protein dispersion

Abstract

A method for preparing a plant-based protein hydrogel slurry is provided. [Solution] The present invention relates to a method for preparing a plant-based protein hydrogel slurry, and a method for preparing a plant-based structured material (e.g., a film, casting, molded product, etc.) from the plant-based protein hydrogel slurry.

Description

[Technical Field]

[0001] The present invention relates to a method for preparing a plant-based protein hydrogel slurry, and to a method for preparing plant-based structured materials (e.g., films, castings, molded products, etc.) from a plant-based protein hydrogel slurry. The present invention also relates to plant-based protein hydrogel slurries and plant-based structured materials themselves, as well as their uses. [Background technology]

[0002] There is an increasingly urgent need to reduce the environmental impact of many of our daily activities and to reduce the amount of non-renewable resources involved in these activities. One example of this is the increasing use of biodegradable or renewable packaging (e.g., edible films for use in food) that replaces conventional plastics such as polyethylene and polypropylene.

[0003] As a result, efforts have been directed towards the use of natural or naturally derived materials, such as cellulose, alginates, starches, collagen, and collagen-derived proteins, as film and packaging-forming materials. However, many of these naturally derived films often have limitations such as limited tensile strength, or sensitivity to moisture, or limited barrier properties, which limit the range of applications in which they are suitable. One option is to chemically modify the film-forming material by typically using crosslinkable chemicals that can crosslink long-chain polymers, but this introduces complexity and additional chemicals that may not be suitable for end-uses such as edible films or products requiring high biodegradability. Similarly, composite materials combining natural and synthetic materials to overcome any limitations of one material also become more complex to produce and are neither edible nor reusable.

[0004] Among different types of biopolymers that can act as building blocks for creating new functional materials such as films, proteins are interesting candidates if they possess the ability to self-assemble into functional structures.

[0005] Currently, the use of these materials for commercial purposes is limited to highly soluble animal-derived proteins. Animal-based proteins commonly used in food, such as whey protein, exhibit good biocompatibility, biodegradability, amphiphilicity, and functional properties such as water solubility, emulsifying, and foaming ability. However, there is a growing demand to replace animal-derived proteins with plant-based alternatives, not only because plant-based alternatives have a lower environmental impact, but also because they are less allergenic and cost-effective.

[0006] The formation of self-assembled hydrogel materials, such as films from plant-based proteins, has been reported, where hydrogels can be obtained from soybean and pea proteins under experimental conditions. However, the mechanical properties obtained from current plant-based materials are generally inferior to those obtained from animal-derived materials, because plant proteins are more difficult to process, at least in part, due to their inherently low water solubility. The same can be said for the optical and barrier properties of current plant-based materials. [Overview of the project] [Problems that the invention aims to solve]

[0007] Therefore, in order to broaden the range of applications in which plant-based materials can be used to further replace synthetic materials, there is a need to develop a simple pathway to plant-based material products with improved properties. [Means for solving the problem]

[0008] In view of the first aspect, the present invention relates to a method for preparing a plant-based protein hydrogel slurry, (a) A step of forming a solution containing one or more plant-based proteins in a solvent system, the solvent system containing a miscible co-solvent, a first co-solvent enhancing the solubility of the plant-based protein, and a second co-solvent reducing the solubility of the plant-based protein; (b) A step of inducing a sol-gel transition of the protein in the solution to form a plant-based protein hydrogel; (c) A step of subjecting the plant-based protein hydrogel to shear treatment to form a plant-based protein hydrogel slurry A method comprising.

[0009] In a further aspect, the present invention provides a plant-based protein hydrogel slurry prepared according to the method previously described herein.

[0010] In a further aspect, the present invention is a method for preparing a plant-based structured material, comprising: (a) A step of preparing a plant-based protein hydrogel slurry according to the method previously described herein; (b) Subjecting the plant-based protein hydrogel slurry to one or more solvent concentration reduction steps to reduce the concentration of the first co-solvent and / or the second co-solvent to obtain the plant-based structured material A method comprising.

[0011] In a further aspect, the present invention provides a plant-based structured material prepared according to the method previously described herein.

[0012] In a further aspect, the present invention provides the use of the plant-based protein hydrogel slurry previously described herein for producing a plant-based structured material.

[0013] In a further aspect, the present invention has a protein solid content of 5 wt% to 25 wt% based on the total mass of the plant-based protein hydrogel slurry, and 50 s -1and a plant-based protein hydrogel slurry having a viscosity in the range of 10 to 10,000 cps at 20°C, wherein the d is 0.5 to 150 microns, determined by laser diffraction. 50 This invention provides a plant-based protein hydrogel slurry containing fragments with particle size.

[0014] In a further aspect, the present invention provides a protein solid content of 5 wt% to 25 wt% based on the total mass of a plant-based protein hydrogel slurry, and 50s -1 and a plant-based protein hydrogel slurry having a viscosity in the range of 10 to 10,000 cps at 20°C, wherein the d is less than 500 nm and is determined by dynamic light scattering. 50 This invention provides a plant-based protein hydrogel slurry containing fragments with particle size.

[0015] In a further embodiment, the present invention provides a film comprising a plant-based protein hydrogel slurry as previously described herein.

[0016] definition As used herein, the term “low shear process” may refer to a process step in which a low level of mechanical energy is applied to a material, preferably by a cutting action, causing it to break or fragment primarily into larger, separated pieces. “Low shear” is a high-speed impact, e.g., 2 ms -1 It typically does not involve any milling process that pulverizes or fragments the material by impact at a differential velocity greater than or equal to the limit. It also typically does not involve milling processes based on cavity formation. In certain embodiments, during a low-shear process, the hydrogel is fragmented such that at least 80% by mass of the hydrogel fragments have a maximum size between 1 mm and 50 mm, as determined by sieving. Sieving of the hydrogel slurry can be carried out according to the methods described herein.

[0017] As used herein, the term “high shear process” may refer to a process step in which energy is applied to reduce a hydrogel into small fragments, for example, to form a colloidal dispersion. During the high shear process, the hydrogel is fragmented, where appropriate, into fragments of 50 nm to 150 microns, determined by dynamic light scattering (DLS) or laser diffraction. 50 Fragments having particle size can be imparted. In certain embodiments, during the high shear process, the hydrogel is fragmented and d is determined by dynamic light scattering (DLS) of less than 500 nm. 50 Fragments with particle size are imparted. In an alternative embodiment, during the high-shear process, the hydrogel is fragmented to 0.5-150 microns, determined by laser diffraction. 50 Fragments having particle size are imparted. DLS and laser diffraction can be carried out according to the methods defined herein.

[0018] To avoid ambiguity, the high-shear step involves subjecting the hydrogel to a higher level of shearing than the low-shear step. If the method includes both low-shear and high-shear steps, the high-shear step must occur after the low-shear step (i.e., they are separate steps that occur in this particular order). [Modes for carrying out the invention]

[0019] This application describes a method for preparing an injectable and pumpable hydrogel slurry that can be dried to form a robust film, coating, molded article, or other structured product. Therefore, the present invention relates to a method for preparing a plant-based protein hydrogel slurry, (a) A step of forming a solution containing one or more plant-based proteins in a solvent system, wherein the solvent system contains a miscible cosolvent, the first cosolvent increasing the solubility of the plant-based protein, and the second cosolvent decreasing the solubility of the plant-based protein. (b) A step of inducing a sol-gel transition of a protein in solution to form a plant-based protein hydrogel, (c) A step of subjecting a plant-based protein hydrogel to shear treatment to form a plant-based protein hydrogel slurry. This provides a method that includes [something].

[0020] In some embodiments, it may be preferable to remove the solvent from the protein solution before step (b) in order to form a more concentrated protein solution prior to step (b). This can be done by applying heat and / or vacuum, among other techniques. This initial solvent reduction can provide advantages such as simplifying the subsequent drying. Suitable equipment includes a scraped-wall evaporator or a twin-screw extruder with the application of vacuum.

[0021] Any suitable plant-based protein can be used in the present invention. Different plant-based proteins can impart hydrogel slurries that confer structures with different properties. For example, soy protein can impart hydrogels (and materials formed from these hydrogels) that are more robust than pea protein and may need to be processed differently for optimal performance. However, suitability for the present invention is determined by more than just the properties of the hydrogels, which are balanced with other factors such as the availability of protein raw materials, lack of competition for food supply, and protein allergenicity. In the preferred method of the present invention, the plant-based protein is selected from soy protein, pea protein, rice protein, potato protein, wheat protein, corn zein protein, or sorghum protein. Preferably, the plant protein is selected from soy protein, pea protein, potato protein, and / or rice protein. More preferably, the plant-based protein is selected from soy protein and / or pea protein.

[0022] In the preferred method of the present invention, the plant-based protein is selected from soy protein, pea protein, rice protein, potato protein, wheat protein, corn zein protein, rapeseed protein, or sorghum protein. Preferably, the plant protein is selected from soy protein, pea protein, potato protein, rapeseed protein, and / or rice protein. More preferably, the plant-based protein is selected from soy protein and / or pea protein.

[0023] Suitable plant-based proteins further include: - Brassica genus: Brassica balearica: Mallorcan cabbage, Brassica carinata: Abyssinian mustard or Abyssinian cabbage, Brassica elongata: Elongated mustard, Brassica fruticulosa: Mediterranean cabbage, Brassica hilarionis: St. Hilarion cabbage, Brassica juncea: Indian mustard, brown and leaf mustard, Salepta mustard, Brassica napus: Rapeseed, canola, rutabaga, Brassica narinosa: Broadbeaked mustard, Brassica nigra: Black mustard, Brassica oleracea This includes: oleracea (kale, cabbage, collard greens, broccoli, cauliflower, kailan, Brussels sprouts, kohlrabi), Brassica perviridis (tender greens, mustard spinach), Brassica rapa (synonymous with B. campestris) (Chinese cabbage, turnip, rapini, komatsuna), Brassica rupestris (brown mustard), and Brassica tournefortii (Asian mustard); - Solanaceae family: Includes tomatoes, potatoes, eggplants, bell peppers, and chili peppers; - Grains: maize, rice, wheat, barley, sorghum, millet, oats, rye, ra Includes wheat and fonio; - Pseudocereals: including amaranth (lovely breeding, red amaranth, prince of walnut feather), breadfruit, buckwheat, kia, celosia (also called quailgrass or soko), pit seed goosefoot, cannua, quinoa, and wattle seed (also called acacia seed); - Leguminous plants: Acacia alata (Winged Wattle), Acacia decipiens, Acacia saligna (commonly known by various names including Coujon, Golden Wreath Wattle, Orange Wattle, and Blue Leaf Wattle), Arachis hypogaea (Peanut), Astragalus galegiformis, Cytisus laburnum (Common Laburnum, Golden Chain or Golden Rain), Cytisus supinus, Dolichios laburnum lablab) (common names include hyacinth bean, hyacinth bean, bonavista bean / pea, dolichos bean, Seym River bean, hyacinth bean, Egyptian kidney bean, Indian bean, batau and Australian pea), Ervum lens (lentil), Genista tinctorial (common names include dyerswin, waxwood and waxwood), Glycine max (soybean), Lathyrus clymenum (pea vine or lamb's weeping grass), Lathyrus odoratus (pea vine or lamb's weeping grass), Lathyrus staivus (pea vine or lamb's weeping grass), Lathyrus silvetris (Lathyrus Silvetris (pea vine or lamb's vine), Lotus tetragonolobus (asparagus - pea or wing pea), Lupinus albus (fan pea), Lupinus angustifolius (fan pea), Lupinus luteus (fan pea), Lupinus polyphyllus (LupinusPolyphyllus (Japanese pea), Medicago sativa (alfalfa), Phaseolus aureus (mung bean), Phaseolus coccineus (runner bean), Phaseolus nanus (green bean / French bean), Phaseolus vulgaris (green bean / French bean), Pisum sativum (pea), Trifolium hybridum (clover), Trifolium pretense (red clover), Vicia faba (broad bean), Vicia sativa (Vicia Includes sativa (Vicia sativa) and Vigna unguiculate (Camellia sinensis); - Non-legum plants: Acanshosicyos horrida, Aesculus hyppocastanum (Conker Tree / Horse Chestnut), Anacardium occidentale (Cashew Tree), Balanites aegyptica, Bertholletia excels (Brazil Nut), Beta vulgaris (Sugar Beet), Brassica napus (Rapeseed), Brassica juncea (Brown Mustard), Brassica nigra (Black Mustard), Brassica hirta (Brassica hirta (Eurasian mustard), Cannabis sativa (marijuana), Citrullus vulgaris (watermelon), Citrus aurantiaca (citrus), Cucurbita maxima (squash), Fagopyrum esculentum (knotweed), Gossypium barbadense (extra-long staple cotton), Heianthus annuus (sunflower), Nicotiana sp.Includes tobacco plants, Prunus avium (cherry), Prunus cerasus (white cherry), Prunus domestica (plum), Prunus amygdalus (almond), Lycinus communis (castor bean / oil plant), Sasamum indicum (sesame), Sinapis alba (white mustard), and Terlfalrea pedata (oyster nut).

