Protein eutectogel, methods of making and uses thereof

Abstract

The present disclosure relates to the field of protein-based materials, and in particular, to a protein eutectogel prepared with natural deep eutectic solvents and one or more plant proteind, methods of making and uses thereof.

Description

PROTEIN EUTECTOGEL, METHODS OF MAKING AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of priority on U.S. provisional patent application no. 63 / 623,874 filed on January 23, 2024, the contents of which are incorporated herein by reference in their entirety.FIELD

[0002] The present disclosure relates to the field of protein-based materials, and in particular, to a protein eutectogel prepared with natural deep eutectic solvents, methods of making and uses thereof.BACKGROUND

[0003] Protein based materials can be applied in a range of areas such as tissue engineering, bioplastics and polymeric coatings for packaging, sensors, medical devices, materials for water purification, energy production and energy storage.fi, 2] Although, animal proteins such as collagen and gelatin have been used as scaffolds for tissue engineering, and plant-based proteins such as soy protein and rice bran protein have high nutritional value and are well suited for creating scaffolds fortissue engineering. [3] Beyond tissue engineering, most of the polymeric substrates used for sensor development, membranes and adsorbents for water purification, medical devices and energy storage devices are constructed with synthetic polymers which are a burden on the environment. Both plant and animal proteins are well-suited for the development of artificial skin devices, wound dressings, membranes and as separators for batteries. [4-7]

[0004] Food waste (30% of food production) significantly impacts the carbon and nitrogen cycles, and global greenhouse gas emissions. [1] Specifically, protein production and waste increase pressure on global freshwater use, land use, and chemical pollution. Converting proteinaceous waste into functional materials can offer a sustainable alternative to synthetic plastics for a range of applications especially for single use, while also reducing the environmental impact of food waste.

[0005] Proteins have a large number of functional groups that can react with various materials and offer unique opportunities to develop composite and functional bioplastics. [8] In recent years, various authors have proposed the development of biobased plastics through food-waste valorization. [9] Proteins are commonly extracted and processed with saline, acidic, alkaline aqueous solutions or in aqueous alcohol. Despitetheir excellent biodegradability, and functional properties, protein-based materials suffer from brittleness and poor mechanical properties.

[0006] Selecting solvents that influence the protein orientation and interactions, and the use of additives such as plasticizers, fdlers and crosslinkers can be utilized to tune the mechanical properties of proteins. However, these solvents and additives influence the overall sustainability of protein-based materials. Typically, the mechanical strength of protein films is increased through the addition of synthetic polymers using volatile organic solvents, or crosslinking. In case of zein (com protein), a film forming protein, mechanically robust films have been obtained by modification with polyethylene maleic anhydride in dimethyl formamide solution or through using dimethyl sulfoxide (DMSO) as a solvent or through crosslinking with glutaraldehyde. Using synthetic polymers, volatile and flammable organic solvents and highly reactive crosslinkers typically lowers the biodegradability of the material produced and increases its environmental impact.

[0007] The use of natural deep eutectic solvents (NADES) such as those based on choline chloride or betaine as hydrogen bond acceptors, and natural or plant derived organic hydrogen bond donors offer an environmentally favorable alternative to overcome the limitations of conventional volatile, toxic and flammable organic solvents especially pertaining to their life cycle costs.

[0010] Deep eutectic solvents (DES) have been extensively applied in extraction of proteins from animal and plant derived resources including agricultural waste and waste from the food industry. [11 -15] A choline chloride and ethylene glycol (1:2 mole ratio) deep eutectic solvents has been applied to produce highly stretchable films of gelatin.

[0016] Similarly, highly stretchable films of gelatin were obtained with glyceline or choline chloride and glycerol (1:2 mole ratio).

[0017] The same study found that zein based materials produced brittle films when blended with glyceline DES, while soy protein showed a modest elongation at break of 20%. Zein has also been electrospun using deep eutectic solvent based on choline chloride and furfuryl alcohol,

[0018] the latter being a potential carcinogen and an irritant. [19, 20]

[0008] The background herein is included solely to explain the context of the disclosure. This is not to be taken as an admission that any of the material referred to was published, known, or part of the common general knowledge as of the priority date.SUMMARY

[0009] The present disclosure includes a method of preparing a plant protein eutectogel comprising combining one or more plant proteins and a deep eutectic solvent (DES) under conditions to form the plant protein eutectogel wherein the one or more plant proteins are present in an amount of about 1 g to about 15 g per mL of DES. In some embodiments, the amount of the one or more plant proteins is about 45% (w / w) to about 95% (w / w) and the amount of DES is about 5% (w / w) to about 55% (w / w), based on the total weight of protein(s) and DES. In some embodiments, one or more plasticizers are included in the above combining step. In some embodiments, the one or more plasticizers are present in an amount of about 1% (w / w) to about 15% (w / w) based on the total weight of the protein(s), DES and plasticizer(s).

[0010] In some embodiments, the present disclosure includes a resin-based method and a solution-based method to fabricate protein eutectogels. For the resin-based method, a protein eutectogel was fabricated by blending plant proteins with a deep eutectic solvent. The protein eutectogel exhibits a high flowability, high tensile strength, and high flexibility at a certain temperature, which allows the protein eutectogel to be shaped by multiple methods such as extrusion printing and injection molding. For the solution-based method, a solution was made by dissolving protein powder in a deep eutectic solvent. The solution was subsequently shaped on anon-stick surface by methods such as electrospinning and coagulated in a cold-water bath. The protein eutectogel was obtained after drying.

[0011] Accordingly, in an aspect of the present disclosure, the resin-based method initially involves preparing a deep eutectic solvent by mixing a first element, a hydrogen bond donor and a second element, a hydrogen bond acceptor with a desired molar ratio. The deep eutectic solvent was subsequently prepared by heating the mixture to a desired temperature. A protein eutectogel was then made by blending protein powder with the deep eutectic solvent with a desired concentration and temperature. In some embodiments, an additional plasticizer is included in the above blending step to further lower viscosity and improve melt processibility. After granulation, the protein eutectogel was shaped on a non-stick surface with a desired method and temperature. In some embodiments, the molar ratio of the first element and the second element ranges from 2: 1 to 1:6.

[0012] In some embodiments, the first element comprises choline chloride or betaine.

[0013] In some embodiments, the second element comprises lactic acid or glucose.

[0014] In some embodiments, the temperature of preparing the deep eutectic solvent ranges from 15 °C to 100 °C.

[0015] In some embodiments, the protein powder comprises zein, gluten, soy protein isolate, rice bran protein, pea protein, or a combination thereof.

[0016] In some embodiments, the concentration of protein powder in deep eutectic solvent ranges from Ig / mL to 10 g / mL.

[0017] In some embodiments, the temperature of blending ranges from 15 °C to 100 °C.

[0018] In some embodiments, the concentration of the plasticizer ranges from about 1% to about 15% by weight in the protein eutectogel formulation.

[0019] In some embodiments, the temperature of shaping protein eutectogel ranges from 70 °C to 160 °C.

[0020] In some embodiments, the method of shaping protein eutectogel comprises die casting, electrospinning, melt-blowing, 2D / 3D extrusion printing, or injection molding.

[0021] In some embodiments, the non-stick surface comprises silicone rubber, wax, parchment, polytetrafluoroethylene, or a combination thereof.

[0022] According to another aspect of the present disclosure, the solution-based method initially involves preparing a deep eutectic solvent by mixing a first element and a second element with a desired molar ratio. A solution was then made by dissolving protein powder in the deep eutectic solvent with a desired concentration and temperature. The solution was subsequently shaped on a non-stick surface in water with a desired method and temperature. A protein eutectogel can be obtained after drying the shaped solution with a desired temperature.

[0023] In some embodiments, the molar ratio of the first element and the second element ranges from 2: 1 to 1 : 6.

[0024] In some embodiments, the first element comprises choline chloride or betaine.

[0025] In some embodiments, the second element comprises lactic acid or glucose.

[0026] In some embodiments, the protein powder comprises zein, gluten, soy protein isolate, rice bran protein, pea protein, or a combination thereof.

[0027] In some embodiments, the concentration of protein powder in deep eutectic solvent ranges from 0.01 g / mL to 0.15 g / mL.

[0028] In some embodiments, the temperature of dissolving ranges from 80 °C to 100 °C.

[0029] In some embodiments, the method of shaping solution on a non-stick surface comprises die casting, electrospinning, or extrusion printing.

[0030] In some embodiments, the non-stick surface comprises silicone rubber, wax, parchment, polytetrafluoroethylene, or a combination thereof.

[0031] In some embodiments, the temperature of water ranges from 1 °C to 5 °C.

[0032] In some embodiments, the temperature of drying shaped solution ranges from 20 °C to 40 °C.

[0033] In some embodiments, the shaped protein eutectogel is crosslinked to in a crosslinker solution / bath or by exposure to crosslinker vapours.

[0034] The present disclosure also includes a plant protein eutectogel comprising one or more plant proteins and a deep eutectic solvent (DES) wherein the one or more plant proteins are present in an amount of about 1 g to about 15 g per mL of DES, or about 45% (w / w) to about 95% (w / w) of the one or more plant proteins and about 5% (w / w) to about 55% (w / w) of the DES, based on the total weight of protein(s) and DES. In some embodiments, one or more plasticizers are included in the above combining step. In some embodiments, the plant protein eutectogel further comprises one or moreplasticizers in an amount of about 1% (w / w) to about 15% (w / w) based on the total weight of the protein(s), DES and plasticizer(s).

[0035] According to another aspect of the present disclosure, the protein eutectogel can be used for food packaging, 3D printing ink and filament, filtration material, or tissue engineering scaffold.

[0036] Other features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the disclosure, are given by way of illustration only and the scope of the claims should not be limited by these embodiments, but should be given the broadest interpretation consistent with the description as a whole.DRAWINGS

[0037] Certain embodiments of the disclosure will now be described in greater detail with reference to the attached drawings in which:

[0038] FIGURE 1 shows schematic of protein eutectogel resin preparation and its conversion into films and 2D extrusion printed mats in exemplary embodiments of the disclosure.

[0039] FIGURE 2 shows schematic of solution-based processing of protein eutectogel in exemplary embodiments of the disclosure.

[0040] FIGURE 3 shows FTIR Spectra of A. Zein films cast from (from top to bottom) ethanol, acetic acid, Choline chloride: lactic acid DES coagulation method and resin method respectively, B. Resins of (from top to bottom) zein, gluten, soy protein with DES as prepared at 100°C, C. mechanically pressed samples of (from top to bottom) zein, gluten and soy protein, all in exemplary embodiments of the disclosure.

[0041] FIGURE 4 shows simultaneous TGA / DSC Thermograms of A. zein, B. gluten, C. soy eutectogels between 25°C to 200°C, and DSC thermogram of D. zein, E. gluten, F. Soy protein eutectogels between -60°C to 200°C, all in exemplary embodiments of the disclosure.

