Instant textured meat substitute

Abstract

A process for producing instant alternative protein products, including plant-based meat, more specifically, plant-based products with the texture, appearance, and taste of meat or dairy products. The instant meat analog or food analog may be based on natural edestin protein isolate (NEPI). NEPI can be combined with water to form a protein hydrosol, followed by the addition of oil and heating in a microwave to harden, to form a hydrogel or meat analog. The protein hydrosol may be mixed in a microwaveable cup, preferably made of a porous material such as paper or plastic, and having suitable dimensions to form a meat analog from the protein-fat hydrosol when heated in a microwave. The materials required for producing meat analogs at home from NEPI may be provided as a convenient kit for producing instant meat analogs.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 318,183, filed on March 9, 2022. The content of each application is hereby incorporated by reference in its entirety into this specification.

[0002] The present disclosure relates to the production of alternative protein products, including plant - based meat analogs and dairy analogs, and more specifically to plant - based products that have the protein content, texture, and appearance of meat or fish and can be flavored as desired.

Background Art

[0003] Consumer standards for instant home - cooked foods have evolved rapidly in recent years, and the demand for products that can be cooked at home has increased. When cooking instant foods at home, consumers have always sought products that can be prepared quickly and taste good. However, recently, consumers are demanding instant foods that are fresh, healthy, clean - label, plant - based, and produced in a sustainable manner.

[0004] One way to produce an instant, texturized plant-based meat alternative at home involves using a dried plant protein, typically a pre-texturized plant protein, as a component or element that contributes to the texture. As used herein, an instant food can be defined as a ready-to-eat food that requires minimal cooking, typically by simply adding one or two ingredients to a pre-cooked base composition. An instant food may require less than five minutes of cooking. Typically, the proteins used as components in the cooking of instant foods that are plant-based meat analogs are derived from soybeans or peas, but products using these proteins have generally not been commercially successful. Hemp-based instant foods, or meat and dairy analogs, are generally not described in the food industry or food science literature. When used as a food ingredient, hemp protein is generally considered inferior to soy and pea proteins, particularly with respect to the properties required for the production of meat analogs.

[0005] Currently, soy protein and pea protein-based products dominate the meat analog market, with companies such as Impossible Foods® and Vision meet® producing a high percentage of meat analog products. The meat analogs produced by these companies are pre-cooked, use a number of ingredients, and are generally pre-packaged and sold in the same way as fresh or frozen animal meat products. Most pre-cooked plant-based meat analogs generally need to be cooked using a stove or oven before consumption. The production of these products generally requires extrusion at relatively high temperatures of about 120°C to 140°C to obtain texturized protein. To form a protein blend having the appearance of fresh ground meat or other animal meat products, it is generally necessary to blend pre-texture protein and post-texture protein with other ingredients such as cellulose, starch, sugar, oil, and binders. These products also generally require immediate microbiological stabilization, for example by continuous refrigeration or freezing, and packaging before sale. Currently, these types of products do not have valid claims regarding sustainability or as clean label materials.

[0006] Isolates of soy, pea, wheat gluten, and other proteins used in the production of conventional meat analogs cannot form fully texturized or meat-like products without prior extrusion of the protein. U.S. Patent No. 3,662,673 to Boyer discloses the use of microwaves to produce texturized protein products. However, texturized protein products produced using proteins such as those described in Boyer, such as isolates of soy protein, or by conventional soy-based tofu processing that requires chemicals such as calcium chloride, result in products with low levels of expansion and are unacceptable to most consumers. The texture of products generally produced by the methods of Boyer or by soy tofu processing is essentially uniform in cross-section and lacks the elasticity, fibrousness, and texture of meat.

[0007] Isolates of soy protein and pea protein cannot be properly texturized unless first subjected to conventional extrusion at temperatures far exceeding 100°C, which limits their usefulness for quick or instant, meat and dairy product-like applications. Further, in an extrusion process at temperatures exceeding 100°C, the inclusion of moisture into the extruded texturized soy and pea proteins is limited, thus requiring pre-hydration of the texturized protein after extrusion before use. Importantly, after extrusion, the protein's ability to hold water within the protein structure itself (instead of between sponge-like protein pockets) is destroyed, and as a result, other ingredients are required to hold moisture in order to mimic the natural moisture held within the structure of meat.

[0008] Accordingly, there is a need for an instant, fresh, texturized plant-based meat analog capable of replicating the texture of meat or fish; being easily manufactured at home without the use of an extruder and industrial equipment or chemicals such as a distiller or calcium chloride, if desired; achieving protein texturization without using temperatures exceeding 100°C; being free of allergenic substances such as soy gluten or wheat gluten; and mimicking the meat texture and nutritional composition of various meat and fish analogs without containing numerous ingredients such as starch, gums, and emulsifiers to hold the required water and oil.

[0009] Hemp-based meat analogs or dairy analogs manufactured using only hemp grains as a protein source are not known to be commercially available and are not described in the food industry or food science literature. With respect to use in food, hemp protein is considered to be inferior to soy and pea proteins, particularly with regard to the properties required for the production of meat analogs and dairy analogs. Meat analogs and dairy analogs, also referred to herein as structured protein foods, require proteins that can form a strong gel matrix, and hemp protein is not recognized as having a strong ability in that regard.

[0010] According to Wang, "The emulsifying and gel-forming properties of hemp protein have generally been found to be inferior to those of soy protein" (Wang et al., 2019). While Malomo showed that salt micellization isolation of hemp protein can improve its gel-forming ability, Shen disclosed that this complex and costly and time-consuming protein isolation method has an adverse effect on the protein structure and that chemical cross-linking agents may be required for sufficient gel-forming ability of hemp protein (Shen et al., 2021, Malomo et al., 2014, Wang et al., 2019).

[0011] As shown by Wang, soy protein is currently preferred over hemp protein for the production of meat analogs. "Currently, mainly soy protein is used to mimic animal protein. This is due to its beneficial gelling properties and the resulting formation of an entangled fibrous matrix" (Schreuders et al., 2019). The latter fibrillation of soy occurs at a typical temperature of 130 °C (266 °F). For example, IMPOSSIBLE FOODS uses soy protein in its IMPOSSIBLE BURGER. However, due mainly to health and nutrition concerns regarding soy products, BEYOND MEAT, the biggest competitor of IMPOSSIBLE FOODS, uses yellow pea protein in its BEYOND BURGER. However, yellow peas "have a much lower gelling ability than soy protein", and "the heat-induced gel of soy protein isolate (SPI) is stronger than the heat-induced gel of pea protein isolate (PPI)" (Schreuders et al., 2019).

[0012] Soy protein and pea protein have known flavor, texture, and phytochemical limitations as protein sources for the manufacture of meat analogs and dairy analogs, but no successful examples of alternatives to plant-based alternative proteins for meat analogs have been found yet. However, hemp protein is being considered as a possible alternative to soy protein in meat analogs. Recently, ZaHari reported that although the excellent nutritional and functional properties of hemp protein have been recognized, it has not yet been used in meat analog production. "Previous studies have shown that especially hemp seed protein has high protein quality and functionality. However, there is no research on using hemp seed protein as a raw material for meat analog production" (ZaHari et al, 2020).

[0013] Zahari further demonstrated that hemp protein concentrate (HPC) can be used in combination with soy protein isolate (SPI) to produce meat analogs by conventional extrusion, but it cannot be used as a single protein source. This study concluded that "therefore, HPC may be a promising novel material included in extruded products, and this study showed that a texture comparable to that of SPI alone was obtained from the resulting meat analogs, and that soy protein can be replaced by hemp protein up to 60%" (ZaHari et al, 2020). Regarding the use of higher concentrations of hemp protein in the formulation, the study showed that this would result in an unacceptable decrease in the hardness and chewiness of the meat analog products. Therefore,

[0014] ZaHari, Wang, and Shen teach against using hemp protein as the sole protein source in meat analog production.

[0015] Despite the need for new and improved plant-based protein sources to meet the growing demand in the plant-based food industry, hemp protein has not gained a sufficient market share in food manufacturing. Soy protein and pea protein continue to dominate the plant-based food market, despite the nutritional and environmental advantages of hemp. Soy protein and pea protein have benefited from decades of research and wide commercial availability, and their use in meat analog production and other foods has significantly improved the quality of ingredients and products and achieved economies of scale.

[0016] Improvements in soy-based and pea-based meat analogues have been brought about through extensive research and development at all stages of the production of meat analogues and dairy analogues. The production of meat analogues generally involves four steps. The first step involves isolating protein from selected plant raw materials. The second step involves combining the isolated protein with water and oil to form a matrix for thermal gelation or extrusion. The third step involves thermally gelating or extruding the raw materials to solidify the protein and impart texture. The final step involves using binders and water binders such as carrageenan, cellulose fibers, starch, gluten, or wheat flour to form meat analogue products, which can then be cooked to mimic products such as minced meat, sliced meat, chicken fillets, pulled pork, etc.

[0017] The first step in meat analogue production involves the isolation of protein from plant raw materials. Conventionally, soy and pea proteins have been used as plant raw materials for protein isolation. However, hemp seed protein has been considered to be a potential protein source for meat analogues due to its excellent digestibility and desirable essential amino acid composition (Tang, Ten, Wang, & Yang, 2006; Wang, Tang, Yang, & Gao, 2008; Russo and Reggiani, 2015a; Callaway, 2004; House et al., 2010; Docimo et al., 2014; Zahari et al., 2020). Recent proteomic characterization of hemp seeds has concluded that hemp seeds are non-leguminous, protein-rich, and underutilized grains (Aiello et al., 2016).

[0018] Hemp protein has high nutritional potential, but the nutritional quality of plant proteins is influenced by many factors as it is measured by amino acid composition and digestibility. Amino acid composition can be affected by genetic variation, or agronomic conditions such as soil fertility, and post-harvest treatments (e.g., dehulling) that change the ratio of cereal components. Protein digestibility can be affected by the protein's structure and the presence of anti-nutritional compounds formed in the plant material or during alkaline or heat treatments (Sarwar, 1997). However, Aiello found that anti-nutritional factors, including condensed tannins, phytic acid, and trypsin inhibitors, are present at low concentrations in hemp grains (Aiello et al., 2016).

[0019] Due to the functional characteristics of hemp protein, its use as a protein source in foods has been hindered. Hemp protein concentrate is commercially available as a result of the production of hemp seed oil. Hemp seeds that have been crushed and pressed for lubricating oil become protein-rich seed cake. The seed cake is green, fibrous, and represents approximately 40% protein concentrate. Unfortunately, it has an overly grassy, earthy flavor and is unacceptable for most foods. By grinding this cake and passing it through a dry sieve, the protein content can be increased to approximately 50%. Many researchers who recognize the nutritional value of this protein-rich source have used the seed cake as a starting material for isolating hemp protein and improving its taste and functionality. Tang found that isolated hemp seed protein (HPI) from the seed cake is not superior to soy protein isolate (SPI) for use in the production of plant-based foods (Tang et al., 2006). Tang showed that in the case of HPI, due to the low water solubility of hemp globulin, its emulsifying and water-holding capacities are considered inferior when compared to soy protein isolate (Tang et al., 2006, Hadnaev et al. 2020). According to Tang, "The data suggest that HPI can be used as a valuable nutritional source for infants and children, but its functionality is insufficient when compared to SPI. The insufficient functionality of HPI is mainly due to the high free sulfhydryl content derived from sulfur-containing amino acids, which results in the formation of disulfide covalent bonds between individual proteins and subsequent aggregation at neutral or acidic pH" (Tang et al., 2006). Furthermore, "Differential scanning calorimetry (DSC) analysis showed that HPI exhibited only one endothermic peak with a denaturation temperature (T(d)) of approximately 95.0 °C due to the edestin component" (Tang et al., 2006).

[0020] Despite the fact that the functional aspects of hemp protein are clearly inferior, its excellent nutritional value has gathered continuous interest in its use in food manufacturing. For this purpose, individual proteins have been isolated from hemp and further studied for their potential functionality. Furthermore, researchers have examined whether different extraction and isolation methods of hemp protein can improve functionality. "The value and use of hemp protein in food are closely related to the structure and functionality of the protein" (Wang et al., 2019).

[0021] To examine the nutritional and functional properties of individual hemp grain proteins, researchers have adopted a method of extracting and separating two of the major proteins present in hemp grains. Hemp grain protein is mainly composed of edestin and albumin protein. Edestin, which is a globulin, accounts for about 60% - 80% of the total protein content (Odani & Odani 1998, Tang et al., 2006), with albumin, a globular protein rather than a globulin, making up the difference. Edestin and albumin have different amino acid compositions and functional characteristics.

[0022] Malamo tested the nutritional differences between edestin and albumin in hemp grains and reached the conclusion that the edestin fraction of hemp protein is nutritionally superior and contains more sulfur-containing (methionine and cysteine), aromatic (AAA), branched-chain, and hydrophobic amino acids (Malomo and Aluko 2015). Malamo separated edestin from albumin and measured the properties regarding their respective nutritional value and functionality. These properties include solubility in water, amino acid content, and digestibility.

[0023] Malmo reported that the albumin fraction is soluble in water, while the edestin fraction is soluble in salt solution. The extracted edestin has extremely low solubility in water at neutral or acidic pH and is only soluble at high ionic strength or alkaline pH (Malomo & Aluko, 2015). "The functions of many proteins, such as surface activity, are correlated with the solubility of the protein" (Jackman & Yada, 1989, Malomo & Aluko, 2015). In hemp seeds, it was found that edestin has good emulsion-forming ability, and the solubility and foaming ability of albumin are higher than those of edestin (Malomo & Aluko, 2015).

[0024] Studies have shown that edestin may only be found in hemp seeds, but edestin-like proteins may be present in grains of the family that includes pumpkin and squash (Vickery, 1940). Therefore, the present disclosure and its applications may relate to edestin, and edestin-like proteins that may have similar or identical properties to edestin. Vickery disclosed that potential alternatives to edestin may be found among plants of the Cucurbitaceae family, including pumpkin seeds. Hirohata investigated the globulins of 38 different species and varieties from 8 genera of this family and noted the close similarity of the globulins from closely related species (Vickery, 1940, Hirotata, 1932). Vickery suggested that the globulins of the Cucurbitaceae family may contain edestin-like proteins that meet the requirements for nutritional substitutes for hemp seed edestin (Vickery 1940).

[0025] Edestin was first isolated and analyzed by Thomas Osborne (Osborne, 1892). In its fully native form, edestin is composed of six identical subunits, each consisting of an acidic (AS) subunit and a basic (BS) subunit linked by one disulfide bond (Farinon 2020, Patel, Cudney, & McPherson 1994). Recently, it has been shown that edestin can exist in several forms even within a single hemp species (Docimo et al., 2014). For example, in one species of Cannabis Sativa, seven genes encode edestin globulin, which results in two branched forms of edestin types. Within a particular strain of hemp, one type of edestin is substantially identical to each other, while the second type of edestin is substantially different from the first type. Ponzoni identified the CsEde3, a type-3 edestin gene, which shows approximately 65% and 58% sequence homology when compared to the genomic forms of CsEde1 and CsEde2, respectively (Ponzoni, Brambia, and Galasso, 2018). The amino acid composition can vary greatly between the two types of edestin, and some types have high nutritional quality (Docimo et al., 2014). Edestin itself has a large particle weight of 309,000, but when denatured, it depolymerizes to 51,000 in concentrated urea solution [Burk & Greenberg, 1930] and to 17,000 in dilute HCl [Adair & Adair, 1934]. These units are, respectively, about 1 / 6 and 1 / 18 the size of the native molecule. In the native state, they have a specific polypeptide pattern and are integrated, in part, perhaps by some chemical bond (e.g., SS bond), but mainly by the lateral attraction between adjacent CO and NH groups and by the interaction between the free acid and basic groups of the side chains. As can be seen from the following analytical data, the number of these latter groups is large: glutamic acid, 19 - 2%, aspartic acid, 10 - 2% [Jones & Moeller, 1928], arginine, 17 - 76% [Vickery, 1940], lysine (2 - 4%), histidine (2 - 03%) [Tristram, 1939], amide-N, 1 - 73% [Bailey, 1937, 2]. Considering the amidated COOH groups, they correspond to 670 charged groups out of 309,000 per molecule. Such a spatial arrangement of charges gives rise to a specific charge symmetry on which the stability of the molecule must ultimately depend and which allows for some variation, as reflected in the change in the dipole moment within definite pH limits. Outside these limits, the ionization of acidic or basic groups is further suppressed, and attractive and repulsive forces occur within the molecule, especially when there are no small mobile ions, distorting and ultimately destroying the native polypeptide arrangement (Beiley, 1940).

[0026] Accordingly, the edestin referred to in this disclosure can incorporate all forms of edestin, whether currently known or potentially unknown at present, having properties similar or identical to the edestin disclosed for the purposes of this disclosure.

[0027] Edestin is readily and rapidly decomposed into edestan under weakly acidic conditions. Edestan, an intermediate product derived from edestin, is formed during the denaturation of edestin and was first identified by Osborne (Osborne, 1901; 1902). Edestan is formed when edestin comes into contact with dilute acid. Edestan results in the liberation of SH groups (Bailey, 1942). Bailey demonstrated that under acidic conditions, edestin can be rapidly converted to edestan in less than 20 minutes (Beiley, 1942). This study showed that the liberation of SH groups is associated with the conversion of edestin to edestan. Bailey also reported that the nitrogen content of edestan decreases compared to edestin, which can be explained by the decrease in tryptophan in edestin. Native, non-denatured edestin has different functionality from denatured or partially denatured edestin or edestan.

[0028] Conventional techniques for isolating hemp protein or separating edestin from albumin can cause structural changes in the protein, some of which may be irreversible. Different methods and conditions for protein extraction and isolation (pH, presence of monovalent and polyvalent salts, ionic strength of the medium used for protein extraction, time, temperature, etc.) can affect the functionality of the protein (Hadnaev et al., 2018). These changes can have an adverse effect on the function of the protein (Hadnaev et al, 2018, Shen et al., 2021). These adverse effects can include changes in digestibility, protein-oil interactions, taste, solubility, and the ability to emulsify and form gels (Shen et al., 2021). Therefore, it is particularly important to maintain the natural structure of the protein as much as possible, especially when extracting and isolating edestin for food use.

[0029] Several different techniques are utilized to isolate hemp protein and edestin. These techniques include the use of mechanical processes, such as compression, milling or sieving of grains or husked grains, or milling of grains and sieving of grain slurries, in aqueous or solvent slurries, at high temperature, alkaline or acidic conditions, isoelectric precipitation, isoelectric focusing, micellization, ultrafiltration, as well as the use of aqueous or solvent slurries. Any of these techniques may change the structure of the protein and reduce its functionality.

[0030] The high temperatures generated by mechanical processes can have an adverse effect on the functionality of proteins. For example, the grinding of grains to produce wheat flour can generate temperatures high enough to change the structure of the proteins. These temperatures can cause the denaturation of edestin and the binding or aggregation of edestin, albumin, or fibers, thereby preventing their independent isolation.

[0031] Dry milling of particles can generate temperatures of at least 80 °C to 100 °C and may denature edestin. Mohammad found that the heat and mechanical forces generated during milling can denature globular proteins (Mohommad, 2015). Mohammad showed that the mechanical stress applied during milling can change the bulk properties of globular proteins.

[0032] Therefore, the high temperatures caused by dry milling to produce wheat flour can change the structure of hemp protein. Farinon calculated the denaturation temperature of hemp grain protein (edestin) to be 92 °C (Farinon et al., 2020). Furthermore, Malamo showed that heat treatment and changes in pH can change the secondary structure of albumin and edestin in hemp grains (Malamo and Aluko, 2015). High temperatures can cause protein unfolding, thereby exposing hydrophobic groups and promoting protein-protein interactions over protein-water interactions in some cases.

[0033] Heating during extraction can be avoided or minimized by using chemical means for protein extraction, but many chemical extraction methods require first mechanically reducing the particle size. Solvent extraction is a common method for separating proteins from plant materials, which involves the use of a liquid solvent to which the protein-containing material is added. The solvent can be water, alcohol, acetone, hexane, or other liquid solvents. Solvent extraction may be combined with mechanical or other extraction means to first break down the plant material to release the protein. Solvent extraction may involve the use of a solvent that breaks down the plant cell wall or fibrous material, thereby releasing the protein.

[0034] Some solvents used in protein extraction have the drawback of denaturing the protein. Furthermore, these solvents are toxic and may not be suitable for ingestion even in small amounts. Additionally, solvents generally require long extraction times and labor-intensive procedures, and solvent residues may remain in the food and be difficult to dispose of safely. Hexane is an example of this type of solvent. Many solvents cannot be used in the production of certified organic foods under the guidelines of the United States Department of Agriculture (USDA) regarding organic food labeling.

[0035] One alternative process for protein extraction that does not require a solvent is water extraction, which involves adding ground or pressed plant material to water and then separating the protein based on its solubility in the aqueous fraction or its solubility in the fat-containing fraction when the fat in the plant material separates from the water. After water extraction, isoelectric precipitation or isoelectric focusing or salt extraction may be performed to isolate the protein.

[0036] Alkali extraction is a common technique in which the cell structure is disrupted by a highly basic solvent, thereby releasing proteins from the cells. However, this process can cause damage to proteins, including racemization of amino acids, formation of lysinoalanine, reduction in digestibility, and loss of essential amino acids (Moure et al., 2006). According to Xu, under alkaline conditions, the polyphenols found in many plant raw materials, including hemp grains, are oxidized and then react with proteins, resulting in a dark green or brown color in the extracted protein solution (Xu and Diosady, 2002).

[0037] When used during the extraction of hemp grain proteins, the pH of the alkali extract generally rises to 9 or 10, higher than that for legume protein extraction (pH 8), because the natural hemp grain proteins are tightly packed and can be closely integrated with other components, such as phenolic compounds (Wang and Xiong, 2019). After alkali extraction, precipitation of the target hemp protein at the isoelectric point generally follows, and after several washing steps, the color derived from the protein isolate often cannot be removed.

[0038] After water extraction or alkali extraction, isoelectric precipitation or salt extraction is generally performed to isolate the protein. Isoelectric precipitation may be used to extract soluble proteins after alkali extraction or solvent extraction and involves adjusting the pH until the charge balance between the target protein and the solvent is reached, thereby precipitating the protein from the solution. Isoelectric precipitation requires a change in pH that can alter the structure of the protein and may thus have an adverse effect on the protein's function.

[0039] Regarding the isoelectric precipitation of edestin, Beiley disclosed that the isoelectric point zone of edestin is at pH 5.5 (Beiley, 1942). In this process, albumin can be removed during the precipitation of edestin at its isoelectric point (Papalamrou et al., 2009). This result may be attributed to the higher solubility (over 75%) of hemp seed albumin at pH 5.0 compared to hemp seed globulin (less than 10%) (Malomo & Aluko, 2015). One of the advantages of isoelectric precipitation compared to other protein isolation methods is that the protein isolate obtained by isoelectric precipitation has been found to have a higher water-binding capacity compared to the same isolate obtained by micellization extraction (Krause et al., 2002). However, the disadvantage of isoelectric precipitation during edestin isolation is that the protein solubility is low compared to edestin isolated by salt extraction, suggesting that the protein is no longer in its native state (Hadnaev, 2018).

[0040] Compared with alkali extraction and isoelectric precipitation, salt extraction, which can involve micellization, represents a milder extraction procedure and does not cause oxidation, polymerization, and co-extraction with polyphenols of proteins. In salt extraction, a group of proteins is "salted in", followed by "salting out" of the target protein. "Salting in" refers to the effect that the solubility of solutes such as proteins increases when the ionic strength of the solution increases. This effect tends to be observed at lower ionic strengths. "Salting out" involves increasing the salt concentration, which causes the solvation force of salt ions to decrease due to the abundance of salt ions, and thus the solubility of the target protein decreases, resulting in precipitation.

[0041] One method of salt extraction involves a micellization step, as described in U.S. Patent No. 6,005,076 to Murray. Salt extraction using micellization initially involves solubilizing the protein in a salt solution having a specific ionic strength. Next, the physiological saline solvent is diluted with the concentrated protein solution to reduce the ionic strength below a specific level, thereby resulting in the formation of individual protein particles, at least partially in the form of protein micelles, in the aqueous phase. The protein micelles then sediment to form a mass of the target protein isolate. The protein isolate can then be separated from the supernatant.