[0024] To avoid any doubt, the plant-based hydrogels and structured materials according to the present invention do not include plants in their natural state; for example, naturally formed plant cells, organelles, or vesicles are not plant-based hydrogels or structured materials of the present invention.

[0025] In the method according to the present invention, a plant-based protein hydrogel is formed by adding a plant-based protein to a solvent system, the solvent system comprising two or more miscible cosolvents as defined herein. The properties of the hydrogel and the associated sol-gel state can be controlled by selecting a solvent system comprising a miscible cosolvent in which the first cosolvent enhances the solubility of the plant-based protein and the second cosolvent reduces the potentiality of the plant-based protein.

[0026] The first co-solvent enhances the solubility of the plant-based protein. The first co-solvent can be considered a solubilizing co-solvent. One or more solubilizing co-solvents may be present, and these can solubilize the plant-based protein entirely or partially.

[0027] Examples of solubilizing cosolvents are organic acids. Organic acids are organic compounds that have acidic properties. Suitable organic acids include acetic acid, formic acid, propionic acid, or α-hydroxy acids. Suitable organic acids include acetic acid, formic acid, propionic acid, α-hydroxy acids, or β-hydroxy acids. Suitable α-hydroxy acids include glycolic acid, lactic acid, malic acid, citric acid, and tartaric acid. Suitable β-hydroxy acids include β-hydroxypropionic acid, β-hydroxybutyl acid, β-hydroxyβ-methylbutyl acid, 2-hydroxybenzoic acid, and carnitine. Particularly preferred organic acids are volatile organic acids, i.e., those with a boiling point below 130°C. This is because volatile organic acids can be easily removed from the plant-based protein hydrogen slurry during the subsequent drying process, so that the final plant-based structural material, if present, contains very few remaining organic acids. Examples of volatile organic acids include acetic acid and formic acid. Preferred organic acids are acetic acid and lactic acid. The use of organic acids allows for the solubilization of plant proteins and also enables their gentle hydrolysis. For example, although we do not wish to be bound by theory, the solubility of plant-based proteins in organic acids is possible due to i) protonation of the protein and ii) the presence of an anionic solvation layer that contributes to a reduction in hydrophobic interactions.

[0028] In the preferred method of the present invention, the first cosolvent is an organic acid.

[0029] In the preferred method of the present invention, the first cosolvent has a boiling point of less than 130°C, more preferably less than 120°C.

[0030] The second cosolvent exhibits reduced solubility of plant-based proteins compared to the first cosolvent. The second cosolvent may be considered a desolubilizing cosolvent. One or more desolubilizing cosolvents may be present.

[0031] An example of a desolubilizable second cosolvent is an aqueous buffer solution. Preferably, the second cosolvent may be water, ethanol, methanol, acetone, acetonitrile, dimethyl sulfoxide, dimethylformamide, formamide, 2-propanol, 1-butanol, 1-propanol, hexanol, t-butanol, ethyl acetate, or hexafluoroisopropanol. Particularly preferably, the second cosolvent is water and / or ethanol. Most preferably, the second cosolvent is water.

[0032] In the preferred method of the present invention, the second cosolvent has a boiling point of less than 130°C, more preferably less than 120°C.

[0033] In the preferred method of the present invention, the concentration of the plant-based protein in the solvent system in step (a) is 50 to 250 mg / ml, preferably 50 to 150 mg / ml. The ratio of the solubilizing cosolvent (typically an organic acid) may vary depending on the protein concentration, for example, by using a higher ratio of organic acid to increasing protein concentration.

[0034] In the preferred method of the present invention, the solvent system has a cosolvent ratio of a first cosolvent to a second cosolvent of about 20-80 v / v%, about 20-60 v / v%, about 25-55 v / v%, about 30-50 v / v%, about 20%, about 30%, about 40%, about 50%, or about 60 v / v%, most preferably about 30-50 v / v%. Such ratios lead to a functionally useful material.

[0035] In a preferred method of the present invention, to form a hydrogel, the protein solution is heated to a first temperature above the sol-gel temperature of one or more plant-based protein solutions, and then cooled to a second temperature below the sol-gel temperature of one or more plant-based protein solutions.

[0036] In the preferred method of the present invention, the degree of protein hydrolysis (i.e., the percentage of cleaved peptide bonds in the protein hydrolysate) is controlled to alter the properties of the resulting hydrogel. For example, increasing the concentration of organic acid present during formation increases the degree of protein hydrolysis. A higher degree of protein hydrolysis leads to the formation of a less rigid hydrogel.

[0037] In the preferred method of the present invention, the degree of protein hydrolysis is 0.1 to 10%, preferably 0.1 to 5%, and more preferably 0.1 to 2.5%.

[0038] To form a solution containing one or more plant-based proteins, it may be necessary to apply physical stimulation to the protein / solvent mixture to enable protein dissolution. Suitable physical stimulations include heating, sonication, stirring, high-shear mixing, or other physical techniques. A preferred technique is heating with optional simultaneous or sequential sonication.

[0039] Preferably, the protein / solvent mixture is subjected to a physical stimulus, which is heating, where the solution is heated to about 70°C or above 70°C. More preferably, the protein / solvent mixture is heated to about 75°C or above 75°C, about 80°C or above 80°C, about 85°C or above 85°C, or about 90°C. Even more preferably, the protein / solvent mixture is heated to 85°C.

[0040] Preferably, the protein / solvent mixture is subjected to physical stimulation, including heating for about 5, 10, 15, 20, 25, 30 minutes, or more than 30 minutes. A preferred heating time is about 30 minutes. The heated protein / solvent mixture is optionally subjected to simultaneous or sequential sonication.

[0041] The resulting protein solution is then heated so that the protein solution is maintained above the sol-gel transition temperature of the protein solution. The sol-gel transition temperature of the protein can be altered by modifying the solvent system (e.g., through the selection of organic acids, the ratio of organic acids to further solvents, or by further means). Through appropriate selection of conditions, the sol-gel transition of the protein can be carefully controlled, thereby controlling the formation of the hydrogel.

[0042] Preferably, the protein solution is heated to about 70°C or above 70°C. More preferably, the protein solution is heated to about 75°C or above 75°C, about 80°C or above 80°C, about 85°C or above 85°C, or about 90°C. Even more preferably, the protein solution is heated to about 85°C.

[0043] The protein solution can be held at a higher temperature for approximately 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 45 minutes, or 1 hour. A preferred time is at least 30 minutes, which allows for complete solubilization of the protein. It is possible to hold the protein solution at a higher temperature for longer periods. This may be useful for commercial batch processes or for use in fluid processing steps where it is necessary to keep the protein solution in liquid form for a longer period.

[0044] When a protein solution is heated above its sol-gel transition temperature, the temperature of the protein solution can be reduced to a second temperature below the sol-gel transition temperature, thereby promoting hydrogel formation. The second temperature may be room temperature. The second temperature may be in the range of 5 to 25°C, preferably 10 to 20°C. The protein solution can be held at a low temperature for a long time, for example, several days or weeks. The protein solution can be held at a low temperature for about 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, or about 30 minutes. A specific reduction time is about 5 minutes. However, the method of the present invention allows the protein to remain in the solution for a long time. As such, if necessary, the protein solution can be held above the sol-gel transition temperature, insofar as it is required to retain the protein in liquid form. This can be several hours, several days, or longer, but preferably in the order of several minutes or hours. Furthermore, because the method is reversible, the solution can be kept at a lower temperature (e.g., room temperature) where the hydrogel will form, and then heated above the sol-gel transition temperature to return the solution to a liquid state for further processing. Protein hydrogels can thus be stored for hours, days, weeks, months, or even years, so that the hydrogel remains stable for an extended period.

[0045] The specific temperature will depend on the properties of the protein source, the solvent conditions used, and therefore the sol-gel transition temperature. Alternatively, the heating and cooling temperatures may be relatively fixed (e.g., to about 85°C, then to approximately room temperature), and the co-solvent mixture conditions may be adjusted to ensure a suitable sol-gel transition temperature for the selected plant-based protein.

[0046] Therefore, the preferred method of the present invention is (ai) A step of forming a protein solution comprising one or more plant-based proteins and a solvent system, wherein the solvent system comprises a miscible co-solvent, wherein a first co-catalyst increases the solubility of the plant-based protein and a second co-catalyst decreases the solubility of the plant-based protein. (aii) A step of subjecting a protein solution to physical stimulation, such as heating and / or ultrasonic treatment, for a certain period of time. (bi) A step of raising the temperature of the protein solution to a first temperature rise above the sol-gel transition temperature for a certain period of time, (bii) A step of lowering the temperature of the protein solution to below the sol-gel transition temperature so that the plant-based protein self-aggregates into a plant-based protein hydrogel, (c) A step of subjecting a plant-based protein hydrogel to shear treatment to form a plant-based protein hydrogel slurry. Includes.

[0047] The protein solution is held at a rising temperature in step (bi), during which time it is formed in a suitable shape, and after the temperature is lowered in step (bii), the protein is allowed to form into a hydrogel.

[0048] While we do not wish to be bound by theory, it is certain that when plant proteins are added to a solvent system and subjected to physical stimuli such as heat and / or sonication, the plant proteins partially unfold, leading to the exposure of hydrophobic amino acids initially embedded within the unfolded protein structure. Once partially unfolded, the cosolvent can interact with the unfolded protein molecules. For example, organic acids have greater access to protonate the amino acid residue and to enable the formation of anionic salt bridges that stabilize hydrophobic interactions. Also, upon heating at higher temperatures, protein-protein non-covalent intermolecular contacts break down.

[0049] Furthermore, it is believed that cooling the protein solution to below the sol-gel temperature enables non-covalent intermolecular contact between proteins, thereby promoting the self-assembly of interconnected protein aggregates of plant protein molecules into a hydrogel.