[0042] FIGURE 5 shows A. Water contact angle of protein eutectogels, B. Stress vs strain curves for protein eutectogels, and C. Dynamic mechanical analysis ofzein, gluten and soy protein eutectogel resins, all in exemplary embodiments of the disclosure.

[0043] FIGURE 6 shows rheology of exemplary zein and DES resin at (from top to bottom) 80°C, 100°C and 120°C, A. viscosity vs shear rate, B. shear Stress vs shear rate.

[0044] FIGURE 7 shows application of exemplary zein based eutectogel in A. Injection molding, B. filament extrusion and C. extrusion 2D printing.

[0045] FIGURE 8 shows SEM images of A. Freestanding wet-electrospun zein eutectogel and B. Melt-electrospun zein eutectogel resin on kimwipe support (larger flat fibers are the fibers of the kimwipe, finer fibers are electrospun layer) in exemplary embodiments of the disclosure.

[0046] FIGURE 9 shows photographs of A. Extrusion printing process and B. Extrusion printed mat using zein eutectogel solution in exemplary embodiments of the disclosure.

[0047] FIGURE 10 shows photographs of 10 layered (3D) extrusion printed exemplary zein eutectogel resin A. Top view B. Cross sectional view C. Side view

[0048] FIGURE 11 shows a scanning electron microscope image of the melt blown exemplary zein eutectogel fibers.

[0049] FIGURE 12 shows fluorescent microscopy images of 3T3 fibroblast cells grown on exemplary zein eutectogels A - Zl, B - Z2, C - Z3, D -ZU, E - Z2’, F - Z3’.DETAILED DESCRIPTIONI, Definitions

[0050] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present disclosure herein described for which they are suitable as would be understood by a person skilled in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[0051] In understanding the scope of the present disclosure, the term “comprising” and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, “including”, “having” and their derivatives.

[0052] The term “consisting” and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other unstated features, elements, components, groups, integers and / or steps.

[0053] The term “consisting essentially of’, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps.

[0054] Terms of degree such as “substantially”, “about” and “approximately” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies. In addition, all ranges given herein include the end of the ranges and also any intermediate range points, whether explicitly stated or not.

[0055] As used in this disclosure, the singular forms “a”, “an” and “the” include plural references unless the content clearly dictates otherwise.

[0056] In embodiments comprising an “additional” or “second” component, the second component as used herein is chemically different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.

[0057] The term “and / or” as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that “at least one of’ or “one or more” of the listed items is used or present.

[0058] The abbreviation, “e.g.” is derived from the Latin exempli gratia and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.”

[0059] The word “or” is intended to include “and” unless the context clearly indicates otherwise.

[0060] It will be understood that any component defined herein as being included may be explicitly excluded by way of proviso or negative limitation, such as any specific compounds or method steps, whether implicitly or explicitly defined herein.

[0061] The term “eutectogel” as used herein refers to a class of soft materials in which traditional eutectic solvents or deep eutectic solvents (DES) are embedded in a three-dimensional network, resulting in a stable, nonflowing, and self-supporting structure.

[0062] The term “resin” as used herein refers to a highly viscous or semi -solid substance.

[0063] The term “biodegradable” as used herein means that a material degrades completely within 45 days under thermophilic compost conditions at 58°C.II, Methods and Compositions of the DisclosureMethods of Preparing a Plant Protein Eutectogel

[0064] The present disclosure includes a method of preparing a plant protein eutectogel comprising combining one or more plant proteins and a deep eutectic solvent (DES) under conditions to form the plant protein eutectogel wherein the one or more plant proteins are present in an amount of about 1 g to about 15 g per mL of DES.

[0065] In some embodiments, the amount of the one or more plant proteins is about 45% (w / w) to about 95% (w / w) and the amount of DES is about 5% (w / w) to about 55% (w / w), based on the total weight of protein(s) and DES. In some embodiments, the amount of the one or more plant proteins is about 60% (w / w) to about 80% (w / w) and the amount of DES is about 20% (w / w) to about 40% (w / w), based on the total weight of protein(s) and DES. In some embodiments, the amount of the one or more plant proteins is about 65% (w / w) to about 75% (w / w) and the amount of DES is about 25% (w / w) toabout 35% (w / w), based on the total weight of protein(s) and DES. In some embodiments, the amount of the one or more plant proteins is about 68% (w / w) to about 72% (w / w) and the amount of DES is about 28% (w / w) to about 32% (w / w), based on the total weight of protein(s) and DES.

[0066] In some embodiments, the combining of the one or more plant proteins and the DES is by combining under conditions to form a resin. Therefore, in some embodiments, the one or more plant proteins and the DES are combined as dry powders and heated to a temperature above the melting point of the DES, for example a temperature of about 15°C to about 100°C, or about 80°C to about 100°C, in a mixer until a resin is formed. In some embodiments, the one or more plant proteins and the DES are mixed for about 10 minutes to about 1 hour, or for about 30 minutes. In some embodiments, the resin is granulated and the resulting granulated resins are shaped into any desired shape, for example by die casting, electrospinning, melt-blowing, 2D / 3D extrusion printing, stamping or injection molding. In some embodiments, the granulated resins are shaped into fdms comprising the plant protein eutectogel, for example by pressing the granulated resin onto a non-stick surface or by extruding the resin in an extruder. In some embodiments, the pressing and extruding are done at a temperature that allows molding of the granulated resin, for example about 70°C to about 160°C, or about 80°C to about 110°C.

[0067] In some embodiments, the combining of the one or more plant proteins and the DES is by combining the one or more plant proteins and a deep eutectic solvent and heating to form a solution and shaping the solution on a non-stick surface and cooling to form the plant protein eutectogel. In some embodiments, the one or more plant proteins and a deep eutectic solvent are heated to a temperature of about 80°C to about 100°C. In some embodiments, the shaping of the solution on a non-stick surface comprises die casting, electrospinning, or extrusion printing. In some embodiments the shaping of the solution is by coagulating in water. In some embodiments, the water is at a temperature of about 1°C to about 5°C. In some embodiments, the water is used at a volume ratio of solutiomwater of about 1:2 to about 1: 10 or about 1:5 for a duration of about 30 minutes to about 2 hours, or about 1 hour. In some embodiments, the water is removed and additional water is added one or more times. In some embodiments, theplant protein eutectogel is air dried and optionally dried by heating. In some embodiments, the temperature of drying ranges from about 20°C to about 40 °C.

[0068] In some embodiments, the non-stick surface is selected from silicone rubber, wax, parchment, polytetrafluoroethylene, and combinations thereof.

[0069] In some embodiments, the method further comprises combining the one or more plant proteins and the DES along with one or more plasticizers. In some embodiments, the one or more plasticizers are used in an amount to improve melt processability and rheology of the plant eutectogel. In some embodiments the one or more plasticizers are used in an amount of from about 1% (w / w) to about 15% (w / w), about 5% (w / w) to about 10%(w / w), or about 7% (w / w), by total weight of the protein(s), DES and plasticizer(s). In some embodiments, the one or more plasticizers are selected from glycerol, triethylene glycol, dibutyl tartrate, levulinic acid, polyethylene glycol, sugars, fatty acids, and combinations thereof. In some embodiments, the one or more plasticizers are selected from glycerol, oleic acid, and a combination thereof. In some embodiments, the plasticizer is glycerol.

[0070] In some embodiments, the method further comprises cross-linking the shaped protein eutectogel to improve rigidity, moisture tolerance and shelflife. In some embodiments, the crosslinker is selected from citric acid, tannic acid, gallic acid, glutaraldehyde and a combination thereof. In some embodiments, the crosslinking is applied to the shaped eutectogel by exposure to a solution / bath or vapor.

[0071] In some embodiments, the one or more proteins are selected from zein, gluten, soy protein, rice bran protein, whey protein, pea protein, and combinations thereof. In some embodiments, the one or more proteins are selected from zein, gluten, soy protein isolate, rice bran protein, pea protein, and combinations thereof. In some embodiments, the one or more proteins are selected from zein, gluten and soy protein.

[0072] The deep eutectic solvents (DES) for use in the present disclosure refer to mixtures of two or more elements (components) that when combined together have a eutectic point, which is the temperature of solidification or freezing (Fp). The eutectic point of the combined elements is generally much lower than either of the elements individually or at any other ratio of mixing and occurs at a single temperature without separation of the individual components on solidification. In some embodiments, theDES is any DES that comprises first element that is a hydrogen bond acceptor and a second element that is a metal salt or a hydrogen bond donor. In some embodiments, the hydrogen bond acceptor is an ionic species. Deep eutectic solvents are well known and have been widely studied. A review of deep eutectic solvents is provided by Smith et al. (Chem. Rev. 2014, 114, 11060) and Zhang et al. (Chem. Soc. Rev. 2012, 41, 7108), which both reference the originating work of Abbott et al. (J. Am. Chem. Soc. 2004, 126, 9142), the contents of all of which are hereby incorporated by reference.

[0073] In some embodiments, the molar ratio of elements in the deep eutectic solvent is selected to provide a mixture that has the lowest melting point, or close to the lowest melting point. In some embodiments, the molar ratio of elements in the deep eutectic solvent is selected to provide a mixture that has the lowest viscosity, or close to the lowest viscosity.

[0074] In some embodiments, the first and second elements are present in the DES in a molar ratio of from 2: 1 to 1 :6. In some embodiments, the molar ratio of the first element to the second element is 1:0.5, 1: 1 or 1:2. In some embodiments, the molar ratio of the first element to the second element is 1:2.

[0075] In some embodiments, the DES has a freezing point, Tf, that is at or close to ambient temperature. However, it is acknowledged that many of the known DESs have a freezing temperature that lies above ambient temperature, such as above 20°C. Thus, in some embodiments, the DES is provided and / or used at a temperature that is above the freezing point of the DES.

[0076] In some embodiments, the DES has a freezing temperature that is less than 90°C, such as less than 80°C, such as less than 70°C, such as less than 60°C.

[0077] In some embodiments, the DES has a freezing temperature that is 10°C or more, such as 20°C or more, such as 30°C or more, such as 40°C or more.

[0078] In some embodiments, the DES has a freezing temperature that is less than the melting temperature of an element of the DES, such as the second element of the DES. In some embodiments, the freezing temperature of the DES is at least 10°C, at least 20°C, at least 30°C, at least 40°C, at least 50°C, or at least 60°C less than the melting temperature of the element.

[0079] In some embodiments, the DES is provided as a liquid. Therefore in some embodiments, the DES is provided or used at a temperature that is above the freezing temperature of the DES, such as a temperature of 1°C or more, 5°C or more, 10°C or more, 20°C or more, 50°C or more above the freezing temperature of the DES.