[0042] Salt-based micellization extracts, as disclosed by Murray, have the advantage of producing protein isolates with higher solubility when compared to isolates obtained by isoelectric precipitation (Karaca et al., 2011; Krause et al., 2002; Paredes-Lovez and Ordrica-Falomir 1986). In addition to the improved solubility, the surface activity was higher for the protein isolates obtained by the micellization technique when compared to isoelectric precipitation. Furthermore, according to Krause and Papalamrou, micellization extraction resulted in protein isolates in which the native protein structure was better preserved when compared to isoelectric precipitation proteins (Krause et al., 2002; Papalamrou et al. 2009). Generally, isoelectric precipitation can denature the extracted protein to some extent, which can lead to hydrophobic interactions between protein molecules and the formation of insoluble protein aggregates. Salt extraction and micellization may be the least damaging among the known methods for isolating hemp protein, but the addition of salt during isolation can have an adverse effect on the structure and function of the protein. "The addition of NaCl also has various effects on the gel structure. Specifically, increasing the NaCl concentration (up to 300 mM) intensively promotes protein-protein interactions and aggregation, resulting in the formation of an HMI [hemp protein micellization isolate] gel structure with larger pore sizes" (Shen et al., 2021).

[0043] Salt extraction was the first method used to isolate edestin from hemp grains (Osborne, 1892). This method was further developed by Malomo, who utilized micellization technology to extract edestin (Malomo and Aluko, 2015). As demonstrated by Malomo, in salt-based micellization extraction, albumin remains in the supernatant after salt removal in the dialysis step, while globulin precipitates and can be collected by centrifugation. In Malomo's study, isolates of globulin were produced by salt extraction of hemp grain flour followed by dialysis against water in a dialysis tube.

[0044] Dialysis of the salt extract of hemp grain flour resulted in albumin remaining in solution while water-insoluble globulin in micellar form precipitated (Malomo and Aluko, 2015). The precipitate was then collected and lyophilized. When comparing the hemp protein albumin and globulin fractions, albumin had significantly higher protein solubility and foaming capacity than globulin, although no difference in emulsion-forming ability was observed between the two protein fractions. Salt extraction and micellization are costly in terms of labor, time, materials, equipment, and waste disposal costs and are not currently considered commercially viable for protein extraction for food use.

[0045] Ultrafiltration is another method that can be used to produce protein isolates with improved functionality compared to other conventional protein extraction techniques. For example, protein isolates obtained by ultrafiltration generally have good emulsifying properties when compared to alkaline extraction. However, one drawback of ultrafiltration is membrane fouling due to the formation of precipitates in the final product, which can increase the extraction cost.

[0046] Newer methods of protein extraction include ultrasound-assisted extraction, enzyme-assisted protein extraction, and electro-protein extraction methods. These methods have drawbacks including high cost, low yield, protein degradation, and protein impurities. Therefore, conventional extraction methods including salt extraction, alkali extraction, and isoelectric precipitation remain the main methods for extracting protein from plant materials such as hemp grains.

[0047] Regarding the disclosed methods of extracting and isolating hemp using the methods described above in this specification, an example of aqueous protein extraction from hemp grains followed by isoelectric precipitation is disclosed in U.S. Patent No. 10,555,542 to Crank. Crank discloses first grinding hemp grains using any suitable means including grinding using a hammer mill, roller mill, or screw-type mill. Grinding by these processes is a high-energy process and generally results in high temperatures, generally reaching about 140°F to 150°F. As is known in the technical field of peanut butter manufacturing, these high temperatures are required to form a paste from solids, and thus are required to obtain a paste. These temperatures can, depending on the end use of the product, in some cases cause undesirable interactions between the protein components of the grain material. In Crank, grinding produces a paste or fine powder (fine powder in the case where the grains are first pressed to remove oil), and water can be added to the ground material at a ratio of about 4 to about 16 parts by weight per part of the plant material. Crank discloses adjusting the pH to about 7.5 by adding a base such as calcium hydroxide to facilitate protein extraction.

[0048] Next, the obtained solution is centrifuged to separate the fat fraction from the aqueous fraction or the low-fat extract. The defatted extract can be used as a defatted plant milk or can be further processed to produce a protein concentrate or isolate. In Crank, the protein in the defatted extract was concentrated by precipitation and separated to produce a plant protein concentrate or isolate from a partially defatted plant raw material. Crank discloses that the protein in the low-fat extract can be precipitated by adding an acid such as citric acid to the isoelectric point of the protein. Crank does not disclose that only the water extract may be used to separate edestin and albumin. Further, Crank mentions in the pending application that hemp seeds can be a source of protein isolated for food according to Crank's process and that hemp seeds contain edestin, but Crank does not disclose the purification and isolation of edestin. Crank discloses discarding the fiber and protein-containing parts of hemp protein after centrifugation.

[0049] Czech Patent No. 33,545 to Beran discloses a method for extracting edestin from hemp grains to produce a protein for human consumption. In the background art section of this patent, Beran discloses that hemp protein is often produced as an isolate of spray-dried hemp protein, which often utilizes high heat and can cause protein denaturation. According to Benan, spray drying may require temperatures of 150°C to 250°C, which are temperatures at which hemp protein may denature. Beran discloses that "thermal denaturation of proteins has an adverse effect on solubility and dispersibility, foaming properties, and emulsifying properties."

[0050] To avoid heat denaturation of edestin during its preparation by spray drying, Beran discloses a method that includes first grinding or pressing hemp grains to remove oil and then purifying edestin by either water extraction and isoelectric precipitation or salt extraction. The preferred method of particle size reduction used by Beran appears to be dry grinding. According to this patent, the ground powder is then added to water at a concentration with a water-to-powder ratio of 5:1. Next, Beran discloses shaking the solution to produce an albumin-containing aqueous fraction and a sediment fraction.

[0051] Beran discloses that the sediment contains edestin, but Beran does not disclose further steps for isolating edestin for use in food. These steps include protein extraction by either isoelectric precipitation, salt extraction, or ultrafiltration. Beran does not disclose the level of purity of edestin in the sediment before such additional steps for isolating edestin, but the need for such additional steps indicates that the purity of edestin in the sediment is not sufficient for Beran's stated purpose of "increasing the protein content of high-protein foods and smooth protein beverages" using edestin. In conclusion, Beran discloses that "due to its beneficial effects on emulsifying properties and sensory acceptability of the final product, this product can be used in these foods."

[0052] Both Crank and Beran generally disclose using ground or pressed hemp grains (to remove oil) and then grinding them to use the hemp fine powder as a starting material for protein extraction. As a result, the hemp fine powder has undergone dry grinding or milling and oil pressing processes that affect the structure and function of the protein. Furthermore, both Crank and Beran disclose isolating edestin by at least isoelectric precipitation, which causes structural changes in the protein and as a result, its function is reduced.

[0053] In the process of extracting protein from grains for use in plant-based meat, oil can also be extracted from the grains. Since plant-based oil has value as food and cosmetics, in some cases, oil extraction can be the main purpose. Common methods of extracting oil from grains, nuts, and seeds include pressing-based extraction methods such as cold pressing and expeller pressing, as well as solvent extraction.

[0054] To extract oil by pressing grains, it involves mechanically compressing the plant material to squeeze out the oil from the solid. Solvent extraction involves putting the plant raw material into a liquid to extract the oil. In some cases, pressing and solvent extraction can be combined. Oil recovery from the expeller pressing method can be relatively inefficient, and a fairly high percentage of fat can remain in the cake. As a result, the pressed cake can be further extracted using an oil solubilizing solvent. Commercially available cakes and flours produced by the pressing method, or the pressing and solvent methods, are thought to have a reduced protein function.

[0055] Conventionally produced hemp grain oil may be green, which may be due to the destruction of protoplasts or chloroplasts during extraction. Hemp grains may contain chlorophyll-containing substances that release chlorophyll when broken. Compared to other types of grains, hemp grains contain more of these substances, so hemp oil extracted by conventional methods tends to turn green.

[0056] According to Leonard et al., "Unrefined hemp seed oil is dark green because it contains chlorophyll" (Leonard et al., 2019). Furthermore, the presence of chlorophyll in the oil can cause fat oxidation and result in off - odors. U.S. Patent No. 9,493,749 to Soe discloses that "Vegetable oils derived from oilseed grains such as soybean oil, coconut oil, canola oil, cottonseed oil, and peanut oil usually contain some chlorophyll. However, the presence of high levels of chlorophyll pigments in vegetable oils is generally not desirable. This is because chlorophyll imparts an undesirable green color, can cause oxidation of the oil during storage, and may lead to oil deterioration."

[0057] Various methods have been used to remove chlorophyll from vegetable oils. These methods include chemical bleaching and ultrasonic bleaching. Chlorophyll can be removed during many stages of the oil manufacturing process, such as grain grinding, oil extraction, degumming, caustic treatment, and bleaching. However, the bleaching step is usually the most important for reducing chlorophyll residues to acceptable levels. During bleaching, the oil is usually heated and passed through an adsorbent to remove chlorophyll and other colored compounds that affect the appearance and / or stability of the finished oil. The adsorbent used in the bleaching process is usually clay.

[0058] Conventional methods for removing chlorophyll from hemp oil are costly and may cause problems in waste disposal. Furthermore, in methods of removing chlorophyll from hemp oil after the chloroplasts are ruptured, the oil may be oxidized due to temporary exposure to chlorophyll. Therefore, an improved method for extracting oil from hemp grains is needed.

[0059] In the production of meat analogues and dairy analogues, after obtaining protein isolates and a preferred oil source, in step 2, the isolated proteins are combined with water and optionally oil to form a material for curing or extrusion. After the protein is isolated from hemp grains or other plant products, it needs to be combined with other components of the meat analogue or dairy analogue to form the final meat analogue. The three basic components for meat analogue production are protein, water, and fat. These components can be combined in various concentrations and processed in various ways to form meat analogues and dairy analogues.

[0060] Considering that meat analogues require structuring to result in gelling or a meat-like texture with chewiness, proteins typically combine with water and optionally oil to form a gel and create a texture by setting with heat. Regarding the gel formation of these components or only using protein and water of hemp protein isolate, studies have shown that hemp protein does not have good gel-forming properties. As disclosed above, Wang, Shen, and Zahari teach that hemp does not have good gel-forming ability and as a result, hemp cannot be a candidate for use as the main protein in meat analogues or dairy analogues (Wang et al., 2019, Shen et al., 2021, Zaari et al., 2020). For example, Wang showed that a combination of hemp protein isolate and water alone does not form a desirable gel even when heated. Wang also showed that even when oil was added to Wang's protein and water mixture and heated again, the mixture of hemp protein, water, and oil did not form a desirable gel upon heating.

[0061] In the production of meat analogues, the third step of thermosetting protein, water, and oil typically involves extrusion, which textures the product to form a more meat-like material. To form textured meat, an extruder can be used to form textured vegetable protein (TVP). TVP is typically a soy-based product, but other vegetable proteins such as pea protein can also be used alone or in combination with soy. To make TVP, plant-based ingredients are fed into an extruder and textured. Conventionally, dry vegetable protein is fed into the extruder, and then water, starch, and optionally fat are added to the protein via separate inlets as the protein is conveyed through the extruder. After extrusion, the extruder discharge may pass through marinating, coating, and / or cooling steps.

[0062] Common problems associated with conventional plant-based meat alternatives that include TVP and HMMA relate to the non-dispersed texture and rubbery mouthfeel when compared to meat. This texture and mouthfeel of conventional meat analogues is due in part to the fact that neither protein, oil, or combinations thereof are incorporated into the molecular structure of the protein's peptide chains or "fibers". In the case of meat derived from animals, fat molecules are incorporated between these muscle fibers that make up the majority of the animal's meat product. This fat is released during chewing and provides consumers with positive and continuous sensory feedback regarding taste and mouthfeel as chewing continues. The sensory feedback provided during chewing of current conventional meat analogues is not equivalent to that obtained in the case of meat, partly because there is no fat between the peptide layers of the protein. In conventional meat analogues, since fat is added after the protein is fully denatured, it may surround fairly large sized cooked protein pieces but is not incorporated within the peptide layer of the protein itself.

[0063] Soybean and pea proteins commonly used to produce fibers of TVP and HMMA may retain only about 10% of their weight in fat. Typically, meat muscle fibers incorporate 5 to about 50% of their weight as fat within the protein fibers, depending on the meat source. Thus, in conventional soybean and pea meat analogs, much of the fat added to the product during extrusion remains outside the fibers, creating a greasy and unappetizing product that does not release fat in a controlled, juicy manner when chewed. As a result, conventional meat analog products mainly appeal to a limited number of dedicated vegan or vegetarian consumers and are not attractive to the majority of meat-eating consumers.

[0064] Different extrusion methods can produce different meat analog textures. Extrusion has been developed over decades to create more meat-like meat analogs. The extrusion of meat analogs, and the preparation for extrusion, involve complex chemical changes and processes within the protein components of the extrusion mixture. During extrusion, the structure of the protein isolate changes significantly, such that the protein can be partially or fully denatured or unfolded, reconfigured, cross-linked with other protein molecules, and chemically bonded to other components of the extrusion mixture. The extruder induces these changes through the application of shear forces by the screw as the mixture moves through the machine, in addition to changes in temperature and pressure. The final texture, flavor, and mouthfeel of the meat analog produced by extrusion are determined by the various types of chemical bonds formed between the components of the extrusion mixture before, during, and after extrusion.

[0065] Regarding the initial development of an extrusion process for manufacturing textured meat analogs, U.S. Patent No. 6,319,539 to Shemer et al. discloses mixing proteins with a high percentage of water and optionally fat, and heating, gelling, and shaping the resulting paste within an extruder. Shemer discloses heating the paste during transfer to the extrusion device, conveying it at a predetermined rate, and then extruding it through an opening. The resulting food product has a fibrous texture that includes substantially aligned axial fibers. However, a problem with this process is that the flow rate is limited, and the types of products are limited because it can only be carried out using specific raw materials, particularly gluten. Gluten is a known allergen, which has also limited the use of soy-based TVP until now.

[0066] A further drawback of the Shemer process and other initial extruder processes is that when the hot product is cooled, water vapor is released, causing the heated product to expand as it is conveyed from the extruder. The water vapor causes disruption of the aligned protein fibers, which is undesirable for an acceptable texture in meat analogs.

[0067] To address this problem, International Publication No. WO2003 / 007729 to Bouvier et al. discloses a twin-screw rotor extruder that has an elongated cooling chamber and enables mixing and extrusion of raw materials at a temperature controlled so that steam does not disrupt the alignment of the protein fibers of the final product, as opposed to a single-screw device. In addition to addressing the cooling and water vapor problems, the '729 application also recognized problems in the existing art regarding incorporating a desired amount of oil and fat into an extruded product using conventional raw material formulations.

[0068] To achieve the desired fat content, the '729 application disclosed a novel extruded mixture containing a fat component mixed with lecithin or casein salts, protein, fiber, starch, and water. This mixture was kneaded to obtain a paste, which was then heated and gelled in an extruder. However, the inclusion of a significant amount of carbohydrates such as starch in the meat analog is undesirable due to taste and nutritional concerns.

[0069] To solve the problem of introducing fat into the meat analog without adding starch and without causing other related problems associated with introducing oil into the extruder, International Publication No. WO2012 / 158023 to Giezen discloses an extrusion process for converting a plant protein composition such as soy protein into a fibrous meat-like structure. Giezen discloses an extrusion outlet temperature higher than the boiling point of water, resulting in an open product structure that allows oil to be injected to reach the desired fat content. The problems with Giezen include adding post-extrusion process steps and the final product being perceived by consumers as being too oily and having too much fat.

[0070] A problem generally recognized in the technical field of meat analog extrusion is that a large amount of oil in the extruded mixture prevents the obtaining of a product having the texture of animal meat. In conventional meat analog extrusion, the presence of oil reduces the high mechanical shear force in the extruder that forms the ideal fibrous structure of the meat analog. Thus, when using conventional processes, adding an optimal amount of oil results in a meat analog with a fiber structure and texture below optimal.

[0071] To overcome the problem that a high oil content results in a sub - optimal texture in a texturized meat analog, U.S. Patent Application No. 20180064137 to Trotter et al. discloses adding oil separately from other raw materials during extrusion. This process involves supplying 40 - 70 wt% water and 15 - 35 wt% vegetable protein to an extruder barrel, and subsequently injecting 2 - 15 wt% oil into the extruder barrel at a location downstream of the supply device. According to this disclosure, the downstream location for injecting the liquid oil is preferably within the second half of the total length of the extruder barrel. On the surface, this configuration allows for high shear forces to be obtained in the first half of the extruder, promoting fiber formation, and enabling oil to be added downstream without hindering fiber formation.

[0072] In Trotter's process, an improved product is obtained compared to the prior art. However, in Trotter, a large amount of oil is not incorporated into the core of the protein fibers. When the oil is not incorporated into the fibers, the resulting product is perceived by consumers as being greasy, and the release of fat during chewing is not controlled. This unsatisfactory result is because people are accustomed to eating animal meat in which a large amount of fat is incorporated into the muscle fibers. The protein fibers of animal meat contain up to approximately 50% of its weight in fat, although this varies depending on the source of the meat. In the Trotter process, fat is incorporated into the fibers only in an amount of approximately 10% of the weight of the vegetable protein fibers. This problem with the Trotter process, which is disclosed in Trotter in terms of soy and wheat, is due to both the type of protein used as a raw material for extrusion and the method by which oil is added to the protein fibers.

[0073] In another patent application addressing the fat content of meat analogs, International Publication No. WO2020 / 208104 was filed with the PTO in 2020. In the application entitled "Meat analogs and meat analog extrusion devices and methods", Pibarot acknowledges the problem of mimicking the fat content of animal meat where adipose tissue is contained both inside and outside the protein matrix. Pibarot suggests that this complex structure may manipulate the appearance of the meat as well as its texture and juiciness.

[0074] To solve this problem, Pibarot discloses injecting fat into the interior of the extrusion mixture while the extrusion mixture is being cooled within the die. In Pibarot, during extrusion, gaps are created between the protein fibers of the extrusion mixture. As the heated and sheared product is conveyed through the cooling die, fat is injected between these gaps, resulting in the deposition of fat between the protein fibers. Pibarot claims that this process yields a marbled appearance similar to lean meat and improves the texture and flavor of the product. Pibarot discloses using this process with soybeans, peas, and other conventional plant-based protein sources. However, Pibarot does not teach a method of incorporating fat into the molecular structure of the protein fibers.

[0075] In summary, the Shemer process could only use a limited number of components and lacked temperature and cooling control, resulting in poor-quality products. Bouvier solved Shemer's cooling problem but blended the extruded material of the raw materials with a large amount of starch to achieve the desired fat content, which led to undesirable taste and nutritional value. Giezen solved the starch problem of the Bouvier process by adding fat after extrusion, but this required additional steps and resulted in products that were greasy and had a poor mouthfeel. With Trotter, both Giezen and Bovier were improved by introducing oil in the later stages of extrusion, but Trotte still has the problem of low oil uptake into the protein fibers of the meat analog. Pibalot discloses injecting fat into the meat analog while cooling, which introduces fat between the protein fibers, but a final product with fat incorporated into the protein fibers is not produced.

[0076] Hemp grains are highly valuable as both a protein source and an oil source. While there are various methods for manufacturing food from hemp grains, there is a clear need for a cleaner, lower-cost, more effective and efficient method of extracting protein and oil from non-oxidized hemp grains with a clean and pleasant taste, in order to produce hemp protein with excellent purity, gelling function, nutritional value, digestibility, and flavor, as well as an oxidatively stable oil with a clean flavor and bright color suitable for cooking and cosmetics. Furthermore, there is a continuing need for processes and raw materials that can be used to create various meat analogs having the appearance, taste, texture, juiciness and chewiness of various animal meats and dairy products. More specifically, there is a need for meat analogs in which an optimal amount of oil or saturated fat is included in the final product, having the appearance, texture and taste of meat, and in which the oil or saturated fat is incorporated into the protein fibers at levels close to those of oil or saturated fat from animal meat or dairy sources. SUMMARY OF THE INVENTION

[0077] This specification discloses instant meat or food analogs based on natural edestin protein isolate (NEPI). According to the process of the present disclosure, NEPI, when combined with water and oil, can form instant texturized meat analogs. In the present disclosure, a process is described that includes using a NEPI concentrate or combining NEPI powder with water to form a hydrated protein suspension, and subsequently adding oil to form a protein-fat hydrogel. When using a NEPI concentrate, after adding any necessary further dilution with water to the NEPI concentrate to obtain the desired moisture level of the formulation, it is important to achieve a hydration temperature of 60°C to 70°C to fully hydrate and open the protein structure. It is important to fully hydrate the NEPI protein and open it at the specified temperature above before adding oil to form the protein-fat hydrogel. If it is necessary to heat the NEPI water suspension before adding oil, it should be noted that it should be gently heated, either directly or preferably with gentle stirring in a water bath at 60°C to 70°C until the temperature of the hydrated NEPI reaches the temperature of the water bath. When using dry powder NEPI to prepare a hydrated protein suspension, it is more preferably to use hot water, and the temperature of the water is preferably 60°C to 80°C. After the hydrated NEPI protein has opened by achieving a minimum temperature of 60°C, the addition of oil can be carried out. After adding and incorporating oil into either the heated NEPI concentrate or the NEPI protein hydrated with hot water, the formed protein-fat hydrogel can then be cooled to refrigeration temperature for at least 48 hours for microbiological stability. If the protein-fat hydrogel is cooled for microbiological stability purposes, it is necessary to gently reheat the protein hydrogel before microwave curing of the protein-fat hydrogel, preferably including gentle intermittent stirring in a water bath at 60°C to 70°C to equalize the heating rate until the temperature of the protein-fat hydrogel reaches the temperature of the water bath.Most preferably, a protein-fat hydrogel is prepared. After reaching the minimum temperature of 60°C as described above, immediately before the protein-fat hydrogel is cooled below 60°C, it is heated using microwaves, and further the protein-fat hydrogel is heated to harden into a solid hydrogel. Then, the protein-fat hydrogel is hardened in a microwave oven, thereby forming a hydrogel or a meat analogue. Water, NEPI protein, and the hydrogel may be blended and mixed in a microwave-heatable container, or separately blended and poured into a microwave-heatable container. The container is preferably made of a microwave-insulating material such as paper, glass, plastic, or ceramic that does not absorb microwaves but allows microwaves to pass through to the materials inside the container. In some embodiments, an irregular surface can help the protein-fat hydrogel and the protein-fat hydrogel adhere to the sidewalls of the container.

[0078] The protein hydrogel is preferably composed of a material having an irregular or rough surface such as paper or coarse plastic and has dimensions suitable for forming a meat analogue from the protein-fat hydrogel when heated by microwaves and can be mixed in a microwave-heatable container. The materials required for manufacturing a meat analogue at home from NEPI powder can be provided as a convenient kit for manufacturing instant meat analogues.

[0079] The present disclosure solves the prior art problems related to the isolation of hemp protein, the preparation of raw material inputs, and the processing of raw material inputs for manufacturing excellent plant-based meat analogues and dairy analogues. The compositions and processes of the present disclosure include the isolation of hemp grain protein, pasteurization, liquid solutions, gel formation, texturing, and processes for manufacturing meat analogues and dairy analogues. By the process of the present disclosure, a structural protein food or a meat analogue having excellent properties is obtained when compared with existing products or similar products manufactured using known techniques.

[0080] The preparation of meat analogs or dairy analogs by the process of the present disclosure can be divided into three broad steps. The first step involves the extraction or isolation of protein from hemp grains. The second step involves combining the isolated protein with water and oil to form a raw material for thermal gelation or extrusion. The third step involves thermally gelating or extruding the raw material to firm up or texture the meat analog. Thereafter, the final meat analog product can be cooked to mimic meats or dairy products such as chicken, fish, and cheese.

[0081] Regarding the isolation of hemp protein, which is the first step, the process of the present disclosure incorporates a known grain processing method disclosed in U.S. Patent No. 7,678,403 to Mitchell (the "Mitchell" or the "‘403 patent") with some modifications. The ‘403 patent is hereby incorporated by reference in its entirety. The Mitchell process discloses performing aqueous wet milling at a low temperature and sieving the resulting product. In the present disclosure, the aqueous wet milling can preferably be performed while maintaining the temperature of the slurry preferably between 33°F and 38°F. At higher temperatures, particularly 42°F or above, there is concern about microbial growth.