[0050] Furthermore, it is believed that mechanical stirring, such as sonication, will break down large colloidal protein aggregates into smaller ones and disrupt intermolecular interactions between proteins. Using this approach, the size of protein aggregates can be significantly reduced to a particle size of less than 100 nm.

[0051] Preferably, the method of the present invention produces a plant-based protein solution containing protein aggregates having an average size of less than 200 nm, preferably less than 150 nm, less than 125 nm, less than 100 nm, less than 90 nm, less than 80 nm, less than 70 nm, less than 60 nm, less than 50 nm, less than 40 nm, or less than 30 nm.

[0052] It is believed that the method of the present invention enables plant proteins to aggregate into supramolecular structures that are held in place by intermolecular hydrogen bonding interactions, particularly between β-strands. The method of the present invention enables materials to be formed such that there are high levels of β-sheet intermolecular interactions.

[0053] A key feature of the method of the present invention is that the plant-based protein self-forms a hydrogel, thus eliminating the need for the use of a crosslinking agent. Therefore, in the preferred method of the present invention, the plant-based protein hydrogel is free of or substantially free of a crosslinking agent.

[0054] However, in an alternative preferred method of the present invention, the plant-based protein hydrogel may contain a crosslinking agent. Suitable crosslinking agents include microbial transglutaminase, glutaraldehyde, formaldehyde, glyoxal, phenol compounds, epoxy compounds, genipin, or dialdehyde starch.

[0055] In the method of the present invention, a plant-based protein hydrogel is subjected to shearing treatment to form a plant-based protein hydrogel slurry. As can be understood by those skilled in the art, the shearing treatment does not include centrifugation. Without wishing to be bound by theory, it is believed that the shearing treatment modifies the composition of the hydrogel such that the resulting slurry consists of small fragments of controlled size. This means that the plant-based protein hydrogel slurry can be injected, pumped, and handled. In addition, the small fragments can then bind well together during subsequent processing to form, for example, a film. The inventors of the present invention have surprisingly found that, generally, the smaller the fragments of the hydrogel slurry, the better the tensile strength and optical properties of the resulting film.

[0056] The inventors have also found that the rheological properties of the hydrogel need to be controlled with certain limitations, ideally to assist in the processing of the hydrogel and the properties of the final film.

[0057] In a preferred method of the present invention, the shearing treatment includes a high-shear step. Preferably, the high-shear step involves fragmenting the plant-based protein hydrogel into fragments.

[0058] In a preferred method of the present invention, the fragments produced in the high-shear step have a d determined by dynamic light scattering (DLS) of less than 500 nm, preferably less than 300 nm, more preferably less than 200 nm, even more preferably less than 50 nm. 50 and have.

[0059] In an alternative preferred method of the present invention, the fragments produced in the high-shear step have a d determined by laser diffraction of 0.5 to 150 microns, preferably 0.6 to 100 microns, more preferably 0.7 to 70 microns, even more preferably 0.8 to 50 microns, more preferably 0.9 to 25 microns, more preferably 1 to 20 microns, more preferably 1 to 10 microns, even more preferably 1 to 5 microns. 50The inventors have found, surprisingly, that when the fragments are within these particle size ranges, the hydrogel slurry can be used to form films with improved properties such as tensile strength or transparency.

[0060] In the preferred method of the present invention, the fragment produced in the high shear step is less than 10 microns, more preferably less than 8 microns, more preferably less than 6 microns, more preferably less than 4 microns, more preferably less than 2 microns, and even more preferably less than 1 micron, as determined by laser diffraction. 10 The present invention has a preferred method, wherein the fragment produced in the high shear step is d, which is 0.05 to 10 microns, more preferably 0.1 to 8 microns, more preferably 0.2 to 6 microns, determined by laser diffraction. 10 It has the following properties. While we do not wish to be bound by theory, it is thought that the presence of a certain proportion of relatively smaller fragments in a hydrogel slurry may help to bind together the relatively larger fragments that are also present therein, which means that any final structured material (e.g., film) made using a plant-based protein hydrogel slurry will have better mechanical properties.

[0061] Therefore, in the preferred method of the present invention, the fragment produced in the high shear step is d, which is 0.5 to 15 microns in size and determined by laser diffraction. 50 , and d, which is determined by laser diffraction, in the range of 0.05 to 10 microns. 10 Preferably d of 0.6 to 100 microns 50 and d of 0.1 to 8 microns 10 More preferably d of 0.7 to 70 microns 50 and d of 0.2 to 6 microns 10 It has.

[0062] In the preferred method of the present invention, the fragment produced in the high-shear step has a d5 determined by laser diffraction of less than 3 microns, more preferably less than 1 micron, and even more preferably less than 0.15 microns.

[0063] In the preferred method of the present invention, the high-shear step includes ultrasonic treatment (using equipment such as Bandelin HD4200, TS 113 probe, or Hielscher UIP1000hdT), high-shear mechanical stirring (using equipment such as Silverson rotor-stator high-shear mixer), or cavitation, preferably ultrasonic treatment.

[0064] In the preferred method of the present invention, the high-shear step includes ultrasonic treatment (using equipment such as Bandelin HD4200, TS 113 probe, or Hielscher UIP1000hdT), high-shear mechanical stirring (using equipment such as Silverson rotor-stator high-shear mixer), high-pressure homogenization, or cavitation, preferably ultrasonic treatment.

[0065] In the preferred method of the present invention, the high-shear step is carried out at a temperature below the sol-gel temperature of the plant-based protein solution. In the preferred method of the present invention, the high-shear step is carried out for at least 5 minutes, more preferably at least 1 minute.

[0066] In a preferred method of the present invention, the low-shear step comprises two steps. Preferably, the shearing treatment comprises a low-shear step followed by a high-shear step.

[0067] In a preferred method of the present invention, the low-shear step comprises fragmenting a plant-based protein hydrogel into fragments. Preferably, at least 50 wt% of the fragments produced in the low-shear step have a particle size in the range of 1 mm to 50 mm, preferably 5 mm to 30 mm, and more preferably 10 mm to 30 mm. More preferably, at least 80 wt% of the fragments produced in the low-shear step have a particle size in the range of 1 mm to 50 mm, preferably 5 mm to 30 mm, and more preferably 10 mm to 30 mm. This can be measured by collecting the fragments on a series of sieves with progressively smaller mesh sizes and calibrating the amount on different meshes.

[0068] The low-shear process is carried out at a temperature below the sol-gel temperature of the plant-based protein solution.

[0069] In the preferred method of the present invention, the low-shear step includes mechanical cutting. "Mechanical cutting" means cutting using a knife edge (e.g., a knife, an extruder blade, etc.).

[0070] In an alternative preferred method of the present invention, the low-shear step includes extrusion. For example, the plant-based protein solution formed in step (a) is extruded into an insolubilizable solvent (e.g., water) to form a plant-based protein hydrogel in the separated large fragment, which may take the form of an extruded product having the form of a thread or string. Thus, the fragment can be directly subjected to a solvent reduction step, as described in more detail below. A low-shear step of this nature is acceptable for large-scale processing. In this case, the low-shear step can reduce at least one dimension of the large fragment to between 1 mm and 50 mm, for example, to the diameter of the extruded product. Preferably, at least 50 wt% of the fragment produced in the low-shear step have at least one internal dimension in the range of 1 mm to 50 mm, preferably 5 mm to 30 mm, and more preferably 10 mm to 30 mm. More preferably, at least 80 wt% of the fragments produced in the low-shear process have at least one internal dimension in the range of 1 mm to 50 mm, preferably 5 mm to 30 mm, and more preferably 10 mm to 30 mm.

[0071] In a preferred method of the present invention, the high-shear step comprises further fragmentation of the plant-based protein hydrogel. Preferably, the fragments produced in the high-shear step have a d size less than 500 nm, preferably less than 300 nm, more preferably less than 200 nm, and even more preferably less than 50 nm, as determined by dynamic light scattering. 50 In an alternative method, the fragment produced in the high shear step is d, which is 0.5 to 150 microns, preferably 0.6 to 100 microns, more preferably 0.7 to 70 microns, even more preferably 0.8 to 50 microns, more preferably 0.9 to 25 microns, more preferably 1 to 20 microns, more preferably 1 to 10 microns, and even more preferably 1 to 5 microns, as determined by laser diffraction. 50 The inventors have found, surprisingly, that when the fragments are within these particle size ranges, the hydrogel slurry can be used to form films with improved properties, such as tensile strength.

[0072] In the preferred method of the present invention, the fragment produced in the high shear step is less than 10 microns, more preferably less than 8 microns, more preferably less than 6 microns, more preferably less than 4 microns, more preferably less than 2 microns, and even more preferably less than 1 micron, as determined by laser diffraction. 10 The present invention has a preferred method, wherein the fragment produced in the high shear step is d, which is 0.05 to 10 microns, more preferably 0.1 to 8 microns, more preferably 0.2 to 6 microns, determined by laser diffraction. 10 It has the following properties. While we do not wish to be bound by theory, it is thought that a certain proportion of relatively smaller fragments in a hydrogel slurry may help bind with relatively larger fragments that are also present therein, which would mean that any final structured material (e.g., a film) made using a plant-based protein hydrogel slurry would have better mechanical properties.

[0073] Therefore, in the preferred method of the present invention, the fragment produced in the high shear step is d, which is 0.5 to 150 microns in size and determined by laser diffraction. 10 , and d, which is determined by laser diffraction, in the range of 0.05 to 10 microns. 10 Preferably, d 50 and d of 0.1 to 8 microns 10 More preferably 0.7 to 7 microns 50 and d of 0.2 to 6 microns 10 It has.

[0074] In the preferred method of the present invention, the fragment produced in the high-shear step has a d5 determined by laser diffraction of less than 3 microns, more preferably less than 1 micron, and even more preferably less than 0.15 microns.

[0075] In a preferred method of the present invention, the particle size distribution of hydrogel fragments in a plant-based protein hydrogel slurry can be adjusted by diversifying the properties and intensity of the high-shear process. In another preferred method, the particle size distribution of hydrogel fragments in a plant-based protein hydrogel slurry can be adjusted by blending or combining two or more different hydrogel slurries that have been subjected to different high-shear processes and have different particle size distributions.

[0076] In the preferred method of the present invention, the high-shear step is carried out at a temperature below the sol-gel temperature of the plant-based protein solution.

[0077] In the preferred method of the present invention, the high-shear step is carried out at a temperature below the protein denaturation temperature of the plant-based protein solution.

[0078] The high-shearing step is performed for at least 5 minutes, more preferably at least 1 minute.

[0079] In the preferred method of the present invention, the high-shear step includes ultrasonic treatment (using equipment such as Bandelin HD4200, TS 113 probe, or Hielscher UIP1000hdT), high-shear mechanical stirring (using equipment such as Silverson rotor-stator high-shear mixer), or cavitation, preferably ultrasonic treatment.

[0080] In the preferred method of the present invention, the high-shear step includes ultrasonic treatment (using equipment such as Bandelin HD4200, TS 113 probe, or Hielscher UIP1000hdT), high-shear mechanical stirring (using equipment such as Silverson rotor-stator high-shear mixer), high-pressure homogenization, or cavitation, preferably ultrasonic treatment.

[0081] In a preferred method of the present invention, step (c) further includes, between the low-shear step and the high-shear step, a step of subjecting a plant-based protein hydrogel slurry to a solvent reduction step, most preferably a solubilizing solvent reduction step.