[0080] In some embodiments, where a deep eutectic solvent is provided as a liquid it is frozen, if required, by appropriate cooling of the DES. Similarly, where a DES is provided as a solid it may be melted, if required, by appropriate heating of the DES.

[0081] In some embodiments, the DES is provided as a solid and is dried before use. In some embodiments, the temperature of drying DES ranges from about 15 °C to about 120 °C, or from about 15 °C to about 100 °C.

[0082] In some embodiments, the DES has a density in the range 1 to 1.7 g / mL, as measured at 25°C, such as a density in the range 1 to 1.5 g / mL, such as 1 to 1.4 g / mL, such as 1 to 1.3 g / mL, such as 1 to 1.2 g / mL.

[0083] In some embodiments, the DES has a viscosity in the range 10 to 100,000 cP, for example as measured at a temperature of 20°C, 25°C, 30°C or 40°C, such as a viscosity in the range 10 to 50,000 cP, such as 100 to 10,000 cP.

[0084] In some embodiments, the ionic species comprises a cation, such as an ammonium, phosphonium or sulfonium cation, and an anion, which is a Lewis base, such as a halide anion.

[0085] In some embodiments, the DES contains, as the second element, a Lewis or Bronsted acid, which may be a metal salt or hydrogen bond donor species. In some embodiments, the second element is a hydrogen bond donor species.

[0086] Deep eutectic solvents are categorized into Types I to IV (see, for example, Smith et al. Chem. Rev. 2014, 114, 11060).

[0087] A Type I deep eutectic solvent comprises an ionic species, such as those described above, and a non-hydrated metal salt, such as a non-hydrated metal halide, such as a non-hydrated metal chloride. In some embodiments, the non-hydrated metal salt metal salt is a Zn, Sn, Fe, Al, Ga or In salt, such as a halide, such as a chloride. In some embodiments, the non-hydrated metal salt metal salt is a Zn salt, such as ZnCL.

[0088] A Type II deep eutectic solvent comprises an ionic species, such as those described above, and a hydrated metal salt, such as a hydrated metal halide, such as a hydrated metal chloride. In some embodiments, the hydrated metal salt is a Cr, Co, Cu, Ni or Fe salt, such as a halide, such as a chloride.

[0089] A Type III deep eutectic solvent comprises an ionic species, such as those described above, and a hydrogen bond donor species, such as lactic acid. A further description of hydrogen bond donor species is given below.

[0090] A Type IV deep eutectic solvent comprises a metal salt, such as a metal halide, such as a metal chloride, and a hydrogen bond donor species. In some embodiments, the metal salt is an Al or Zn salt, such as a halide, such as a chloride.

[0091] In some embodiments, the DES is Type III.

[0092] In some embodiments, the first element is an ionic species. In some embodiments, the first component is a quaternary ammonium salt, such as a quaternary ammonium chloride.

[0093] In some embodiments, the first element is selected from choline chloride (ChCl), betaine (trimethylglycine), betaine hydrochloride, carnitine, choline bitartrate, histidine, glycine, nicotinic acid, phytic acid sodium, proline, glutamic acid, serine ethylammonium chloride (EtNFECl), N-ethyl-2-hydroxy-N,N-dimethylethanaminium chloride, 2-(chlorocarbonyloxy)-N,N,N-trimethylethanaminium chloride, N-benxyl-2- hydroxy-N,N-dimethylethanaminium chloride, tetramethylaminium chloride (TMAC1), tetrabutylaminium chloride (TBAC1), methyltriphenylphosphonium bromide (MeP(Ph)3Br), methyltriphenylphosphonium chloride (MeP(Ph)3Cl), benzyltriphenylphosphonium bromide, tetraethylaminium bromide (TEABr), tetrabutylaminium bromide (TBABr), 2-hydroxy-N,N,-diethylethanaminium chloride (Et2(EtOH)ACl), 2-chloro-N,N,N-trimethylethanaminium chloride (CIChCl) and acetyl choline chloride (AcChCl).

[0094] In some embodiments, the first element is choline chloride or betaine. Choline chloride is advantageous as it is unreactive to water, and it is biodegradable and biocompatible. For example, choline chloride is used as a feedstock for chicken food, and it is produced on an industrial scale of this purpose, and others.

[0095] In some embodiments, the second element is any suitable hydrogen bond donor. For example, the second component is selected from urea, p- toluenesulfonic acid (TsOH), acetamide, 1-methyl-urea, 1,3-dimethylurea, 1,1- dimethylurea, 1,2-propanediol, thymol, cyclodextrin, imidazole, 2,2,2- trifluoroacetamide, thiourea, benzamide, glycerol, ethylene glycol, lactic acid, maleic acid, malic acid, malonic acid, benzoic acid, ascorbic acid, caffeic acid, adipic acid, oxalic acid, succinic acid, citric acid, phenylacetic acid, phenylpropionic acid, tricarbahylic acid, levulinic acid, itaconic acid, gallic acid, glucose, erythritol, xylitol, sorbitol, tartaric acid, fructose, isomaltose, isosorbide, lactose, maltose, mannitol, ribitol, sucrose, trehalose, xylose, tartaric acid, urea, decanoic acid, dodecanoic acid, isosorbide, 4-hydroxybenzoic acid, caffeic acid, coumaric acid, cinnamic acid, suberic acid, gallic acid and resorcinol. In some embodiments, the second component is lactic acid or glucose.

[0096] In some embodiments, the DES is a mixture of choline chloride or betaine and lactic acid or glucose. In some embodiments, the DES is a mixture of choline chloride and lactic acid. In some embodiments, the DES is a mixture of betaine and lactic acid. In some embodiments, the DES is a mixture of betaine and glucose.

[0097] In some embodiments, each of the first and second elements has a molecular weight that is at least 50, at least 75, at least 100, at least 150, or at least 200.

[0098] In some embodiments, each of the first and second elements has a molecular weight that is at most 300, at most 500, at most 1,000, at most 2,000 or at most 5,000.

[0099] In some embodiments, each of the first and second elements has a molecular weight that is in a range with the lower and upper limits selected from those given above. For example, in some embodiments, each of the first and second elements has a molecular weight that is in the range 50 to 1,000.

[0100] A DES may be prepared by mixing together the elements that make up the solvent, such as the first element and the second element. The elements may be heated during the mixing process, for example to a temperature of 80°C, such as 70°C.

[0101] In some embodiments, the method further comprises combining the one or more plant proteins, the DES, and optionally one or more plasticizers, with one ormore other ingredients that would be useful in the plant protein eutectogels of the disclosure. In some embodiments, the one or more other ingredients are selected from gelatin, collagen, albumin, starch, carbohydrates, and combinations thereof. In some embodiments the one or more other ingredients are added in an amount of about 0.1% (w / w) to about 20% (w / w), about 0.5% (w / w) to about 10% (w / w), or 1% (w / w) to about 5% (w / w), by total weight of the one or more plant proteins, the DES, and optionally the one or more plasticizers and the one or more other ingredients.

[0102] In some embodiments, in the method of the present disclosure:- the amount of the one or more plant proteins is about 45% (w / w) to about 95% (w / w);- the amount of the DES is about 5% (w / w) to about 55% (w / w);- the amount of the of one or more plasticizers is about 1% (w / w) to about 15% (w / w); and optionally the amount of the of one or more other ingredients is about 0.1% (w / w) to about 20% (w / w), based on the total weight of the plant protein eutectogel.

[0103] In some embodiments, in the method of the present disclosure:- the amount of the one or more plant proteins is about 60% (w / w) to about 80% (w / w);- the amount of the DES is about 15% (w / w) to about 35% (w / w);- the amount of the of one or more plasticizers is about 1% (w / w) to about 15% (w / w); and optionally the amount of the of one or more other ingredients is about 0.1% (w / w) to about 20% (w / w), based on the total weight of the plant protein eutectogel.

[0104] In some embodiments, in the method of the present disclosure:- the amount of the one or more plant proteins is about 65% (w / w) to about 85% (w / w);- the amount of the DES is about 20% (w / w) to about 30% (w / w);- the amount of the of one or more plasticizers is about 5% (w / w) to about 10% (w / w); and optionally the amount of the other ingredients is about 0.5% (w / w) to about 10% (w / w), based on the total weight of the plant protein eutectogel.

[0105] In some embodiments, in the method of the present disclosure:- the amount of the one or more plant proteins is about 70% (w / w);- the amount of the DES is about 23% (w / w); and- the amount of the of one or more plasticizers is about 7% (w / w), based on the total weight of the plant protein eutectogel.

[0106] In some embodiments, in the method of the present disclosure: the amount of the one or more plant proteins is about 70% (w / w) and the one or more plant proteins selected from zein, gluten and soy protein.;The amount of the DES is about 23% (w / w) and the DES comprises a first element comprising choline chloride or betaine and a second element comprising lactic acid or glucose, wherein the first and second elements are present in a molar ratio of 1 :1 or 1:2; and- the amount of plasticizer is about 7% (w / w) and the plasticizer is glycerol, based on the total weight of the plant protein eutectogel.Plant Protein Eutectogels

[0107] The present disclosure also includes a plant protein eutectogel comprising one or more plant proteins and a deep eutectic solvent (DES) wherein the one or more plant proteins are present in an amount of about 1 g to about 15 g per mL of DES.

[0108] In some embodiments, the plant protein eutectogel comprises about 45% (w / w) to about 95% (w / w) of the one or more proteins and about 5% (w / w) to about 55% (w / w) of the DES, based on the total weight of the plant protein eutectogel. In some embodiments, the plant protein eutectogel comprises about 60% (w / w) to about 80%(w / w) of the one or more proteins and about 20% (w / w) to about 40% (w / w) of the DES, based on the total weight of the plant protein eutectogel. In some embodiments, the plant protein eutectogel comprises about 65% (w / w) to about 75% (w / w) of the one or more proteins and about 25% (w / w) to about 35% (w / w) of the DES, based on the total weight of the plant protein eutectogel. In some embodiments, the plant protein eutectogel comprises about 68% (w / w) to about 72% (w / w) of the one or more proteins and about 28% (w / w) to about 32% (w / w), based on the total weight of the plant protein eutectogel.

[0109] In some embodiments, the plant protein eutectogel further comprises one or more plasticizers. In some embodiments, the one or more plasticizers are present in an amount to improve melt processability and rheology of the plant eutectogel. In some embodiments, the one or more plasticizers are selected from glycerol, triethylene glycol, dibutyl tartrate, levulinic acid, polyethylene glycol, sugars, fatty acids, and combinations thereof. In some embodiments the one or more plasticizers are present in an amount of from about 1% (w / w) to about 15% (w / w), about 5% (w / w) to about 10%(w / w), or about 7% (w / w), based on the total weight of the plant protein eutectogel. In some embodiments, the one or more plasticizers are selected from glycerol, oleic acid, and a combination thereof. In some embodiments, the plasticizer is glycerol.