[0082] In some embodiments, the grinding can be performed using whole - grain hemp kernels or dehulled hemp kernels (also called hulled hemp kernels). Depending on whether whole - grain hemp kernels are used or dehulled hemp kernels are used, the color of the final meat - like or dairy - like product may be different. Using whole - grain hemp kernels results in a darker, more beef - like color, while using dehulled hemp kernels results in a lighter, more chicken - or fish - like color. The use of some whole - grain hemp kernels and some dehulled hemp kernels, in one embodiment, the whole - grain hemp kernels are used at a concentration of about 20 - 30% by weight relative to the amount of dehulled hemp kernels, resulting in a color similar to beef in the final product. In one embodiment, the hull previously removed by the dehulling of hemp kernels can be re - introduced into the dehulled hemp kernels to add color, and in one embodiment, to achieve a beef - like color, the hull can be added to the dehulled hemp kernels in an amount of approximately 10 - 15% by weight relative to the dehulled hemp kernels.

[0083] After aqueous wet - grinding, Mitchell teaches in the '403 patent to screen at a different mesh size than the present disclosure. In the present disclosure, it is preferred to screen at a mesh size of 170 - 200. Mitchell considered screening rice grains for milk production in the '403 patent and disclosed a mesh size of 150 or less, which is appropriate for certain grains but not for the removal of chloroplasts from hemp kernels. The present disclosure surprisingly discovers that a mesh size of 170 - 200, preferably or approximately 160 - 200, while preventing the filter - passage of chloroplasts or chlorophyll - containing particles, allows sufficient protein particles to pass through the filter without significantly reducing the protein or nutrient yield.

[0084] When hemp grains are processed according to the modified Mitchell process, an insoluble protein-containing precipitate by-product is produced. Mitchell discovered that this protein-containing material has unique and valuable properties and is particularly well-suited for the manufacture of meat analogs and dairy analogs. This hemp grain protein-containing material has not been previously disclosed. Further research determined that Mitchell that this material is primarily composed of edestin and, importantly, is substantially free of albumin, another major protein component of hemp grains. Due to the processing parameters of the Mitchell process, edestin appears to be substantially maintained in its natural state. Because of the high concentration of substantially natural edestin, this material is hereinafter referred to as natural edestin protein isolate (NEPI). NEPI is composed of approximately 80% protein and also contains oil, fiber, carbohydrates, and ash.

[0085] After milling and screening, NEPI can be separated from the aqueous oil albumin emulsion (AOAE) by centrifugation and decantation. The aqueous oil albumin emulsion may optionally be further processed to produce hemp oil and albumin. NEPI extracted according to the present disclosure can be used in a variety of different plant-based foods that replicate meat or dairy products. NEPI may optionally be combined with oil to form protein hydrogels and protein-fat hydrogels and may be processed to produce evaporated or spray-dried products. The hydrogels can be used to manufacture plant-based meat analogs.

[0086] The present disclosure is based on methods and materials for creating plant-based products that more closely replicate meat products, including the texture, juiciness, fibrousness, and texture uniformity of animal meat. A process for manufacturing meat analogs is described herein, including selecting proteins based on protein unfolding or denaturation, properties, and fat retention capabilities. Further, the process described herein includes a method of preparing an extrusion mixture or feed, which may be a protein-fat hydrogel prior to extrusion that incorporates water and fat into the selected protein in such a way that the water and fat form a liquid matrix, which may also be referred to as a protein-fat hydrogel containing the protein. In some embodiments, the liquid matrix may have additional components, while the protein-fat hydrogel may contain nothing other than protein, fat, and water. Still further, the process described herein includes a method of extruding or otherwise heating the liquid matrix, which may be referred to herein as an extrusion feed or extrusion mixture, and the liquid matrix may be a fat-protein hydrogel. The process of extruding the liquid matrix includes supplying the liquid matrix into a pump at a first end of an extrusion chamber. The liquid matrix is supplied to an extruder and the extruder is set to parameters adapted to the liquid matrix.

[0087] The present disclosure relates to compositions containing edestin or edestin-like proteins, and methods for isolating edestin from hemp grains. As disclosed herein, edestin can be isolated from hemp grains or other grains and seeds containing edestin or edestin-like proteins. In one embodiment, the hemp grains are wet milled during aqueous protein extraction to obtain an edestin-containing fraction and an aqueous oil albumin emulsion.

[0088] In one aspect, the present disclosure can utilize an aqueous wet grinding method to separate the fats stored within hemp grains without destroying the chloroplasts and releasing chlorophyll into the oil. When the seeds are ground by this process, the resulting ground product is sieved through meshes of various sizes. By sieving at approximately 170 mesh, or in some embodiments 160 - 200 mesh, or in some embodiments 200 - 270 mesh, the husk, chloroplasts, and fibers are removed. More preferably, a mesh size of 160 - 200 can be used. In a preferred embodiment, a mesh size of 170 can be used. A mesh size of 150 has openings that are too large, and undesirable materials, including fibers and chlorophyll-containing materials, may enter the filtrate. Surprisingly, the chlorophyll-containing particles remain larger than the pore openings of a 170-mesh, and most of the protein-containing particles pass through a mesh of this size. According to the process of the present invention, by sieving through meshes of different sizes, chloroplasts, protoplasts, or other chlorophyll-containing particles are separated from the hemp oil and protein-containing fractions, thereby obtaining a pale yellow final oil product.

[0089] In the process of the present disclosure, after sieving, an insoluble fraction containing NEPI and an aqueous oil albumin emulsion may be present in the filtrate. The AOAE can be decanted after centrifugation. The insoluble fraction and the pellet-containing portion can be washed to remove residual oil. In some embodiments, washing with cold water may be performed twice.

[0090] In some embodiments, the AOAE can be cooled to about 33°F to 38°F until the albumin begins to separate from the oil in the emulsion and precipitate, 35°F is preferred, and in some embodiments, this can be assisted by centrifugation. According to the process of the present disclosure, albumin binds strongly to hemp seed oil, thereby improving the separation of the insoluble edestin fraction, i.e., the oil and albumin from NEPI. By this process, albumin can be separated from hemp seed oil. Gel electrophoresis shows that substantially all of the albumin can be removed from NEPI by this process, and mainly, edestin remains in NEPI. The AOAE can be removed from NEPI by centrifugation and decantation, leaving NEPI as a solid material, which can be washed to remove residual material.

[0091] In one embodiment, the NEPI can then be heated to a temperature of about 145°F for about 30 minutes to pasteurize the product. In some jurisdictions, 145°F may be the legal lower limit for pasteurization. Here, to prevent granulation occurring at the high temperatures observed in the present disclosure, the temperature should be maintained at about 145°F, or 145°F to 155°F. In NEPI, granulation can occur at a temperature much lower than the denaturation of edestin, for example, about 158°F. Therefore, it is very important to pasteurize at a temperature lower than the temperature commonly used by those skilled in the art for pasteurization. Those skilled in the art conventionally pasteurize protein isolates at temperatures that would cause significant granulation in the present disclosure to process the product quickly. After pasteurization is complete, the NEPI can be spray dried or stored as a concentrate for use in meat analogs and dairy analogs. To prevent granulation or aggregation of the protein, spray drying should also be performed at a lower temperature, preferably about 145°F to 155°F.

[0092] In some embodiments, particularly for commercial use, NEPI enters a tank heated to 145°F as it exits the production line, incubates the product at this temperature for 30 minutes, and then sends it to a cooling tank where it is cooled to approximately 35°F. After cooling, NEPI is shipped as needed for spray drying, freezing, freeze drying, or vacuum microwave drying and then used in the production of meat analogs and dairy analogs, or structured protein foods.

[0093] In the case of the production of meat analogs, the pasteurized product can be prepared by first hydrating NEPI if it is dry, or by maintaining an appropriate degree of hydration in other ways. In one embodiment, the amount of water can be about 3 parts of water per 1 part of NEPI. Before addition to NEPI, the water may preferably be preheated to about 135°F to form a protein hydrocolloid before hardening. Since the hydrocolloid structure of the protein may be disrupted, salt should not be added during this process. Salt can be added immediately before or after hardening, but not before hardening. In some embodiments, the hydration and loosening of the protein can be carried out at 100°F - 135°F, or in some embodiments, at 100°F - 155°F. Or in other embodiments, the formation of the protein hydrocolloid may be carried out at a lower temperature, but that temperature must be higher than the low temperature at which the protein cannot be loosened. Preferably, the temperature during the hydration and protein preparation steps should remain as close as possible to the minimum temperature considered for pasteurization, 145°F, without reaching the temperature at which granulation of the product occurs.

[0094] When the protein is hydrated, in one embodiment, oil can be added and mixed with the protein hydrogel to form a protein-fat hydrogel. Oil should not be added until the NEPI is fully hydrated so that the protein hydrogel has a smooth appearance. If oil is added before hydration and preparation of the protein, granulation may occur. Further, according to the process of the present disclosure, it is necessary to add oil before the hardening of the material, where hardening means that protein bonds are formed to create a more solidified gel product, and when protein aggregation occurs, it generally means that the protein aggregates at a high temperature where protein denaturation occurs. In the case of the present disclosure, there is no free oil during the hardening process, and all oil is incorporated into the emulsion or protein structure before curing the product with an extruder or other thermosetting means. In conventional extrusion, there is free oil with the material that partially or fully cures within the extruder. Therefore, in the present disclosure, it is important to add water to fully hydrate and loosen the NEPI before adding oil so that there is no free oil during extrusion or curing. This protein-fat hydrogel can then be heated or extruded to form a meat analog. In conventional extrusion, for example, using soy or pea protein, oil is added to the protein material after hardening has started at a high temperature within the extruder for lubrication rather than incorporation of the oil.

[0095] Once the protein hydrogel is fully hydrated, oil can be added to form a protein-fat hydrogel. The oil can preferably be preheated, and the temperature of the oil can preferably be about 130°F - 135°F. In other embodiments, the oil can be preheated to 100°F - 135°F, or 100°F - 155°F, while in other embodiments, the oil can be added at a lower temperature, but the oil should not be added at a low temperature that would disrupt the structure of the protein hydrogel and prevent the oil from being incorporated into the protein hydrogel to form a protein-fat hydrogel. The material can also be set in a retort system, but in a retort, it may not be possible to produce a fibrous product like extrusion.

[0096] The texture of the retort-processed NEPI meat analog was surprisingly good, having a texture including hardness and chewiness far superior to commercially available hemp protein concentrates and isolates under the same conditions. The process of the present disclosure unexpectedly resulted in the thermal gelation and extrusion of a high-quality fibrillated meat analog produced using only hemp as the protein source. Due to the properties of conventionally used meat analog and dairy analog proteins, including soy and hemp, conventional meat analogs and dairy analogs cannot reproduce the texture of processed meat similar to animal meat products, such as sliced chicken breast. The unexpectedly advantageous properties and results resulting from the use of NEPI described in the present disclosure and the process using it have created a much superior structured meat analog using only hemp protein as the protein source when compared to other commercially available products. At this point, hemp protein is only known to be used in combination with soy or other plant proteins to produce meat analogs.

[0097] In one aspect, the present specification features a meat analog extrusion input, or a liquid matrix, in which the protein-to-fat ratio can range from about 4:1 to 0.5:1.

[0098] In one aspect, the present specification features a process in which water is added to the protein isolate at the ratios disclosed hereinbelow, or water is maintained in the protein isolate at a specific ratio, and after adding or maintaining water in the protein isolate, fat is added to the mixture of protein and water at a nearly constant ratio of water-to-fat and protein-to-fat.

[0099] In one aspect, the present specification features a product with a target water content of 35 wt% to 75 wt%.

[0100] In any of the methods or compositions described herein, the isolated plant protein in the liquid matrix can include seed oil proteins such as edestin, albumin, globulin, or mixtures thereof.

[0101] In any of the methods or compositions described herein, the isolated protein can first be isolated from all other plant proteins within the plant.

[0102] In any of the methods or compositions described herein, the isolated protein used can be isolated in its native or non-modified state. Native can mean completely natural, substantially natural, partially natural, or identified as substantially natural by conventional methods of detecting protein structure or as natural as understood by one of ordinary skill in the art.

[0103] In any of the methods or compositions described herein, a protein isolated from a seed protein preferably has a higher cysteine content than that normally found in soy or casein.

[0104] In any of the methods or compositions described herein, the liquid matrix can contain a flavoring agent, starch, fiber, or other carbohydrate source.

[0105] In some embodiments, the meat analogs and milk analog products provided herein may not contain animal products, wheat gluten, soy protein, or pea protein.

[0106] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials similar or equivalent to those described herein can be used in the practice of the invention, but the preferred methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, this specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. References to percentages are by weight throughout.

[0107] Details of one or more embodiments of the present invention are described in the accompanying drawings and the following description. Other features, objects, and advantages of the present invention will become apparent from the description, drawings, and examples, as well as from the claims.

Brief Description of the Drawings

[0108]

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[0109] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. All references to percentages are by weight unless otherwise specified. Details of one or more embodiments of the invention are described in the accompanying drawings and in the following description. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0110] Generally, the present disclosure provides methods and materials for producing plant-based meat analogs or dairy analogs, herein also referred to as structured protein foods, from hemp hull proteins. In either the methods or compositions described herein, and in some embodiments, the extracted protein-containing product can be separated from other hemp grain proteins. In either the methods or compositions described herein, edestin can be substantially isolated from some or all of the other proteins in the hemp grain. In either the methods or compositions described herein, the protein isolated from the grain proteins preferably has a higher cysteine content than that typically found in soy or casein.

[0111] The plant proteins used in accordance with the present disclosure can be isolated plant proteins. For the purposes of the present disclosure, a "native" protein is a protein that can have the same tertiary and quaternary structure as a living, active cell. In some embodiments, a "native" protein can be substantially native. In any of the methods or compositions described herein, the isolated protein can be isolated in a nearly native state, a substantially native state, or a non-denatured state. In any of the methods or compositions described herein, the isolated protein used can be isolated in a native or non-denatured state. Native can mean completely native, substantially native, partially native, or identified as substantially native by conventional methods of detecting protein structure, or native as understood by one of ordinary skill in the art. Changes and disruptions to the subunit structure and tertiary structure can occur due to changes in temperature (usually above 41 °C), or contact with an acidic or alkaline aqueous solution, an oxidizing or reducing agent, or an organic solvent. Disruption of the quaternary structure can render the protein biologically inactive, or potentially so, in a living cell. However, the tertiary structure of the released subunits, which have a specific shape created by hydrogen bonds, van der Waals forces, and disulfide bonds, can be functionally active and exhibit functions similar to those in a living cell. One example of this is the lock-and-key function of an enzyme due to the tertiary shape of the protein.

[0112] Accordingly, for the purposes of the present disclosure, these can be considered "native" proteins if the quaternary or tertiary structure is substantially maintained in the same state as in a living cell after extraction. The present disclosure has found that certain oilseed globular proteins, which can be considered native in the sense that their tertiary structure has not been denatured by changes in temperature (usually above 41 °C), an acidic or alkaline aqueous solution, an oxidizing or reducing agent, or an organic solvent, have unique and excellent functionality.

[0113] Conventional plant protein extraction processes are known to disrupt the quaternary and tertiary structures of proteins. In some cases, this disruption can cause the loss or reduction of the function of the quaternary or tertiary structure. The tertiary structure can be denatured by the disruption of functional bonds and forces, including hydrogen bonds, van der Waals forces, or disulfide bonds, which all act together to form a specific tertiary protein structure. Changes in the protein environment and the mode of denaturation of the tertiary structure can cause changes in the tertiary structure or shape of the protein, as well as its bonds, forces, and linkages.

[0114] As used herein, the term "isolated plant protein" can include plant proteins such as edestin, glutelin, albumin, legumin, vicilin, convicilin, glycinin, and protein isolates from any seed or bean, including but not limited to soybeans, peas, lentils, or combinations thereof, or plant protein fractions (e.g., 7S fraction), that are separated from other components of the source material (e.g., proteins of other animals, plants, fungi, algae, or bacteria), such that the protein or protein fraction contains, on a dry weight basis, less than at least 2% (e.g., less than at least 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%) of the other components of the source material. For example, an isolated native globular protein having a high cysteine content can be used alone, or in combination with one or more other proteins (e.g., albumin), or from any other protein source such as soybeans, peas, whey, etc.

[0115] In any of the methods or compositions described herein, the fat can be a non-animal fat, an animal fat, or a mixture of non-animal and animal fats. The fat can be algal oil, fungal oil, corn oil, olive oil, soybean oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, sunflower oil, flaxseed oil, palm oil, palm kernel oil, coconut oil, argan oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, lycii oil, blackcurrant oil, sea buckthorn oil, macadamia oil, babassu oil, conjugated linoleic oil, arachidonic acid-enriched oil, docosahexaenoic acid (DHA)-concentrated oil, eicosapentaenoic acid (EPA)-concentrated oil, palm stearic acid, sea buckthorn berry oil, macadamia oil, babassu oil, or rice bran oil; or it can be margarine or other hydrogenated fat. In some embodiments, for example, the fat is algal oil. The fat can contain flavoring agents and / or isolated plant proteins (e.g., conglycinin protein). The composition of the fat or oil in the liquid matrix can be made to preferentially match the saturated and unsaturated composition of the target source material of the analog.

[0116] Thus, in some embodiments, the isolated protein can be a protein such as substantially natural edestin isolated from hemp grains or other grains that may have edestin or edestin-like proteins. In some embodiments, the proteins can be separated based on their molecular weight, for example, by size exclusion chromatography, ultrafiltration through a membrane, or density centrifugation. In some embodiments, the proteins can be separated based on their surface charge, for example, by isoelectric precipitation, anion exchange chromatography, or cation exchange chromatography. Proteins can also be separated by solvent extraction based on their solubility, including, for example, ammonium sulfate precipitation, isoelectric precipitation, surfactants, detergents, or water extraction. Proteins can also be separated by affinity to another molecule, for example, using hydrophobic interaction chromatography, reactive dyes, or hydroxyapatite. Affinity chromatography can also include the use of antibodies with specific binding affinity for heme-containing proteins, nickel nitrilotriacetic acid (NTA) for His-tagged recombinant proteins, lectins that bind to sugar moieties on glycoproteins, or other molecules that specifically bind to the protein. In some embodiments, the plant-based meat described herein is composed substantially or entirely of components derived from non-animal sources, such as plant-based, fungus-based, or microbe-based sources. In some embodiments, the plant-based meat or plant-based dairy product can include one or more animal-based products. For example, meat replicas can be made from a combination of plant-based and animal-based sources.

[0117] References: The following documents are hereby incorporated by reference in their entirety: U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis, U.S. Patent No. 7,678,403 to Mitchell and Mitchell.

[0118] Definitions: Hemp seeds (HS) are generally defined herein as seeds with germination ability that are normally used for further propagation and sowing. HS may be of food grade or not, based on the cleaning implementation method and the agricultural storage implementation method of the seeds.

[0119] Whole hemp grains (WHG) are generally defined herein as hemp grains that include both hemp grains with germination ability and pasteurized hemp grains.

[0120] Viable hemp grains (VHG) are generally defined herein as viable hemp seeds from which all dust and foreign matter have been further removed, are suitable for food grade, and in which the core and shell are completely intact.

[0121] Pasteurized hemp grains (PHG) are generally defined herein as hemp grains that have been treated by heat or irradiation to destroy the germination ability of the seeds.

[0122] Defatted hemp grain cake (DHGC) is generally defined herein as the dry solid residue resulting from the non-aqueous removal of oil from hemp grains.

[0123] Hemp grain oil (HGO) is generally defined herein as the raw green oil obtained from the non-aqueous extraction of hemp grains.

[0124] Hemp grain oil sludge (HGOS) is generally defined herein as the crude oil sludge slurry obtained from the non-aqueous extraction of oil from hemp grains.

[0125] Hulled hemp grains (HHG) are generally defined herein as the core of hemp or hemp nuts; equivalent to hemp grains with the outer shell removed.

[0126] Defatted hulled hemp grain cake (DHHGC) is generally defined herein as the dry solid residue resulting from the non-aqueous removal of oil from hulled hemp grains.

[0127] Dehulled hemp grain oil (HHGO) is generally defined herein as a yellow oil resulting from the non-aqueous extraction of dehulled hemp grains.

[0128] Hemp protein isolate (HPI) is generally defined herein as an isolate of albumin, edestin, or aggregates thereof.

[0129] Aqueous oil albumin emulsion (AOAE) is generally defined herein as an aqueous emulsion of oil and soluble albumin protein.

[0130] Native edestin protein isolate (NEPI) is generally defined herein as the product of the protein isolation process disclosed herein and can refer to NEPI in liquid, slurry, and powder forms, as would be understood by one of ordinary skill in the art in the appropriate context of its use.

[0131] All products described in the flowchart can exist in various physical forms, including liquids, gels, or solids, as would be understood by one of ordinary skill in the art in the appropriate context of their use.

[0132] The present disclosure may relate to compositions containing native edestin protein isolate (NEPI) containing edestin or edestin-like proteins, and methods for extracting and using NEPI to produce meat analogs and dairy analogs. The present disclosure solves the prior art problems related to the isolation of hemp protein, the preparation of raw material inputs, and the processing of raw material inputs for producing excellent plant-based meat analogs and dairy analogs. The compositions and processes of the present disclosure include the isolation of hemp grain protein, pasteurization, sol formation, gel formation, texturization, and processes for producing meat analogs and dairy analogs. The processes of the present disclosure result in meat analogs or dairy analogs having excellent properties when compared to existing or similar products produced using known techniques.

[0133] In addition to the protein isolate, the present disclosure is based on methods and materials for making plant-based products that more closely reproduce meat products, including the texture, juiciness, fibrousness, and texture uniformity of animal meat. Processes for manufacturing meat analogs are described herein that may include selecting a protein based on protein unfolding or denaturation, properties, and fat holding capacity. Further, the processes described herein include a method of preparing an extrusion mixture prior to extrusion, which incorporates water and fat with a selected protein in such a manner that the water and fat together with the protein form a liquid matrix (also referred to herein as a liquid-fat hydrogel, hydrogel, extruder or extrusion feed, and feed material). Still further, the processes described herein include methods of extruding or heating the liquid matrix. The process of extruding the liquid matrix includes feeding the liquid matrix into a pump at a first end of an extrusion chamber. The liquid matrix is fed into the extrusion chamber of an extruder and the extruder is set to parameters appropriate for the liquid matrix.

[0134] As disclosed herein, NEPI can be extracted from hemp grains or other grains, nuts or seeds containing edestin or edestin-like proteins. However, at present, hemp grains are considered to be the sole source of edestin. In one embodiment, the hemp grains are wet-milled and subjected to water extraction, thereby producing an insoluble edestin-containing extract, herein referred to as NEPI, and an aqueous oil albumin emulsion.

[0135] In the process according to the present disclosure, a pasteurized functional hemp grain protein concentrate can be produced, which concentrate can be the concentrated liquid obtained from the production line or from centrifugation and decantation, or NEPI powder, which, in some embodiments, has a low or no amount of trypsin inhibitor and has high nutritional value and functionality. This process may not use isoelectric point extraction, alkaline solubilization methods or CO2 solubilization methods. The texturable protein NEPI concentrate or NEPI powder is thought to be produced by utilizing the natural pH and oil content of hemp grains in combination with water and by oil extraction and albumin separation. Due to the emulsion-forming ability of soluble albumin, an emulsion can be formed that can be easily separated from insoluble edestin by centrifugation. Freeze-drying, pH readjustment, and separation by ultrafiltration are not necessary. Furthermore, fiber and chlorophyll can be removed during the NEPI process. Maintaining a low temperature, preferably between 33°F and 38°F, promotes the insolubility of globulin and also promotes the coagulation of albumin.

[0136] One aspect of the present disclosure relates to the isolation of edestin and edestin-like proteins from plant material containing hemp grains. Edestin is found in hemp plants, particularly in hemp grains. While hemp grains are thought to be the most common or the sole source of edestin, other plants may also contain edestin.

[0137] A protein-containing composition can be produced using an edestin extract composition prepared according to the method of the present disclosure, or a native edestin protein isolate (NEPI). NEPI can preferably be composed of about 80% protein on a dry basis. In some embodiments, NEPI can contain at least 65% protein on a dry basis, and in some embodiments, at least 90% protein on a dry basis. Thus, NEPI can be defined as an edestin-containing composition manufactured according to the method described in the present disclosure that results in a product having the functional characteristics described in the present disclosure. The aqueous oil albumin emulsion (AOAE) described in the present disclosure can be further processed to produce other plant-based products containing hemp oil or grain oil and albumin.