[0082] The term "solubilizing solvent" refers to a solvent or solvent mixture in which a plant-based protein hydrogel slurry is dissolved. Examples include organic acids, such as acetic acid, formic acid, propionic acid, and / or α-hydroxy acids. The α-hydroxy acid can preferably be selected from glycolic acid, lactic acid, malic acid, citric acid, and / or tartaric acid. The β-hydroxy acid can preferably be selected from β-hydroxypropionic acid, β-hydroxybutyl acid, β-hydroxyβ-methylbutyl acid, 2-hydroxybenzoic acid, and carnitine.

[0083] In the preferred method of the present invention, the solvent reduction step is performed as follows: (i) A step of contacting a fragment of a plant-based hydrogel slurry with an insoluble solvent, (ii) A step of separating fragments of the plant-based hydrogel slurry from an insolubilizing solvent to obtain a washed plant-based protein hydrogel, (iii) A process in which process (i) and process (ii) are repeated at the discretion of the user. Includes.

[0084] Step (i) involves contacting fragments of the plant-based protein hydrogel slurry with an insolubilizing solvent. “Insolubilizing solvent” means a solvent or solvent mixture in which the plant-based protein hydrogel slurry does not dissolve. Examples include water, or a mixture of water and ethanol.

[0085] In a preferred method of the present invention, step (ii) comprises mesh filtration. More preferably, the mesh filtration comprises the use of multiple meshes with decreasing size.

[0086] As can be understood by those skilled in the art, if the fragments produced in the low-shear process are too small, the solvent reduction process can be made difficult because the fragments can finish blocking the mesh. However, if the fragments produced in the low-shear process are too large, the solvent reduction process may take an excessive amount of time due to the slow mass transport of the solvent from the core of the fragments.

[0087] While we do not wish to be bound by theory, due to the porous nature of hydrogels, it is conceivable that the solvent reduction process can remove some or all of the solvent (e.g., organic acids) from the hydrogel through solvent exchange.

[0088] It is important that the hydrogel fragments are weak, deformable, and small enough to fit together well during subsequent processing to form the final film or other structured material. If the hydrogel fragments are not sufficiently deformable, the strength and integrity of any film or structured material will be reduced. In addition, strong hydrogels are more difficult to disperse and form a slurry.

[0089] It is also important that the hydrogel fragments are neither too soft nor too deformable. If they are too soft, any intermediate processing steps (e.g., washing and solvent exchange) can become difficult. Excessively soft hydrogel fragments typically result from an insufficient level of pre-formed macrostructure in the hydrogel, which typically leads to a weaker film or structured material.

[0090] The strength of the protein hydrogel can be altered by varying the concentrations of protein and organic acid, among other factors.

[0091] Therefore, it is important that the strength of the hydrogel used to form the hydrogel slurry is within a certain limit. This can be measured by vibrational rheometry. A suitable measure of hydrogel strength is the storage modulus G' of the hydrogel. Suitable test conditions are 1% strain at a vibration frequency of 1 Hz at 20°C. A suitable instrument is the Anton Paar MCR 92 Rheometer, which has a conical and flat plate measuring shape with a diameter of 50 mm and an angle of 1°.

[0092] Therefore, in the preferred method of the present invention, before washing, the plant-based protein hydrogel has a storage modulus (G') at 10 rad / sec of greater than 1000 Pa, preferably greater than 2000 Pa, more preferably greater than 5000 Pa, even more preferably greater than 6000 Pa, and most preferably greater than 8000 Pa. As can be understood by those skilled in the art, 2π rad / sec is equivalent to 1 Hz.

[0093] In the preferred method of the present invention, before washing, the plant-based protein hydrogel has a storage modulus (G') at 10 rad / sec of less than 20,000 Pa, preferably less than 15,000 Pa, and more preferably less than 10,000 Pa.

[0094] Furthermore, in the preferred method of the present invention, the washed plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of greater than 500 Pa, preferably greater than 1000 Pa, more preferably greater than 2500 Pa, even more preferably greater than 3000 Pa, and most preferably greater than 4000 Pa.

[0095] In the preferred method of the present invention, the washed plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of less than 20,000 Pa, preferably less than 15,000 Pa, and more preferably less than 10,000 Pa.

[0096] A preferred method of the present invention further comprises the step of (d) changing the pH of a plant-based protein hydrogel slurry.

[0097] In the preferred method of the present invention, step (d) is performed after step (c). In the alternative preferred method of the present invention, step (d) is performed sequentially with step (c).

[0098] During pH adjustment of a plant-based protein hydrogel slurry, the slurry can pass the isoelectric point of the protein. Due to the lack of charge repulsion at the isoelectric point, dispersed protein fragments in the plant-based protein hydrogel slurry can rapidly coagulate. To avoid this, pH-changing materials are used to rapidly change the pH, thus minimizing the time the slurry is at the isoelectric point.

[0099] Therefore, in the preferred method of the present invention, step (d) includes adding a pH-modifying material to a plant-based protein hydrogel slurry. Preferably, the pH-modifying material is an aqueous solution containing monovalent metal ions, divalent metal ions such as calcium, or aluminum ions, and more preferably, an alkaline aqueous solution containing monovalent metal ions, divalent metal ions, or aluminum ions. More preferably, the pH-modifying material is a hydroxide aqueous solution, such as sodium hydroxide, potassium hydroxide, or ammonium hydroxide.

[0100] In the preferred method of the present invention, the pH of the plant-based protein hydrogel slurry after step (d) is at least 1 pH unit below the isoelectric point of the plant-based protein.

[0101] In the preferred method of the present invention, the pH of the plant-based protein hydrogel slurry after step (d) is at least 1 pH unit above the isoelectric point of the plant-based protein. The preferred method of the present invention further comprises adding additional components to the plant-based protein hydrogel slurry. Preferably, the additional components are selected from plasticizers, opacifiers, preservatives, pigments, and other inorganic nanoparticles (e.g., clay), or mixtures thereof.

[0102] Particularly preferably, the additional component is a plasticizer. In the preferred method of the present invention, the plasticizer is selected from ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, polyethylene glycol, propylene glycol, sorbitol, mannitol, xylitol, lactic acid, glycolic acid, triethyl citrate, fatty acids, glucose, mannose, fructose, sucrose, ethanolamine, urea, triethanolamine, vegetable oil, lecithin, wax, and amino acids.

[0103] Particularly preferably, the additional component is a plasticizer. In the preferred method of the present invention, the plasticizer is selected from glycerol, ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, polyethylene glycol, propylene glycol, sorbitol, mannitol, xylitol, lactic acid, citric acid, glycolic acid, triethyl citrate, fatty acids, glucose, mannose, fructose, sucrose, ethanolamine, urea, triethanolamine, vegetable oil, lecithin, wax, and amino acids.

[0104] The amount of plasticizer to be incorporated will depend on the final material, such as the use of a film. Preferably, the plant-based protein hydrogel slurry may contain about 1 wt% of plasticizer based on the total mass of protein present in the plant-based protein hydrogel slurry, and may contain about 2 wt%, about 5 wt%, about 10 wt%, about 20 wt%, about 30 wt%, about 40 wt%, about 50 wt%, about 60 wt%, or more. More preferably, the plant-based protein hydrogel slurry may contain between about 5 wt% and 50 wt%, about 10-50 wt%, about 20-40 wt%, about 15-35 wt%, or about 20 wt% of plasticizer based on the total mass of protein present in the plant-based protein hydrogel slurry.

[0105] The addition of plasticizers can affect the mechanical properties of a material. Typically, the addition of plasticizers increases the elasticity of the material, which usually, conversely, reduces the strength of the resulting material, such as its tensile strength.

[0106] Composite films having improved physical and / or barrier particles can be formed by adding inorganic particles, such as clay plates, to a protein hydrogel slurry.

[0107] In the preferred method of the present invention, the plant-based protein hydrogel slurry is measured using an Anton Paar MCR 92 Rheometer employing a 50 mm flat plate and a flat plate-cone measurement shape with a 1° angle at 50s⁻¹. -1 The viscosity is in the range of 10 to 10,000 cps, preferably 10 to 8,000 cps, preferably 12 to 6,000 cps, preferably 15 to 5,000 cps, and preferably 20 to 2,000 cps. Controlling the viscosity is beneficial for the sequential processing of plant-based protein hydrogel slurries. For example, if the viscosity is too high, using the slurry to form a film or coating becomes more difficult, for example, when spreading the slurry to form a film. If the viscosity is too low, the slurry spreads too easily, which can also make it difficult to form structured materials such as films. Unless otherwise stated, viscosity is measured at 20°C.

[0108] In the preferred method of the present invention, the plant-based protein hydrogel slurry has a protein solid content in the range of 5 wt% to 25 wt%, preferably 6 wt% to 20 wt%, more preferably 7 wt% to 15 wt%, and more preferably 7.5 wt% to 12.5 wt%, based on the total mass of the plant-based protein hydrogel slurry. The protein concentration affects the rheology of the hydrogel. Hydrogels containing less than 5 wt% protein solid content are generally not robust enough to obtain handleable fragments. While hydrogels containing more than 25 wt% protein solid content are more robust, they have been found to be more difficult to disperse and therefore form weaker structured materials (e.g., films).

[0109] The plant-based hydrogel slurry described herein enables the formation of a range of useful plant-based biomaterials. The use of plant-based materials offers numerous advantages over previously used animal or petrochemical resources. Firstly, plant resources are renewable and can be obtained efficiently in an environmentally effective manner. Secondly, plant resources are biodegradable and therefore environmentally sound alternatives to other plastics. Thirdly, in contrast to animal-derived proteins, plant-based proteins have the significant advantage of not introducing animal-derived proteins into humans. This advantage has positive implications from both a pharmacological and pharmaceutical perspective, where animal-derived materials must undergo rigorous checks and processes (e.g., prion removal) to ensure the absence of harmful elements, as well as because the products are suitable for vegetarians / vegans.

[0110] Because plant-based proteins are naturally present in the diet of humans (or other animals), the biomaterials produced according to the present invention exhibit a higher degree of digestibility compared to other biopolymers such as polysaccharides (e.g., alginates or chitosan). This makes them particularly suitable for use in pharmaceuticals, foods, and / or cosmetics.

[0111] Preferably, the plant-based hydrogel slurry of the present invention can be used to form films, such as thin films. Plant protein-derived films have many applications, including the formation of biodegradable flexible films for food packaging applications.

[0112] An advantage of the plant-based materials of the present invention over animal-based or starch-based / cellulose materials is their inherent insolubility. Most biopolymer films dissolve immediately in water, thus rendering them unusable for food packaging applications on their own and requiring an extra coating layer with synthetic polymers. These challenges can be overcome with the present invention. The films of the present invention can also remain soluble under alkaline conditions, or in the presence of proteolytic enzymes, or in the presence of chaotropic agents.

[0113] The present invention also provides a plant-based protein hydrogel slurry prepared according to the method previously described herein.

[0114] The present invention also relates to a method for preparing plant-based structured materials, (a) a step of preparing a plant-based protein hydrogel slurry according to a method previously described herein, (b) A step of subjecting a plant-based protein hydrogel slurry to one or more solvent concentration reduction steps to reduce the concentration of the first co-solvent and / or the second co-solvent in order to obtain the plant-based structured material. This provides a method that includes [something].

[0115] In a preferred method of the present invention, one or more solvent concentration reduction steps reduce the concentration of the first co-solvent (e.g., an organic acid).

[0116] In a preferred method of the present invention, one or more solvent concentration reduction steps involve reducing the concentration of a second co-solvent (e.g., an alcohol such as ethanol).

