their excellent rheological properties (116). Li et al. (117) used natural cod proteins to stabilize HIPPEs and improved the stability of HIPPEs by forming cross-linked structures. The HIPPEs stabilized with 50 mg·mL−1 of cod protein showed good print resolution. Sea bass protein microgel particles stabilized HIPPEs could also exhibit extrudability and printability, and the bioavailability of astaxanthin in HIPPEs reached 51.17% (118). WPI and soybean oil could be prepared into gel emulsion by microfluidic technology. The gel emulsion could be made into various shapes with 3D printing process, showing potential as cake decorations or custom functional foods (119). Gelatin cross-linked by TG could stabilize HIPPEs with high viscosity and low frequency dependence. The 3D printed models with many different colors could be obtained (120). In addition to various shapes, the 3D printing extrusion process should also delay the release of flavor substances. Cinnamaldehyde has high volatility and low solubility. HIPPEs containing cinnamaldehyde showed superior gel strength and texture properties after 3D printing process. More important, the fragrance component could be retained to a large extent. The loss of cinnamaldehyde was as low as 11.29 ± 0.01% (121).

4 Conclusion and prospective

With the development of ultrafine particle preparation and surface modification technologies, Pickering emulsification has ushered in new opportunities and become a new hot spot in the field of emulsification research. Emulsions with high stability and broad applicability have been developed using a variety of Pickering particles. Protein-based particles, gels, and fibrils generally give secure and nutritious properties to emulsions and therefore can be advantageous for food and medicine applications. Different flavors of food products need to be further developed to meet popular demand. In addition, PEs have a shear thinning behavior and can recover high viscosity after spraying on wounds, thus allowing the development of spray PEs for medicine applications such as wound dressings. Protein-based PEs templates for the preparation of porous materials have the advantage of controlled size and high biocompatibility, and future research should be devoted to the construction of porous materials encapsulating bioactive ingredients, drugs, and catalysts. Currently, most biodegradable food packaging incorporates antimicrobial or antioxidant agents into the film’s base fluid, where the curing effect of protein-based PEs can significantly prolong the antimicrobial and antioxidant effects, although the issue of whether the antimicrobial and antioxidant agents will leach into the food remains a concern. In sewage treatment applications, new protein composite particles should be developed to create membrane materials with tunable pores to improve membrane permeability, fouling resistance, and separation rates. The rheological properties of protein-based PEs are critical for 3D printing, and future research will also need to look at the rheological properties and stability of HIPPEs, as well as flavor and shape, in order to develop 3D printed products with special shapes. PEs have droplet size distributions ranging from a few nanometers to several hundred micrometers, and solid particles are also able to participate in shell formation with high encapsulation efficiency and fast thermal conductivity. Research work should be carried out to explore more applications of protein-based PEs, such as emulsion polymerization and catalyst/emulsifier cycling.

In this review, we focused on the formation and assembly methods of different morphologies of protein particles, and briefly describe the interfacial stability properties of protein polymer particles (nanoparticles, microgels, fibrils, nanogels, etc.). The applications of protein particle-based PEs in various fields, including food, biomedicine, porous materials, biodegradable packaging films, sewage treatment, and 3D printing, were also reviewed and prospected. We hope to provide more knowledge and facilities for scholars studying the application of natural protein-based PEs.

  1. Funding information: This work was supported by the National Natural Science Foundation of China (U2004211) and China Tobacco Henan Industrial Co., Ltd, CNTC, China (Technology Project A202014).

  2. Author contributions: Qianqian Ma: resources, writing – original draft; Sensen Ma: resources, writing – original draft; Jie Liu: visualization, supervision; Ying Pei: visualization; Keyong Tang: supervision; Jianhua Qiu: project administration; Jiqiang Wan: project administration; Xuejing Zheng: conceptualization, writing – review and editing, funding acquisition; Jun Zhang: conceptualization, writing – review and editing, funding acquisition. All authors have read and agreed to the published version of the manuscript.

  3. Conflict of interest: The authors state no conflict of interest.

  4. Data availability statement: Data are available only upon request to the authors.

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Received: 2023-01-06
Revised: 2023-03-05
Accepted: 2023-03-06
Published Online: 2023-04-25

© 2023 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  78. Adsorption and recovery of Cr(vi) from wastewater by Chitosan–Urushiol composite nanofiber membrane
  79. Comprehensive performance evaluation based on electromagnetic shielding properties of the weft-knitted fabrics made by stainless steel/cotton blended yarn
  80. Review Articles
  81. Preparation and application of natural protein polymer-based Pickering emulsions
  82. Wood-derived high-performance cellulose structural materials
  83. Flammability properties of polymers and polymer composites combined with ionic liquids
  84. Polymer-based nanocarriers for biomedical and environmental applications
  85. A review on semi-crystalline polymer bead foams from stirring autoclave: Processing and properties
  86. Rapid Communication
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https://doi.org/10.1515/epoly-2023-0001
Keywords for this article
Pickering emulsion; natural polymer; morphology; synthesis; application
Creative Commons
BY 4.0