[0138] The present disclosure can be practiced using suitable cereal grains, seeds or plant materials containing edestin or edestin-like proteins, such edestin-like proteins being homologous or having similar structures and functions.

[0139] The cereal grains used in the present disclosure can be substantially whole-fat cereal grains, i.e., cereal grains that have not been defatted or pressed prior to milling. In some embodiments, the cereal grains may be partially defatted. Partially defatted cereal grains include any plant material from which at least a portion of the fat has been removed.

[0140] As is known to those skilled in the art, substantially all-fat hemp grains can have a fat (or oil) content of 10% by weight or more. In the present disclosure, the terms fat and oil can be used interchangeably. Preferably, the fat content of grains that are substantially all-fat can be at least about 10%, 15%, 20%, 30%, 40%, or even 50% by weight. The fat content of hemp grains is usually at least 30%. The fat content of partially defatted plant materials can be fat in excess of about 5%, 10%, or 15% by weight. The edible portion of hemp grains after removing the husk contains on average 46.7% oil and 35.9% protein.

[0141] As shown in FIG. 1, hemp grains 102 can be selected for use in the structured protein food process 100. Whole hemp grains 101 and dehulled hemp grains 105 can be used. Pasteurized whole hemp grains 103, produced by the pasteurization process 107, can also be used. The hemp grains 102 used in accordance with the present disclosure can be prepared for treatment by suitable means including, but not limited to, drying, conditioning to achieve an equilibrium moisture level, dehulling, cracking, and cleaning by countercurrent air aspiration, sorting methods, pasteurization that does not damage viable seeds, or other methods known in the art. The hemp grains 102 can be selected from any type of hemp plant, but in the present disclosure, Cannabis Sativa containing 0.3% or less THC is preferably used. The hemp grains 102 can be whole hemp grains or dehulled (peeled) hemp grains 102, and the hemp grains 102 can be dehulled prior to processing in the structured protein food process 100, thereby producing dehulled hemp grains 150, as shown in FIG. 4.

[0142] Referring now to FIG. 1, hemp grains 102 in the structured protein food process 100 are subjected to a native edestin protein isolation process 200 (shown in FIG. 2) to extract a native edestin protein isolate (NEPI) 250. A structured protein food 120, which can be a meat analog or a dairy analog, can be produced using the native edestin protein isolate (NEPI) slurry 252 or powder 254, or the NEPI 250. In conventional methods of extracting hemp protein or edestin from hemp grains, or producing hemp protein isolates, aggregation of edestin and albumin or protein denaturation may occur, and satisfactory structured protein foods or meat analogs may not be producible. However, the NEPI 250, as described by Zahari, can produce excellent novel meat analogs when used as the sole protein source in meat analogs without combining with soy or other types of plant-based protein isolates (Zahari et al., 2020).

[0143] As shown in FIG. 1, in some embodiments, the NEPI 250 can be pasteurized (104) and combined with water 106 to form a protein hydrogel 108. The NEPI 250 can be combined with preheated water 106 to form a protein hydrogel 108 (as shown in FIG. 6). The NEPI 250 needs to be present at least 20 wt / wt% with respect to water to up to 80 wt / wt% or more with respect to water to fully hydrate the protein. The hydration time depends on the conditions. To facilitate hydration, it is preferred to mix at high shear.

[0144] Next, oil can be added to the protein hydrogel 110, followed by high-shear mixing 112. In some embodiments, after the high-shear mixing 112, the mixture can be optionally incubated 113 without mixing. The addition 110 of oil and mixing 112 produce a protein-fat hydrogel 114.

[0145] The protein-fat hydrogel 114 is used as an input for means 116 of heating the protein-fat hydrogel to harden the product. Coagulation may involve heating by means including microwave, steam tunnel, oven, retort, and extrusion (as shown in FIGS. 7 and 8). The means for heating and hardening may include other means of heating protein- or starch-based foods to form hardened products, as would be known to those skilled in the art. By hardening the protein-fat hydrogel 114, a structured protein food 120 is produced. The structured protein food 120 can be a meat analog or a dairy analog.

[0146] As shown in FIG. 2, to produce NEPI250, hemp grains 102 can be added to cold water 202 to form a hemp grain mixture 204. The extraction temperature during grinding and throughout the native edestin protein isolation process 200 can be more preferably 35°F, or 33°F - 38°F, or less than about 120°F, and is added to the hemp grains 102 to form the hemp grain mixture 204. The hemp grains can be extracted with an aqueous solution, preferably water. As used herein, the term "aqueous solution" includes water substantially free of solutes (e.g., tap water, distilled water, or deionized water) and water containing solutes. According to the present disclosure, the aqueous solution may not contain additives such as salts, buffers, acids, bases, and demulsifiers. In some embodiments, the aqueous solution can have an ionic strength lower than the ionic strength that changes the structure of the protein. More or less water can be used.

[0147] In the process of the present invention, there is no need to adjust the pH to isolate NEPI. Preferably, throughout the structured protein food process 100, the pH is maintained at approximately neutral, between 6.5 and 7. In one embodiment, the pH of the solution does not change substantially during the grinding of the grains.

[0148] The hemp grain mixture 204 may be wet-milled 206 substantially as described in U.S. Patent No. 7,678,403 to Mitchell. In one embodiment, the milling 206 of the hemp grains can be performed using a Silverson rotor stator type mill. The wet milling 206 can be carried out as part of a water extraction process. Preferably, the aqueous wet milling 206 can be carried out for a suitable period, and more preferably, the wet milling 206 is carried out for a suitable period. As will be understood by those skilled in the art, a longer extraction period may be used. In some embodiments, enzymes can be used to assist in the treatment. For example, liquefaction can be achieved using an α-amylase enzyme having dextrinizing activity to obtain a liquefied slurry. Such enzymes can include amylase, or other carbohydrases known in the art of food processing. In one aspect, the present disclosure utilizes a method of aqueous wet milling to separate the fats stored within the hemp grains 102 without disrupting the chloroplasts and releasing chlorophyll into the oil. After centrifugal decantation 222, calcium chloride can be added to the NEPI 250 to improve the flavor.

[0149] After aqueous wet milling the hemp grains 206, the extract can be separated from at least a portion of the insoluble by-products or fibrous slurry 210 (e.g., the insoluble fiber fraction) using a mesh. In some embodiments, the hemp grain slurry 208 can be screened in two steps. Screening can remove unwanted impurities that impart an unpleasant color or taste to the edestin. The insoluble fibers can be removed by passing through a first screen. Another unwanted product that can be removed by screening without substantially affecting the protein yield is, surprisingly, chlorophyll derived from the chloroplasts in the hemp grains and dehulled hemp, which can cause an unwanted color, taste, and oxidation of the fats in the oil fraction or protein fraction. In some embodiments, the chlorophyll-containing particles can be removed in the step 212 of passing through a second screen. After screening 212, the chloroplasts and fiber sludge can be in the holding liquid together with the raw hemp milk having a fat to protein ratio of about 1:3:1 in the DSB in the filtrate.

[0150] In some embodiments, in the step of sieving through a first sieve, the hemp grain slurry can be sieved through a 30-mesh sieve and the husk can be removed. The by-product of the step of sieving through the first sieve can be the fibrous slurry 210. In the step 212 of sieving through a second sieve, the hemp grain slurry is sieved through about 170 mesh, or in some embodiments 160 - 200 mesh, or in some embodiments 200 - 220 mesh 212 to remove chloroplasts or chlorophyll-containing materials, and the remaining fibers. A mesh size of 150 may generally have openings that are too large, such that undesirable substances, including fibers and chlorophyll-containing particles, may pass through into the filtrate. Surprisingly, the chlorophyll-containing particles remain larger than the pore openings of 170 mesh, and most of the protein-containing particles pass through a mesh of this size. By sieving through different sizes of meshes, chloroplasts, protoplasts, or other chlorophyll-containing particles are separated from the hemp oil and the protein-containing fraction, thereby obtaining a pale yellow final oil product.

[0151] The chloroplasts isolated by the edestin extraction process 100 can be used as dietary supplements in some embodiments. According to the process of the present disclosure, the chlorophyll-containing particles 214 are selectively removed from the hemp grain slurry 208 while allowing the protein-containing particles to pass through into the filtrate. This method is effective for both whole grain hemp with the husk not removed prior to aqueous wet milling and dehulled hemp grains.

[0152] The hemp grain slurry is sieved through a 170 mesh screen to remove the chlorophyll-containing particles 212, and the resulting product is an aqueous oil albumin emulsion (AOAE) and an edestin mixture 220, which may also contain to a greater or lesser extent other components of the hemp grain 102. The AOAE and edestin mixture 220 can be centrifuged and decanted 222 to obtain NEPI 250 and AOAE 230. After being separated from NEPI 250, AOAE 230 can be further processed to produce albumin 550 and hemp oil 560, as shown in FIG. 5.

[0153] In some embodiments, NEPI 250 can be composed of about 76% protein, 2% oil, 4% fiber, 1% carbohydrate, and 17% ash. AOAE 220 can be composed of about 14% protein, 76% oil, 3% fiber, 4% carbohydrate, and 3% ash. In some embodiments, NEPI can preferably be composed of about 80% protein on a dry basis. In some embodiments, NEPI can contain at least 65% protein on a dry basis, and in some embodiments, at least 90% protein on a dry basis. Thus, NEPI can be defined as an edestin-containing composition manufactured according to the methods described in the present disclosure that results in a product having the functional characteristics described in the present disclosure. In some embodiments, NEPI can contain at least about 65%, 75%, 85%, or 90% protein on a dry weight basis.

[0154] Table 2 shows the approximate analytical data of the nutritional compositions of NEPI 250 and commercially available hemp protein products. Table 2 shows that the NEPI 250 product has a high protein content and a high ratio of protein to fat, similar to VICTORY HEMP. Other commercially available products have a much lower protein content and a much lower ratio of protein to fat. This indicates that among the products tested, NEPI 250 and VICTORY HEMP may be far superior to other products.

[0155] Figures 9 to 11 show SDS PAGE gel data of NEPI250 products and commercial products showing protein composition, structure, and integrity (non-reducing conditions are shown in Figure 9 and reducing conditions are shown in Figure 10). Regarding Figure 9, 910 is the edestin dimer and 920 is albumin. Regarding Figure 10, 930 is the acidic subunit of edestin, 940 is the basic subunit of edestin, and 950 is albumin. Figure 11 shows SDS PAGE data of the prior art showing the known molecular weights of edestin and edestin products under similar conditions. The lanes are identified below and apply to Figures 9 and 10. M = Molecular weight standard 1 = DP-276 HempLife powder SD HPI 2 = DP-276 HempLife powder SD HPI 3 = DC-344 HempLife liquid concentrate HPI 4 = GH-350 Good Hemp powder HPI 5 = A-560 Anthony’s powder HPC 6 = LP-643 dehulled HempLife SD powder HPI 7 = VH-794 Victory Hemp powder V70 HPI 8 = N-950 Nutiva powder HPC 9 = N-950 Nutiva powder HPC

[0156] Figure 11A shows SDS PAGE of the prior art from hemp proteins published by Mamone and Wang (Mamone et al., 2019, from Wang and Xiong, 2019). Figure 11B shows SDS PAGE of the prior art from hemp proteins published by Shen (Shen et al., 2020).

[0157] In summary, Figures 9-10 show that the NEPI250 product has a different protein composition from other commercially available products, is generally structurally intact, and VICTORY HEMP is the closest in terms of natural edestin content and non-degraded protein products. Interestingly, as expected, the NEPI250 product contained substantially no albumin. In the present disclosure, a hypothesis is proposed that albumin hinders the ability of hemp protein isolate to form a good structured protein food 120 with excellent texture. This theory is supported by the texture profile analysis data shown in Table 2. When compared with commercially available hemp protein products, the NEPI products have much higher hardness and chewiness. The excellent natural structural characteristics of edestin in NEPI250 may also contribute to the unexpectedly excellent texture formation of NEPI250 shown in Table 2. The excellent structural preservation of edestin in its natural state is further shown in Table 3. Figures 12 and 13 show the differential scanning calorimetry data of the products.

[0158] Table 3 shows a differential scanning calorimetry thermogram providing structural information on the edestin contained in NEPI250 and commercially available products. The DSC thermographs of two NEPI products (Figure 12), as well as the DSC thermographs of two commercially available hemp protein powders, VICTORY HEMP and NUTIVA (Figure 13). Based on the DSC results, the NEPI products are superior in terms of structure when compared with commercially available products, indicating that the edestin in NEPI250 is in a more natural state than commercially available products.

[0159] As described above in the background art, the quality of edestin in NEPI250 is excellent compared to hemp protein isolates produced by conventional means. Furthermore, when compared with the process of the present disclosure, the prior art protein extraction methods have significant drawbacks and limitations. For example, salt extraction and dialysis in the HMI process do not remove residual phenol from the final product. Furthermore, HMI is not commercially viable.

[0160] The process of the present disclosure has many advantages compared to the prior art. This process can release phenol and tocopherol from NEPI250 and AOAE230. The process of the present disclosure can make hemp oil 560 more oxidatively stable. In the process of the present disclosure, during aqueous wet grinding, phenol is separated from hemp oil 560, thereby providing stability.

[0161] The process of the present disclosure is different from the conventional methods of extracting protein from hemp grains in that the conventional methods generally involve pressing grains to extract oil and producing hemp grain cakes, and then milling and sieving to produce fine powder. The resulting cakes or fine powder can contain aggregated edestin and albumin along with oil, carbohydrates, phenols, and minerals. In some cases, the seeds can also be dry-milled to directly produce fine powder.

[0162] Mechanical processes that result in high heat or high pressure, such as pressing grains, can lead to the formation of chemical bonds between edestin and albumin. Aggregation of edestin and albumin may occur when whole hemp grains or dehulled hemp grains are pressed.

[0163] High pressure can change the structure of proteins and cause protein aggregation. According to Yang, high-pressure modification of proteins involves changes in the secondary, tertiary, and quaternary structures of proteins from the native state through an intermediate state to a fully denatured state (Yang et al., 2016). High pressure mainly changes the structure of proteins through changes in non-covalent bond - electron interactions, hydrophobic interactions, and hydrogen bonds. High pressure can also form new disulfide bonds, thereby stabilizing the denatured protein or resulting in protein aggregation (Yang et al., 2016).

[0164] Heat is also known to change the structure of proteins. Heat generated by friction during particle grinding can cause changes in the structure of proteins. Heat can cause protein denaturation and the formation of protein aggregates. Aggregation between edestin and albumin can occur during dry grinding where the temperature can reach 100 °C or higher.

[0165] In one embodiment, NEPI is then heated to a temperature of about 145 °F for about 30 minutes, and the product can be pasteurized. In some jurisdictions, 145 °F may be the legal lower limit for pasteurization. In one embodiment, the temperature can be maintained at about 145 °F, or 145 °F - 155 °F, to prevent granulation. It has been observed in the present disclosure that granulation occurs at a temperature of about 158 °F. Granulation may occur in NEPI at a temperature well below the denaturation temperature of edestin, for example, about 158 °F, and the denaturation temperature of edestin has been shown to be about 95 °C. It is important to pasteurize NEPI at a temperature lower than the temperature normally used by those skilled in the art for pasteurizing plant-based proteins for food use. Those skilled in the art conventionally pasteurize protein isolates at a temperature that causes significant granulation in the present disclosure. Pasteurized NEPI 270 is the result of washing and diluting with cold water 232.

[0166] As shown in FIG. 3, after the pasteurization 104 is completed, the NEPI 250 can be spray dried by the NEPI spray drying process 300 or stored at low temperature as a concentrate for use in the manufacture of the structured protein food 120. Immediately after centrifugal decanter separation, the solids content of the NEPI concentrate is in the range of about 35% to 45% and is a thick paste that is difficult to pump. At this point, cold water is added to preferably reduce the solids content of the NEPI 250 concentrate to about 30% and facilitate rapid pumping of the slurry through a heating pipe maintained at a temperature not exceeding 158°F. Dilution results in more turbulent flow, improves the heat distribution for heating to 145°F, and enables pasteurization without forming undesirable protein aggregates and granules in the finished dried edestin product. Prior to spray drying, the NEPI concentrate can be held in a tank at about 145°F or at the temperature of the pasteurization 104 before spray drying. The spray drying 306 can then be carried out at a higher spray drying 306 temperature or at a temperature at which the existing product can reach about 158°F or higher, which can cause protein aggregation and result in functionally inferior NEPI 250. This protein aggregation can be visible at about 100 kDa on a non-reducing SDS-PAGE gel (shown in FIG. 9), and bands other than the bands of the expected edestin or hemp granule protein are visible. Bands present at high molecular weights above the band of about 50 kDa expected for the edestin dimer may represent aggregation caused by excessive heat during spray drying 300 under non-reducing conditions. Thus, in some embodiments, one potential way to measure whether the maximum temperature of the spray drying 300 is below the temperature at which significant protein aggregation occurs may be to identify unexpected high molecular weight bands on a non-reducing SDS-PAGE gel. Microwave drying is another method that can be used in the present disclosure, during which the NEPI 250 is kept at a low temperature such as 130°F to 140°F while moisture is removed under a vacuum pressure.

[0167] Figure 4 shows a process for adding color to the structured protein food 120. The white and dark meat analog processes 400 can produce either a white meat NEPI 412 that can replicate chicken or fish meat, or a dark meat NEPI 422 that can replicate beef or dark meat chicken. To produce the white NEPI 422, the dehulled hemp grain 105 can be used. In one embodiment, the dehulled hemp grain 105 can be subjected to the natural edestin protein isolation process 200, whereby the white meat NEPI 412 is obtained and can be used in the structured protein food process 100 to produce a white meat replica. To produce the dark meat NEPI 412, the whole hemp grain 101 can be used. In one embodiment, the dehulled hemp grain 101 can be subjected to the natural edestin protein isolation process 200, whereby the dark meat NEPI 412 is obtained and can be used in the structured protein food process 100 to produce a dark meat replica. In one embodiment, the use of some whole hemp grains and some dehulled hemp grains, where the whole hemp grains are used at a concentration of about 20-30% by weight relative to the amount of dehulled hemp grains, may result in the dark NEPI 412 or an intermediate color NEPI 432. In one embodiment, the husk previously removed by the decortication of the hemp grain can be reintroduced into the dehulled hemp grain 105 to add the color of dark meat, and in one embodiment, to achieve a color similar to dark meat, the husk can be added to the dehulled hemp grain 105 in an amount of approximately 10-15% by weight relative to the dehulled hemp grain to produce the intermediate color NEPI 422.

[0168] Figure 5 shows the process of extracting oil and albumin 500. AOAE230, which is the product of the natural edestin protein isolation process 200, can be a process for producing albumin 550 and hemp oil 560. In the oil and albumin extraction process 500, AOAE230 can be evaporated to form a concentrate 506. The product can be homogenized 504, heated, and pasteurized at a low temperature 530. Clarifying 502 the AOAE can be useful. Heating to 180°F 520 can cause the emulsion to break down. Evaporate to preferably have more oil than water 506. Cool to near freezing or until frozen 508. Centrifuge with a cream separator 510 to obtain albumin 550 or hemp oil 560.

[0169] Figure 6 shows the hydrogel formation process 600, in which NEPI250 can be combined with preheated water to form a protein hydrogel 108, which is substantially described in Figure 2. In the hydrogel formation process 600, preheated water at about 135°F can be added to NEPI250 and mixed under high shear 106 to form a protein hydrogel 108. The protein hydrogel can be pasteurized at 145°F before, during, or after the formation of the protein hydrogel. After the production of NEPI250, the pasteurization conditions need to be maintained or created. The pasteurized product 104 can be prepared by first hydrating NEPI250 when spray drying 306 to form NEPI powder 308, or by maintaining an appropriate degree of hydration of NEPI250 and maintaining the pasteurization conditions as much as possible. In one embodiment, the amount of preheated water added to NEPI250 can make the solution about 3 parts water per 1 part NEPI by dry solids weight. In some embodiments, NEPI can be frozen 310 with a cooler and lyophilized 312 to produce NEPI powder 308. Heating the hydrogel to 130°F 111 can be useful. Heating the oil to 110 - 115°F can be useful.

[0170] In some embodiments, the preheated water may be tap water, and in some embodiments, the preheated water may be tap water supplied from Lake Erie and may be substantially free of solutes (e.g., tap water, distilled water, or deionized water). Salt should not be added to the solution during the hydration and protein preparation processes because it may disrupt the structure of the protein hydrogel 108 or the protein-fat hydrogel 114. Salt may be added after curing, but not before curing. In some embodiments, the hydration and relaxation of the protein can be carried out at 100°F to 135°F (although not bound by theory, to allow for proper interaction with the oil during the formation of the protein-fat hydrogel 114, the structure of the protein is slightly changed or relaxed), or in some embodiments, at 100°F to 155°F, or in other embodiments, the formation of the protein hydrogel may be carried out at a lower temperature, but the temperature must be higher than the low temperature at which the protein cannot be hydrated and relaxed. Preferably, the temperature during the hydration and protein preparation steps should be kept as close as possible to 145°F or the temperature of pasteurization 104 without reaching a temperature that can cause protein aggregation and granulation. Once the protein hydrogel is formed, the preheated oil 109 may be heated to 110°F to 115°F in some embodiments, to 100°F to 155°F in other embodiments, or in some cases, to a temperature higher than a temperature considered to be so low that the addition of the oil disrupts the hydrogel structure of the protein, but lower than the temperature at which the granules 114 of the protein-fat hydrogel are formed.

[0171] In some embodiments, the protein-fat hydrogel 114 can be produced by combining fat with a warmed suspension of hydrated protein having a pH of 6.5 to pH 7.8 (e.g., pH 7.5) (e.g., a protein isolate containing edestin). An emulsion is formed by rapid stirring, such as with a waring blender or a hand-held homogenizer, or by homogenization of this mixture. The physical properties of the protein-fat hydrogel 114 can be controlled by varying the type of protein, protein concentration, pH level during homogenization, rate of homogenization, and fat-to-water ratio.

[0172] To form the protein-fat hydrogel 114, preferably, a polyunsaturated fatty acid (PUFA) oil or fat, which can be coconut oil or fat, is heated to just above the melting point of the fat and added to the protein hydrogel 108. Without being bound by theory, the fat forms a layer surrounding the hydrated native edestin, thereby forming a liquid matrix that essentially encapsulates the hydrated protein, i.e., the protein-fat hydrogel 114, and can form an oil-in-hydrated-protein emulsion that effectively creates a thick and stable gel. Effectively, the oil can seal and protect the hydrated protein structure. Hydrated proteins can hold significantly more fat in the gel state than dry proteins. Generally, as contemplated in this application, it has been found that native globular proteins that are first hydrated and then gently heated below their denaturation temperature can hold up to twice their weight in fat. The water content of the protein-fat hydrogel can range from about 30 wt% to about 70 wt% in some embodiments. Water content refers to the amount of water in the material measured by an analytical method and is calculated as the percentage change in mass after water has evaporated from the sample.

[0173] In any of the methods or compositions described herein, the protein-fat hydrogel 114 may contain a flavoring agent or other additional components. The following components: a fat-soluble or other flavor system, a salt containing sodium chloride, a plant-based albumin source, a plant-based insoluble or soluble fiber, can be optionally added typically at less than 2% by weight, based on the finished protein-fat hydrogel 114. Starch can be added alone or in combination with other soluble carbohydrates, optionally including complex carbohydrates or sugars, at a level of up to about 10% by weight, more preferably less than 5% by weight. Auxiliary components can be added to the protein-fat hydrogel 114 before hardening for the purpose of improving and varying the flavor or texture. Fibers can be added to reduce the "grittiness" of the structured protein food 120.

[0174] In one embodiment, the protein-fat hydrogel 114 may, in one aspect, contain from about 15% to about 25% by weight, or more preferably from about 18% to about 22% by weight, of protein, which may be a natural oilseed protein. In one embodiment, about 75% to about 85% by weight of the protein isolate contains globular protein, and preferably the protein isolate contains less than 15% albumin, more preferably less than 5% albumin. More importantly, the globular protein may be in its native state and preferably has a significant content of the amino acid cysteine in a greater amount than casein or soy protein isolate. The balance of the protein composition may, in some embodiments, mainly be minerals such as calcium and phosphorus. The natural oilseed globular protein may preferably have a substantial amount of cysteine.