[0110] In some embodiments, the one or more proteins are selected from zein, gluten, soy protein, rice bran protein, whey protein, pea protein, and combinations thereof. In some embodiments, the one or more proteins are selected from zein, gluten, soy protein isolate, rice bran protein, pea protein, and combinations thereof. In some embodiments, the one or more proteins are selected from zein, gluten and soy protein.

[0111] In some embodiments, the DES is any DES that comprises first element that is a hydrogen bond acceptor and a second element that is a metal salt or a hydrogen bond donor. In some embodiments, the hydrogen bond acceptor is an ionic species. Deep eutectic solvents are well known and have been widely studied. A review of deep eutectic solvents is provided by Smith et al. (Chem. Rev. 2014, 114, 11060) and Zhang et al. (Chem. Soc. Rev. 2012, 41, 7108), which both reference the originating work of Abbott et al. (J. Am. Chem. Soc. 2004, 126, 9142), the contents of all of which are hereby incorporated by reference.

[0112] In some embodiments, the molar ratio of elements in the deep eutectic solvent is selected to provide a mixture that has the lowest melting point, or close to the lowest melting point. In some embodiments, the molar ratio of elements in the deep eutectic solvent is selected to provide a mixture that has the lowest viscosity, or close to the lowest viscosity.

[0113] In some embodiments, the first and second elements are present in the DES in a molar ratio of from 2: 1 to 1 :6. In some embodiments, the molar ratio of the first element to the second element is 1:0.5, 1: 1 or 1:2. In some embodiments, the molar ratio of the first element to the second element is 1:2.

[0114] In some embodiments, the DES has a freezing point, Tf, that is at or close to ambient temperature. However, it is acknowledged that many of the known DESs have a freezing temperature that lies above ambient temperature, such as above 20°C. Thus, in some embodiments, the DES is provided and / or used at a temperature that is above the freezing point of the DES.

[0115] In some embodiments, the DES has a freezing temperature that is less than 90°C, such as less than 80°C, such as less than 70°C, such as less than 60°C.

[0116] In some embodiments, the DES has a freezing temperature that is 10°C or more, such as 20°C or more, such as 30°C or more, such as 40°C or more.

[0117] In some embodiments, the DES has a freezing temperature that is less than the melting temperature of an element of the DES, such as the second element of the DES. In some embodiments, the freezing temperature of the DES is at least 10°C, at least 20°C, at least 30°C, at least 40°C, at least 50°C, or at least 60°C less than the melting temperature of the element.

[0118] In some embodiments, the DES has a density in the range 1 to 1.7 g / mL, as measured at 25°C, such as a density in the range 1 to 1.5 g / mL, such as 1 to 1.4 g / mL, such as 1 to 1.3 g / mL, such as 1 to 1.2 g / mL.

[0119] In some embodiments, the DES has a viscosity in the range 10 to 100,000 cP, for example as measured at a temperature of 20°C, 25°C, 30°C or 40°C, such as a viscosity in the range 10 to 50,000 cP, such as 100 to 10,000 cP.

[0120] In some embodiments, the ionic species comprises a cation, such as an ammonium, phosphonium or sulfonium cation, and an anion, which is a Lewis base, such as a halide anion.

[0121] In some embodiments, the DES contains, as the second element, a Lewis or Bronsted acid, which may be a metal salt or hydrogen bond donor species. In some embodiments, the second element is a hydrogen bond donor species.

[0122] Deep eutectic solvents are categorized into Types I to IV (see, for example, Smith et al. Chem. Rev. 2014, 114, 11060).

[0123] A Type I deep eutectic solvent comprises an ionic species, such as those described above, and a non-hydrated metal salt, such as a non-hydrated metal halide, such as a non-hydrated metal chloride. In some embodiments, the non-hydrated metal salt metal salt is a Zn, Sn, Fe, Al, Ga or In salt, such as a halide, such as a chloride. In some embodiments, the non-hydrated metal salt metal salt is a Zn salt, such as ZnCL.

[0124] A Type II deep eutectic solvent comprises an ionic species, such as those described above, and a hydrated metal salt, such as a hydrated metal halide, such as a hydrated metal chloride. In some embodiments, the hydrated metal salt is a Cr, Co, Cu, Ni or Fe salt, such as a halide, such as a chloride.

[0125] A Type III deep eutectic solvent comprises an ionic species, such as those described above, and a hydrogen bond donor species, such as lactic acid. A further description of hydrogen bond donor species is given below.

[0126] A Type IV deep eutectic solvent comprises a metal salt, such as a metal halide, such as a metal chloride, and a hydrogen bond donor species. In some embodiments, the metal salt is an Al or Zn salt, such as a halide, such as a chloride.

[0127] In some embodiments, the DES is Type III.

[0128] In some embodiments, the first element is an ionic species. In some embodiments, the first component is a quaternary ammonium salt, such as a quaternary ammonium chloride.

[0129] In some embodiments, the first element is selected from choline chloride (ChCl), betaine (trimethylglycine), betaine hydrochloride, carnitine, choline bitartrate, histidine, glycine, nicotinic acid, phytic acid sodium, proline, glutamic acid, serine,ethylammonium chloride (EtNHsCl), N-ethyl-2-hydroxy-N,N-dimethylethanaminium chloride, 2-(chlorocarbonyloxy)-N,N,N-trimethylethanaminium chloride, N-benxyl-2- hydroxy-N,N-dimethylethanaminium chloride, tetramethylaminium chloride (TMAC1), tetrabutylaminium chloride (TBAC1), methyltriphenylphosphonium bromide (MeP(Ph)3Br), methyltriphenylphosphonium chloride (MeP(Ph)3Cl), benzyltriphenylphosphonium bromide, tetraethylaminium bromide (TEABr), tetrabutylaminium bromide (TBABr), 2-hydroxy-N,N,-diethylethanaminium chloride (Et2(EtOH)ACl), 2-chloro-N,N,N-trimethylethanaminium chloride (CIChCl) and acetyl choline chloride (AcChCl).

[0130] In some embodiments, the first element is choline chloride or betaine. Choline chloride is advantageous as it is unreactive to water, and it is biodegradable and biocompatible. For example, choline chloride is used as a feedstock for chicken food, and it is produced on an industrial scale of this purpose, and others.

[0131] In some embodiments, the second element is any suitable hydrogen bond donor. For example, the second component is selected from urea, p- toluenesulfonic acid (TsOH), acetamide, 1-methyl-urea, 1,3-dimethylurea, 1,1- dimethylurea, 1,2-propanediol, thymol, cyclodextrin, imidazole, 2,2,2- trifluoroacetamide, thiourea, benzamide, glycerol, ethylene glycol, lactic acid, maleic acid, malic acid, malonic acid, benzoic acid, ascorbic acid, caffeic acid, adipic acid, oxalic acid, succinic acid, citric acid, phenylacetic acid, phenylpropionic acid, tricarballyhc acid, levulinic acid, itaconic acid, gallic acid, glucose, erythritol, xylitol, sorbitol, tartaric acid, fructose, isomaltose, isosorbide, lactose, maltose, mannitol, ribitol, sucrose, trehalose, xylose, tartaric acid, urea, decanoic acid, dodecanoic acid, isosorbide, 4-hydroxybenzoic acid, caffeic acid, coumaric acid, cinnamic acid, suberic acid, gallic acid and resorcinol. In some embodiments, the second component is lactic acid or glucose.

[0132] In some embodiments, the DES is a mixture of choline chloride or betaine and lactic acid or glucose. In some embodiments, the DES is a mixture of choline chloride and lactic acid. In some embodiments, the DES is a mixture of betaine and lactic acid. In some embodiments, the DES is a mixture of betaine and glucose.

[0133] In some embodiments, each of the first and second elements has a molecular weight that is at least 50, at least 75, at least 100, at least 150, or at least 200.

[0134] In some embodiments, each of the first and second elements has a molecular weight that is at most 300, at most 500, at most 1,000, at most 2,000 or at most 5,000.

[0135] In some embodiments, each of the first and second elements has a molecular weight that is in a range with the lower and upper limits selected from those given above. For example, in some embodiments, each of the first and second elements has a molecular weight that is in the range 50 to 1,000.

[0136] In some embodiments, the plant protein eutectogel is further cross-linked to improve rigidity, moisture tolerance and shelf life. In some embodiments, the crosslinker is selected from citric acid, tannic acid, gallic acid, glutaraldehyde and a combination thereof.

[0137] In some embodiments, the plant protein eutectogel further comprises one or more other ingredients that would be useful in the plant protein eutectogels of the disclosure. In some embodiments, the one or more other ingredients are selected from water, gelatin, collagen, albumin, starch, carbohydrates, and combinations thereof. In some embodiments the one or more other ingredients are present in an amount of about 0.1% (w / w) to about 20% (w / w), about 0.5% (w / w) to about 10% (w / w), or 1% (w / w) to about 5% (w / w), by total weight of the plant protein eutectogel.

[0138] In some embodiments, the plant protein eutectogel of the present disclosure comprises: about 45% (w / w) to about 95% (w / w) one or more plant proteins; about 5% (w / w) to about 55% (w / w) of a DES; about 1% (w / w) to about 15% (w / w) of one or more plasticizers; and optionally about 0.1% (w / w) to about 20% (w / w) of one or more other ingredients, based on the total weight of the plant protein eutectogel.

[0139] In some embodiments, the plant protein eutectogel of the present disclosure comprises: about 60% (w / w) to about 80% (w / w) one or more plant proteins; about 15% (w / w) to about 35% (w / w) of a DES; about 1% (w / w) to about 15% (w / w) of one or more plasticizers; and optionally about 0.1% (w / w) to about 20% (w / w) of one or more other ingredients, based on the total weight of the plant protein eutectogel.

[0140] In some embodiments, the plant protein eutectogel of the present disclosure comprises: about 65% (w / w) to about 85% (w / w) one or more plant proteins; about 20% (w / w) to about 30% (w / w) of a DES; about 5% (w / w) to about 10% (w / w) of one or more plasticizers; and optionally about 0.5% (w / w) to about 10% (w / w) of one or more other ingredients, based on the total weight of the plant protein eutectogel.

[0141] In some embodiments, the plant protein eutectogel of the present disclosure comprises: about 70% (w / w) one or more plant proteins; about 23% (w / w) of a DES; and about 7% (w / w) of one or more plasticizers, based on the total weight of the plant protein eutectogel.

[0142] In some embodiments, the plant protein eutectogel comprises: about 70% (w / w) one or more plant proteins selected from zein, gluten and soy protein.; about 23% (w / w) of a DES comprising a first element comprising choline chloride or betaine and a second element comprising lactic acid or glucose,wherein the first and second elements are present in a molar ratio of 1 :1 or 1 :2; and about 7% (w / w) of glycerol, based on the total weight of the plant protein eutectogel.