[0117] In a preferred method of the present invention, step (b) comprises placing a plant-based protein hydrogel slurry on a surface before carrying out one or more solvent concentration reduction steps.

[0118] In a preferred method of the present invention, the solvent concentration reduction step includes heating. Preferably, the solvent concentration reduction step includes heating at a temperature in the range of 50 to 100°C, more preferably in the range of 55 to 95°C. In such a solvent concentration reduction step, the solvent is therefore removed by evaporation.

[0119] In a preferred method of the present invention, the solvent concentration reduction step includes forced convection of dry air.

[0120] In the preferred method of the present invention, the plant-based structured material is a film.

[0121] In an alternative preferred method of the present invention, the plant-based structured material is a casting (i.e., a plant-based structured material formed by molding, preferably non-thermally reversible molding, more preferably injection molding).

[0122] In an alternative preferred method of the present invention, the plant-based structuring material is a coating. Preferably, the coating is a food coating, seed coating, pharmaceutical coating, or surface coating (e.g., paper coating).

[0123] The coatings of the present invention are fully biodegradable and therefore provide an environmentally friendly alternative to conventional coatings made from synthetic materials (e.g., chemically modified natural polymers or fossil fuel-derived polymers). For example, the food coating of the present invention provides a fully biodegradable coating that conforms to food standards and can extend the shelf life of coated food items. The pharmaceutical coating of the present invention provides a fully biodegradable coating that can be used as a replacement for conventional enteric coatings and can mask any unpleasant taste associated with coated pharmaceutical ingredients.

[0124] In a preferred method of the present invention, the plant-based structured material comprises a plant-based protein having a secondary structure containing at least 40% intermolecular β-sheets, at least 50% intermolecular β-sheets, at least 60% intermolecular β-sheets, at least 70% intermolecular β-sheets, at least 80% intermolecular β-sheets, or at least 90% intermolecular β-sheets. Preferably, the % intermolecular β-sheet content is measured by FTIR.

[0125] In the preferred method of the present invention, the plant-based structured material has a Young's modulus greater than 20 MPa, preferably greater than 50 MPa, greater than 80 MPa, greater than 100 MPa, greater than 200 MPa, greater than 300 MPa, greater than 400 MPa, greater than 500 MPa, or greater than 600 MPa. Young's modulus is a measure of the strength of the structured article.

[0126] In the preferred method of the present invention, the plant-based structured material is a film.

[0127] Preferably, the film has a thickness in the range of 1 to 1000 μm, preferably 10 to 150 μm, more preferably 20 to 100 μm, even more preferably 30 to 70 μm, and most preferably 35 to 60 μm. This can be measured in micrometers.

[0128] Preferably, the film has a break elongation percentage of more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 100%, or higher.

[0129] Films produced according to the method of the present invention can be finely patterned with structures ranging from 100 nm to 1000 μm, enabling novel functional properties such as superhydrophobicity (lotus leaf effect) or structural color (contributing to Mie scattering).

[0130] Functional composite films can be manufactured by embedding inorganic nanoparticles, such as gold or silver nanoparticles, in a protein matrix. Applications include flexible electronics or films with antibacterial properties. Composite films with improved physical and / or barrier particles can be formed by embedding particles, such as clay plates, in a protein matrix.

[0131] The present invention also provides plant-based structuring materials prepared according to the methods previously described herein.

[0132] Preferred plant-based structuring materials according to the present invention are films, castings, or coatings. Preferably, coatings are food coatings, seed coatings, pharmaceutical coatings, or surface coatings (e.g., paper coatings). Preferred properties of the plant-based structuring materials of the present invention are described above. The plant-based structuring materials of the present invention are useful in a variety of applications, including food, beverages, cosmetics, formulations (e.g., paints), and packaging. The transparency and high strength of the films of the present invention make them particularly well suited for packaging applications.

[0133] The present invention also provides the use of a plant-based protein hydrogel slurry, as previously defined herein, for producing a plant-based structured material. Preferably, the plant-based structured material is a film, casting, molded product, or coating. Preferably, the coating is a food coating, seed coating, pharmaceutical coating, or surface coating (e.g., paper coating).

[0134] The present invention also provides a protein solid content of 5 wt% to 25 wt% based on the total mass of a plant-based protein hydrogel slurry, and 50s -1 The invention provides a plant-based protein hydrogel slurry having a viscosity in the range of 10 to 10,000 cps at 20°C, wherein the plant-based protein hydrogel slurry has a diameter of 0.5 to 150 microns, determined by laser diffraction. 50 It contains fragments having particle size.

[0135] The preferred plant-based protein hydrogel slurry of the present invention is 0.6 to 100 microns, more preferably 0.7 to 70 microns, even more preferably 0.8 to 50 microns, more preferably 0.9 to 25 microns, more preferably 1 to 20 microns, more preferably 1 to 10 microns, and even more preferably 1 to 5 microns, as determined by laser diffraction. 50 It contains fragments having particle size.

[0136] The preferred plant-based protein hydrogel slurry of the present invention has a protein solid content of 6 wt% to 20 wt%, more preferably 7 wt% to 15 wt%, and even more preferably 7.5 wt% to 12.5 wt%, based on the total mass of the plant-based protein hydrogel slurry.

[0137] Preferably, the viscosity of the plant-based protein hydrogel slurry is such that it is suitable for a 50 mm plate and 50 s. -1The measurement is performed using an Anton Paar MCR 92 Rheometer employing a flat plate and cone measuring shape with a 1° angle. The preferred plant-based protein hydrogel slurry of the present invention is measured in 50s -1 And at 20°C, within the range of 10 to 8000 cps, more preferably 50s -1 And at 20°C, within the range of 12 to 6000 cps, more preferably 50s -1 It also has a viscosity in the range of 15 to 5,000 cps at 20°C.

[0138] The present invention also provides a protein solid content of 5 wt% to 25 wt% based on the total mass of a plant-based protein hydrogel slurry, and 50s -1 The invention provides a plant-based protein hydrogel slurry having a viscosity in the range of 10 to 10,000 cps at 20°C, wherein the plant-based protein hydrogel slurry is determined by dynamic light scattering of less than 500 nm. 50 It contains fragments having particle size.

[0139] The preferred plant-based protein hydrogel slurry of the present invention is less than 300 nm, more preferably less than 200 nm, and even more preferably less than 50 nm, as determined by dynamic light scattering. 50 It contains fragments having particle size.

[0140] The preferred plant-based protein hydrogel slurry of the present invention has a protein solid content of 6 wt% to 20 wt%, more preferably 7 wt% to 15 wt%, and even more preferably 7.5 wt% to 12.5 wt%, based on the total mass of the plant-based protein hydrogel slurry.

[0141] Preferably, the viscosity of the plant-based protein hydrogel slurry is such that it is suitable for a 50 mm plate and 50 s. -1 The measurement is performed using an Anton Paar MCR 92 Rheometer employing a flat plate and cone measuring shape with a 1° angle. The preferred plant-based protein hydrogel slurry of the present invention is measured in 50s -1and at 20°C for 10 to 8000 cps, more preferably 50s -1 and at 20°C for 12 to 6000 cps, more preferably 50s -1 It also has a viscosity in the range of 15 to 5,000 at 20°C.

[0142] The present invention also provides the use of the plant-based protein hydrogel slurry previously described herein for producing plant-based structured materials. Preferably, the plant-based structured material is a film, casting, molded product, or coating. Preferably, the coating is a food coating, seed coating, pharmaceutical coating, or surface coating (e.g., paper coating).

[0143] The present invention also provides a film comprising a plant-based protein hydrogel slurry as previously described herein.

[0144] A preferred film of the present invention is (a) Preferably, the % intermolecular β-sheet content is measured by FTIR and the plant-based protein contains a secondary structure with at least 40% intermolecular β-sheets, at least 50% intermolecular β-sheets, at least 60% intermolecular β-sheets, at least 70% intermolecular β-sheets, at least 80% intermolecular β-sheets, or at least 90% intermolecular β-sheets, and / or (b) Having a tensile strength of 4 to 25 MPa, preferably 6 to 15 MPa, more preferably 8 to 12 MPa, and / or (c) Having a break elongation percentage of more than 10%, preferably more than 20%, preferably more than 30%, preferably more than 40%, preferably more than 50%, preferably more than 60%, preferably more than 70%, preferably more than 80%, preferably more than 90%, preferably more than 100%, or higher.

[0145] The present invention also provides films obtained from plant-based protein hydrogel slurries as previously described herein. For example, films can be obtained by subjecting a plant-based protein hydrogel slurry to one or more solvent concentration reduction steps. Preferred features of the solvent concentration reduction steps are as described above. Industrial methods for producing structured materials such as films from hydrogel slurries include casting, in which the hydrogel slurry is injected or extruded onto a moving surface, such as a belt or drum, in a carefully controlled manner and subjected to controlled drying conditions. [Brief explanation of the drawing]

[0146] [Figure 1] The image below shows a transmission electron microscope (TEM) image of the diluted pea protein-based slurry prepared in Example 1. [Figure 2a] This is a photograph demonstrating the transparency of the film prepared in Example 2. [Figure 2b] This is a photograph demonstrating the transparency of the film prepared in Example 3. [Figure 3] The image shows a scanning electron microscope (SEM) image of the coating layer on a paper substrate prepared from the pea protein-based slurry described in Example 5. The uncoated paper portion is also shown. [Figure 4] Figure 4a shows a scanning electron microscope (SEM) image of the coated layer on strawberries prepared from the pea protein-based slurry described in Example 6. Figure 4b shows a scanning electron microscope (SEM) image of uncoated strawberries prepared from the pea protein-based slurry described in Example 6. [Figure 5] Figure 5a shows a scanning electron microscope (SEM) image of the coating layer of paracetamol tablets prepared from the pea protein-based slurry described in Example 7. Figure 5b shows a scanning electron microscope (SEM) image of uncoated paracetamol tablets prepared from the pea protein-based slurry described in Example 7. [Figure 6] Figure 6a shows a scanning electron microscope (SEM) image of the coating layer on wheat seeds prepared from the pea protein-based slurry described in Example 8. Figure 6b shows a scanning electron microscope (SEM) image of uncoated wheat seeds prepared from the pea protein-based slurry described in Example 8. [Examples]

[0147] material Pea protein isolate (PPI) (80% protein) was purchased from Cambridge Commodities Ltd. Soy protein isolates were purchased from Cambridge Commodities Ltd. Lactic acid (food grade, ≥80%) was purchased from Cambridge Commodities Ltd. (Glacial)acetic acid was purchased from Fisher Scientific.

[0148] Measurement method Viscosity was measured on a 50mm flat plate at a 1° angle and 20°C for 50 seconds. -1 The measurements were performed using an Anton Paar MCR 92 Rheometer, which employs a flat plate and cone shape with constant shear.

[0149] The storage modulus (G') can be measured using an Anton Paar MCR 92 Rheometer, which employs a measurement shape consisting of a 50 mm flat plate and a flat plate and cone with a 1° angle and a 1% strain at a vibration frequency of 1 Hz.