[0175] The protein-fat hydrogel 114 may, in one aspect, contain from about 40% to about 70% by weight, or more preferably from 40% to 60% by weight, of water.

[0176] Protein-fat hydrogel 114, in one embodiment, may contain from about 0 wt% to about 35 wt% of fat, and the ratio of saturated to polyunsaturated fatty acids (PUFAs) is between 100 wt% saturated fat and 100 wt% PUFAs. The combination of these two amounts of fat provides various unique textures not previously reported, depending on the amount of protein used in combination with the fat.

[0177] Protein-fat hydrogel 114, in some embodiments, may optionally contain from about 0 wt% to about 5 wt% of starch. The amount of starch added may depend on the amount of water added to the protein, exceeding the amount of water required for protein hydration.

[0178] Protein-fat hydrogel 114 can be formed by manually or mechanically mixing the components for forming protein-fat hydrogel 114. Preferably, the hydrated protein is first warmed to just below the granulation temperature of the protein, oil and / or molten fat is added, and preferably the mixture is gently homogenized.

[0179] In one embodiment, protein-fat hydrogel 114 can be mixed at a temperature of 120°F to 150°F. It has been found that the temperature range for placing the protein in a heating environment without breaking the formed gel or matrix is 70°C to 100°C. These temperatures are significantly lower than the extrusion temperatures typically required for extrusion of conventional meat-like proteins such as soybeans. The temperature for denaturation and fibrillation of soy protein under conditions normally used in extruders is in the range of about 130°C to 140°C. According to the present disclosure, good texture processing can be obtained by heating protein-fat hydrogel 114 in an oven and / or by pressure cooking (retorting) protein-fat hydrogel 114 to actively cure the protein.

[0180] The physical properties of the protein-fat hydrogel 114 are those of a hydrogel. Viscosity depends on the content of oil, fat, water, and protein. When there is a high water content, the viscosity will substantially decrease even if the protein-to-fat ratio is low. Similarly, when the protein-to-fat ratio is very low and the water content is low, the viscosity can become very high. The quality and selection of the fat-based and protein-based components also significantly affect the viscosity.

[0181] The formation of the protein-fat hydrogel 114 can be carried out below the denaturation point of the native protein. However, according to the present disclosure, storing the protein-fat hydrogel 114 at that temperature is not desirable as it is not microbiologically stable. It is preferable to immediately heat-treat it to harden the shape of the protein. The liquid matrix can be cooled to below 6 °C by a heat exchanger or other means and stored before further processing.

[0182] Figure 7 shows the retort process of NEPI700 that results in the structured protein food 120. The NEPI protein-fat hydrogel 114 is dispensed into the formed TETRAPAK 200 mL containers 702, and in one embodiment, each container is filled with 180 g. The top can be sealed 704 using a TETRA RECART machine. The packed NEPI protein-fat hydrogel can be placed in a retort machine 706. The NEPI protein-fat hydrogel is then heated under retort conditions 708 and can be hardened 710. In some embodiments, this process results in the structured protein food 120.

[0183] Regarding the retort according to the present disclosure, Figures 14 to 18 show photographs of the results of retorting various NEPI products and commercially available hemp protein powders. Each figure includes an enlarged view of the retorted product. Boiled chicken was used as a standard. Table 6 below shows the results of texture profile analysis of the retorted hemp products. Tables 7 and 8 show the colorimetric data of each product manufactured and tested by the retort process. Boiled chicken breast was used as a standard.

[0184] Figures 14 to 18 show photographs of the retort-treated NEPI dehulled powder 250, where the solid has a protein-to-fat (NEPI250 to coconut oil) ratio of approximately 2:1 and a solids-to-liquid (water) ratio of approximately 2:3. After the preparation of the protein-fat hydrogel, a retort-treated product was manufactured as known to those skilled in the art.

[0185] A in Figure 14 is a photograph of a cross-section of boiled chicken breast. B is an enlarged photograph of the cross-section of the boiled chicken breast in A. C is a photograph of an enlarged cross-section of the boiled chicken breast in B.

[0186] A in Figure 15 is a photograph of a cross-section of a retort meat analogue using NEPI dehulled hemp grain concentrate. B is an enlarged photograph of the cross-section of the retort meat analogue using NEPI dehulled hemp grain concentrate in A. C is an enlarged photograph of the cross-section of the retort meat analogue using NEPI dehulled hemp grain concentrate in B according to the present disclosure.

[0187] A in Figure 16 is a photograph of a cross-section of a retort meat analogue using NEPI dehulled hemp grain powder. B is an enlarged photograph of the cross-section of the retort meat analogue using NEPI dehulled hemp grain powder in A. C is an enlarged photograph of the cross-section of the retort meat analogue using NEPI dehulled hemp grain powder in B according to the present disclosure.

[0188] A in Figure 17 is a photograph of a cross-section of a retort meat analogue using VICTORY HEMP dehulled hemp grain powder. B is an enlarged photograph of the cross-section of the retort meat analogue using VICTORY HEMP dehulled hemp grain powder in A. C is an enlarged photograph of the cross-section of the retort meat analogue using VICTORY HEMP dehulled hemp grain powder in B according to the present disclosure.

[0189] A in Figure 18 is a photograph of a cross-section of a retort meat analogue using HEMPLAND dehulled hemp grain powder. B is an enlarged photograph of the cross-section of the retort meat analogue using HEMPLAND dehulled hemp grain powder in A. C is an enlarged photograph of the cross-section of the retort meat analogue using HEMPLAND dehulled hemp grain powder in B according to the present disclosure.

[0190] Figure 8 shows a process for producing a structured protein food 120 obtained by extruding NEPI250 and subjecting it to texturing. Figure 8 shows the step of providing an extruder having a heating auger, preferably, in one embodiment, a hollow steam heating auger 801, or another type of heating auger extruder. In one embodiment, the extruder can be a POWERHEATER PH100 provided by Source TECHNOLOGY. The technology used in this machine that can be utilized in the present disclosure is described in U.S. Patent Nos. 10,893,688, 10,624,382, 10,149,484, 10,092,013, 10,028,516, 9,931,603, U.S. Patent Application Nos. 2010 / 0062093, 2011 / 0091627, 2019 / 0299179, 2020 / 0113222, 2020 / 012095, and 2020 / 02680205, which are hereby incorporated by reference in their entirety. The POWERHEATER PH100, in the present disclosure, can more appropriately control the temperature of the auger and the inner wall of the extrusion pipe or chamber by a hollow auger design that can introduce steam into the auger to heat the auger and provide a more uniformly heated protein-fat hydrogel, which is very important for appropriately setting the present disclosure. Conventional extruders such as those developed by CLEXTRAL or WENGER were tested in the present disclosure but did not provide a satisfactory final product. In the conventional extruders, the protein-fat hydrogel of the present disclosure caused adhesion to the inner wall of the extruder pipe.

[0191] The POWERHEATER PH100 is known to be used with a fibrillated input material, but generally is known to be used to set the input material to starch rather than protein. The extrusion of protein curds is generally carried out at temperatures far exceeding 100°C, and thus the input material for protein hardening is not considered to be used with the POWERHEATER PH100. However, the protein-fat hydrogel of the present disclosure is effectively textured and fibrillated by the POWERHEATER PH100 at 75°C when fibrillating the protein-fat hydrogel of the present disclosure, which is achieved at a relatively low temperature of about 75°C to 85°C, the auger and extruder can be preheated to 75°C to 85°C802, and the extrusion can occur in the range of about 70°C to 95°C. In one embodiment, in the protein-fat hydrogel extruder, the POWERHEATER PH100 is used at 75°C and an 8 mm screw size is used instead of a 3 mm screw size. The protein-fat hydrogel can be introduced into the POWERHEATER PH100 using a suction pump or a stuffing pump, and the starting temperature can be about 85°C802. After pumping the protein-fat hydrogel into the extruder 804, the extrusion can proceed at about 75°C to 85°C, and the protein-fat hydrogel does not adhere to the inner wall of the extrusion pipe 806. This process produces a textured structured protein food 120. The textured structured protein food 120 extruded according to the present disclosure has been demonstrated in tests to have a texture, fiber and color similar to that of cooked chicken breast, and has excellent unexpected properties considering the prior art and the knowledge of those skilled in the art.

[0192] Figure 19 is a photograph of NEPI and poached chicken breast slices extruded from threshing powder, showing the similarity of texture and fiber structure according to the present disclosure, extruded onto POWERHEATER PH100 as described above. The poached chicken breast 1910 is shown next to the processed extruded NEPI250 chicken product 1920, which is manufactured from hemp grains NEPI spray-dried according to the present disclosure. This result is unexpected from hemp grains using only three ingredients, NEPI250, coconut oil, and water, in a ratio of 2:1:3, respectively.

[0193] In almost all extrusions involving the extrusion of soy-based meat analogs, it has been found that the ratio of protein to fat is typically greater than 10:1. Therefore, extruded, denatured, and fiberized soybeans can hardly retain fat. However, according to the present disclosure, the hydrated gel of natural globular proteins such as edestin can retain up to twice its weight of fat even after the formation of the hardened or solid form of the gel produced by the application of radiation, microwaves, or other forms of heating including direct heating or extrusion.

[0194] According to the process of the present disclosure, the protein-fat hydrogel 114 can be hardened into a solid state at a temperature of about 70 °C to 100 °C depending on the concentration of the protein in the system. The low hardening temperature is consistent with the denaturation of the natural protein in NEPI250.

[0195] According to the present disclosure, the solid structure formed during extrusion can be cooled and is representative of the cured product, but is accompanied by incomplete denaturation similar to uncooked protein or "raw" meat. Further heating of the "uncooked" protein causes further denaturation of the protein, strengthening the shape, elasticity, texture, etc., and in this process, a certain amount of water is finally released. According to the process of the present disclosure, it is not desirable to heat the product to the extent that a significant amount of water is released from the cured product in the extruder. Rather, it is desirable to simply solidify the gel and the shape or texture of the protein. In one embodiment, the present disclosure describes a process for preparing a raw meat analogue or dairy analogue, or a structured protein food 120, in an extruder, similar to raw animal meat. When this raw meat analogue is further cooked by traditional or commercial means, the meat is strengthened and cured.

[0196] The process according to the present disclosure is in contrast to existing techniques where a meat-like texture is created by using fully denatured proteins and then co-blending them with other binders including fat, starch, and other proteins to form the appearance of a ground meat type material. According to existing techniques, this type of cured product is mainly achieved during cooking through the gelation of added raw proteins such as starch or gluten.

[0197] The final texture of the structured protein food 120 can depend on the properties of the liquid matrix including the ratio of protein, fat, and water, as well as the extrusion conditions. As described herein, an extrusion mixture of isolated plant-based protein may be referred to as a structured protein food 120, which can be a meat analogue, and the fibrousness and tensile strength of the meat analogue can be controlled by the co-variation of extrusion parameters such as temperature, pressure, throughput, and die size. For example, a combination of lower extrusion temperature, medium / low throughput, and smaller die is advantageous for the production of a high-fibrous tissue with low tensile strength, while higher extrusion temperature, higher throughput, and larger die are advantageous for the production of a replica of a low-fibrous tissue with very high tensile strength.

[0198] The fibrousness and tensile strength of the meat analog can also be adjusted by changing the composition of the extrusion mixture. For example, by increasing the ratio of isolated plant protein to fat and water, or by decreasing the water content in the extrusion mixture, a meat analog with thinner fibers and higher tensile strength can be produced.

[0199] The extrusion of the liquid matrix involves feeding the liquid matrix into an extruder. In some embodiments, the extruder can be a SOURCE TECHNOLOGY POWERHEATER PH100. Although twin-screw extruders from CLEXTRAL and WENGER were tested, satisfactory results were not obtained. According to the process of the present disclosure, cooling is important in extrusion to achieve a temperature below 21°C, whereby saturated fat can easily harden within the structure and the product can be cooled more efficiently to refrigeration or freezing temperatures.

[0200] For each product, the wet ingredient blend is transferred to a feeder that can meter the liquid matrix from the inlet of the extruder at a constant feed rate. In conventional extrusion, a dry protein product is fed into the inlet of the machine. As the dry product moves through the machine, water and fat are introduced from separate inlets. In contrast, in the process according to the present disclosure, the hydrated protein and oil are first mixed as described herein to tightly control the chemical reactions that occur during the formation of the protein-fat hydrogel 114. Thus, in some embodiments, additional water, starch, or fat may or may not be added to the extruder during extrusion. In some embodiments, fibers can also be added.

[0201] In conventional extrusion of plant-based meat analogs, if water and fat are added before starting the extrusion, as the temperature rises, water will flow out of the product, resulting in undesirable steam release. Therefore, in the present disclosure, the process of adding water and fat is strictly controlled during extrusion. In the process according to the present disclosure, the liquid matrix extrusion mixture is designed, in particular, to prevent the release of water from the product by forming a gel. During the preparation of the liquid matrix according to the present disclosure, by adding oil to the hydrated protein, an emulsion gel is formed that prevents the release of water from the product during extrusion, although normally the water would be released as steam from the machine. The formation of the gel also enables the maintenance of high moisture content in the liquid matrix during extrusion and in the final product, which is desirable for the excellent texture of the structured protein food 120.

[0202] The temperature during extrusion is important for the resulting product. It is necessary to gradually increase the temperature and maintain it at about 70 °C to 100 °C, or 100 °C to 110 °C. In conventional extrusion, the temperature inside the extruder generally exceeds 130 °C. In the process of the present disclosure, the low temperature can prevent the destruction of the protein-fat hydrogel 114, thereby enabling the molecular structure of the compound to remain substantially or partially intact. The temperature of the protein-fat hydrogel 114 is preferably maintained at about 75 °C to 85 °C to harden the protein-fat hydrogel 114 and then cooled during the extrusion process to reduce the temperature below 21 °C. In the process of the present disclosure, it is important to maintain a temperature lower than that used during conventional extrusion. Here, the temperature rises only to the point where the setting of disulfide bonds is possible, and as a result, the fat is completely incorporated between all peptide layers of the protein. The residence time inside the extruder or in any heating environment should be sufficient for the input temperature of the liquid matrix to reach 70 °C to 110 °C, or preferably 75 °C to 85 °C.

[0203] Preferably, the extruder rotates the protein-fat hydrogel 114 at a relatively low screw speed, measured in revolutions per minute (rpm), during extrusion to maintain the gel structure and form a meat analog product that maintains a high level of moisture in the product. The screw speed can be carefully monitored to prevent a temperature increase and prevent disruption of the chemical structure of the liquid matrix.

[0204] To prevent disruption of the structure of the loose protein-fat hydrogel 114 formed by hydration of protein and fat encapsulation, it may be essential to move the gel slowly through a heating system to form shape and some degree of fibrillation while maintaining the initial gel hardening (partial protein denaturation). Fermentation (such as occurs in cheese making), or complete cooking and denaturation, will ultimately occur during subsequent use of the product. In some embodiments, a finished extruded product having a water content of 35 wt% to 75 wt% can then be fully cooked at an elevated temperature by a normal or commercial cooking process, optionally for microbiological stability, fermented, refrigerated or frozen for a time until the desired finished texture is obtained prior to consumption. Additional relevant extrusion parameters can include die diameter, die length, product temperature at the end of the die, and feed rate.

[0205] After extrusion, the final product can have a structure more similar to animal meat than conventional or known structured protein foods such as meat analogs and dairy analogs. Without being bound by theory, extrusion of the protein-fat hydrogel 114 according to the present disclosure can cause the protein to form substantially aligned protein fibers, which are discrete lengths of continuous filaments composed of proteins held together by intermolecular forces such as disulfide bonds, hydrogen bonds, electrostatic bonds, hydrophobic interactions, entanglement of peptide chains, and Maillard reaction chemistry that forms covalent crosslinks between the side chains of the proteins. The strength of the cured product after the first extruder is not complete or not as strong as expected. In fact, it may be desirable to remove the finished thermoset product and subject it to further heating by direct or indirect heat, common cooking methods such as boiling, baking, frying, roasting, microwaving, fermenting, and pressing (as in the production of cheese including the addition of salt and acid) to complete the strength and shape of the initial cured product.

[0206] The preparation and extrusion conditions of the protein-fat hydrogel 114 according to the process of the present disclosure can, in some embodiments, allow substantially aligned protein fibers to retain up to about 50 wt% fat within the protein. Thus, the final product is not greasy and has a mouthfeel and fat release during chewing that more closely matches animal meat than existing meat analogs. Mouthfeel can refer to a combination of properties including moistness, chewiness, biting force, breakdown, and fattiness that together provide a satisfying sensory experience.

[0207] The expected final structure of the structured protein food 120 can vary based on the composition of the protein-fat hydrogel 114. In one embodiment of the present disclosure, the expected final composition of the structured protein food 120 is represented in Table 4 in terms of the weight of the protein, the weight of the carbohydrate (if present), the weight of the lipid, and the weight of the water, along with the weight of any other possible components. Table 5 shows the physical properties of the structured protein food 120 shown in Table 4. After extrusion is complete, the product can be cooled, shaped, or cut. A post-treatment step may be performed on the extruded product.

[0208] The meat analog, also referred to herein as the structured protein food 120, can be produced from the protein-fat hydrogel 114 by methods other than extrusion. Additional methods for producing the meat analog from the protein-fat hydrogel 114 include the application of mechanical energy (e.g., shear, pressure, friction), radiant energy (e.g., microwave, electromagnetic), and thermal energy (e.g., heating, steam texturization).

[0209] The present invention will be further illustrated by the following examples, which do not limit the scope of the present invention as set forth in the claims.

Examples

[0210] Example 1 Preparation of an isolate of natural edestin protein (NEPI) Hemp grains were obtained from Hemp Oil Canada (Manitoba, Canada) and River Valley Specialty Farms (Manitoba, Canada). Dehulled hemp grains were obtained from River Valley Specialty Farms and whole hemp grains were obtained from Hemp Oil Cananda.

[0211] HHG contained 5.5 wt% moisture, 46 wt% Kjeldahl protein on a dry basis, 35 wt% fat on a dry basis, and a protein-to-fat ratio of 1.3:1 by weight. WHG contained 8.8 wt% moisture, 22 wt% Kjeldahl protein on a dry basis, 30 wt% fat on a dry basis, and a protein-to-fat ratio of 0.7:1 by weight.

[0212] In an 800-gallon agitated tank, 1000 pounds of HHG was mixed with 5000 pounds of water at 34°F. The HHG was wet-milled while maintaining the temperature between 34°F and 38°F. The hemp slurry was milled in a SILVERSON rotor-stator tank at a rate of 56 gallons per minute for 30 minutes to wet-mill the HHG. The diluted slurry was held for an average time of 30 minutes. A size 120-mesh SWECO 60-inch screen was used to separate the extract from the insoluble by-products, removing most of the solids. Next, the material passing through the 120-mesh screen was passed through a 200-mesh screen of another SWECO vibratory screening device to obtain a slurry, which was then transferred to a 500-gallon jacketed tank and the temperature of the slurry was maintained between 34°F and 38°F. Then, the slurry was fed to a DELAVAL centrifugal decanter at a rate of 13 gpm to obtain a separation of edestin solids from the AOAE emulsion. Next, the AOAE emulsion was pasteurized through a tubular heat exchange system at a maximum temperature of 185°F for 10 minutes. Next, the AOAE was held in a 900-gallon tank for processing. The 40% solids edestin solids were diluted with cold water to 30% solids and pumped through a preheated tubular system set at less than 150°F, which exited the system at 146°F and entered a jacketed holding tank with a temperature of 145°F in the jacket. After 30 minutes, the material was cooled to 35°F through a heat exchanger and placed in a tote in a refrigerator for further processing and drying by a spray dryer.

[0213] 1,000 pounds of WHG was mixed with 5,000 pounds of water at 34°F in an 800 - gallon agitated tank. HHG was wet - milled while maintaining the temperature at 34°F to 38°F. The hemp slurry was milled in a SILVERSON rotor - stator tank at a rate of 48 gallons per minute for 30 minutes to wet - mill the WHG. The diluted slurry was held for an average of 30 minutes. Using a size 60 mesh on a two - stage SWECO 60 - inch screen, the extract was separated from the insoluble by - products to remove the hulls. A 200 - mesh screen was installed in the second stage of the SWECO. The slurry from the SILVERSON first passed through the 60 - mesh to remove the hulls and immediately fell onto the 200 - mesh screen, thereby removing the chloroplasts and micro - fibers. The flow rate through the SWECO was approximately 6 gpm. The slurry screened was fed directly to a 500 - gallon jacketed tank, and the temperature of the slurry was maintained at 34°F to 38°F. When the tank was full, the slurry free of hulls, fibers, or chloroplasts was then fed to a DELAVAL centrifuge decanter at a rate of 13 gpm to obtain a separation of edestin solids from the AOAE emulsion. Next, the AOAE emulsion was pasteurized at a maximum temperature of 185°F for 10 minutes through a tubular heat - exchange system. Then, the AOAE was held in a 900 - gallon tank for processing. The 40% solids light - brown edestin solids coming out of the decanter were diluted with cold water to 30% solids and pumped through a pre - heated tubular system set at less than 150°F. It exited the system at 146°F and entered a jacketed holding tank with a temperature of 145°F in the jacket. After 30 minutes, the material was cooled to 35°F through a heat exchanger and placed in a tote in the refrigerator for further processing and drying by a spray dryer. The dry - matter - based yield of NEPI based on the weight of the starting material WGH was 15% or 79% of the theoretical value. The AOAE yield was 25.3% on a DSB basis, and the hull, fiber, and chloroplast fractions were 46.9% on a DSB basis. The overall recovery rate was 92%. The NEPI yield from HHG was 30% or 86% of the theoretical value. The AOAE yield was 40.9% on a DSB basis, and the hull, fiber, and chloroplast fractions were 22.5% on a DSB basis. The overall recovery rate was 98%.The analysis of the NEPI products obtained from WGH and HHG is shown in Tables 1 and 2 below. TIFF2025508562000002.tif119135

[0214] The NEPI concentrates prepared by the process of the present disclosure exhibit high microbiological activity even before pasteurization and spray drying into powder while maintaining the process temperature below 38°F (see Table 1). The incoming raw materials, whether hemp grains or dehulled hemp, typically have a total plate count (TPC) in the range of 2,000 TPC to 250,000 TPC. In a protein-rich aqueous medium, it is essential to maintain the temperature below 42°F, preferably below 38°F. Despite the low temperature, if pasteurization is not carried out immediately after the start of aqueous grinding, the TPC will continue to increase, resulting in protein damage. The short duration of the process and the ability to pasteurize both the AOAE and edestin slurry immediately after separation by a centrifuge decanter are essential factors in the process. The edestin product obtained as a result of pasteurization at a low temperature of 145°F maintains its gelling function as described above. The AOAE can be heated briefly at a much higher temperature above 145°F, more preferably above 195°F, which is advantageous for further processing to remove residual insoluble solids by centrifugation and then separate the aqueous albumin phase and the oil phase by emulsion breaking. The success of pasteurization of the final powdered NEPI product is reflected in the TPC of the products in Table 1. TIFF2025508562000003.tif133135

[0215] Table 3 shows the DSC thermogram. The structure of NEPI measured by the DSC thermograph (partially shown in A - B of Figure 12 and A - B of Figure 13) can be compared with commercially available products below. TIFF2025508562000004.tif152135

[0216] Further structural and compositional analysis of NEPI and commercially available hemp protein products measured by SDS-PAGE gel electrophoresis are shown in FIGS. 9 and 10.

[0217] Example 2 Spray-dried NEPI The NEPI cooling slurry obtained from Example 1 was sent to a commercially available spray dryer for drying. An ALFA LAVAL type spray dryer equipped with a nozzle having a water removal capacity of 1200 lb per hour was used to dry the powder. The refrigerated product was pumped into a 250-gallon jacketed tank, and the water temperature set to maintain the jacket at 155°F was used. The tank had a low-speed agitator and it took several hours to heat approximately 200 gallons of 30% concentrated edestin slurry. Once the product reached that temperature, it was sent to another tank for feeding into the dryer. It should be noted that NEPI dried very easily without adhering to the walls of the dryer. The final outlet temperature of the dried product was 85°F. For each of the NEPI (WG and HHG) products obtained from Example 1, the composition of the dried product is shown in Table 2 below.