Articles in the same Issue

  1. Research Articles
  2. Chitosan nanocomposite film incorporating Nigella sativa oil, Azadirachta indica leaves’ extract, and silver nanoparticles
  3. Effect of Zr-doped CaCu3Ti3.95Zr0.05O12 ceramic on the microstructure, dielectric properties, and electric field distribution of the LDPE composites
  4. Effects of dry heating, acetylation, and acid pre-treatments on modification of potato starch with octenyl succinic anhydride (OSA)
  5. Loading conditions impact on the compression fatigue behavior of filled styrene butadiene rubber
  6. Characterization and compatibility of bio-based PA56/PET
  7. Study on the aging of three typical rubber materials under high- and low-temperature cyclic environment
  8. Numerical simulation and experimental research of electrospun polyacrylonitrile Taylor cone based on multiphysics coupling
  9. Experimental investigation of properties and aging behavior of pineapple and sisal leaf hybrid fiber-reinforced polymer composites
  10. Influence of temperature distribution on the foaming quality of foamed polypropylene composites
  11. Enzyme-catalyzed synthesis of 4-methylcatechol oligomer and preliminary evaluations as stabilizing agent in polypropylene
  12. Molecular dynamics simulation of the effect of the thermal and mechanical properties of addition liquid silicone rubber modified by carbon nanotubes with different radii
  13. Incorporation of poly(3-acrylamidopropyl trimethylammonium chloride-co-acrylic acid) branches for good sizing properties and easy desizing from sized cotton warps
  14. Effect of matrix composition on properties of polyamide 66/polyamide 6I-6T composites with high content of continuous glass fiber for optimizing surface performance
  15. Preparation and properties of epoxy-modified thermosetting phenolic fiber
  16. Thermal decomposition reaction kinetics and storage life prediction of polyacrylate pressure-sensitive adhesive
  17. Effect of different proportions of CNTs/Fe3O4 hybrid filler on the morphological, electrical and electromagnetic interference shielding properties of poly(lactic acid) nanocomposites
  18. Doping silver nanoparticles into reverse osmosis membranes for antibacterial properties
  19. Melt-blended PLA/curcumin-cross-linked polyurethane film for enhanced UV-shielding ability
  20. The affinity of bentonite and WO3 nanoparticles toward epoxy resin polymer for radiation shielding
  21. Prolonged action fertilizer encapsulated by CMC/humic acid
  22. Preparation and experimental estimation of radiation shielding properties of novel epoxy reinforced with Sb2O3 and PbO
  23. Fabrication of polylactic acid nanofibrous yarns for piezoelectric fabrics
  24. Copper phenyl phosphonate for epoxy resin and cyanate ester copolymer with improved flame retardancy and thermal properties
  25. Synergistic effect of thermal oxygen and UV aging on natural rubber
  26. Effect of zinc oxide suspension on the overall filler content of the PLA/ZnO composites and cPLA/ZnO composites
  27. The role of natural hybrid nanobentonite/nanocellulose in enhancing the water resistance properties of the biodegradable thermoplastic starch
  28. Performance optimization of geopolymer mortar blending in nano-SiO2 and PVA fiber based on set pair analysis
  29. Preparation of (La + Nb)-co-doped TiO2 and its polyvinylidene difluoride composites with high dielectric constants
  30. Effect of matrix composition on the performance of calcium carbonate filled poly(lactic acid)/poly(butylene adipate-co-terephthalate) composites
  31. Low-temperature self-healing polyurethane adhesives via dual synergetic crosslinking strategy
  32. Leucaena leucocephala oil-based poly malate-amide nanocomposite coating material for anticorrosive applications
  33. Preparation and properties of modified ammonium polyphosphate synergistic with tris(2-hydroxyethyl) isocynurate for flame-retardant LDPE
  34. Thermal response of double network hydrogels with varied composition
  35. The effect of coated calcium carbonate using stearic acid on the recovered carbon black masterbatch in low-density polyethylene composites
  36. Investigation of MXene-modified agar/polyurethane hydrogel elastomeric repair materials with tunable water absorption
  37. Damping performance analysis of carbon black/lead magnesium niobite/epoxy resin composites
  38. Molecular dynamics simulations of dihydroxylammonium 5,5′-bistetrazole-1,1′-diolate (TKX-50) and TKX-50-based PBXs with four energetic binders
  39. Preparation and characterization of sisal fibre reinforced sodium alginate gum composites for non-structural engineering applications
  40. Study on by-products synthesis of powder coating polyester resin catalyzed by organotin
  41. Ab initio molecular dynamics of insulating paper: Mechanism of insulating paper cellobiose cracking at transient high temperature
  42. Effect of different tin neodecanoate and calcium–zinc heat stabilizers on the thermal stability of PVC
  43. High-strength polyvinyl alcohol-based hydrogel by vermiculite and lignocellulosic nanofibrils for electronic sensing
  44. Impacts of micro-size PbO on the gamma-ray shielding performance of polyepoxide resin
  45. Influence of the molecular structure of phenylamine antioxidants on anti-migration and anti-aging behavior of high-performance nitrile rubber composites
  46. Fiber-reinforced polyvinyl alcohol hydrogel via in situ fiber formation