[0143] In some embodiments, the plant protein eutectogel is biodegradable, meaning that the eutectogel degrades completely within 45 days under thermophilic compost conditions at 56°C to 60°C, or 58°C. In some embodiments, thermophilic compost condition are as set out in ASTM 5338 without CO2 dissolution.

[0144] In some embodiments, the plant protein eutectogel is hydrophilic.

[0145] In some embodiments, the plant protein eutectogel has an elongation at break that is greater than about 20%, or from about 20% to about 140%, as measured using a Shimadzu Autograph AG-X tensile tester with a crosshead speed of 2 mm / min and force recorded using a 0.5 kN load cell on samples that are conditioned for 24 hours at 50 ± 5% RH at 25 °C for 1 hour using a temperature-humidity / environmental chamber (ESPEC North America Inc., ESL-2CW).

[0146] In some embodiments, the plant protein eutectogel can be injection molded or extrusion printed at temperatures of about 80°C to about 170°C, about 80°C to about 120°C, or about 110°C.

[0147] In some embodiments, the plant protein eutectogel is a solid or is rigid at room temperature and ductile when heated above about 80°C.Uses of the Plant Protein Eutectogels

[0148] In some embodiments, the plant protein eutectogels of the present disclosure are used for food packaging, 3D printing inks and filaments, filtration materials, or as tissue engineering scaffolds. In some embodiments, the plant protein eutectogels are used in single use plastic containers, cups or other utensils, as insulation materials, in clothing or as filler materials for packaging.

[0149] Accordingly, the present disclosure also includes a food packaging, 3D printing ink or filament, filtration material, tissue engineering scaffold, single useplastic container, cup or utensil, insulation material, clothing, or filler material for packaging comprising a plant protein eutectogels of the present disclosure.Numbered Embodiments of the Disclosure

[0150] The present disclosure further comprises the following embodiments:1. A method of making a protein eutectogel, comprising: preparing a deep eutectic solvent by mixing a first element and a second element with a desired molar ratio; drying the deep eutectic solvent with a desired temperature; making a protein eutectogel by blending protein powder with the deep eutectic solvent with a desired concentration and temperature;Granulating the protein eutectogel; andShaping the protein eutectogel on a non-stick surface with a desired method and temperature.2. The method of embodiment 1 , wherein the molar ratio of the first element and the second element ranges from 2:1 to 1:6.3. The method of embodiment 1, wherein the first element comprises choline chloride or betaine.4. The method of embodiment 1, wherein the second element comprises lactic acid or glucose.5. The method of embodiment 1, wherein the temperature of drying deep eutectic solvent ranges from 15 °C to 100 °C.6. The method of embodiment 1, wherein the protein powder comprises zein, gluten, soy protein isolate, rice bran protein, pea protein, or a combination thereof.7. The method of embodiment 1, wherein the concentration of protein powder in deep eutectic solvent ranges from 1 g / mL to 10 g / mL.8. The method of embodiment 1, wherein the temperature of blending ranges from 15 °C to 100 °C.9. The method of embodiment 1, wherein the temperature of shaping protein eutectogel ranges from 70 °C to 150 °C.10. The method of embodiment 1, wherein the method of shaping protein eutectogel comprises die casting, electrospinning, melt-blowing, 2D / 3D extrusion printing, or injection molding.11. The method of embodiment 1, wherein the non-stick surface comprises silicone rubber, wax, parchment, polytetrafluoroethylene, or a combination thereof.12. A method of making a protein eutectogel, comprising: preparing a deep eutectic solvent by mixing a first element and a second element with a desired molar ratio; making a solution by dissolving protein powder in the deep eutectic solvent with a desired concentration and temperature; shaping the solution on a non-stick surface in water with a desired method and temperature; and drying the shaped solution with a desired temperature.13. The method of embodiment 12, wherein the molar ratio of the first element and the second element ranges from 2:1 to 1:6.14. The method of embodiment 12, wherein the first element comprises choline chloride or betaine.15. The method of embodiment 12, wherein the second element comprises lactic acid or glucose.16. The method of embodiment 12, wherein the protein powder comprises zein, gluten, soy protein isolate, rice bran protein, pea protein, or a combination thereof.17. The method of embodiment 12, wherein the concentration of protein powder in deep eutectic solvent ranges from 0.01 g / mL to 0.15 g / mL.18. The method of embodiment 12, wherein the temperature of dissolving ranges from 80 °C to 100 °C.19. The method of embodiment 12, wherein the method of shaping solution on a non-stick surface comprises die casting, electrospinning, or extrusion printing.20. The method of embodiment 12, wherein the non-stick surface comprises silicone rubber, wax, parchment, polytetrafluoroethylene, or a combination thereof.21. The method of embodiment 12, wherein the temperature of water ranges from 1 °C to 5 °C.22. The method of embodiment 12, wherein the temperature of drying shaped solution ranges from 20 °C to 40 °C.23. Use of the method of any one of embodiments 1-22 for food packaging, 3D printing ink and filament, fdtration material, or tissue engineering scaffold.EXAMPLES

[0151] The following non-limiting examples are illustrative of the present disclosure:Methods

[0152] Materials: Zein (powder), gluten (powder), choline chloride (powder), lactic acid (85%, liquid) and ethanol (anhydrous, liquid) were purchased from Sigma Aldrich Canada. Additionally, soy protein isolate (powder) was purchased from Now Foods.

[0153] Preparation of protein and eutectogel resin: The deep eutectic solvent used in this study was prepared by mixing choline chloride and lactic acid in a mole ratio of 1:2 (hereafter referred to as DES) in a glass jar using a magnetic stir bar, and dried under vacuum at 70°C. To prepare the protein resin, 100 g of the respective proteins, namely zein, gluten and soy protein isolate was blended with 35 mL (zein) or 40 mL (gluten and soy protein isolate) of DES in a twin-screw mixer (Haake, Rheomix3000) at 100°C and 100 rpm for 30 minutes for zein, and at 80°C for 30 minutes for gluten and soy protein isolate. The blended resins were granulated using a 66SRE-Extreme granulator (Rapid Granul ators Inc.). They were then pressed intofilms using a heated hydraulic press and a 50 mm x 50 mm x 0.5 mm square aluminum mold (Carver, 10 tons), or extruded thereafter at 100-110°C. The above procedures have been shown in Figure 1.

[0154] Preparation of protein eutectogel solutions: As shown in Figure 2, first, 15% w / v solution of zein was prepared by blending zein in the DES using an overhead stirrer mounted with an anchor paddle mixer (polytetrafluoroethylene construction, 300 rpm) at 80 °C for 12 hours. The zein solution was then cast on a nonstick silicone rubber mat and coagulated in a cold-water bath (4°C) at a 1:5 volume ratio over 1 hour duration. The cold water was then drained and replaced an additional two times, each time retaining the coagulated mass in water for 30 minutes. The coagulated mass was then air dried at room temperature overnight and further dried in an oven for 4 hours at 80°C.

[0155] Fourier transform infrared spectroscopy with attenuated reflectance (FTIR-AR): The resin film samples were prepared for analysis of surface functional groups. Additional samples of zein were prepared by casting it from a 70% alcohol solution and glacial acetic acid for comparison with the samples prepared from the DES. Additionally, flakes of each protein were prepared by pressing the powders between two aluminum sheets at 6-ton pressure on a hydraulic press (Carver). The surface functional groups were characterized using a Bruker Hyperion 3000 FTIR spectrometer operating in attenuated total reflectance (ATR) mode over 64 scans in a range from 4000-400 cm-1 with a 4 cm-1 resolution.

[0156] Thermal characterization using thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC): Thermal stability of the resins was studied by thermogravimetric analysis (TGA) (Mettler Toledo, Model TGA / DSC3+ STARe System) under atmospheric air and a second under argon. Testing was done under non-isothermal conditions over a temperature range from room temperature to 200°C, with a heating rate of 10°C / min. Additionally, thermal characteristics of the zein resin were also evaluated by using a TA Instruments Q200 differential scanning calorimeter (DSC) (New Castle, DE, USA). Samples of 5-10 mg of resin were sealed in an aluminum hermetic pan, and subsequently heated under a nitrogen environment from 20°C to 140-150°C, then cooled to -80°C, followed by finally reheating it to 200°C at a rate of 10°C / min to characterize the processed eutectogel.

[0157] Tensile testing: Prior to mechanical testing, the samples were conditioned for 24 hours at 50 ± 5% RH at 25 °C for 1 hour using a temperature- humidity / environmental chamber (ESPEC North America Inc., ESL-2CW). Following exposure to a humidified environment, tensile properties were measured on strips with a dimension of 50 mm * 13 mm x 0.45 mm using a Shimadzu Autograph AG-X tensile tester. The tensile testing was performed at a crosshead speed of 2 mm / min and force was recorded using a 0.5 kN load cell. Mechanical properties were reported as an average of three repeats.

[0158] Water contact angle measurement: 1 g sample of the protein resins was hot-pressed onto a glass microscope slide between two flat aluminum sheets (1 / 8 inch thick) with minimal pressure for 5 min at 100°C and cooled to ambient temperature before the measurement was performed. Contact angle was measured using optical contact angle measurement using OCA 25 (DataPhysics Instruments USA Corp, Charlotte, NC, USA) immediately after a 6 pL droplet of distilled water was placed on the solid polymer surface. The test was repeated a minimum of three times for each sample.

[0159] Rheology of polymer resin: The flowability of the resin samples were measured with a Discovery HR-2 hybrid rheometer (TA Instruments, U.S.) by conduction. Approximately 1-2 g of the resin was evenly loaded onto the plate of the rheometer and compressed with a 10 N axial force. A generalized flow sweep was conducted using the parallel plate geometry at 80°C, 100°C and 120°C respectively, over a shear rate range of 0.1-1000 s-1. Further, the melt flow index (MFI) of the zein and DES resin was determined using the protocol described in ASTM D1238 with a weight of 2 kg at a temperature of 100 °C and Kayeness Inc. The MFI measurement was taken in triplicate for the resin.

[0160] Dynamic mechanical analysis: Thermal stability of the resin films was investigated using dynamic mechanical analysis (DMA) to assess the materials compatibility for commonly used applications and products. Films prepared in a heated hydraulic press (Carver) with the resin material pressed at 10-ton pressure using aluminum molds with a thickness of 500±50 pm were characterized using DMA tester (TA instruments, DMA 850) with a film tension clamp. The tests were conducted over -50°C to 60°C temperature range to analyze the stability of the mechanical propertiesover a temperature range from freezing temperatures to those on a very hot day. The oscillation temperature ramp test was performed for all the samples over the above- mentioned temperature range at ramp rate of 5°C / min, preload force of 10-4 N, amplitude of 5 pm and frequency 1 Hz.