[0218] Experimental Example 3 Production of protein-fat hydrogels from NEPI and commercially available hemp powder Protein hydrogels are easily made in a 5-gallon plastic bucket by adding 14 lbs of water preheated to 140°F. While stirring this water using a 1 / 4 horsepower hand-held industrial homogenizing rod, 14 lbs of NEPI dry powder is slowly added. Homogenization is maintained until all the powder has been added. The temperature at this time is 130°F and after holding for about 15 minutes, 7 lbs of canola oil is added all at once and the mixture is blended briefly with the homogenizing rod for about 1 minute or until the slurry appears well blended to incorporate the oil as a uniform emulsion. Example 4 Protein-fat hydrogel formulations and properties for various types of meat analogs and milk analogs

[0219] Example 4 discloses a formulation containing a liquid matrix used to produce various types of meat analogs. According to the present disclosure, different types of meat analogs can be obtained, including plant-based meat analog targets that reproduce seafood, white meat, dark meat, eggs, and cheese, depending on the ratio of protein, fat, and water. TIFF2025508562000005.tif112135

[0220] Regarding Table 4, the target water content is 35 wt% to 75 wt%. At least 70 wt% of globular natural plant protein has an albumin content of less than 15 wt%, preferably less than 5 wt%. The liquid matrix temperature needs to be maintained at 140°F from the mixing blend to the processing. Due to the ability of natural seed oil proteins, which can be natural edestin in Table 4, the amount of fat can be varied to obtain different types of meat analogs. The structural characteristics of the obtained products are similar to those of the materials they replicated. For example, the texture of seafood was white and had a very elastic structure similar to raw shrimp or scallops. The white meat was white and had a similar texture expected from partially cooked chicken breast. The dark meat was slightly light brown in color, and its texture was also similar to chicken thigh meat, with more fat and moisture compared to white meat. The eggs were similar to what was expected from scrambled eggs and were also white in color. The cheese was similar to a cheese card and actually made a squeezing sound when chewed, similar to a fresh cheese card. JPEG2025508562000006.jpg78135

[0221] Example 5 Manufacture of Structured Protein Foods by Retort The retort conditions were carried out over 15 minutes from a temperature of 77°F to a peak of 270°F and reduced to 95°F in 15 minutes. The pressure was 0.20 bar in 1 minute, increased to 3.0 bar in 4 minutes, and decreased to 0.8 bar in 15 minutes. The machine used was of the SUNDRY retort type: AP-95, serial number: 705. TIFF2025508562000007.tif193135TIFF2025508562000008.tif21135TIFF2025508562000009.tif132135 TIFF2025508562000010.tif124135 TIFF2025508562000011.tif69135

[0222] Example 6 Manufacture of Structured Protein Foods by Extrusion The protein-fat hydrogel of Example 3 was used in a Power100 Source Technology extruder set at a screw auger diameter of 3 MM at a flow rate of 6 lbs per minute and 185°F to create a structured gel with the appearance and texture of white chicken. For an image comparison of white chicken and the extruded hydrogel-structured protein food, refer to Figure 19.

[0223] The present disclosure surprisingly demonstrates that an excellent hemp-based structured protein product can be manufactured using only three components: hemp grains, oil, and water. It is shown herein that the hemp meat analogs manufactured according to the present disclosure surprisingly reproduce chicken in terms of color, texture, and taste. Some commercially available protein products claim to produce excellent meat analogs, but when used for this purpose, they did not compare favorably to natural edestin protein isolates in terms of taste, color, or texture.

[0224] No commercially available products that use only hemp protein to produce meat analogs have been found. Furthermore, the prior art teaches that hemp protein alone is not a promising protein for producing structured protein foods such as meat analogs and dairy analogs. This disclosure demonstrates otherwise.

[0225] Definition: As used herein, the articles "a" and "an" are understood to mean one or more of the materials claimed or described, for example, as used herein, "an anionic surfactant" or "a fiber".

[0226] "Basis weight" means the weight per unit surface area (in the machine direction / cross direction plane) of a sample of web-like material (one-sided), expressed in grams per square meter (gsm). The basis weight may be specified in the manufacturing specification or measured, and reflects the weight of the material before adding any liquid composition.

[0227] As used herein, "web-like structure" means a web or sheet-like hydrogel that includes at least elements of yarns, sheets, and the container sidewall adjacent or bottom adjacent portions.

[0228] As used herein, "container" means an object that can hold a liquid protein-fat hydrosol and can be used in a microwave oven.

[0229] As used herein, "container material" may include materials capable of holding liquids, preferably may be composed of food-grade materials that can be used in a microwave oven, including but not limited to plastics such as acrylic or polymethyl methacrylate (PMMA), polycarbonate (PC), polyethylene (PE), polypropylene (PP), polyethylene terephthalate (PETE or PET), polyvinyl chloride (PVC), acrylonitrile-butadiene-styrene (ABS); paper, mixed paper with materials such as plastics that allow heat treatment or can hold boiling water, in contrast to conventional plastics, and paper containers containing polylactic acid (PLA); ceramic materials including glazed and unglazed ceramics; glass including microwave-heatable glass; and other materials known to those skilled in the art.

[0230] As used herein, "expansion ratio" means the V of the protein hydrogel or protein-fat hydrogel after microwave heating max divided by the V of the protein hydrogel or protein-fat hydrogel. As used herein, the volume measurement of the expanded product includes the voids 2124 formed by gas bubbles, unless otherwise specified. i As used herein, the volume measurement of the expanded product includes the voids 2124 formed by gas bubbles, unless otherwise specified.

[0231] As used herein, the final volume (V f ) means the volume of the protein-fat hydrogel in the container after microwave heating measured from the final height (H f ). As used herein, the final height means the highest point on the container to which the material is bonded after collapse.

[0232] As used herein, the final meniscus center volume (V mcmeans the volume of the protein-fat hydrogel in the container after microwave heating, measured after it has collapsed from the upper surface of the collapsed material in the container after the end of microwave heating. This calculation is an estimate and may not necessarily result in an accurate measurement of the volume as there can be significant variation in the shape from run to run even if all conditions are the same.

[0233] As used herein, "hydrogel meniscus" means a complete or partial meniscus, or recess, formed from a hydrogel material that may be present after microwave heating according to the present disclosure. The hydrogel meniscus can be formed from the uppermost layer of the hydrogel material. This uppermost layer, without being bound by theory, may be referred to as a protein film or a protein-oil film. This definition can include a complete or partial meniscus that can be formed when the bubbles and hydrogel material collapse when microwave heating is stopped. At this point, a specific amount of the hydrogel material can be bound to the sidewalls of the container while a portion of the hydrogel in the center of the container can drop to a level lower than the maximum height of the material bound to the sidewalls of the container, thereby forming a meniscus or partial meniscus having a crescent or concave shape. Generally, the center of this meniscus, which can be approximately in the center of the container, may be the bottom of the meniscus or the bottom of the partial meniscus. The meniscus can have a meniscus depth measured from the bottom of the recess of the meniscus to the final height (H f ) of the hydrogel measured at the highest point where the hydrogel binds to the sidewalls of the container after microwave heating.

[0234] As used herein, "inclusion" means an edible material that can be included in the preparation of a hydrogel.

[0235] As used herein, "layer" means one thickness, row, or fold of a protein-containing material that is placed on top of or beneath another, or lying horizontally.

[0236] As used herein, the "meniscus ratio" means the depth of the meniscus divided by the final height (H f ) of the protein-fat hydrogel in the container.

[0237] As used herein, "microwave oven" means a form of electromagnetic radiation having wavelengths in the range of approximately 1 meter to 1 millimeter corresponding to frequencies of 300 MHz to 300 GHz, respectively. In either case, the microwave may include at least the entire SHF band (3 - 30 GHz, or 10 - 1 cm). Usually, consumer ovens operate at a nominal 2.45 gigahertz (GHz) - 2.4 GHz to 2.5 GHz in the ISM band with a wavelength of 12.2 centimeters (4.80 inches), while large industrial or commercial ovens often use 915 megahertz (MHz) - 32.8 centimeters (12.9 inches). For the purposes of this disclosure, microwave radiation of any wavelength is contemplated, but preferably, microwave radiation commonly used for cooking food in a home, commercial, or industrial environment can be utilized.

[0238] As used herein, "particulate" means particulate matter or powder.

[0239] With respect to the proportion of components of a structure or composition, "predominant" or its form means that the component constitutes the majority of the weight of the structure or composition.

[0240] As used herein, "visually distinguishable" means visible to the naked eye.

[0241] In this specification, when the amount of a component of a fibrous web-like structure is expressed as "X weight percent" or "X percent by weight", or an abbreviation or short form thereof, the amount means that the weight of the component constitutes X percent of the total weight of the material in which it is contained.

[0242] The "z - direction" with respect to a web or fibrous web structure means a direction perpendicular to the general plane defined by the web - like structure.

[0243] Figure 20 discloses a flowchart illustrating a process for manufacturing an instant meat analog. In one embodiment, heating water to boiling, i.e., 110 °C, or to a sufficiently high temperature, can be the first step in the microwave texturization process 2000 of the present disclosure. As is known to those skilled in the art, water can be heated and boiled 2010. The heating method can include the upper surface of a stove, a microwave oven, a hot plate, etc. In one embodiment, hot water 2002, which can be 90 °C or less in some embodiments, is poured into a measuring container and subsequently added to container 2012 (shown in FIG. 21). In one embodiment, the temperature of the water preferably does not exceed 85 °C after being added to container 2100. In some embodiments, the amount of NEPI250, hot water 2002, or a similar liquid, and oil 110 added to container 2100 can be 25 grams of NEPI250, or a similar material, 110 grams of hot water 2002, which can be homogenized before adding 12.5 grams of oil 110 to produce a chicken breast meat analog, or in some embodiments, other meat analogs 120 that can be similar to chicken breast meat. Container 2100 may have the dimensions of a 16 - ounce paper Chinet® Comfort Cup®, and heating the protein - aliphatic hydrocolloid 114 to produce a protein - aliphatic hydrogel 120 (which may also be referred to herein as a structured protein food 120) can, in one embodiment, be carried out using a Bosch® microwave (model number HMC54151UC / 05, manufactured in May 2018, input 1700W, output 1000W) at 50% output for about 1.5 minutes to obtain a preferred protein - fat hydrogel 120. The microwave cycle time is 30 seconds. In one embodiment, only 20% of water 200 is lost during heating in the microwave oven, and the remaining water 200 can be incorporated into the protein - fat hydrogel 120.

[0244] Container 2100 must be able to adequately hold the hot soup 2002. In some embodiments, container 2100 may be similar to Chinet® Comfort Cup®, a paper that is food grade and suitable for use in a microwave. In other embodiments, paper container 2100 may contain polylactic acid (PLA), and paper container 2100 may contain polylactic acid (PLA) and may be an Amazon® Basics compostable 20-ounce hot paper cup containing PLA. Container 2100 may be disposable, reusable, or compostable. In some embodiments, container 2100 may be composed of various types of paper, as known to those skilled in the art. In other embodiments, container 2100 may be composed of plastic. In some embodiments, this plastic may be an Oster® 24-ounce plastic polycarbonate measuring cup pitcher with a lid for an immersion stick blender, and the inner wall of the Oster® container can be roughened sufficiently by regular use of the blender over a period of time so that the protein-fat hydrogel can bind to the container sidewall 2102. The new Oster® 24-ounce plastic pitcher also binds. Other microwave-heatable materials may include porcelain, ceramic, glass, heat-resistant glass, glass ceramic, paper, silicone, and thermoplastic. In some embodiments, container 2100 may have a rough interior so that protein-fat hydrogel 120 can bind or adhere to its surface.

[0245] In some embodiments, the container 2100 is made of a material suitable for heating food with microwaves, and at the same time, when the internal temperature of the cooked food reaches 150°F - 212°F, the external temperature of the container 2100 is less than 140°F, or the container can be insulated so as to maintain a comfortable handling temperature. Preferred materials may include ceramics, HPE, polypropylene, double- or triple-walled paper containers, or similar materials. The material is preferably reusable and may be environmentally sustainable. In some embodiments, the preferred container 2100 may be a double- and triple-walled paper container 2100. An example of a ceramic container 2100 for use in accordance with the present disclosure is the W&P PORTER® ceramic mug with a protective silicone sleeve. An example of a polypropylene container that is microwave-heatable but not insulated for use in the present disclosure may be the 32-ounce microwave-heatable contact translucent round deli container from CHOICE. An example of a microwave-heatable and insulated three-layer paper container is the Chinet Comfort Cup®.

[0246] In some embodiments, the container 2100 may preferably be composed of a rough or textured surface material such as paper. The fibrous, or texturized or roughened nature of the container 2100 may, in some embodiments, as shown in FIG. 21C, enable both the binding of the expanded protein-fat hydrocolloid 2110 to the container sidewall 2102 and the rapid cooling of the protein-fat hydrogel 120. This may facilitate the formation of a more textured final product, which may, in some embodiments, resemble chicken breast or partially separated or minced chicken breast. The use of a smooth material such as glass or coated ceramics may not be as effective in binding the protein-fat hydrogel 120 to the container sidewall 2102, maintaining the heat of fusion, and retarding the cooling and hardening of the hydrogel, and as a result, may lead to different final products that may be more similar to different types of meat and may be inferior for a particular desired product such as chicken breast.

[0247] In some embodiments, the container 2100 can be transparent, such that during heating with microwaves, it can be observed that the protein - aliphatic hydrogel 114 expands and rises within the container 2100. Visual observation of the complete expansion and rise of the expanded protein - fat hydrogel 2110 during heating in the microwave, as well as the cessation of heating at a visual cue such as the peak of the visual rise or the rise to a desired level, can, in some embodiments, be included as part of the microwave texturization process 2000.

[0248] In some embodiments, the container 2100 may have different shapes for manufacturing specific types of meat analogs or dairy analogs, such as the shape of a chicken breast or the shape of a chicken nugget. In some embodiments, the container 2100 can have specific dimensions that are desirable for a particular type of meat product. For example, in some embodiments, if the amount of protein - fat hydrogel 114 added to the container 2100 is 110 grams of water, the container 2100 preferably has a base diameter to height ratio of about 1:2.5 and a base diameter to top diameter ratio of about 1:1.5, and the volume of the container is preferably about 16 ounces. In some embodiments, the container 2100 is generally cylindrical, open at the top, and the diameter can be about 2 inches to 3.5 inches.

[0249] Other embodiments can produce various types of meat analogs, including seafood, white meat, and dark meat, as presented and described in U.S. Patent Application No. 17 / 551,163 to Matchell Ellis, which is hereby incorporated by reference in its entirety, and more specifically, Example 4 of U.S. Patent Application No. 17 / 551,163 to Matchell Ellis, which is hereby incorporated by reference in its entirety.

[0250] In some embodiments, the preferred size of the container 2100 can be a 16-ounce or 32-ounce container 2100, with a diameter of 2 to 4 inches, which aids in the use of a conventionally sized immersion blender, and a height of about 5 inches to 8 inches. The size and shape of the container 2100 can vary according to the amount of protein-fat hydrogel 114 used in the microwave texturization process 2000. The amount of protein-fat hydrogel 114 preferably used can vary and is related to the microwave time, settings, and size of the container 2100. Preferably, a minimum microwave time is desirable to solidify or harden the protein-fat hydrogel 120, which may also be referred to herein as a structured protein food or meat analog. An even heating, or even distribution of heat, within the protein-fat hydrogel 114 is desirable. The expansion of the melted protein-fat hydrogel 114, or gaseous rise, is important to achieve an elongation of the hydrogel that allows the protein-fat hydrogel 120 to have a fibrous appearance upon hardening. Without being bound by theory, in this microwave texturization process 2000, expansion and solidification can occur simultaneously, and the expanded product can be hardened or further hardened by the rapid addition of cold water 128, thereby cooling the protein-fat hydrogel 120 to a more rigid and malleable state. After hardening of the protein-fat hydrogel 120, preferably no residual liquid or material remains in the container, which indicates a complete and uniform distribution of heat during heating.

[0251] After adding hot water or hot water 2002 to container 2100 for 112, NEPI250 or a similar substrate (which may be flavored NEPI250) can be added to the hot water 2002 or similar liquid in container 2100. NEPI250 can be manufactured as described in U.S. Patent Application No. 17 / 551,163 entitled "Native Edestin Protein Isolate and Use As A Cellidient" to Matchell Ellis (which is incorporated herein by reference in its entirety). The extraction method of NEPI250 is described in more detail in paragraphs

[0071] -

[0080] ,

[0151] -

[0162] , and Figure 2 of U.S. Patent Application No. 17 / 551,163 (which is incorporated herein by reference in its entirety).

[0252] When NEPI250 is added to hot water 2002, the hot water 2002 can be, in some embodiments, preferably at a temperature of about 60°C to 80°C, or 75°C to 85°C. Generally, when NEPI250 is added, the hot water 2002 should be at a temperature that allows for rapid hydration of the protein and interaction with the hot water 2002, but the temperature should not be so high as to cause granulation of NEPI250. NEPI250 may not function properly in the microwave texturing process 2000 when heated to a temperature above about 70°C at which granulation can occur. Therefore, it is desirable that the temperature of the hot water 2002 in container 2100 before adding NEPI250 is 80°C or less. If the hot water 2002 is at 80°C when NEPI250 is added, the temperature of the mixture can rapidly drop to about 70°C when NEPI250 is added, thereby preventing interference with the function of NEPI250 in the microwave texturing process 2000.

[0253] In some embodiments, it may be important to add NEPI 250 to the hot water 2002 after the hot water 2002 is already in the container 2100. The ratio of hot water 2002 to NEPI 250 can vary depending on the desired texture and type of meat analog, as may be described in U.S. Patent Application No. 17 / 551,163, entitled "Native Edestin Protein Isolate and Use As A Cellavidient" to Matchell Ellis, which is hereby incorporated by reference in its entirety.

[0254] In one embodiment, first, water is added to the container 2100 and only the water is heated in the microwave. Additional water is added to the NEPI 250 concentrate, and then the mixture is gently heated to a target temperature of 60°C. Next, the oil 110 can be added to the mixture. Optionally, the protein-fat hydrogel 114 mixture can then be cooled and then gently preheated to 60°C before heating in the microwave.

[0255] The present disclosure manufactures a protein-fat hydrogel 120 in a microwave oven using a protein-containing solution, a protein hydrogel 108, or more preferably, a protein-fat hydrogel 114. The protein-fat hydrogel 114 forms a unique protein-fat hydrogel 120 when heated in a microwave oven under the conditions described in the present disclosure. In some embodiments, the oil 110 may not be added to the protein hydrogel 108, and the process is performed without the addition of the oil 110. Important conditions for the manufacture of the claimed protein-fat hydrogel 120 may include the content of the protein-fat hydrogel 114, the amount or volume of the protein-fat hydrogel 114 in the container 2100, the size and shape of the container, the material of which the container is composed, the output setting of the microwave oven, and the type or structure of the microwave oven. Other conditions that may be important for the manufacture of the protein-fat hydrogel 120, also referred to as the structured protein food 120, may include the addition of additives, including particulates or inclusions, to the protein-fat hydrogel 114, the presence or absence of a lid on the container during heating, the temperature of the protein-fat hydrogel 114 prior to microwave heating, and different types of homogenizers for preparing the protein-fat hydrogel 114, but are not limited thereto. The addition of salts to the protein-fat hydrogel 114 may also affect the final product in some embodiments by increasing the swelling rate. These elements, which may be important for the manufacture of the protein-fat hydrogel 120, are not exclusive, and additional elements may be included or considered, as would be understood by one of ordinary skill in the art.

[0256] An important element of the products and processes of the present disclosure is that when the protein-fat hydrogel 114 is heated in a microwave oven, voids 2124 are formed within the protein-fat hydrogel 114 as the protein-fat hydrogel 114 expands in volume. These voids can have a diameter or width of at least 1 mm, or at least 2 mm, or at least 5 mm. The volume expansion of the protein-fat hydrogel 114, which may also be referred to simply as expansion herein, is defined for the purposes of the present disclosure as at least a portion of the protein-fat hydrogel 114 that vertically rises within the chamber of the container 2100. Without being bound by theory, the expansion of the protein-fat hydrogel 114 during microwave heating may be related to the formation of pockets of vapor within the melting hydrogel protein membrane, which in turn results in the formation of air bubbles within the protein-fat hydrogel 114. Under certain conditions, according to the present disclosure, the protein-fat hydrogel 114 expands or rises within the container 2100 to at least about 2 times, or about 3 times, or about 4 times, or about 5 times, or about 6 times, or about 7 times its original volume.

[0257] During microwave heating, the protein-fat hydrogel 114 first begins to “melt” and forms a membrane structure that entraps water molecules. Next, when the water molecules reach a temperature of 100° C. and form a gas, the protein membrane then begins to expand until the protein membrane can harden or until a temperature is reached at which substantially all of the protein-fat hydrogel 114 can harden. Hardening can be defined as a transition from a liquid state to a solid state, and the solid state may initially be moldable while it is hot or warm. According to the present disclosure, when hardened in a moldable or non-moldable solid state, the product is referred to as a protein-fat hydrogel 120 or, if no fat is added, a protein hydrogel 108. If further heated beyond the point at which the protein-fat hydrogel 120 hardens, the quality of the protein-fat hydrogel 120 or the structured protein food 120 may deteriorate.

[0258] Referring now to FIG. 20, after NEPI250 is added to the hot water in container 2100 at 116, NEPI250 and the hot water 2002 can be blended at 2018 to form a smooth and uniform protein hydro sol 108. The blending can be performed by an immersion blender 2600, as shown in FIG. 30. The immersion blender 2600 may have a handle and a blade end. In some embodiments, the diameter of the blade may generally match the diameter of the bottom 2104 or base of the container (shown in FIG. 30). The handle 2602 can be used to rotate the blade end 2604 to facilitate complete mixing of the components. An immersion blender 2600 suitable for the present disclosure can be a Vtamix® immersion blender. In some embodiments, the blender can be a hand mixer, an immersion blender or a stick blender, a single - serve blender, a portable blender, a countertop blender, a stand mixer, or a commercial blender.

[0259] In some embodiments, a flavoring can be added to the NEPI250 either before or after the addition of NEPI250 to the hot water 2002 at 114. In some embodiments, the bulk preparation of the protein - fat hydro sol 114 can be performed in any blender (Waring® or others) according to the preferred blend components, amounts, and blending procedures. However, to cure the protein - fat hydro sol 114 to form the protein - fat hydro gel 120, a preferred amount of the protein - fat hydro sol 114 should be added to a container 2100 of a preferred size and shape at the preferred microwave output settings and times to achieve the desired results described herein.

[0260] Blended in a minimum time that allows for complete hydration and relaxation of the protein, as demonstrated by a smooth and uniform emulsion with no visible particulates or granules, in 2018, thereby forming an appropriate protein hydrogel 108, after which oil 110 can be added to the protein hydrogel 108 in 2020. Oil 110 is taught in U.S. Patent Application No. 17 / 551,163 to Matchell Ellis and can be added in various amounts depending on the desired product type, as in the following examples. Many different types of oils 160 can be used, including sunflower oil, coconut oil, olive oil, and other vegetable or animal oils. The type of oil 110 may depend on the type of desired meat product analog. For example, if a pork analog is desired, bacon fat may be added to the protein hydrogel 108.

[0261] After the addition of oil 110, the mixture of protein hydrogel 108 and oil 110 can be blended by the aforementioned method in 2022 to form a protein-fat hydrogel 114. The protein-fat hydrogel 114 may have the appearance of a thick pudding in some embodiments, prior to refrigeration and hardening of the pudding. Over-blending of the protein-fat hydrogel 114 may reduce the viscosity of the protein-fat hydrogel 114 to that of a loose milkshake. After blending, the protein-fat hydrogel 114 can be held at a temperature of preferably about 65°C to 70°C, or 0.5°C to 23°C in some embodiments. Holding the protein-fat hydrogel 114 at a temperature of 0.5°C to 23°C, or 65°C to 70°C, delays the growth of microorganisms.

[0262] Next, the protein-fat hydrogel 114 can be heated within the microwave oven 2026, and optionally, the container 2100 may be covered by an indicator lid 2024. The microwave oven can be a standard household microwave oven. In some embodiments, the microwave oven may be set to 50% output. In some embodiments, the run time of the microwave oven can be about 1 - 2 minutes, or more preferably, 1 minute and 30 seconds.