[0150] Particle size measurements were performed using dynamic light scattering (DLS) or laser diffraction techniques. DLS measurements were taken using a Zeta Sizer Nano S from Malvern Panalytical, following the manufacturer's instructions. It is important that the slurry is sufficiently diluted to avoid confusing results due to particle coagulation during testing. The hydrogel slurry was diluted with deionized water at a coefficient of 100. It is also important that the pH of the sample is far from the isoelectric point of the sample to avoid confusing results due to coagulation. The isoelectric point of the pea protein isolate material tested herein was 4.5, and the pH of the slurry was adjusted to 3 with acetic acid or lactic acid before measurement. Typically, several diluents and / or buffer solutions should be tested to ensure proper dispersion. 200 μL of the diluted slurry was placed in a cuvette and positioned in the instrument. The test was then performed according to standard instrument procedures. d 50 The distribution was about wt / volume. DLS can generally be used to measure particle size up to 500 nm. The upper limit is mainly controlled by the onset of sedimentation. For particles exceeding this, particle size measurement was performed using laser diffraction with an Anton Paar PSA 1190. The measurement was performed by diluting the protein slurry in an aqueous solution containing acetic acid or lactic acid adjusted to the same pH. The slurry was diluted to the required concentration to have the desired optical density for measurement (usually 5-15% obscuration). Cited d 50 This concerns volume distribution. 10 The values ​​and d5 values ​​can also be obtained by this method using laser diffraction.

[0151] Other devices, such as the NANO-flex II® system from Colloid Metrix, can be used.

[0152] The particle size of hydrogel fragments from the low-shear process can be determined by sieving. A preferred technique involves taking 200 g of the hydrogel mixture from the low-shear process and rapidly dispersing it in 1000 mL of DI water. The dispersed mixture is then rapidly injected through a series of stacked sieves with mesh sizes decreasing from 50 mm to 1 mm. Sieves from Endecotts are preferred. The percentage of slurry within a specific size range can be calculated by combining the masses of fragments on different meshes and calculating this as a percentage of the total slurry volume. Errors caused by any additional solvent exchange are minimal due to the short test period.

[0153] Transmission electron microscopy (TEM) measurements were performed using a Thermo Scientific FEI Talos F200X G2 TEM. A preferred technique involves preparing a test sample by diluting a fine plant-based hydrogel slurry with a 3% acetic acid solution in a 1:100 dilution, precipitating the sample on a TEM grid (C400Cu, EM resolution), and subsequently staining it with uranyl acetate. The largest fragment size is then determined by optical testing of at least 30 fragments randomly selected from within the field of view of the test sample. The maximum length is the maximum length of a line drawn between any two opposing boundaries of the fragment that do not intersect with any external boundaries.

[0154] Scanning electron microscope (SEM) images were taken using a MIRA 3 FEG-SEM with a 10nm platinum coating, manufactured by TESCAN.

[0155] Structural analysis was performed using an FTIR-Equinox 55 spectrometer (Bruker). Samples were used without further pretreatment and loaded into the FTIR holder. An air-corrected spectrum was subtracted from the original FTIR spectrum, and secondary derivatives were applied for further analysis. Each FTIR measurement was repeated three times. The instrument's sensitivity was detected as 5%. Vibrational changes in amide I, which are closely correlated with the protein secondary structure, were followed to degrade the raw structural conversions of the pea protein isolate into supramolecular aggregates. Structural analysis was performed either in solution (e.g., directly using a plant-based protein hydrogel slurry) or on the resulting dried film. In the latter case, film samples were prepared for structural analysis by drying 200 μL of the relevant plant-based protein hydrogel slurry at 37°C for 6 hours.

[0156] The Young's modulus and tensile strength of structured materials such as films can be measured using a 5ST electromechanical universal tester from Tinius Olsen. A 10cm x 1cm strip was placed between grips and stretched at 12.5mm / min, and the force and elongation were recorded. The thickness of the film before testing can be measured in micrometers.

[0157] The light transmittance of structured materials such as films can be measured using a Cary 500 UV-vis spectrometer. Measurements were performed at wavelengths in the range of 300 to 800 nm, with a scanning speed of 600 nm / min and an average time of 0.1 seconds.

[0158] (Example 1) Preparation of pea protein gel structures (a) Formation of protein hydrogels A 500 ml mixture consisting of 12.5 (w / v) pea protein isolate in a 40 (v / v) lactic acid solution was prepared.

[0159] Next, the mixture was heated in a water bath at 80°C for 30 minutes, followed by a short ultrasonic treatment step to break down large colloidal aggregates (Hielscher UIP1000hdT (1000W, 20kHz)), after which a clear solution was obtained. The applied energy was 16Wh over 7 minutes.

[0160] Next, the solution was poured into a 220 mm Petri dish and left to cool at 10°C for 72 hours to obtain a self-standing protein hydrogel.

[0161] (b) Application of shearing to protein hydrogels Next, shearing was applied to the hydrogel formed in step (a) as follows: The protein hydrogel was cut into cubes of approximately 1 cm in size via a low-shear cutting process. These cubes were placed in a 75 μm filter bag and then immersed in a bucket containing 5 L of deionized water. This formed a coarse protein hydrogel slurry in the filter bag. The hydrogel cubes were left immersed for 1 hour, with occasional gentle stirring. This step was performed to reduce the concentration of lactic acid in the hydrogel by diffusion into a continuous aqueous phase, and was repeated five more times until the final pH of the aqueous solution was 3.28.

[0162] The strained gel cube was transferred to a 500 ml bottle and subjected to probe sonication for 10 minutes (approximately 0.2 kJ / ml) in a high-shear process to form a homogeneous, low-viscosity protein dispersion. The viscosity of the slurry was 12 cps. Fine slurry fragments were then processed. 50 The size was measured by DLS and was 90 nm.

[0163] (c) Preparation of pea protein film The fine hydrogel slurry prepared in step (b) was adjusted to pH 2.6 by adding a small amount of lactic acid, then injected onto a heated surface (maintained at 80°C), and dried for 1 hour to form a structured film with an average thickness of 18.1 μm. The resulting film was transparent and had a Young's modulus of 361 MPa and a tensile strength of 14 MPa.

[0164] (Example 2) Preparation of pea protein gel structures (a) Formation of protein hydrogels 800 ml of a mixture consisting of 10 w / v pea protein isolate in a 40 v / v acetic acid solution was prepared.

[0165] Next, the mixture was heated in a water bath at 85°C for 20 minutes, followed by a short ultrasonic treatment step to break down large colloidal aggregates (Hielscher UIP500hdT (500W, 20kHz)), after which a clear solution was obtained. The applied energy was 200kJ for 30 minutes.

[0166] Next, the solution was poured into two 220 mm Petri dishes. The dishes were sealed and allowed to cool by storage in a refrigerator at 4°C for 20 hours to obtain a self-standing protein hydrogel. The storage modulus of this hydrogel was 2640 Pa.

[0167] (b) Application of shearing to protein hydrogels Next, shearing was applied to the hydrogel formed in step (a) as follows: The protein hydrogel was cut into approximately 1 cm cubes via a low-shear cutting process. These cubes were placed in a 75 μm filter bag and then immersed in a bucket containing 7 L of deionized water. This formed a coarse protein hydrogel slurry in the filter bag. The hydrogel cubes were left immersed for 1.5 hours, with occasional gentle stirring. This step was performed to reduce the concentration of acetic acid in the hydrogel by diffusion into a continuous aqueous phase, and was repeated once more until the final pH of the aqueous solution reached 3.1.

[0168] The strained gel cube was transferred to a 1L bottle and exposed to high shear with a rotor stator (15,000 rpm for 5 minutes), followed by probe sonication on ice for 30 minutes (approximately 0.5 kJ / ml) to form a homogeneous, low-viscosity protein dispersion. The viscosity of the slurry was 23 cps. Fine slurry fragments were then extracted. 50 The size was measured by DLS and was 103 nm.

[0169] (c) Preparation of pea protein film The fine hydrogel slurry prepared in step (b) was mixed with 20 w / w% glycerol, poured into a plastic petri dish, and dried at room temperature for 24 hours to form a structured film with an average thickness of 78.5 μm. The PSD and optical properties of the pea protein film of Example 2 were investigated. The results are shown in Table 1. The obtained film was transparent, as clearly shown in Figure 2a and with a transmittance of 81.1% at 600 nm.

[0170] [Table 1]

[0171] The results indicate that highly transparent functional films can be formed using the plant-based protein hydrogel slurry of the present invention. While we do not wish to be constrained by theory, it is believed that achieving a controlled particle size distribution will enable the observation of high levels of transparency.

[0172] (Example 3) Preparation of soy protein gel structures (a) Formation of protein hydrogels A mixture consisting of 7.0 (w / w) soy protein isolate in a 30 (v / v) acetic acid solution was prepared in 430 g.

[0173] Next, the mixture was heated in a water bath at 85°C for 30 minutes, followed by a short ultrasonic treatment step to break down large colloidal aggregates (Bandelin HD4200, TS 113 probe), after which a clear solution was obtained. The applied energy was 200 kJ over 30 minutes.

[0174] Next, the solution was poured into two 220 mm Petri dishes. The dishes were sealed and allowed to cool by storage in a refrigerator at 4°C for 20 hours to obtain self-standing protein hydrogels.

[0175] (b) Application of shearing to soy protein hydrogel Next, shearing was applied to the hydrogel formed in step (a) as follows: The protein hydrogel was cut into cubes of approximately 1 cm in size via a low-shear cutting process. These cubes were placed in a 75 μm filter bag and then immersed in a bucket containing 5 L of deionized water. This formed a coarse protein hydrogel slurry in the filter bag. The hydrogel cubes were left immersed for 1.5 hours, with occasional gentle stirring. This step was performed to reduce the concentration of acetic acid in the hydrogel by diffusion into the continuous aqueous phase, and was repeated once more until the final pH of the aqueous solution was 3.07.

[0176] A strained gel cube (469g) was transferred to a 0.5L bottle and mixed with 40g of deionized water. A colloidal suspension of large gel particles was obtained, which was divided into 50g aliquots. These were then subjected to high shear at different levels to obtain samples with varying particle size distributions, as detailed in Table 2.

[0177] [Table 2]

[0178] (c) Preparation of soy protein film The fine hydrogel slurry of sample B prepared in step (b) was mixed with 20 w / w% glycerol, poured into a plastic petri dish, and dried at room temperature for 24 hours to form a structured film with an average thickness of 61 μm. The PSD and optical properties of the soy protein film of Example 3 were examined. The results are shown in Table 3. The obtained film was transparent, as clearly shown in Figure 2b and with a transmittance of 89.8% at 600 nm.

[0179] [Table 3]

[0180] The results indicate that functional films with high transparency can be formed using the plant-based protein hydrogel slurry of the present invention. While we do not wish to be constrained by theory, it is thought that achieving a controlled particle size distribution will enable the observation of high levels of transparency.

[0181] (Example 4) Preparation of pea protein gel structures (a) Formation of protein hydrogels A mixture consisting of 11.11 (w / w) pea protein isolate was prepared in a 30 (v / v) acetic acid solution, totaling 450 g.

[0182] Next, the mixture was heated in a water bath at 85°C for 30 minutes, followed by a short ultrasonic treatment step to break down large colloidal aggregates (Bandelin HD4200, TS 113 probe), after which a clear solution was obtained. The applied energy was 200 kJ over 30 minutes.

[0183] Next, the solution was poured into two 220 mm Petri dishes. The dishes were sealed and allowed to cool by storage in a refrigerator at 4°C for 20 hours to obtain self-standing protein hydrogels.

[0184] (b) Application of shearing to protein hydrogels Next, shearing was applied to the hydrogel formed in step (a) as follows: The protein hydrogel was cut into cubes of approximately 1 cm in size via a low-shear cutting process. These cubes were placed in a 75 μm filter bag and then immersed in a bucket containing 5 L of deionized water. This formed a coarse protein hydrogel slurry in the filter bag. The hydrogel cubes were left immersed for 1.5 hours, with occasional gentle stirring. This step was performed to reduce the concentration of acetic acid in the hydrogel by diffusion into the continuous aqueous phase, and was repeated once more until the final pH of the aqueous solution was 3.02.