[0161] Injection molding: Injection molding is one of the common polymer processing methods to manufacture commonly used products and compatibility of the formulations to this processing method can be a good indicator of future possibilities to be used in mass manufacturing. To showcase processability of the products, Z-DES (zein based resin) was injection molded into a stainless-steel cylinder / screw mold. The injection was performed at 110°C and pressure of 4.25 MPa.

[0162] Extrusion printing using zein and DES based solution and resin: Extrusion printing is a commonly used 3D printing process that uses spools of resin fdament and extrudes it through a heated nozzle layer by layer to create a 3D part. Extrusion is also applied to produce the 3D printing filament and as such utilized a custom-made 3D printer designed to extrude and print the zein eutectogel solution and resin was used. The printer consisted of several components including an XYZ positioner, a printhead, a pneumatic control system and an integrated computer control to synchronize the movement of the printhead with the pneumatic system. The printhead comprised of a heated stainless-steel syringe (Allevi US, 10 mL) with a 400 pm metal nozzle and was connected to a solenoid valve using pneumatic lines connected to a 6.2 bar (90 psi) pneumatic source. A toolpath for a 2D mesh network was created for continuous extrusion with overlapping lines. As a first demonstration, a 2D mesh of 96 cells with overall dimensions of 26 mm x 26 mm was printed with each mesh cell printed over a region of 2.6 mm x 2.6 mm on a non-stick surface of silicone rubber. The printing speed in x and y directions was set to 250 mm / min. The syringe containing the protein eutectogel resin was heated to 120 °C using a syringe heater (Braintree scientific, Model BS-SYR). Extrusion pressure of 4 bar was applied to the zein resin and the total printing time was approximately 2 min. Similarly, room temperature and a pressure of 5.5 bar was utilized in printing the zein eutectogel solution.

[0163] Electrospinning of zein and DES solution and molten resin: Electrospinnability of the zein solution in DES was assessed using a benchtopelectrospinning set up. A syringe pump (Harvard Apparatus, Canada) was used to deliver a heated solution or heated resin at 80°C to an 18-gauge blunt tip needle at a feed rate of 3 mL / hour. For solution electrospinning, the syringe was mounted vertically facing down at an aluminum foil collector submerged under 2 cm of cold water at 4°C. The needle tip was separated by 10 cm from the current collector. A voltage of 18 - 25 kV was applied between the tip of the syringe and the collector until a stable Taylor cone was observed and a fdm began to form on the surface of water. After 15 minutes of electrospinning, the fdm was collected onto a glass slide for drying in air overnight before characterization. In case of the melt electrospinning, a separation of 10 cm was maintained between the syringe tip and the collector (stainless steel sheet, 12” x 12”). A kimwipe was attached to the collector with tape to serve as a backing substrate on which the electrospun fibers can be collected. A voltage of 20 -27 kV was applied between the tip of the syringe and the collector. The applied voltage was determined by visually assessing whether a stable Taylor cone was formed at the tip of the syringe. The electrospinning was continued for 15 minutes after which the kimwipe samples were collected for characterization.

[0164] Biodegradability: Biodegradability tests were conducted in an aerobic soil compost mixture and were carried out under thermophilic conditions at 58 ± 2°C (ASTM 5338 conditions without CO2 dissolution), over a period of 28 days. Three flat samples (2 cm x 2 cm x 0.5 mm) of each resin held in polyethylene mesh bags that allow contact with the compost, but retain the samples within the enclosure, were distributed in 240 g of moist compost. Finally, similar size squares of Whatman Grade 2 fdter paper were used as a cellulosic standard material and were distributed between several layers of 240 g of moist compost in similar enclosures. In each case, consecutive layers of compost and pieces of resin or fdter paper were spread out in a sealable glass container. 10-20 mL of water were added to each glass container to ensure that the contents remained moist. The container was then left open with a small gap to allow for air flow, and placed in an oven maintained at 58 ± 2°C. The disintegration of the samples was observed at the end of every week by extracting the pieces of each material type. The extracted samples were air dried, and the soil was brushed off gently from the extracted samples using a brush. Photographs of the cleaned samples were also taken to demonstrate the degradation of the samples under composting conditions. Thisprocess was repeated at weekly intervals until the samples fully disintegrated upon visual inspection.

[0165] Zein and DES based scaffolds for mammalian cell culture: Zein fdms were prepared as described before using three different DES - namely choline chloride and lactic acid (1:2 molar ratio, termed as Zl), betaine and lactic acid (1:2 molar ratio, termed as Z2) and betaine and glucose (1:1 molar ratio, termed as Z3). To prepare the fdms, 5 g of zein was blended in a mortar and pestle with the DES, and then pressed between 2 aluminum sheets using a 0.5 mm square mold at 110 °C and 10 ton pressure using a heated hydraulic press. One batch of the 3 variants of the fdms were saved as-is and the other were rinsed with deionized water and soaked in it for 30 minutes. After the soak, these fdms were removed and air dried overnight.

[0166] The as-prepared (Zl, Z2, Z3) and rinsed (ZE, Z2’, Z3’) fdm samples of the 3 respective variants of zein and DES eutectogels were then used as scaffold materials to culture murine fibroblast cells. GFP / NIH-3T3 fibroblast cells were cultured in their growth media, Dulbecco's Modified Eagle Medium (DMEM) (high glucose, L- glutamine, sodium pyruvate, Gibco), supplemented with 10% v / v Fetal Bovine Serum (FBS, Gibco), and 1 % v / v penicillin / streptomycin (10,000 U / ml, ThermoFisher). Once they reached 80-90% confluency, they were trypsinized and seeded on the scaffolds. The fdms were cut into 1 x1 cm2squares and placed in a polystyrene 24-well plate. After exposing them to UV light for 1 hour (30 minutes each side), they were immersed in 1 mL complete culture media overnight, followed by washing the samples once with PBS, and once with complete culture medium. Then, 5xl05cells were seeded on the scaffolds, and after 30 minutes of incubation at 37°C and 5% CO2, 2 mL extra complete culture media was added to each sample and incubated for an additional 10 days. The culture media were refreshed every other day. Fluorescence microscopy (inverted fluorescent microscope, Zeiss) using an FITC fdter was performed at a 5x magnification on days 1, 4, 7, and 10 after seeding to monitor cellular adhesion and proliferation.Results and Discussion

[0167] Surface characterization and chemical properties: The chemical structure of the protein eutectogel was characterized using FTIR-ATR and the spectrafor characterized films are shown in Figure 3. In Figure 3A, a significant peak can be observed due to N-H stretching around 3298 cm-1 for zein films cast from ethanol, acetic acid and DES solutions. However, in the case of the films cast from DES, a superimposed broad O-H stretching peak was also observed at 3303 cm-1 from the DES that results in the broadening of the peak. Additionally, another significant peak was also observed at 1742 cm-1 from the C=O stretching from the carbonyl groups in the DES. Three distinct peaks at 1128 cm-1, 1095 cm-1, 1048 cm-1 and 950 cm-1 in the DES have been reported in previous characterizations of choline chloride and lactic acid DES.

[0021] In Figure 3A, when the zein eutectogel is compared against the zein films prepared with ethanol or acetic acid solvent, the intense bands at 2875 cm-1 and 2960 cm-1 are similar for the DES and acetic acid based films. The asymmetric and symmetric C-H stretching between the ethanol-based films and other films are likely different due to the induced dipole moment caused by the change in solvent / plasticizer polarity. The FTIR spectra of each protein eutectogel prepared using DES and those of pure protein flakes have been illustrated in Figure 3B and Figure 3C, respectively.

[0168] Thermal properties of zein eutectogels: Thermal properties of zein- DES resins were characterized using thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). Figure 4A, B and C show the simultaneous TGA-DSC thermogram of the eutectogels under argon. Under an argon environment, a distinct mass loss was observed until 105 °C for all protein eutectogels, which can be attributed to moisture loss. Above 140 °C, an increase in heat evolved and sharp decrease in weight were observed, suggesting a likely chemical reaction. When the sample is further heated, an increasing loss in weight is observed under a constant rate of temperature change, which is likely due to decomposition of lactic acid and / or zein, or potentially the formation of an ester of lactic acid. [22-24] Additionally, DSC thermogram of the protein eutectogels was characterized between 50 to 200 °C and these are illustrated in Figure 4 D, E and F. No glass transition point was observed in this temperature range and no distinct melting / freezing point was observed over 2 cycles between -50 to 150 °C for Z-DES, and -50 to 140 °C for G-DES and S-DES, suggesting that the eutectogels are amorphous and chemically stable over this temperature range. The protein eutectogels start to degrade above 150 °C (170 °C for S-DES) as shown by the sharp increase in heat evolved (slope of DSC curve increases).

[0169] Water contact angle measurement: The water contact angle for each protein eutectogels is shown in Figure 5 A. All films were found to be hydrophilic, and the degree of hydrophilicity of eutectogels increased in the order of S-DES>Z-DES>G- DES.

[0170] Mechanical properties: The tensile properties of the protein eutectogels prepared with the resin-based process have been evaluated and are shown in Figure 5B. Z-DES films show high ductility with elongation at break values of 128% and ultimate tensile strength exceeding 500 kPA. G-DES films have slightly better tensile strength than Z-DES but excellent elongation at break values of 170%. Finally, S-DES films show significant strength at 4.5 MPa with elongation at break values of 165%. However, their tensile strength is lower than the samples prepared with the solution process due to loss of DES from the eutectogel during the coagulation step. Based on these results, while not wishing to be limited by theory, it is possible that the DES acts as a plasticizer for the zein and higher the amount of DES results in a more flexible eutectogel.

[0171] Dynamic mechanical analysis: The storage modulus and Tan-Delta (ratio of the loss to the storage modulus) are reported in Figure 5C over the temperature range of -50°C to 60°C. Soy samples exhibited the highest temperature stability over the test temperature range as no noticeable drop in the storage modulus was experienced and no peak or variation was detected in Tan-Delta. These results indicate the acceptable capability of the soy samples to preserve their elasticity and mechanical properties over this temperature range, which is optimal for commonly used polymeric products. Gluten Tan-Delta showed two peaks, one at almost -35°C and one at close to 40°C, that shows the material had the highest proportion of viscous response compared with elastic response under conditions of deformation at these two temperatures. Therefore, gluten can be stable in a smaller temperature range than soy, which still can be acceptable for a certain range of products. Zein had shown the highest elasticity (lowest Tan-Delta) among the samples (soy was the second best and gluten was the least elastic sample) under 20°C and exhibited an acceptable mechanical stability under 40°C (Tan-Delta peak). Considering all three formulations, based on the required working temperature range and preferred elasticity for a product, each of these materials can be functional for a certain application.