[0263] In conventional microwaves, variable output levels provide flexibility in microwave cooking. Each output level provides microwave energy for a specific percentage of time. For example, output level 7 provides microwave energy for 70% of the time. Output level 3 is 30% of the time's energy.

[0264] In one embodiment, complete cooking is desirable so that no residual material remains in the container 2100 after heating. In some embodiments, a double-walled container 2100 may be desirable to prevent the user from burning their hand when removing the container 2100 after heating. In some embodiments, a cozy or container jacket, such as can be used with a beer can, can be used to hold the container 2100 after heating.

[0265] A - D of FIG. 21 show the protein-fat hydrogel 114 within the container 2100, the container sidewall 2102, the container bottom 2104, the container base 214, the expanded protein-fat hydrogel 2110 of FIG. 21C, and the protein-fat and protein-fat hydrogel 120 of FIG. 21D. When the protein-fat hydrogel 114 is heated, the protein-fat hydrogel 114 rises and, as shown in FIG. 21C, an expanded protein-fat hydrogel 2110 can be formed. The expanded protein-fat hydrogel 2110 rises vertically to a specific height and can then collapse or degas when microwave heating is stopped, and the degassed protein-fat hydrogel 120 is shown in FIG. 21D.

[0266] The protein-fat hydrogel 120, or protein hydrogel, in some embodiments, may also be referred to herein as a meat analog 120 if no fat is included in the protein-fat hydrogel 120. Heating in a microwave oven can preferably be stopped at the point where the expanded protein-fat hydrosol 2110 reaches a specific height, and a specific amount of the hydrosol material is in an expanded state before collapsing when the heating stops. The expanded material may be defined as the material that is not in the vicinity of the bottom of the container 2400, which can be defined as a material that is essentially uniform, in contact with the bottom of the container, and not part of the vicinity of the side wall of the container. The vicinity of the side wall of the container can generally be defined as the material that is bonded or attached to the side wall but not part of the threads and sheets located more centrally within the container 2100. When a predominant amount of the protein-fat hydrosol 114 is cured, in some embodiments, at least about 99% of the material is in an expanded state, in other embodiments 98%, in other embodiments 97%, in other embodiments 96%, in other embodiments 95%, in other embodiments 94%, in other embodiments 93%, in other embodiments 92%, in other embodiments 91%, in other embodiments 90%, in other embodiments 89%, in other embodiments 88%, in other embodiments 87%, in other embodiments 86%, in other embodiments 85%, in other embodiments 84%, in other embodiments 83%, in other embodiments 82%, in other embodiments 81%, in other embodiments 80%, in other embodiments 79%, in other embodiments 78%, in other embodiments 77%, in other embodiments 76%, in other embodiments 75%, in other embodiments 74%, in other embodiments 73%, in other embodiments 72%, in other embodiments 71%, in other embodiments 70%, in other embodiments 65%, in other embodiments 60%, in other embodiments 55%, in other embodiments 50%, and in other embodiments at least approximately 40% of the protein-fat hydrosol 114 is in an expanded state.

[0267] Upon further heating, the protein-fat hydrogel 120 may dehydrate, resulting in a poor-quality final product. By having appropriate parameters of the material and the shape of the container 2100, a preferred product may be possible at the maximum height of the expanded protein-fat hydrosol 2110. The adhesion of the expanded protein-fat hydrosol 2110 to the container sidewall 2102 may be important for the production of a properly structured or textured protein-fat hydrogel 120, or a structured protein food 120. If there is too much oil 110 in the protein-fat hydrosol 114, the expanded protein-fat hydrosol 2110 cannot adhere to the container sidewall 2102, and as a result, a plug-like, denser, solid, unstructured mass of protein-fat hydrogel 120 lacking the desired texture is formed.

[0268] In one embodiment, a preferred microwave texturing process 2000 may include a minimum heating time to create a fully cooked protein-fat hydrogel 120, with no residual material remaining after removal of the protein-fat hydrogel 120, which is indicated by an increase in the maximum volume, or a vertical rise, of the expanded protein-fat hydrosol 2110. When used with the microwave texturing process 2000, other protein materials, including soy or pea protein isolates, and other hemp protein isolates tested as disclosed in U.S. Patent Application No. 17 / 551,163 to Matchell-Ellis, were also unable to produce an equivalent expanded protein-fat hydrosol 2110, or a significant vertical rise, and did not result in an acceptable protein-fat hydrogel 120. Other hemp protein isolates tested, such as those manufactured by VICTORY HEMP and HEMPAND, formed products with a loose texture and low elasticity and were not considered functional meat or dairy analogs.

[0269] Examples are included in this specification for illustrative purposes only and the process can be varied as would be understood by one of ordinary skill in the art. In Example 8, 100 mL of the protein-fat hydrogel 114 was heated with the output of the microwave oven set to 5. When observed through a transparent plastic container during heating at output 5 in a Bosch microwave oven, after the first 30-second cycle, at the 10-second mark when entering the second cycle, the protein-fat hydrogel 114 began to significantly expand and rise within the container 2100 and then collapsed at the 45-second mark when the magnetron was turned off. In the third 30-second cycle, after about 5 seconds, at the 1 minute 5-second mark, the protein-fat hydrogel began to expand again. At about 1 minute 20 seconds, the protein-fat hydrogel hardened and collapsed.

[0270] In the present disclosure, the maximum volume (V max ) of the protein-fat hydrogel 114 is generally defined by the approximate maximum volume to which the protein-fat hydrogel 114 expands before collapsing that occurs when microwave heating is stopped. In some cases, it is possible to increase V max by additional heating, but in the present disclosure, V max is not defined as the absolute maximum volume achievable during heating expansion but as the maximum volume achieved in a particular process. V max may be measured using the approximate maximum height (H max ) of the protein-fat hydrogel 114 to which the protein-fat hydrogel 114 rises within the container 2100 during heating in the microwave oven.

[0271] Generally, the protein-fat hydrogel 114 can expand on the upper surface while maintaining somewhat of a uniform height across the entire container. Due to air bubbles, the upper surface becomes partially non-uniform. However, the height when the upper surface rises to reach the peak height can be observed and estimated. As used herein, the final height has a different meaning than the maximum height, and the final height (H f) is at the highest level where the protein-fat hydrogel 120 binds to the container side wall 2102 of the container. The volume (V i ) is the volume of the protein-fat hydrosol 114 in the container before microwave heating. The initial height of the protein-fat hydrosol 114 in the container before microwave heating is denoted as H i .

[0272] When the protein-fat hydrosol 114 hardens after reaching V max , it has a non-uniform web-like structure 2160 as shown in FIG. 24A. The fibrous web-like structure 2160 may include additives other than NEPI250, oil 110, and hot water 2002, which may include the core material of the products of the present disclosure, such as softeners or strengthening agents, flavoring agents, inclusions, nutritional additives, fibers, and other additives.

[0273] As shown in FIG. 24A, the web-like structure 2160 of the protein-fat hydrogel 120 may have threads 2130 that can span the width of the container 2100. The threads 2130 may be joined to a sheet 2128 of material, and the sheet 2128 may be substantially thinner than the threads 2130. Also, as shown in FIGS. 26A and 26B, there may be a container adjacent side wall portion 2300 composed of the protein-fat hydrogel 120, which may, in some cases, be thicker and larger than the threads 2130 and the sheet 2128. The threads 2130, the sheet 2128, and the container adjacent side wall portion 2300 may be interconnected and may form a semi-continuous structure similar to chicken breast slices or other meat analogs. These structures are shown in FIGS. 23 to 26, which are produced from the protein-fat hydrosol 114 shown in FIGS. 22A and 22B. FIGS. 22 to 26 are composed of the same protein-fat hydrosol 114 processed through the steps described in the present disclosure.

[0274] FIGS. 23A and 23B show the protein-fat hydrogel 120 in the container 2100. Also shown are the container side wall 2102 and the container bottom 2104. The protein-fat hydrogel 120 has its final height (H f)It is shown by 2122. The voids 2124 within the protein-fat hydrogel 120 are shown in A and B of FIG. 23.

[0275] In one embodiment of the present disclosure, upon curing of the protein-aliphatic hydrosol 114, substantially all of the protein-aliphatic hydrosol 114 had expanded from the bottom of the container and was present in either the container-adjacent side wall portion 2300, the thread 2130, or the sheet 2128. As a result, the lower portion of the protein-fat hydrogel 120 did not significantly exhibit the shape of the bottom of the container 2100. In some embodiments, in contrast to pooling at the bottom of the container during setting, the percentage of protein-aliphatic hydrosol 114 within the container-adjacent side wall portion 2300, the thread 2130, or the sheet 2128 can be about 5%, or more preferably 10%, or more preferably 15%. The mass of the protein-fat hydrogel 120 at the bottom of the container 2100 can generally be considered undesirable for most purposes of the present disclosure.

[0276] In some embodiments, the voids 2124 can be present between portions of the hydrogel 120. These voids 2124 can be caused by the presence of air bubbles within the protein-fat hydrosol 114 during curing. Without being bound by theory, these air bubbles can be composed of vapor caused by microwave heating of the protein-fat hydrosol 114. In some embodiments, the voids 2124 can have non-uniform shapes and sizes and can provide a desirable non-uniform structure to the hydrogel 120. The voids 2124 can create a layer 2200 within the hydrogel 120 as shown in A and B of FIG. 25. In one aspect, the layer 2200 can be formed by the thread 2130 and the sheet 2128 over the voids 2124 on either side.

[0277] As shown in A and B of FIG. 24, V maxUpon reaching and after the protein-fat hydro sol 114 has hardened, the protein-fat hydrogel 120 may collapse and form a hydrogel meniscus 2121 (as shown in FIG. 28) having a meniscus center 2119 as shown in FIGS. 24A and 24B along the meniscus center line 2126 (as shown in FIG. 28). In FIGS. 24A and 24B, a plurality of voids 2124 can be seen. These voids 2124 can be non-uniform and can range in size from at least about 2 mm, or from 5 mm to 1 cm, or more. The voids 2124 can be formed from bubbles formed during microwave heating. The sheet 2128 may be formed between the threads 2130 of the web-like structure 2160 shown in FIGS. 24A and 24B. The web-like structure 2160 can include threads 2130 and sheets 2128 throughout the collapsed protein-fat hydrogel 120. In some embodiments, the hydrogel meniscus 2121 may be formed from a protein-oil film 2131 that forms the upper portion of the protein-fat hydro sol 120 during hardening. Bubbles can flow through the protein-oil film 2131, enabling the protein-fat hydrogel 120 to collapse and form the hydrogel meniscus 2121.

[0278] As shown in FIGS. 25A and 25B, the layer 2200 within the protein-fat hydrogel 120 can be formed within the hydrogel 120 when the threads 2130 and the sheet 2128 collapse within the protein-fat hydrogel 120. Immediately after collapse and after microwave heating has been stopped, the protein-fat hydrogel 120 can still be malleable and at a high temperature and, in some embodiments, can be formed into different shapes before cooling with cold water. In some preferred embodiments, at least 85%, or at least 80%, or at least 75% of the protein-fat hydrogel is present in the container sidewall adjacent portion, the threads, and the sheet and is not present in the container bottom adjacent portion 2400. This can correspond to the fact that in an Oyster 24 ounce pitcher having a height of about 7 inches, a bottom bottom diameter of about 2.75 inches, and an upper diameter of about 3.75 inches, 100 ml of starting protein-fat hydro sol is approximately optimal, and a width of about 6 mm to 10 mm of the material in the container bottom adjacent portion 2400 is not acceptable for meat analogs.

[0279] A and B in FIG. 26 show the sheet 2128 and layer 2200 of the protein-fat hydrogel 120, as well as the container sidewall adjacent portion 2300 and the container sidewall contact surface 2302. These portions of the protein-fat hydrogel 120 are formed when adhesion to the sidewall of the container 2100 occurs during the expansion and hardening of the product as a result of microwave heating. The container sidewall contact surface 2302 can be substantially flat, similar to the container sidewall 2102, which can be an aesthetic advantage of the meat analog 120 prepared according to the present disclosure in that certain portions and surfaces of conventional meats such as chicken breast can be naturally smooth and flat.

[0280] A - C in FIG. 27 show the container adjacent sidewall portion 2300 and the container sidewall adjacent surface 2302, as well as the container adjacent bottom 2400 and the container adjacent bottom surface 2402. In some embodiments, it is desirable to prevent the protein-fat hydrogel 120 from conforming to the shape of the bottom of the container 2100. In the present disclosure, the conditions for manufacturing the bottom of the protein-fat hydrogel 120 that is not shaped to the shape of the bottom of the container 2100 are described. A in FIG. 27 shows a front perspective view of the protein-fat hydrogel removed from a plastic container, and the bottom of the hydrogel is not substantially shaped to the shape of the bottom of the container. B shows a front perspective view of the protein-fat hydrogel removed from a plastic container, and the bottom of the hydrogel is partially shaped to the shape of the bottom of the container. C shows a front perspective view of the protein-fat hydrogel removed from a plastic container according to the present disclosure, and the bottom of the hydrogel is substantially shaped to the shape of the bottom of the container.

[0281] Figure 28 shows a cross-sectional view of the protein-fat hydrogel 120 within the container 2100 after collapse. It can be observed that a hydrogel meniscus 2121 can be formed within the protein-fat hydrogel 120. This hydrogel meniscus 2121 may result from the degassing of the protein-fat hydrogel 120 after heating is stopped, and since the outer portion of the material is fibered and bound to the sidewall of the container 2100, it leads to a significant reduction in the height of the hydrogel 120 in the central portion of the material. In some embodiments, the depth of the hydrogel meniscus 2121 may be correlated with the quality of the protein-fat hydrogel 120 as a product.

[0282] The hydrogel meniscus ratio can be a useful tool for evaluating the quality of a protein-fat hydrogel for specific applications, such as for manufacturing acceptable meat analogs. The hydrogel meniscus ratio range may be from 0 to 1. Also, there may be cases where no hydrogel meniscus 2121 is formed at all. The hydrogel meniscus ratio is calculated as the ratio of the hydrogel meniscus depth 2127 divided by the hydrogel final height (Hf) 2122.

[0283] At least a part of the bottom of the meniscus is flat with respect to the bottom surface of the container, and if the shape of the meniscus curve at the bottom end is disrupted, meniscus formation may not exist.

[0284] When the bottom tip of the meniscus just touches the bottom of the container 2100, the hydrogel meniscus ratio is 1. When the top layer, or the protein oil film 2131, is flat and has a height equal to H f the hydrogel meniscus ratio is 0. Here, the depth of the meniscus is 0, and the hydrogel meniscus ratio is 0 / 1, where 1 is H f is.

[0285] According to the present disclosure, a hydrogel meniscus ratio of about 0.3 to 0.7 may be preferable for the production of meat analogs. This ratio was calculated using the process of the present disclosure with an Oster® plastic 24-ounce pitcher and a substantially optimal protein-fat hydrogel 114 composition. As discussed elsewhere in the present disclosure, some variation can be expected under different conditions. When the hydrogel meniscus ratio is higher than 0.7, generally, products that are undercooked for meat analogs are shown, while when the hydrogel meniscus ratio is less than 0.3, generally, overcooked products for meat analogs are shown. However, in certain cases, overcooked products may be desirable.

[0286] Hydrogel meniscus formation appears to be unique to the process of the present disclosure in that other protein isolates tested, including soy protein isolate, potato protein isolate, and other commercially available hemp protein isolates, were unable to form hydrogel menisci under the conditions tested and described herein.

[0287] In some embodiments, a protein-fat hydrogel meniscus range of about 0.3 to 0.7 may be preferable. In some embodiments, a range of about 0.4 to 0.6 may be preferable, in some embodiments, a range of about 0.45 to 0.55 may be preferable, and in some embodiments, a hydrogel meniscus of about 0.5 may be preferable. In some embodiments, a hydrogel meniscus ratio of 0.2 to 0.8 may be preferable. In some embodiments, the hydrogel meniscus ratio is 0.3 to 0. In some embodiments, it may be preferable for a hydrogel meniscus ratio of 0 to 1 to exist. In some embodiments, there may be no hydrogel meniscus.

[0288] Furthermore, certain chemical compounds containing carbonate compounds and salts may increase expansion and potentially change the hydrogel meniscus ratio in the present disclosure. Such changes are considered to be within the scope of the invention, even if they may change the claimed scope. In some embodiments of the present disclosure, when using spray-dried NEPI powder, for example, 1% calcium carbonate was added to NEPI250 up to a concentration of 1%. The addition of calcium carbonate while increasing the concentration can promote the expansion of the protein-fat hydrocolloid. This can result in the formation of carbon dioxide gas in the material during microwave heating and an increase in expansion. In some embodiments, calcium carbonate may be added for flavor purposes.

[0289] For example, protein isolates such as soybeans and peas may not form a hydrogel meniscus 2121 under the conditions of the present disclosure, although not bound by theory, due to expansion and the lack of accompanying fibrosis and texturization along the container sidewall. The formation of a hydrogel meniscus 2121, or a significant or substantial hydrogel meniscus 2121, requires significant binding to the container sidewall 2102 in combination with a high degree of gas bubble formation within the protein-fat hydrogel 120. This may not be achievable with prior art materials. As used herein, the meniscus ratio means the ratio of the depth of the meniscus to the final height (H f ) of the protein-fat hydrogel 120, as evidenced by the height of the protein-fat hydrogel 120 bound to the container sidewall 2102.

[0290] After heating in the microwave oven has stopped, the protein-fat hydrogel 120 can be quickly immersed in cold water (if the temperature is in the range of 50°F to 70°F typical of household cold water, it may be sufficient to rapidly cool the product by quickly reducing the product temperature to 100°F or less). Usually, cold water such as that coming out of a household kitchen sink is generally sufficient in terms of temperature. Thereafter, in some embodiments, the meat analog 120 can be separated from the container 2100 by using a spatula to scrape along the inner perimeter of the container sidewall 2102. The meat analog 120 can then be removed from the container 2100 by the spatula or by hand.

[0291] In some embodiments, the meat analog 120 can have a texture similar to that of warmed chicken breast. An advantage of the present disclosure is that, in some embodiments, voids 2124 can be present at or immediately adjacent to the bottom of the container, such that the final product does not have an appearance shaped like the container. Avoiding the shaped appearance significantly improves the aesthetic appeal of the protein-fat hydrogel 120.

[0292] The conditions for manufacturing a microwave-textured protein-fat hydrogel 120 that is not fully shaped to the shape of the container bottom are not limited to these, but can vary depending on many variables including the amount of starting material, the size and shape of the container, the material of which the container is composed, the type of microwave oven, the output of the microwave oven, the temperature of the starting material, and other variables that would be understood by one of ordinary skill in the art, where with routine optimization, it is possible to produce a product that does not appear to be fully shaped to the container bottom. In some embodiments, partial shaping of the material to the container bottom may be acceptable.

[0293] Using the processes described in this disclosure with protein isolates other than those effective in this disclosure generally results in a final product different from the protein-fat hydrogel 120 of this disclosure that is shaped to the shape of the bottom of the container and cannot be shaped after microwave heating. When using a protein isolate prepared according to this disclosure, but where the starting material of the protein isolate is ineffective, it has been observed with soy protein isolate, potato protein isolate, commercially available hemp protein isolate, and other protein isolates that other protein isolate materials will fully conform to the bottom of the container. In some embodiments, a thin or brittle layer of the protein-fat hydrogel 120 material may be present at the bottom of the material that can be shaped to the shape of the bottom of the container, but is thin or brittle such that at least a portion of its shape is lost when removed from the container.

[0294] In some embodiments, the meat analog 120 can be, for example, sliced and eaten in a salad, for example, without further cooking. The meat analog 120 can also be sautéed in a frying pan and browned such that the chicken breast turns brown and can be sautéed.

[0295] As shown in FIGS. 29A-C, the container 2100 may have different shapes or may include an insert 2500. In some embodiments, the insert 2500 may have a central cylindrical protrusion 2510a, as shown in FIG. 29B. The central cylindrical protrusion 2510a can create a donut-shaped space that can be occupied by liquid. In some embodiments, the insert 2500 can be added after the blending step to avoid interference with the immersion blender 2600. The insert 2500 may be composed of a solid microwave-heatable material or a hollow material such that hot water 2002 cannot enter the insert 2500. In some embodiments, the insert 2500 may be foldable and can be folded from the sidewall of the container or folded up from the container base.

[0296] In some embodiments, the insert protrusion 2510 may have a peninsula shape with respect to the container sidewall, and the sidewall protrusion 2510b can be formed, such that the protrusion 2510 extends from the sidewall to the center of the container 2100 and can form a horseshoe-shaped chamber that the hot water 2002 can occupy. The insert 2500 may have other geometric shapes including rectangular or triangular. The insert 2500 can be composed of a microwave-heatable material as previously described herein. The insert 2500 can have an insert base 320 with a shape corresponding to the container base. The insert base 2520 can provide stability to the insert 2500. The insert 2500 can be removable or foldable and can be connected to the container sidewall 2102 or the container base 2104 to enable access to the immersion blender 2600 (shown in FIG. 30). In some embodiments, the insert base 2520 can have an opening or hole to allow liquid flow. In some embodiments, the container 2100 can have ridges, cavities, sidewalls, or grooves in the container sidewall 2102 and the container bottom 2104, or can have paper inserts of various shapes and sizes.

[0297] In some embodiments, as shown in FIG. 31, the indicator lid 2700 may be placed on top of the protein-fat hydrogel 114 in the container 2100 before heating and can be used to more easily visualize the point at which the swollen protein-fat hydrogel 2110 reaches its maximum height within the container 2100. In some embodiments, the indicator lid 2700, or another lid, can be used to prevent steam from leaking out of the container 2100 during heating, provide a more hydrated product, or provide some pressure as the swollen protein-fat hydrogel 2110 expands and rises.

[0298] With respect to FIG. 32, the materials required for the production of the instant meat analog 120 by the microwave texturing process 2000 can be provided as part of a kit 2800. The kit can include a container 2100, a NEPI packet 2802 (which includes a protein isolate and, in some embodiments, may also include a desired chicken flavor or other meaty flavor), bottled oil 2804, and optionally, a container jacket 2806, an indicator lid 2700, any inserts 2810, or a flavor packet 2812 (which may contain additional flavorings or spices in some embodiments).

[0299] Regarding the source of the NEPI 250 material from which the microwave-textured protein-fat hydrogel 120 is produced, as previously described herein, spray-dried powder or concentrate can be used. In some embodiments, when texturing the protein-fat hydrogel 120 in a microwave oven, the NEPI 250 powder may be more effective than the NEPI 250 concentrate.

[0300] Without being bound by theory, the shorter the microwave heating time, the better the final product produced. Thus, it may be desirable to produce a sufficient amount of the protein-fat hydrogel 120 with the shortest heating time possible. Excellent texturing may correlate with the degree of expansion of the material during microwave heating.

[0301] In some embodiments, as shown in FIGS. 26A and 26B, after the protein-fat hydrogel 120 is removed from the container, it may be desirable to fold or roll it. In some embodiments, the protein-fat hydrogel 120 is foldable. Folding can result in an appearance similar to that of chicken breast or other types of meat analogs. In one embodiment, the protein-fat hydrogel 120 may be rolled such that the smooth container sidewall contact surface is on the outer surface of the folded or rolled meat analog 120, and the more textured portions of the meat analog 120 include the inner portions of the folded or rolled meat analog 120 as shown in FIGS. 26A and 26B.

[0302] In some embodiments, the present disclosure contemplates the use of protein isolates made from seeds or grains that can have properties similar to edestin as described in this application. This can include, among others, globulins from pumpkin and squash (Cucurbita moschita and Cucurbita maxima), watermelon (Citrullus vulgaris), cucumber (Cucumis sativus), tobacco, and cottonseed.

[0303] In some embodiments, after microwave heating and before cooling with water, the protein-fat hydrogel 120 may be shaped as shown in FIG. 20. Shaping may be performed using a tool such as a spatula. The minimum temperature for shaping can be about 150°F, or 160°F, more preferably 170°F, up to about 212°F. Since the protein-fat hydrosol 114 is an aqueous liquid and does not exceed the boiling temperature, the temperature inside the microwave oven generally does not get higher than boiling for the protein-fat hydrosol 114. However, generally, the product may be shaped between about 150°F and 212°F, and 150°F can make it more difficult to shape because the layers or portions of the protein-fat hydrogel 120 may not adhere to each other when pressure is applied.