[0185] A strained gel cube (400g) was transferred to a 0.5L bottle and mixed with 100g of deionized water. A colloidal suspension of large gel particles was obtained, which was divided into 50g aliquots. These were then subjected to high shear at different levels to obtain samples with diverse particle size distributions, as detailed in Table 4.

[0186] [Table 4]

[0187] (c) Preparation of pea protein film The fine hydrogel slurry prepared in step (b) was mixed with 20 w / w% glycerol, poured onto a PTFE evaporating dish, and dried at room temperature for 24 hours to form a structured film. The characteristics of the obtained film are described in Table 5.

[0188] [Table 5]

[0189] The results demonstrate that the method of the present invention enables the preparation of plant-based protein hydrogel slurries having a controlled particle size distribution. The results also demonstrate that the plant-based protein hydrogel slurry of the present invention can also be used to prepare structured materials such as films having excellent tensile properties that can be controlled through control of slurry particle size. The prepared films have secondary structures with a high level of intermolecular β-sheets (e.g., at least 50% intermolecular β-sheets). This high degree of intermolecular interaction is thought to contribute to the observed enhanced mechanical properties.

[0190] (Example 5) Use of pea protein dispersion as a coating layer for paper The colloidal pea protein dispersion obtained from Example 2 was spray-coated onto an uncoated cardboard substrate using an airbrush. One layer was applied first, followed by drying in an oven at 80°C for 1 minute to evaporate any remaining solvent. This procedure was repeated 15 times until a uniform, transparent coating was achieved. The results are shown in Figure 3.

[0191] (Example 6) Use of pea protein dispersion as a food coating layer The colloidal pea protein dispersion obtained from Example 2 was applied as a coating to unused strawberries via a dip coating process. The strawberries were first immersed in 50 ml of the pea protein dispersion for 5 seconds, followed by a brief application of compressed air to remove excess dispersion. The coated strawberries were then left to dry at room temperature for 10 minutes. This procedure was repeated five times until a uniform, transparent coating was achieved. The results are shown in Figure 4.

[0192] (Example 7) Use of pea protein dispersion as a coating layer for pharmaceuticals The colloidal pea protein dispersion obtained from Example 2 was spray-coated onto uncoated paracetamol tablets using an airbrush. One layer was applied first, followed by drying in an oven at 80°C for 1 minute to evaporate any remaining solvent. This procedure was repeated 15 times until a uniform, transparent coating was achieved. The results are shown in Figure 5.

[0193] (Example 8) Use of pea protein dispersion as a seed coating layer. The colloidal pea protein obtained from Example 2 was applied as a coating to wheat seeds via a dip coating process. The wheat seeds were first immersed in 50 ml of pea protein dispersion for 5 seconds, followed by a brief application of compressed air to remove excess dispersion. The coated wheat seeds were then left to dry at room temperature for 10 minutes. This procedure was repeated twice until a uniform, transparent coating was achieved. The results are shown in Figure 6. Note Note 1. A method for preparing a plant-based protein hydrogel slurry, (a) A step of forming a solution containing one or more plant-based proteins in a solvent system, wherein the solvent system contains a miscible cosolvent, the first cosolvent increasing the solubility of the plant-based protein, and the second cosolvent decreasing the solubility of the plant-based protein. (b) A step of inducing a sol-gel transition of a protein in solution to form a plant-based protein hydrogel, (c) A step of subjecting the plant-based protein hydrogel to a shearing treatment to form a plant-based protein hydrogel slurry. Methods that include... Note 2. The method according to Appendix 1, wherein the plant protein is selected from soybean protein, pea protein, rice protein, potato protein, wheat protein, corn zein protein, or sorghum protein. Note 3. The method according to Appendix 1 or Appendix 2, wherein the first cosolvent is an organic acid, preferably acetic acid, formic acid, propionic acid and / or α-hydroxy acid, where the α-hydroxy acid can be preferably selected from glycolic acid, lactic acid, malic acid, citric acid and / or tartaric acid, and the organic acid that is particularly preferred is acetic acid and / or lactic acid. Note 4. The method according to any one of Appendix 1 to 3, wherein the second or further cosolvent is an aqueous buffer solution, preferably selected from water, ethanol, methanol, acetone, acetonitrile, dimethyl sulfoxide, dimethylformamide, formamide, 2-propanol, 1-butanol, 1-propanol, hexanol, t-butanol, ethyl acetate, or hexafluoroisopropanol, particularly preferably selected from water and / or ethanol, and even more preferably selected from water. Note 5. The method according to any one of the appendices 1 to 4, wherein the solvent system has a cosolvent ratio of the first cosolvent to the second cosolvent of about 20-80 v / v%, preferably about 20-60 v / v%, about 25-55 v / v%, about 30-50 v / v%, about 20%, about 30%, about 40%, about 50%, or about 60 v / v%, most preferably about 30-50 v / v%. Note 6. The method according to any one of the appendices 1 to 5, comprising heating the protein solution to a first temperature above the sol-gel temperature of one or more plant-based protein solutions, and then lowering it to a second temperature below the sol-gel temperature of one or more plant-based protein solutions to form a hydrogel. Note 7. The method according to any one of the appendices 1 to 6, wherein the shearing treatment includes a high shearing step. Note 8. The method according to Appendix 7, wherein the high-shear step comprises fragmenting the plant-based protein hydrogel into fragments. Note 9. The fragment produced in the high shear process has a wavelength of less than 500 nm, preferably less than 300 nm, more preferably less than 200 nm, and even more preferably less than 50 nm, as determined by DLS. 50 The method described in Appendix 8, having the characteristics described herein. Note 10. The method according to any one of Appendix 7 to 9, wherein the high shear step includes ultrasonic treatment, high shear mechanical stirring, or cavitation. Note 11. The method according to any one of the appendices 1 to 6, wherein the shearing treatment comprises two steps. Note 12. The method according to Appendix 11, wherein the shearing treatment includes a low-shear step followed by a high-shear step. Note 13. The method according to Appendix 12, wherein the low-shear step comprises fragmenting the plant-based protein hydrogel into fragments. Note 14. The method according to Appendix 13, wherein at least 80% of the fragments produced in the low-shear step have a particle size determined by sieving, in the range of 1 mm to 50 mm, preferably 1 mm to 30 mm, more preferably 10 mm to 30 mm, more preferably 15 mm to 30 mm, and even more preferably 20 mm to 30 mm. Note 15. The method according to any one of appendices 12 to 14, wherein the low-shear step includes mechanical cutting. Note 16. The method according to any one of appendices 13 to 15, wherein the high-shear step comprises further fragmentation of the plant-based protein hydrogel. Note 17. The method according to Appendix 16, wherein the fragment produced in the high shear step has a D50 determined by DLS, which is less than 500 nm, preferably less than 300 nm, more preferably less than 200 nm, and even more preferably less than 50 nm. Note 18. The method according to any one of appendices 12 to 17, wherein the high shear step includes ultrasonic treatment, high shear mechanical stirring, or cavitation. Note 19. The method according to any one of Appendix 12 to 18, wherein step (c) further comprises subjecting the plant-based protein hydrogel slurry to a solvent reduction step, preferably an organic solvent reduction step, between the low-shear step and the high-shear step. Note 20. The solvent reduction step is (i) A step of contacting a fragment of the plant-based hydrogel slurry with an insoluble solvent, (ii) A step of separating the fragments of the plant-based hydrogel slurry from the insolubilizing solvent to obtain a washed plant-based protein hydrogel, (iii) A process in which process (i) and process (ii) are repeated at the discretion of the user. The method described in Appendix 19, including the method described in Appendix 19. Note 21. The method described in Appendix 20, wherein step (ii) includes mesh filtration. Note 22. The method according to Appendix 20 or 21, wherein, before washing, the plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of more than 1000 Pa, preferably more than 2000 Pa, more preferably more than 5000 Pa, even more preferably more than 6000 Pa, and most preferably more than 8000 Pa. Note 23. The method according to Appendix 20 or 22, wherein, before washing, the plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of less than 20,000 Pa, preferably less than 15,000 Pa, more preferably less than 10,000 Pa, and optionally, before washing, the plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of between about 1,000 Pa and 20,000 Pa, preferably between about 2,000 Pa and 15,000 Pa, more preferably between about 2,000 Pa and 10,000 Pa. Note 24. The method according to any one of appendices 20 to 23, wherein the washed plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of greater than 500 Pa, preferably greater than 1000 Pa, more preferably greater than 2500 Pa, even more preferably greater than 3000 Pa, and most preferably greater than 4000 Pa. Note 25. The method according to any one of the appendices 20 to 24, wherein the washed plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of less than 20,000 Pa, preferably less than 15,000 Pa, and more preferably less than 10,000 Pa; optionally, the washed plant-based protein hydrogel has a storage modulus (G') at 10 rad / second of about 500 Pa to 20,000 Pa, about 500 Pa to 15,000 Pa, about 500 Pa to 10,000 Pa, about 1,000 Pa to 20,000 Pa, about 1,000 Pa to 15,000 Pa, or about 1,000 Pa to 10,000 Pa. Note 26. (d) The method according to any one of Appendix 1 to 25, further comprising the step of changing the pH of the plant-based protein hydrogel slurry so that it differs from the isoelectric point of the protein hydrogel by more than 1 pH unit. Note 27. The method described in Appendix 26, wherein step (d) is performed after step (c). Note 28. The method described in Appendix 26, wherein step (d) is carried out sequentially with step (c). Note 29. The method according to any one of Appendix 26 to 28, wherein step (d) comprises adding a pH-changing material to the plant-based protein hydrogel slurry. Note 30. The method according to Appendix 29, wherein the pH-changing material is a solution containing monovalent metal ions, divalent metal ions, or ammonium ions, and preferably an alkaline aqueous solution containing monovalent metal ions, divalent metal ions, or ammonium ions. Note 31 The method according to Appendix 30, wherein the pH-changing material is an aqueous hydroxide solution, preferably sodium hydroxide, potassium hydroxide, or ammonium hydroxide. Note 32. The method according to any one of the appendices 26 to 31, wherein the pH of the plant-based protein hydrogel slurry after step (d) is at least 1 pH unit below the isoelectric point of the plant-based protein. Note 33. The method according to any one of the appendices 26 to 31, wherein the pH of the plant-based protein hydrogel slurry after step (d) is at least 1 pH unit above the isoelectric point of the plant-based protein. Note 34. The method according to any one of Appendix 1 to 33, further comprising adding an additional component to the plant-based protein hydrogel slurry. Note 35. The method according to Appendix 34, wherein the additional components are selected from plasticizers, opacifiers, preservatives, pigments, and nanoparticles, or mixtures thereof. Note 36. The method according to Appendix 35, wherein the additional component is a plasticizer. Note 37. The method according to Appendix 36, wherein the plasticizer is selected from ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, polyethylene glycol, propylene glycol, sorbitol, mannitol, xylitol, fatty acids, glucose, mannose, fructose, sucrose, ethanolamine, urea, triethanolamine; vegetable oils, lecithin, waxes, and amino acids. Note 38. The aforementioned plant-based protein hydrogel slurry, 50s -1 In the range of 10 to 10000 cps, preferably 50s -1 The method according to any one of the appendices 1 to 37, wherein the viscosity is in the range of 15 to 5000 cps. Note 39. The method according to any one of Appendix 1 to 38, wherein the plant-based protein hydrogel slurry comprises protein aggregates having an average size of less than 200 nm, preferably less than 150 nm, less than 125 nm, less than 100 nm, less than 90 nm, less than 80 nm, less than 70 nm, less than 60 nm, less than 50 nm, less than 40 nm, or less than 30 nm. Note 40. The method according to any one of Appendix 1 to 39, wherein the plant-based protein hydrogel slurry has a protein solid content in the range of 5 wt% to 25 wt% based on the total mass of the plant-based protein hydrogel slurry. Note 41. A plant-based protein hydrogel slurry prepared according to the method described in any one of Appendix 1 to 40. Note 42. A method for preparing plant-based structured materials, (a) A step of preparing a plant-based protein hydrogel slurry according to the method described in any one of Appendix 1 to 40, (b) A step of subjecting the plant-based protein hydrogel slurry to one or more solvent concentration reduction steps to reduce the concentration of the first co-solvent and / or the second co-solvent in order to obtain the plant-based structured material. Methods that include... Note 43. The method according to Appendix 42, wherein step (b) comprises placing the plant-based protein hydrogel slurry on a surface before forming one or more solvent concentration reduction steps. Note 44. The method according to Appendix 42 or Appendix 43, wherein the solvent concentration reduction step includes heating. Note 45. The method according to Appendix 44, wherein the solvent concentration reduction step includes heating at a temperature in the range of 50 to 100°C. Note 46. The method according to Appendix 42 or Appendix 43, wherein the solvent concentration reduction step includes forced convection of dry air. Note 47. The method according to any one of appendices 42 to 46, wherein the plant-based structured material is a film. Note 48. The method according to any one of the appendices 42 to 46, wherein the plant-based structural material is a casting. Note 49. The method according to any one of appendices 42 to 46, wherein the plant-based structured material is a coating. Note 50. The method according to any one of Appendix 42 to 49, wherein the plant-based structured material comprises a plant-based protein having a secondary structure comprising at least 40% intermolecular β-sheets, at least 50% intermolecular β-sheets, at least 60% intermolecular β-sheets, at least 70% intermolecular β-sheets, at least 80% intermolecular β-sheets, or at least 90% intermolecular β-sheets. Note 51. The method according to any one of the appendices 42 to 50, wherein the plant-based structured material has a Young's modulus of more than 20 MPa, preferably more than 50 MPa, more than 80 MPa, more than 100 MPa, more than 200 MPa, more than 300 MPa, more than 400 MPa, more than 500 MPa, or more than 600 MPa. Note 52. The method according to any one of appendices 42 to 51, wherein the plant-based structured material is a film having a thickness in the range of 1 to 1000 μm, preferably 1 to 100 μm, more preferably 10 to 100 μm, even more preferably 20 to 60 μm, and most preferably 30 to 50 μm. Note 53 The method according to any one of appendices 42 to 52, wherein the plant-based structured material is a film having a tensile strength of more than 1 MPa, preferably more than 5 MPa, preferably more than 10 MPa, and most preferably more than 25 MPa. Note 54. The method according to any one of the appendices 42 to 53, wherein the plant-based structured material is a film having a break elongation percentage of more than 10%, more than 20%, more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 80%, more than 90%, more than 100%, or greater. Note 55. A plant-based structuring material prepared according to the method described in any one of the appendices 42 to 54. Note 56. Use of a plant-based protein hydrogel slurry described in any one of Appendix 1 to 40 for the production of plant-based structured materials. Note 57. The use as described in Appendix 56, wherein the plant-based structuring material is a film, casting, or coating.