[0172] Rheology of zein eutectogel: The rheology of zein eutectogels (resin) was characterized at different temperatures below 140 °C above which the eutectogel is expected to degrade. The generalized flow curves illustrating the change in viscosity and shear stress at increasing shear rates are reported in Figure 6. At 80 °C, the eutectogel was extremely viscous and demonstrated a shear thinning behavior. Due to the high viscosity, it was unable to apply shear rates above 10 s-1. At 100 °C, the viscosity of the resin decreased by one order of magnitude, and it was able to apply higher shear rates to the eutectogel melt. As the temperature increased to 120 °C, the melt became free flowing. A shear thinning behavior was observed for all temperatures. Moreover, the melt flow index of the eutectogel (resin) was estimated to be 40.8±5, further confirming the high melt flowability of the eutectogel, which makes it much more amenable to cast, coat and extrude. The gluten and soy protein based eutectogels showed more of a thermoset behavior owing to the higher degree of self-crosslinking between gluten and soy polymer chains respectively, and their melts rheology could not be tested.

[0173] Injection moldability and extrusion printability of thermoplastic zein eutectogel: The ability to produce injection molded 3D components using the zein eutectogel was demonstrated. Z-DES was successfully injection molded at 110 °C to a threaded screw with a cylindrical shaft as illustrated in Figure 7A. The ability to extrude fibers was demonstrated in Figure 7B and using extrusion printed zein eutectogel to produce a 2D mesh of 26 mm x 26 mm has been shown in Figure 7C. Although a protein eutectogel solution can also be extrusion printed and coagulated in an aqueous bath as illustrated in Figure 2, this process involves loss of DES and further air drying of the printed structure. It may also be prone to swelling in water depending on the proteins of the properties. Utilizing a protein eutectogel resin that is solid or rigid at room temperature and ductile when heated above 80 °C, addressed these limitations of printing with a eutectogel solution, and offered smooth printability of the above described 2D mesh in an extrusion printer. The printed formulation solidified as it cooled to room temperature and a coagulation process was not required.

[0174] Effect of eutectogel processing on electrospinning: Wetelectrospinning the zein-DES solution over a collector submerged under a cold-water bath and melt electrospinning the zein-DES resin using a metallic collector in air wasdemonstrated. A significant fiber deposition was observed while electrospinning zein solution in DES and the SEM image of the fibers produced is illustrated in Figure 8A. The electrospun zein mat is microfibrous with several regions that are fused together suggesting that the residual DES may result in some redissolution and further coagulation of the zein into non-fibrous structures. In contrast, as illustrated in Figure 8B, melt electrospinning the resin produced nanofibrous mats. However, the throughput of the fibers is low likely due to high viscosity of the polymer melt, and a significant amount of viscous resin solidifies and is wasted at the nozzle tip. In both solution and resin-based approaches, the addition of oleic acid made it more challenging to produce any fibers.

[0175] Extrusion printing of zein eutectogel solution: In Figure 9, the ability to extrusion print the zein eutectogel solution prepared with choline chloride and lactic acid was demonstrated by producing a 2D mesh of 26 mm x 26 mm on a non-stick surface of silicone rubber. In the solution-based formulation, the extruded eutectogel demonstrated smooth flow properties and was able to make the turns on the mesh pathway with minimal drag. When the 2D mesh prepared with the solution was submerged in cold water to remove the DES, initially no noticeable changes were observed in the dimensions of the mesh. However, after prolonged exposure over 1 hour in water, substantial swelling and eventual detachment from substrate suggested that loss of DES made the printed mesh mechanically weak.

[0176] The ability to extrusion print the protein eutectogel resin without the need for a coagulation bath to retain the mechanical properties of the eutectogel resin in the printed structure was also demonstrated. An additional plasticizer, glycerol, was used in the formulation to prepare the resin with an overall formulation containing 70% (w / w) zein, 23% (w / w) DES (1 mole choline chloride: 2 moles of lactic acid) and 7% (w / w) glycerol. The resin was prepared in the Haake batch mixer as discussed previously. In Figure 10 A-C, a 3D printed layer by layer square of 3 cm x 3 cm with 10 layers and a layer height of 300±10 pm is demonstrated. It was observed that the lower 3 layers were significantly compressed due to the temperature and weight of the upper layers, likely due to poor dissipation of heat from the layer onto the polymer film on the print bed. However, the upper layers retained their integrity, thereby showing promise in using this formulation to print 3D structures.

[0177] Biodegradability of protein eutectogel resins: The resin samples were tested for biodegradability in compost soil under thermophilic conditions. It was noticed that the S-DES (soy eutectogel) samples biodegraded completely in 7 days and all other resin samples (zein and gluten eutectogels) and cellulosic paper were fully biodegraded in 14 days. These results indicate that the protein eutectogels fully biodegrade within days in thermophilic compost conditions.

[0178] Melt blowing of zein-DES resin: The ability to melt blow the thermoplastic zein eutectogels prepared with choline chloride and lactic acid has been shown in Figure 11. A lab scale melt blower (Areka, E-25 / 25-D) was used to prepare melt blown mats with the Z-DES resin at 140-160 °C. The extruder was operated at 100 rpm and temperatures between 100-130 °C, and the die was maintained at 140 °C. At die temperatures above 170 °C, the resin fibers were still formed but started to appear slightly charred and such high temperatures were avoided.

[0179] Zein and DES based scaffolds for mammalian cell culture: Cell adhesion on the 6 scaffold variants after 1 day is illustrated in Figure 12. A low degree of cell adhesion was observed on the unwashed Z1 films (A), but all other films samples showed reasonable cell adhesion after 1 day of seeding the medium. However, the washed ZE (D) samples showed several dead cells (dots) alongside the live cells (spindles). The proportion of live cells to dead cells was found to be lower for the Zl’ samples likely due to chloride inhibiting cellular growth. Washed and unwashed samples of the betaine-based DES (B - Z2, E - Z2’, C - Z3, F - Z3’) showed reasonable cell adhesion and proliferation of the 3T3 fibroblast cells.

[0180] While the present disclosure has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples but should be given the broadest interpretation consistent with the description as a whole.

[0181] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present disclosure is found to be defineddifferently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.FULL CITATIONS FOR DOCUMENTS REFERENCED IN THE DISCLOSURE1. Peydayesh, M., et al., Turning Food Protein Waste into Sustainable Technologies. Chemical Reviews, 2023. 123(5): p. 2112-2154.2. Abascal, N.C. and L. Regan, The past, present and future of protein-based materials. Open Biology, 2018. 8(10): p. 180113.3. Xiang, N., et al., Edible fdms for cultivated meat production. Biomaterials, 2022. 287: p. 121659.4. Ma, L., et al., Nanoporous and lyophilic battery separator from regenerated eggshell membrane with effective suppression of dendritic lithium growth. Energy Storage Materials, 2018. 14: p. 258-266.5. Chen, M., et al., A Janus protein-based nanofabric for trapping polysulfides and stabilizing lithium metal in lithium-sulfur batteries. Journal of Materials Chemistry A,2020. 8(15): p. 7377-7389.6. Lin, C.-W., et al., Design and Investigation of an Eco-Friendly Wound Dressing Composed of Green Bioresources- Soy Protein, Tapioca Starch, and Gellan Gum. Macromolecular Bioscience, 2022. 22(12): p. 2200288.7. Gogurla, N., B. Roy, and S. Kim, Self-powered artificial skin made of engineered silk protein hydrogel. Nano Energy, 2020. 77: p. 105242.8. Bayer, I.S., Zein in Food Packaging, in Sustainable Food Packaging Technology.2021. p. 199-224.9. Bhargava, N., et al., Active and intelligent biodegradable packaging films using food and food waste-derived bioactive compounds: A review. Trends in Food Science & Technology, 2020. 105: p. 385-401.10. Zaib, Q., et al., Are deep eutectic solvents really green?: A life-cycle perspective. Green Chemistry, 2022. 24(20): p. 7924-7930.11. Kalhor, P. and K. Ghandi, Deep Eutectic Solvents for Pretreatment, Extraction, and Catalysis of Biomass and Food Waste. Molecules, 2019. 24(22): p. 4012.12. Jablonsky, M., et al., Extraction of value-added components from food industry based and agro-forest biowastes by deep eutectic solvents. Journal of Biotechnology, 2018. 282: p. 46-66.13. Lin, Z., et al., Optimization of protein extraction from bamboo shoots and processing wastes using deep eutectic solvents in a biorefinery approach. Biomass Conversion and Biorefinery, 2021. 11(6): p. 2763-2774.14. Wahlstrom, R., et al., High Yield Protein Extraction from Brewer's Spent Grain with Novel Carboxylate Salt - Urea Aqueous Deep Eutectic Solvents. ChemistrySelect, 2017. 2(29): p. 9355-9363.15. Zhou, Y., et al., Deep eutectic solvents as new media for green extraction of food proteins: Opportunity and challenges. Food Research International, 2022. 161: p. 111842.16. Qin, H., et al., Highly stretchable and nonvolatile gelatin-supported deep eutectic solvent gel electrolyte-based ionic skins for strain and pressure sensing. Journal of Materials Chemistry C, 2019. 7(3): p. 601-608.17. Qu, W., et al., Globular and Fibrous Proteins Modified with Deep Eutectic Solvents: Materials for Drug Delivery. Molecules, 2019. 24(19): p. 3583.18. Khatri, M., et al., Zein nanofibers via deep eutectic solvent electrospinning: tunable morphology with super hydrophilic properties. Scientific Reports, 2020. 10(1): p. 15307.19. Sachse, B., et al., A hemoglobin adduct as a biomarker for the internal exposure to the rodent carcinogen furfuryl alcohol. Archives of Toxicology, 2017. 91(12): p. 3843- 3855.20. Toxicology and Carcinogenesis Studies of Furfuryl Alcohol (CAS No. 98-00-0) in F344 / N Rats and B6C3F1 Mice (Inhalation Studies). Natl Toxicol Program Tech Rep Ser, 1999. 482: p. 1-248.21. Fanali, C., et al., Choline Chloride-Lactic Acid-Based NADES As an Extraction Medium in a Response Surface Methodology -Optimized Method for the Extraction of Phenolic Compounds from Hazelnut Skin. Molecules, 2021. 26(9): p. 2652.22. Rodriguez Rodriguez, N., et al., Degradation of Deep-Eutectic Solvents Based on Choline Chloride and Carboxylic Acids. ACS Sustainable Chemistry & Engineering, 2019. 7(13): p. 11521-11528.23. Skulcova, A., et al., Long-term isothermal stability of deep eutectic solvents based on choline chloride with malonic or lactic or tartaric acid. Int J Sci Eng Res, 2017. 8(7): p. 2249-2252.24. Delgado-Mellado, N., et al., Thermal stability of choline chloride deep eutectic solvents by TGA / FTIR-ATR analysis. Journal of Molecular Liquids, 2018. 260: p. 37- 43.