[0304] The shaping may be performed within the container 2100. In some embodiments, shaping may enable the user to eliminate the molded appearance of the hydrogel 120 that initially presents the shape of the bottom of the container 2100. The protein-fat hydrogel 120 may be shaped to have, for example, a structure similar to chicken breast, or other shapes that may be more aesthetically appealing or more convenient for consumption.

[0305] In some embodiments, calcium carbonate may be added to the NEPI250, which may increase the expansion ratio of the protein-fat hydrosol 114 under some conditions. Without being bound by theory, calcium carbonate may act as a catalyst to promote the expansion of the protein-fat hydrosol 114. Calcium carbonate, sodium carbonate, and sodium hydroxide may increase expansion and fibrillation when combined with NEPI250. Calcium chloride does not appear to affect the expansion ratio. In some embodiments, by adding about 0.5% calcium carbonate to the protein-fat hydrosol 114, the expansion ratio may increase significantly. Depending on the amount of a particular carbonate compound or other compounds that increase expansion, a relative increase may occur in relation to the amount of compound added to the protein-fat hydrosol 114.

[0306] Using a NEPI250 concentrate or dry NEPI250 that is rehydrated with hot water and combined with oil 110 to form a thick hydrosol, placed in a microwaveable container, dish, or tube and exposed to microwaves, first heats enough water to "dissolve" or transfer the hydrosol-gel and forms a protein-oil film sufficient to hold the water within the membrane so that the water turns into gas, it can be confined within the membrane and forced to expand and then cool, as a result, the hydrogel solidifies. As a result, heating the hydrosol blend of NEPI250, water, and oil 110 to a temperature higher than the boiling point of water can create a fiber structure similar to cooked meat.

[0307] This is disclosed in the prior U.S. patent application Ser. No. 17 / 551,163, specifically, when NEPI is hydrated, relaxed, and blended with oil to form a hydrosol, the hydrosol is cured to a hydrogel by a direct or indirect external heat source such as a stove, oven, frying pan, steam, pressure extrusion, or IR heating. Surprisingly, the applicant has discovered that when the protein-fat hydrosol 114 suspension is heated with microwaves (e.g., in a microwave oven), a unique and unexpected fibrous structure is formed. It is hypothesized that the water encapsulated within the hydrated protein-fat hydrosol 114 suspension heats the protein-fat hydrosol 114 by microwave activation until the water molecules reach a transition point at which the curing or "melting" of the protein-fat hydrosol 114 is initiated.

[0308] As the water molecules continue to change to vapor (gas), the water molecules can be uniquely trapped within the curing hydrogel. The gas can expand the curing or molten hydrosol 114 until the protein-fat hydrogel 120 is fully cured. A similar example can be found in the glassblowing industry, where air is blown into a sphere of liquid molten glass to expand the glass before it cools to a solid. As a result, an infinite number of shapes and forms can be obtained. However, similar to the case of glassblowing, the temperature of the glass, the amount and rate of gas addition can be important to achieve the desired result.

[0309] If the protein-fat hydrogel 114 is heated too quickly (between the temperature at which the transition melt forms and the temperature at which vapor forms), the transition of the hydrogel 114 to the hydrogel 120 becomes too rapid, and a "melt" of the hydrogel 114 that could capture or influx existing water during the hardening to the hydrogel 120 may not be formed. Stirring may exacerbate this effect. The slower this process is, the more water may influx and the greater the initial expansion may be. If the temperature of the internal water remains slightly below 100 °C, i.e., the boiling point of water, the maximum amount of water may influx. When the temperature of the water exceeds the boiling point of water, the resulting gas begins to expand the "melt" of the transitioning protein-fat hydrogel 114, which here may be partially composed of the protein-fat hydrogel 120 until the hardening of the protein-fat hydrogel 120 is complete. Cooling the protein-fat hydrogel 120 can terminate and stabilize the hardening from the melt.

[0310] Described herein are conditions using a microwave oven that allow the influx of water when transitioning from the protein-aliphatic hydrogel 114 to the protein-aliphatic hydrogel, while at the same time allowing the gas formed to forcefully expand the melted protein-fat hydrogel 114, thereby creating a unique structure that simulates the texture and fibers typically associated with meat.

[0311] What is unique to the NEPI hydrogel is that the melting temperature of the protein-oil hydrogel is lower than the boiling point of water. Without being bound by theory, it is only because of the low melting temperature that water can first fully flow into the protein-oil film 2131. Soybean or pea isolate products extruded at high temperatures are usually carried out between 130 °C and 140 °C, "melting" the protein, generating fibers, and exiting the extruder with less than 10% moisture. In this case, water cannot be retained within the structure of the hydrogel 114 type. Therefore, soy or pea type texturized vegetable protein (TVP) initially has very little encapsulated moisture and must be rehydrated while adding many ingredients such as starch and gum to suspend and retain moisture.

[0312] Microwaves may not destroy proteins (proteins and fats are zero or unaffected by microwaves). Due to the increase in heat of water molecules, the protein-oil film 2131 may first begin to harden as a hydrogel. However, before the gel fully solidifies and hardens, the activated water molecules are converted into gas, and almost simultaneously with being encapsulated in the formed hardened gel, the hydrogel expands when it is converted to and hardened into a hydrogel 120. Eventually, a portion of the vapor is released, and the released vapor destroys a portion of the open protein membrane. As a result, the structure collapses, and subsequent condensation can cool the hydrogel, which enables the hydrogel to fully and irreversibly harden after being stretched in its unique formation. What is important and decisive is that water continues to be incorporated into the hydrogel. The unique moisture activity characteristics of the protein-fat hydrogel 120 can be quantified using a moisture activity assay with a moisture activity meter that can measure the moisture activity of solid or semi-solid materials such as hydrogels or meat slices.

[0313] It is hypothesized that a similar situation may occur with NEPI hydrogels heated with microwaves. Unlike many other proteins, including soy or pea isolates that require temperatures above 140 °C to melt and elongate, the fact that NEPI250 protein - aliphatic hydrogel 114 can "melt and elongate" at temperatures below the boiling point of water allows water to be incorporated as it turns into gas and the resulting web - like structure 2160 expands.

[0314] In this case, the Applicant has found that the melting and hardening of the protein at a temperature lower than the gas formation of water at 100 °C allows the enclosed water to turn into gas, thereby making the simultaneous melting and expansion of the protein - oil film more or less possible, and thereby releasing the leaked water, condensing the steam back into water, or confining the water within the protein - oil hydrogel structure to expand and structure the hydrogel before the hydrogel is cooled to its final hardening.

[0315] Additional tests may be conducted to identify additional effective conditions for manufacturing the protein - fat hydrogel 120 according to the present disclosure. For different hemp protein isolates, it can be carried out with commercially available hemp protein isolates, such as those described in US Patent Application No. 17 / 551,163 to Victory Hemp and Matchell Ellis, to further characterize the differences in capabilities. The tests may be conducted under the same conditions as those described in the present disclosure, observing the resulting texture and comparing it with the products obtained from the use of NEPI250.

[0316] Further tests using both plant - based and animal - based saturated to unsaturated oils may be conducted under the same conditions as those described in the present disclosure, observing the texture of the resulting products and comparing them with the products obtained from the use of NEPI.

[0317] Further test the influence of container sizes ranging from 65 ml to 550 ml. By testing the influence on the texture at intervals within this range, different influences can be identified. Preferably, the shape of the container is consistent, but only the diameter or width of the container can vary. Such tests according to the methods described in this disclosure can identify additional effects and advantages of this disclosure. The ratio of the size of container 2100 to the starting liquid volume of protein-fat hydrogel 114 is important for the structure of protein-fat hydrogel 120. To optimize the results, the microwave output may also be changed as the container size changes.

[0318] In some embodiments of this disclosure, it may be important to maximize the encapsulation of water within the hydrogel for favorable results. In some embodiments of this disclosure, the water content of protein-fat hydrogel 120 may correspond to the water content of normal meat.

[0319] Additional tests of the container material may also result in different effects based on different container materials. In some embodiments of this disclosure, the adhesion or binding of protein-fat hydrogel 114, or protein-containing material, to the inner sidewall of the container is important for texturization. If the binding is too strong, it may not be possible to effectively remove the hydrogel from the sidewall of the container, and washing may become difficult. Ceramic materials, especially unglazed ceramic materials, may enable effective microwave heating, expansion, and binding of protein-fat hydrogel 114 to the sidewall of the container. However, due to the strength of the binding of poly-fat hydrogel 114 to the sidewall and the presence of pores in the ceramic material, the container is difficult to wash after use. Using known or unknown methods, such as immersing container 2100 in milk, heating to caramelize the milk, and filling the pores, an unglazed ceramic material with filled pores can be used to produce a more effective container for in vivo use. This disclosure.

[0320] To potentially identify different effects on the texture of the protein-fat hydrogel 120, additional tests regarding the indicator lid 2700 may be performed. Without being bound by theory, the lid 2700 enables the inflow of water within the protein-fat hydrogel 120 and the heating rate within the microwave oven and can have an impact. The weight, size, position, and material of the lid can be varied in accordance with the present disclosure to potentially change the texture by affecting the heating rate and potentially allow different amounts of water to flow into the material. In some embodiments, the lid 2700 can be composed of any microwave heatable material including, but not limited to, plastic, paper, glass, and ceramic. In some embodiments, the lid 2700 can be disposed within the container 2100, and in other embodiments, the lid 2700 can be disposed on top of the container 2100. The lid 2700 can be adapted to rise as the protein-fat hydrosol 114 expands and rises. In some embodiments, the lid 2700 can be adapted to increase the pressure within the container 2100.

[0321] To achieve different effects regarding the addition of flavorants to the protein-containing material, further tests may be performed. In some embodiments, the flavorant can be added to the water during the process before adding NEPI250 to the water. In other embodiments, the flavorant may be blended with the NEPI250 powder or concentrate. In some embodiments, the flavorant can be mixed with the oil 110. In some embodiments, the flavorant can be blended after the formation of the protein-fat hydrosol 114.

[0322] The presence of albumin or albumin-containing complexes in hemp grain protein isolates may prevent the hemp grain protein from forming a protein-fat hydrogel 120 having an appropriate texture. Our data show that when albumin, or albumin-containing complexes are separated from the protein-fat hydrogel and then reintroduced into the NEPI250, an acceptable protein-fat hydrogel is not produced using the process described in this disclosure. Protein-fat hydrogel 120 with reintroduced albumin becomes softer and less elastic when cured in a microwave oven with 25% albumin and 75% edestin. In some embodiments, it is also contemplated that very low levels of albumin that may be present after production of NEPI according to this disclosure may improve the texture. However, substantial amounts, or too much albumin or albumin-containing complexes result in a low-quality product in terms of texture.

[0323] Example 7 With respect to Example 1, a plant-based chicken analog was produced according to the process of this disclosure. As shown in Table 1, for test sample 1, hot water was first added to a 1 / 3 measuring container and then to a plastic container with dimensions of 2.5 inches × 8 inches and a volume of 20 ounces. For test sample 2, as shown in Table 1, 65.0 grams of boiling water was first added to the measuring container and then to a straight-wall Pyrex glass beaker with dimensions of 2.5 inches in diameter and 10 inches in height and a volume of 400 ml. For test samples 1 and 2, the protein hydrosol was then mixed with an immersion blender (Braun Multiquick MQ7025x) for about 1 minute.

[0324] Next, as shown in Table 1, vegetable oil (Crisco) was added to the protein hydrogels of both samples to produce protein-fat hydrogels. The protein-to-fat ratio of Test Sample 1 was 1.96:1. The protein-to-fat ratio of Test Sample 2 was 2.04 to 1. Next, the samples were mixed using an immersion blender (Braun Multiquick MQ7025x) for about 1 minute for the protein and water, and about 30 seconds when adding oil to the protein hydrogel. Each sample was heated in a Bosch (registered trademark) microwave (model number HMC54151UC / 05, manufactured in May 2018). The heating time of Test Sample 1 was 1 minute and 25 seconds. The heating time of Test Sample 2 was 1 minute and 58 seconds. TIFF2025508562000012.tif55135

[0325] Based on visual observation during heating in the microwave, Test Sample 1 expanded sufficiently and rose as it was heated in the microwave. When heating was stopped and the product was immediately rinsed with cold water, the product adhered to the side of the container. After immersing in cold water to cool and scraping off to remove the hydrogel, the hydrogel showed fibrous curls, and the hydrogel had a depth exceeding 1 inch and a length of about 3 inches. When sliced, the hydrogel resembled a chicken breast slice of about 1 / 4 inch × 2 inches and had good tensile strength and chewiness due to characteristics similar to warm chicken.

[0326] Based on visual observation during heating in the microwave, Test Sample 2 expanded sufficiently and rose as it was heated in the microwave. When heating was stopped, the product adhered to the side of the container. After immersing in cold water to cool and scraping off to remove the hydrogel, the hydrogel was hardened firmly so that its shape did not change further. Its shape had several outward curls and protrusions similar to those expected from shredded chicken.

[0327] Other test samples did not yield satisfactory results when appropriate process parameters were not used. For example, for cooking the hydrogel, in contrast to container materials having inner sidewalls of paper and plastic, glazed ceramic containers having very smooth inner sidewalls, or preferably, microwave-heatable materials having a rough or irregular surface were used, and it was revealed that the protein-fat hydrogel swelled but did not bind to the container sidewalls as desired. However, when the walls were lined with parchment paper after adding the protein-fatty hydrogel, the protein-fatty hydrogel swelled very rapidly and collapsed after reaching a height above the edge of the ceramic container. The meat analogue obtained from the use of parchment paper was thin but very fibrous and had excellent tensile strength. However, in this embodiment, there was no "sliced" type interior for the product; instead, there was a web of irregularly shaped, very thin, fibrous meat slices.

[0328] Example A In Example A, test samples A-1 to A-10 regarding the protein-fat hydrogel were prepared by U.S. Patent Application No. 17 / 551,163 to Matchell Ellis. This process can be used to prepare chicken analogues. The composition of the test materials is shown in Table A.

[0329] Example 8 Table 10 was created using the same process as Table 10A as described above. TIFF2025508562000013.tif57135 TIFF2025508562000014.tif127135 TIFF2025508562000015.tif198135TIFF2025508562000016.tif185135TIFF2025508562000017.tif69135 TIFF2025508562000018.tif150135

[0330] Example 9 Example 9 shows that about 17% - 38% NEPI in a solution generally prepared according to the description of Example 1 is effective in producing an acceptable meat analog according to the present disclosure. Example 9 shows that 0% - 50% oil in a solution generally prepared according to the description of Example 1 is effective in producing an acceptable meat analog according to the present disclosure.

[0331] At least sufficient water must be present in the protein - fat hydrogel to fully hydrate and unwind the NEPI. The amount of water required to fully hydrate and unwind the NEPI can be about 30% of the protein hydrogel. The maximum concentration of water in the protein - fat hydrogel is about 80%. TIFF2025508562000019.tif93135 TIFF2025508562000020.tif153135 TIFF2025508562000021.tif74135TIFF2025508562000022.tif68135 TIFF2025508562000023.tif129135 TIFF2025508562000024.tif81135

[0332] Example 10 Test of the effect of albumin complex on protein - fat hydrogel The formulations used in Table 20 were made using products prepared according to the procedure described in Example 7. In this experiment, a total of 5 meat analog samples with 6 replicates were used for each time point. The prepared meat analog pieces were cut to dimensions of 50×15×15 mm. Values were measured using a texture analyzer (TA.XTplus, Sable Microsystems) equipped with a Warner - Bratzler blade, a 30 kg load cell adjusted with a descent and entry speed of 2.00 mm / second, an entry depth of 30 mm, and a contact force of 10 g.

[0333] Device Texture Analyzer - TA.XTPlus Connect Texture Analyzer 650H s / n 2 - P6_Z11140 - 01 - V003C98CB Texture Analyzer Probe - TA - 007

[0334] These results indicate that when the concentration of the albumin - containing complex is high, the intensity of texture analysis decreases. This result shows that meat analogs with increased albumin complex concentration have reduced strength and toughness, become softer, and have a texture that is no longer acceptable as a meat analog. TIFF2025508562000025.tif75135 TIFF2025508562000026.tif192135 TIFF2025508562000027.tif32135JPEG2025508562000028.jpg74135

[0335] Example 11 Evaluation of the Effect of Chemical Additives on Protein - Fat Hydrogels The following chemical additives can affect the expansion ratio of protein - fat hydrogels. In some cases, they increase the expansion ratio of protein - fat hydrogels, and in other cases, they decrease the expansion ratio of protein - aliphatic hydrogels. TIFF2025508562000029.tif187135

[0336] Example 11 Evaluation of the influence of container material composition and container structure, including the size and shape of the container. Certain container materials were effective in binding or adhering protein-fat hydrogels to the container sidewalls during and after the swelling of the protein-fat hydro sols and hydrogels. These materials were specific plastic containers including polycarbonate plastics, and the binding was enhanced by polishing the inner surface of the sidewalls of the container. Paper containers were also effective in binding protein-fat hydro sols and hydrogels.

[0337] The shape of the container was important to produce the desired swelling. An increase in the diameter of the container resulted in a negative synergistic effect on the swelling ratio of the protein-fat hydro sol, while reducing the diameter of the plastic container had a synergistically positive effect on the swelling ratio. In the examples tested, a diameter of 3.25 inches was not effective in producing swelling, while a diameter of 2.75 was effective in producing acceptable swelling and fibrillation. TIFF2025508562000030.tif165135 TIFF2025508562000031.tif127135

[0338] List of symbols: IpH = Initial pH, WOpH = pH after oil addition, IV = Initial volume, V = Viscosity, IH = Initial height, IW = Initial weight, IWT = Initial water temperature, SMT = Starting material temperature, MPS = Microwave output setting, MCT = Microwave cooking time, MV = Maximum volume, MR = Meniscus ratio, FH = Final height, FMT = Final material temperature, FW = Final weight, WL = Water loss, FD = Final density, BD = Bottom diameter, TD = Top diameter, ITS = Initial TS, FTS = Final TS, F = Failure, A = Acceptable, P = Preferred, FA / TF = Functionally acceptable / Taste non - conforming, VH = Victory Hemp competing product, VH = Victory Hemp competing product, product, HL = Hemp Land competing product, PLA = Polylactic acid, PMP = Polymethylpentene, PP = Polypropylene, PE = Polyethylene, UKR = Ukrainian, NEPI = Native edestin protein isolate, E = Edestin, AC = Albumin complex E1 = 100% NEPI EA2 = 90% NEPI + 10% AC (Albumin complex) EA3 = 80% NEPI + 20% AC (Albumin complex) EA4 = 70% NEPI + 30% AC (Albumin complex) EA5 = 60% NEPI + 40% AC (Albumin complex) TS = Total solids NEPI in Tables 10 - 25 = Dehulled NEPI NEPI in Tables 10 - 25 = Industrially manufactured, except for Example 10 where NEPI was manufactured on a bench top The higher the microwave output, the finer the fibers produced

[0339] In Tables 10 - 25, using the "x" unit for the description of expansion means that the height of the 24 - ounce Oster container has increased by "x" times. This means that if 1x is 1 inch in height, the Oster container at 3x will be approximately 3 inches in height. In the Oster container, a height of 4 inches in the liquid - holding chamber corresponds to approximately 500 mL. 2 inches corresponds to approximately 225 mL. 3 inches corresponds to 350 mL. 4 inches corresponds to 500 mL. 5 inches corresponds to 675 mL

[0340] Examples 7 to 11 utilized a process of blending generally the same NEPI as in Tables 10A and 10B, unless otherwise specified.

[0341] Other Embodiments Although the invention has been described in conjunction with its detailed description, it should be understood that the foregoing description is intended to be illustrative and not limiting of the scope of the invention, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. A process comprising: adding a protein-fat hydrosol to a container, said protein-fat hydrosol comprising native edestin protein isolate, said container having a bottom and at least one sidewall; placing the container in a microwave oven; heating the protein-fat hydrosol with microwaves; forming a plurality of gas bubbles in said protein-fat hydrosol; Swelling the protein-fat hydrosol; and curing the protein-fat hydrosol to form a web-like protein-fat hydrogel, the web-like protein-fat hydrogel being heterogeneous and comprising at least one thread, at least one sheet, at least one container adjacent sidewall portion, and a plurality of voids; and separating the web-like protein-fat hydrogel from the container.

2. 10. The process of claim 1, further comprising shaping the protein-fat hydrogel after curing the protein-fat hydrogel and prior to substantially cooling the web-like protein-fat hydrogel.

3. 2. The process of claim 1, wherein the concentration of the native edestin protein isolate in the protein-fat hydrosol is at least 15% by weight.

4. 10. The process of claim 1, wherein the microwave oven is set at a power of 4 to 7 for a conventional domestic microwave oven.

5. The process of claim 1 , wherein a hydrogel meniscus is formed in the web-like protein-fat hydrogel.

6. The process of claim 1, wherein the hydrogel meniscus ratio is between 0.3 and 0.

7.

7. The process of claim 1 , wherein the at least one sidewall comprises at least one continuous sidewall.

8. The process of claim 1 , wherein the at least one sidewall allows for significant expansion of the protein-fat hydrosol.

9. The process of claim 1 , wherein at least one of the voids has a diameter of at least 3 mm.

10. The process of claim 1 , wherein at least one of the threads has a width of at least 5 mm.

11. A process comprising: adding a protein hydrosol to a container, the protein hydrosol comprising a protein isolate, the container having a bottom and at least one sidewall; allowing significant expansion of the protein hydrosol; placing the container in a microwave oven; adjusting the time and power of the microwave oven; heating the protein hydrosol; forming gas bubbles in said protein hydrosol; Swelling the protein hydrosol; and curing the protein hydrosol to form a web-like protein hydrogel, the web-like protein hydrogel being heterogeneous and comprising at least one thread, at least one sheet, at least one container-forming sidewall portion, and a plurality of voids; cooling the web-like protein hydrogel; and and separating the web-like protein hydrogel from the container.

12. 12. The process of claim 11, wherein fat is added to the protein hydrosol to produce a web-like protein-fat hydrosol prior to placing the container in the microwave oven.

13. 13. The process of claim 12, further comprising cooling the web-like protein-fat hydrogel with an aqueous liquid.

14. 12. The process of claim 11, wherein the protein isolate is a native edestin protein isolate.

15. 10. The process of claim 1, further comprising cooling the web-like protein-fat hydrogel with an aqueous liquid.

16. A process comprising: 、 adding a protein-fat hydrosol to a container, the protein-fat hydrosol comprising a protein isolate, the container having a bottom and at least one sidewall; placing the container in a microwave oven; heating the protein-fat hydrosol in microwaves; forming a plurality of gas bubbles in said protein-fat hydrosol; Swelling the protein-fat hydrosol; and curing the protein-fat hydrosol to form a web-like protein-fat hydrogel, wherein a hydrogel meniscus is formed in the web-like protein-fat hydrogel; cooling the web-like protein-fat hydrogel; and and separating the web-like protein-fat hydrogel from the container.

17. 17. The process of claim 16, further comprising cooling the web-like protein hydrogel.

18. 17. The process of claim 16, further comprising cooling the web-like protein hydrogel with an aqueous liquid.

19. 17. The process of claim 16, wherein the protein isolate is a native edestin protein isolate.

20. 17. The process of claim 16, wherein the web-like protein-fat hydrogel is heterogeneous and comprises at least one thread, at least one sheet, at least one container adjacent sidewall portion, and a plurality of voids.

21. 1. A composition comprising a web-like hydrogel, the web-like hydrogel is produced by microwave heating a substantially liquid material selected from the group consisting of a protein hydrosol containing a protein isolate and a protein-fat hydrosol containing the protein isolate; the web-like protein-fat hydrogel is heterogeneous and comprises at least one thread, at least one sheet, at least one container-adjacent sidewall portion, and a plurality of voids; The composition.

22. 22. The composition of claim 21, wherein the protein isolate is a native edestin protein isolate.

23. 1. A composition comprising a web-like hydrogel, the web-like hydrogel is produced by microwave heating a substantially liquid material selected from the group consisting of a protein hydrosol containing a protein isolate and a protein-fat hydrosol containing the protein isolate; The web-like hydrogel comprises a hydrogel meniscus. The composition.

24. 24. The composition of claim 23, wherein the protein isolate is a native edestin protein isolate.

25. The composition of claim 23, wherein the hydrogel meniscus ratio is from 0.3 to 0.7.

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