Claims

1. A method for preparing a plant-based protein hydrogel slurry, (a) A step of forming a solution containing one or more plant-based proteins in a solvent system, wherein the solvent system contains a miscible cosolvent, the first cosolvent increasing the solubility of the plant-based protein, and the second cosolvent decreasing the solubility of the plant-based protein. (b) A step of inducing a sol-gel transition of the protein in the solution to form a plant-based protein hydrogel, (c) A step of subjecting the plant-based protein hydrogel to a shearing treatment to form a plant-based protein hydrogel slurry. Methods that include...

2. The method according to claim 1, wherein the plant protein is selected from soybean protein, pea protein, rice protein, potato protein, wheat protein, corn zein protein, or sorghum protein.

3. The method according to claim 1 or 2, wherein the protein solution is heated to a first temperature above the sol-gel temperature of the one or more plant-based protein solutions, and then reduced to a second temperature below the sol-gel temperature of the one or more plant-based protein solutions to form a hydrogel.

4. The shearing process includes a high shear step, preferably the high shear step encompasses fragmentation of a plant-based protein hydrogel into fragments, and more preferably the fragments produced in the high shear step are less than 500 nm, preferably less than 300 nm, more preferably less than 200 nm, and even more preferably less than 50 nm, as determined by DLS. 50 The method according to any one of claims 1 to 3, comprising:

5. The shearing process includes a high-shear step, preferably the high-shear step encompasses fragmentation of a plant-based protein hydrogel into fragments, and more preferably the fragments produced in the high-shear step are 0.5 to 150 microns, preferably 0.6 to 100 microns, more preferably 0.7 to 70 microns, more preferably 0.8 to 50 microns, more preferably 0.9 to 25 microns, more preferably 1 to 20 microns, more preferably 1 to 10 microns, and more preferably 1 to 5 microns, as determined by laser diffraction. 50 The method according to any one of claims 1 to 3, comprising:

6. The method according to any one of claims 1 to 5, wherein the shearing process includes a low-shear step followed by a high-shear step.

7. The method according to claim 5, wherein at least 80% of the fragments produced in the low-shear step have a particle size determined by sieving, in the range of 1 mm to 50 mm, preferably 1 mm to 30 mm, more preferably 10 mm to 30 mm, more preferably 15 mm to 30 mm, and even more preferably 20 mm to 30 mm.

8. Step (c) further includes, between the low-shear step and the high-shear step, subjecting the plant-based protein hydrogel slurry to a solvent reduction step, preferably a solubilizing solvent reduction step, wherein the solvent reduction step is (i) A step of contacting a fragment of the plant-based hydrogel slurry with an insoluble solvent, (ii) A step of separating the fragments of the plant-based hydrogel slurry from the insolubilizing solvent to obtain a washed plant-based protein hydrogel, (iii) A process in which process (i) and process (ii) are repeated at the discretion of the user. The method according to claim 6 or claim 7, including the method described in claim 6 or 7.

9. The method according to claim 8, wherein, before washing, the plant-based protein hydrogel has a storage modulus (G') at 10 rad / sec between approximately 1,000 Pa and 20,000 Pa, between approximately 1,000 Pa and 15,000 Pa, between approximately 1,000 Pa and 10,000 Pa, between approximately 2,000 Pa and 20,000 Pa, between approximately 2,000 Pa and 15,000 Pa, and between approximately 2,000 Pa and 10,000 Pa.

10. The method according to claim 8 or 9, wherein the washed plant-based protein hydrogel has a storage modulus (G') at 10 rad / sec between approximately 500 Pa and 20,000 Pa, between approximately 500 Pa and 15,000 Pa, between approximately 500 Pa and 10,000 Pa, between approximately 1,000 Pa and 20,000 Pa, between approximately 1,000 Pa and 15,000 Pa, or between approximately 1,000 Pa and 10,000 Pa.

11. (d) The method according to any one of claims 1 to 10, further comprising the step of changing the pH of the plant-based protein hydrogel slurry to differ from the isoelectric point of the protein hydrogel by more than 1 pH unit.

12. The method according to any one of claims 1 to 11, further comprising adding an additional component to the plant-based protein hydrogel slurry, wherein the additional component is selected from plasticizers, opacifiers, preservatives, pigments and nanoparticles, or mixtures thereof.

13. The aforementioned plant-based protein hydrogel slurry, 50s -1 In this case, 10 to 10,000 cps, preferably 50s -1 In this case, 10 to 8000 cps, preferably 50s -1 In this case, 12 to 6000 cps, preferably 50s -1 The method according to any one of claims 1 to 12, wherein the viscosity is in the range of 15 to 5000 cps.

14. A plant-based protein hydrogel slurry prepared according to the method described in any one of claims 1 to 13.

15. A method for preparing plant-based structured materials, (a) a step of preparing a plant-based protein hydrogel slurry according to the method of any one of claims 1 to 14, (b) A step of subjecting the plant-based protein hydrogel slurry to one or more solvent concentration reduction steps to reduce the concentration of the first co-solvent and / or the second co-solvent in order to obtain the plant-based structured material. Methods that include...

16. The method according to claim 15, wherein the plant-based structuring material is a film, a casting, or a coating.

17. The method according to claim 16, wherein the plant-based structuring material is a coating that is a food coating, a seed coating, a pharmaceutical coating, or a surface coating (e.g., a paper coating).

18. The method according to any one of claims 15 to 17, wherein the plant-based structured material comprises a plant-based protein having a secondary structure comprising at least 40% intermolecular β-sheets, at least 50% intermolecular β-sheets, at least 60% intermolecular β-sheets, at least 70% intermolecular β-sheets, at least 80% intermolecular β-sheets, or at least 90% intermolecular β-sheets.

19. A plant-based structured material prepared according to the method described in any one of claims 15 to 18.

20. Use of the plant-based protein hydrogel slurry according to claim 14 for producing a plant-based structured material.

21. The use according to claim 20, wherein the plant-based structuring material is a film, casting, molded product, or coating.

22. The use according to claim 21, wherein the plant-based structured material is a coating that is a food coating, a seed coating, a pharmaceutical coating, or a surface coating (e.g., a paper coating).

23. Based on the total mass of the plant-based protein hydrogel slurry, the protein solid content is 5 wt% to 25 wt%, and 50s -1 and a plant-based protein hydrogel slurry having a viscosity in the range of 10 to 10,000 cps at 20°C, wherein d is 0.5 to 150 microns, determined by laser diffraction. 50 A plant-based protein hydrogel slurry containing fragments with particle size.

24. The plant-based protein hydrogel slurry has fragments having a particle size d determined by laser diffraction of 0.6 to 100 microns 50 The plant-based protein hydrogel slurry according to claim 23, comprising fragments having a particle size.

25. Based on the total mass of the plant-based protein hydrogel slurry, the protein solid content is 5 wt% to 25 wt%, and 50s -1 and a plant-based protein hydrogel slurry having a viscosity in the range of 10 to 10,000 cps at 20°C, wherein the d is less than 500 nm and is determined by dynamic light scattering. 50 A plant-based protein hydrogel slurry containing fragments with particle size.

26. d, which is determined by dynamic light scattering below 300 nm 50 A plant-based protein hydrogel slurry according to claim 25, comprising fragments having particle size.

27. The plant-based protein hydrogel slurry according to any one of claims 23 to 26, wherein the protein solid content is 6 wt% to 20 wt% based on the total mass of the plant-based protein hydrogel slurry.

28. The viscosity is 50s -1 A plant-based protein hydrogel slurry according to any one of claims 23 to 27, wherein the cps at 20°C is in the range of 10 to 8000 cps.

29. A film comprising a plant-based protein hydrogel slurry according to any one of claims 23 to 28.

30. (a) comprising a plant-based protein having a secondary structure with at least 40% intermolecular β-sheets, and / or (b) Having a tensile strength of 4 to 20 MPa, and / or (c) Having a break elongation percentage of more than 10%, The film according to claim 29.

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