Claims

Claims:

1. A plant protein eutectogel comprising one or more plant proteins and a deep eutectic solvent (DES) wherein the one or more plant proteins are present in an amount of about 45% (w / w) to about 95% (w / w) and the DES is present in an amount of about 5% (w / w) to about 55% (w / w), based on the total weight of the plant protein eutectogel.

2. The plant protein eutectogel of claim 1, further comprising one or more plasticizers.

3. The plant protein eutectogel of claim 2, wherein the one or more plasticizers are present in an amount of from about 1% (w / w) to about 15% (w / w), about 5% (w / w) to about 10%(w / w), or about 7% (w / w), based on the total weight of the plant protein eutectogel and are selected from glycerol, triethylene glycol, dibutyl tartarate, levulinic acid, polyethylene glycol, sugars, fatty acids, and combinations thereof.

4. The plant protein eutectogel of any one of claims 1 to 3, wherein the one or more proteins are selected from zein, gluten, soy protein, rice bran protein, whey protein, pea protein, and combinations thereof.

5. The plant protein eutectogel of any one of claims 1 to 4, wherein the DES comprises a first element and a second element, wherein the first element is selected from choline chloride (ChCl), betaine (trimethylglycine), betaine hydrochloride, carnitine, choline bitartrate, histidine, glycine, nicotinic acid, phytic acid sodium, proline, glutamic acid, serine, ethylammonium chloride (EtNEECl), N-ethyl-2-hydroxy-N,N-dimethylethanaminium chloride, 2- (chlorocarbonyloxy)-N,N,N-trimethylethanaminium chloride, N-benxyl-2- hydroxy-N,N-dimethylethanaminium chloride, tetramethylaminium chloride (TMAC1), tetrabutylaminium chloride (TBAC1), methyltriphenylphosphonium bromide (MeP(Ph)3Br), methyltriphenylphosphonium chloride (MeP(Ph)3Cl), benzyltriphenylphosphonium bromide, tetraethylaminium bromide (TEABr), tetrabutylaminium bromide (TBABr), 2-hydroxy-N,N,-diethylethanaminium chloride (Et2(EtOH)ACl), 2-chloro-N,N,N-trimethylethanaminium chloride (CIChCl) and acetyl choline chloride (AcChCl), and the second element isselected from urea, p-toluenesulfonic acid (TsOH), acetamide, 1-methyl-urea, 1,3-dimethylurea, 1,1 -dimethylurea, 1,2-propanediol, thymol, cyclodextrin, imidazole, 2,2,2-trifluoroacetamide, thiourea, benzamide, glycerol, ethylene glycol, lactic acid, maleic acid, malic acid, malonic acid, benzoic acid, ascorbic acid, caffeic acid, adipic acid, oxalic acid, succinic acid, citric acid, phenylacetic acid, phenylpropionic acid, tricarballyhc acid, levulinic acid, itaconic acid, gallic acid, glucose, erythritol, xylitol, sorbitol, tartaric acid, fructose, isomaltose, isosorbide, lactose, maltose, mannitol, ribitol, sucrose, trehalose, xylose, tartaric acid, urea, decanoic acid, dodecanoic acid, isosorbide, 4- hydroxybenzoic acid, caffeic acid, coumaric acid, cinnamic acid, suberic acid, gallic acid and resorcinol.

6. The plant protein eutectogel of claim 5, wherein the first and second elements are present in the DES in a molar ratio of from 2: 1 to 1 :6, optionally 1:0.5, 1:1 or 1:2.

7. The plant protein eutectogel of any one of claims 1-6, wherein the DES is a mixture of choline chloride or betaine and lactic acid or glucose.

8. The plant protein eutectogel of any one of claims 1-7, wherein the plant protein eutectogel is further cross-linked, optionally with a crosslinker selected from citric acid, tannic acid, gallic acid, glutaraldehyde and a combination thereof.

9. The plant protein eutectogel of any one of claims 1-8, wherein the plant protein eutectogel further comprises one or more other ingredients selected from water, gelatin, collagen, albumin, starch, carbohydrates, and combinations thereof optionally wherein the one or more other ingredients are present in an amount of about 0.1% (w / w) to about 20% (w / w), about 0.5% (w / w) to about 10% (w / w), or 1% (w / w) to about 5% (w / w), by total weight of the plant protein eutectogel.

10. A plant protein plant eutectogel comprising: about 45% (w / w) to about 95% (w / w) one or more plant proteins; about 5% (w / w) to about 55% (w / w) of a DES; about 1% (w / w) to about 15% (w / w) of one or more plasticizers; and optionally about 0.1% (w / w) to about 20% (w / w) of one or more other ingredients,based on the total weight of the plant protein eutectogel.

11. The plant protein plant eutectogel of claim 10 comprising, about 70% (w / w) one or more plant proteins selected from zein, gluten and soy protein.; about 23% (w / w) of a DES comprising a first element comprising choline chloride or betaine and a second element comprising lactic acid or glucose, wherein the first and second elements are present in a molar ratio of 1 :1 or 1 :2; and about 7% (w / w) of glycerol, based on the total weight of the plant protein eutectogel.

12. The plant protein plant eutectogel of any one of claims 1 to 11, wherein the plant protein eutectogel is biodegradable, is hydrophilic, has an elongation at break that is greater than about 20%, can be injection molded or extrusion printed at temperatures greater than about 80°C to about 170°C and / or is a solid or is rigid at room temperature and ductile when heated above about 80°C.

13. A method of preparing a plant protein eutectogel comprising combining one or more plant proteins and a deep eutectic solvent (DES) under conditions to form the plant protein eutectogel wherein the one or more plant proteins are in an amount of about 45% (w / w) to about 95% (w / w) and the DES is in an amount of about 5% (w / w) to about 55% (w / w), based on the total weight of protein(s) and DES.

14. The method of claim 13, wherein the combining of the one or more plant proteins and the DES is by combining under conditions to form a resin, optionally wherein the one or more plant proteins and the DES are combined as dry powders and heated to a temperature above the melting point of the DES in a mixer until a resin is formed.

15. The method of claim 14, wherein the resin is granulated and the resulting granulated resins are shaped into any desired shape, for example by die casting, electrospinning, melt-blowing, 2D / 3D extrusion printing, stamping or injection molding, or the granulated resins are shaped into films comprising the plantprotein eutectogel by pressing the granulated resin onto a non-stick surface or by extruding the resin in an extruder.

16. The method of claim 13, wherein the combining of the one or more plant proteins and the DES is by combining the one or more plant proteins and a deep eutectic solvent and heating to form a solution and shaping the solution on a non-stick surface and cooling to form the plant protein eutectogel.

17. The method of claim 16, wherein the shaping of the solution on a non-stick surface comprises die casting, electrospinning, or extrusion printing or the shaping of the solution is by coagulating in water.

18. The method of any one of claims 13 to 17, further comprising combining the one or more plant proteins and the DES along with one or more plasticizers.

19. The method of claim 18, wherein the one or more plasticizers are used in an amount of from about 1% (w / w) to about 15% (w / w), about 5% (w / w) to about 10%(w / w), or about 7% (w / w), by total weight of the protein(s), DES and plasticizer(s) and are selected from glycerol, tri ethylene glycol, dibutyl tartarate, levulinic acid, polyethylene glycol, sugars, fatty acids, and combinations thereof.

20. The method of any one of claims 13 to 19, further comprising cross-linking the shaped protein eutectogel to improve rigidity, moisture tolerance and shelflife.

21. The method of any one of claims 13 to 20, wherein the one or more proteins are selected from zein, gluten, soy protein, rice bran protein, whey protein, pea protein, and combinations thereof.

22. The method of any one of claims 13 to 20, wherein the DES comprises a first element and a second element, wherein the first element is selected from choline chloride (ChCl), betaine (trimethylglycine), betaine hydrochloride, carnitine, choline bitartrate, histidine, glycine, nicotinic acid, phytic acid sodium, proline, glutamic acid, serine, ethylammonium chloride (EtNEECl), N-ethyl-2-hydroxy- N,N-dimethylethanaminium chloride, 2-(chlorocarbonyloxy)-N,N,N- trimethylethanaminium chloride, N-benxyl-2-hydroxy-N,N- dimethylethanaminium chloride, tetramethylaminium chloride (TMAC1),tetrabutyl aminium chloride (TBAC1), methyltriphenylphosphonium bromide (MeP(Ph)3Br), methyltriphenylphosphonium chloride (MeP(Ph)3Cl), benzyltriphenylphosphonium bromide, tetraethylaminium bromide (TEABr), tetrabutyl aminium bromide (TBABr), 2-hydroxy-N,N,-diethylethanaminium chloride (Et2(EtOH)ACl), 2-chloro-N,N,N-trimethylethanaminium chloride (CIChCl) and acetyl choline chloride (AcChCl), and the second element is selected from urea, p-toluenesulfonic acid (TsOH), acetamide, 1-methyl-urea, 1,3-dimethylurea, 1,1 -dimethylurea, 1,2-propanediol, thymol, cyclodextrin, imidazole, 2,2,2-trifluoroacetamide, thiourea, benzamide, glycerol, ethylene glycol, lactic acid, maleic acid, malic acid, malonic acid, benzoic acid, ascorbic acid, caffeic acid, adipic acid, oxalic acid, succinic acid, citric acid, phenylacetic acid, phenylpropionic acid, tricarballyhc acid, levulinic acid, itaconic acid, gallic acid, glucose, erythritol, xylitol, sorbitol, tartaric acid, fructose, isomaltose, isosorbide, lactose, maltose, mannitol, ribitol, sucrose, trehalose, xylose, tartaric acid, urea, decanoic acid, dodecanoic acid, isosorbide, 4- hydroxybenzoic acid, caffeic acid, coumaric acid, cinnamic acid, suberic acid, gallic acid and resorcinol.

23. The method of claim 22, wherein the first and second elements are present in the DES in a molar ratio of from 2:1 to 1:6, optionally 1:0.5, 1:1 or 1:2.

24. The method of any one of claims 13 to 23, wherein the DES is a mixture of choline chloride or betaine and lactic acid or glucose.

25. A use of a plant protein eutectogel of any one of claims 1-12 for food packaging, 3D printing inks or filaments, filtration materials, tissue engineering scaffolds, single use plastic containers, cups or utensils, insulation materials, clothing or filler materials for packaging.

26. A food packaging, 3D printing ink or filament, filtration material, tissue engineering scaffold, single use plastic container, cup or utensil, insulation material, clothing, or filler material for packaging comprising a plant protein eutectogels of any one of clams 1-12.

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