  47. Preparation and performance of homogenous braids-reinforced poly (p-phenylene terephthamide) hollow fiber membranes
  48. Synthesis of cadmium(ii) ion-imprinted composite membrane with a pyridine functional monomer and characterization of its adsorption performance
  49. Impact of WO3 and BaO nanoparticles on the radiation shielding characteristics of polydimethylsiloxane composites
  50. Comprehensive study of the radiation shielding feature of polyester polymers impregnated with iron filings
  51. Preparation and characterization of polymeric cross-linked hydrogel patch for topical delivery of gentamicin
  52. Mechanical properties of rCB-pigment masterbatch in rLDPE: The effect of processing aids and water absorption test
  53. Pineapple fruit residue-based nanofibre composites: Preparation and characterizations
  54. Effect of natural Indocalamus leaf addition on the mechanical properties of epoxy and epoxy-carbon fiber composites
  55. Utilization of biosilica for energy-saving tire compounds: Enhancing performance and efficiency
  56. Effect of capillary arrays on the profile of multi-layer micro-capillary films
  57. A numerical study on thermal bonding with preheating technique for polypropylene microfluidic device
  58. Development of modified h-BN/UPE resin for insulation varnish applications
  59. High strength, anti-static, thermal conductive glass fiber/epoxy composites for medical devices: A strategy of modifying fibers with functionalized carbon nanotubes
  60. Effects of mechanical recycling on the properties of glass fiber–reinforced polyamide 66 composites in automotive components
  61. Bentonite/hydroxyethylcellulose as eco-dielectrics with potential utilization in energy storage
  62. Study on wall-slipping mechanism of nano-injection polymer under the constant temperature fields
  63. Synthesis of low-VOC unsaturated polyester coatings for electrical insulation
  64. Enhanced apoptotic activity of Pluronic F127 polymer-encapsulated chlorogenic acid nanoparticles through the PI3K/Akt/mTOR signaling pathway in liver cancer cells and in vivo toxicity studies in zebrafish
  65. Preparation and performance of silicone-modified 3D printing photosensitive materials
  66. A novel fabrication method of slippery lubricant-infused porous surface by thiol-ene click chemistry reaction for anti-fouling and anti-corrosion applications
  67. Development of polymeric IPN hydrogels by free radical polymerization technique for extended release of letrozole: Characterization and toxicity evaluation
  68. Tribological characterization of sponge gourd outer skin fiber-reinforced epoxy composite with Tamarindus indica seed filler addition using the Box–Behnken method
  69. Stereocomplex PLLA–PBAT copolymer and its composites with multi-walled carbon nanotubes for electrostatic dissipative application
  70. Enhancing the therapeutic efficacy of Krestin–chitosan nanocomplex for cancer medication via activation of the mitochondrial intrinsic pathway
  71. Variation in tungsten(vi) oxide particle size for enhancing the radiation shielding ability of silicone rubber composites
  72. Damage accumulation and failure mechanism of glass/epoxy composite laminates subjected to repeated low velocity impacts
  73. Gamma-ray shielding analysis using the experimental measurements for copper(ii) sulfate-doped polyepoxide resins
  74. Numerical simulation into influence of airflow channel quantities on melt-blowing airflow field in processing of polymer fiber
  75. Cellulose acetate oleate-reinforced poly(butylene adipate-co-terephthalate) composite materials
  76. Radiation shielding capability and exposure buildup factor of cerium(iv) oxide-reinforced polyester resins
  77. Recyclable polytriazole resins with high performance based on Diels-Alder dynamic covalent crosslinking
  78. Adsorption and recovery of Cr(vi) from wastewater by Chitosan–Urushiol composite nanofiber membrane
  79. Comprehensive performance evaluation based on electromagnetic shielding properties of the weft-knitted fabrics made by stainless steel/cotton blended yarn
  80. Review Articles
  81. Preparation and application of natural protein polymer-based Pickering emulsions
  82. Wood-derived high-performance cellulose structural materials
  83. Flammability properties of polymers and polymer composites combined with ionic liquids
  84. Polymer-based nanocarriers for biomedical and environmental applications
  85. A review on semi-crystalline polymer bead foams from stirring autoclave: Processing and properties
  86. Rapid Communication
  87. Preparation and characterization of magnetic microgels with linear thermosensitivity over a wide temperature range
  88. Special Issue: Biodegradable and bio-based polymers: Green approaches (Guest Editors: Kumaran Subramanian, A. Wilson Santhosh Kumar, and Venkatajothi Ramarao)
  89. Synthesis and characterization of proton-conducting membranes based on bacterial cellulose and human nail keratin
  90. Fatigue behaviour of Kevlar/carbon/basalt fibre-reinforced SiC nanofiller particulate hybrid epoxy composite
  91. Effect of citric acid on thermal, phase morphological, and mechanical properties of poly(l-lactide)-b-poly(ethylene glycol)-b-poly(l-lactide)/thermoplastic starch blends
  92. Dose-dependent cytotoxicity against lung cancer cells via green synthesized ZnFe2O4/cellulose nanocomposites
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