Ready-to-eat textured meat substitute

Abstract

A method of producing a ready-to-eat alternative protein product, including plant-based meat, and more particularly plant-based products having the texture, appearance and taste of meat or dairy products. Ready-to-eat meat or food mimicking materials may be based on natural hemp seed globulin isolate (NEPI). The NEPI may be combined with water to form a proteinaceous hydrosol, then added with oil, and heated to solidify in a microwave oven to form a hydrogel or meat imitation product. The protein hydrosol may be mixed in a microwaveable cup, which is preferably composed of a porous material, such as paper or plastic, and which is sized to facilitate the formation of a meat imitation product from the protein-fat hydrosol upon microwave heating. Materials required for the production of meat mimics from the NEPI at home can be provided as a convenient kit for the production of ready-to-eat meat mimics.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Patent Application Serial No. 63 / 318,183, filed on March 9, 2022. The contents of each application are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure relates to the production of alternative protein products, including plant-based meat and dairy analogs, and more particularly to plant-based products that have the protein content, texture and appearance of meat or fish and the ability to be seasoned as desired. Background Art

[0004] Consumer standards for ready-to-eat home-cooked meals have evolved rapidly in recent years, and demand for products that can be prepared at home has continued to increase. When cooking ready meals at home, consumers have always expected meals to be quick to prepare and delicious. However, more recently, consumers are demanding ready meals that are fresh, healthy, clean label, plant-based and sustainably produced.

[0005] One method of producing a ready-to-eat, textured plant-based meat substitute at home involves using dry vegetable protein (typically pre-texturized vegetable protein) as an ingredient or component that provides texture. Ready-to-eat foods, as used herein, can be defined as convenient foods that require minimal preparation and typically involve only adding one or two components to a pre-prepared base composition. The preparation time for a ready-to-eat food may be less than five minutes. Typically, the proteins used as ingredients for preparing plant-based imitation meat instant foods are derived from soy or peas, however, products using these proteins are generally not commercially successful. Ready-to-eat products or imitation meat and imitation dairy products based on hemp seeds are generally not described in the food industry or food science literature. When used as an ingredient in food, hemp seed protein is generally considered inferior to soy and pea proteins, especially in terms of the properties required for producing imitation meat products.

[0006] Currently, soy and pea protein-based products dominate the meat-like product market. and Beyond Companies such as and others produce a large number of imitation meat products. The imitation meat products produced by these companies are all pre-prepared and use a large number of ingredients, and are usually sold in pre-packaged form like fresh or frozen animal meat products. Most pre-prepared plant-based imitation meat products must be cooked before consumption, usually using a stove or oven. The manufacture of these products usually requires extrusion at relatively high temperatures of about 120°C to 140°C to obtain texturized proteins. Pre-texturized and post-texturized proteins are usually blended with other ingredients (such as cellulose, starch, sugar, oil and adhesives) to form a protein blend with the appearance of fresh hamburgers or other animal meat products. These products also usually require immediate microbial stabilization through continuous refrigeration or freezing, and are packaged before sale. Currently, these types of products have no effective claims in terms of sustainability and as clean label materials.

[0007] Soy, pea, wheat gluten and other protein isolates used to produce conventional imitation meat products cannot form a product with good texture or similar to meat without pre-extruding the protein. U.S. Patent No. 3,662,673 issued to Boyer discloses the use of microwaves to produce texturized protein products. However, the expansion of texturized protein products produced using proteins described in Boyer (e.g., soy protein isolates) or even by traditional soy tofu processing requiring chemicals such as calcium chloride is low, and the resulting products are not accepted by most consumers. The texture of products typically produced according to the Boyer method or soy tofu processing is essentially uniform in cross-section and lacks the elasticity, fibrosis and texture of meat.

[0008] Soy and pea protein isolates cannot be properly textured without first being conventionally extruded at temperatures well above 100°C, which limits their usefulness in fast or ready-to-eat meat and dairy-like applications. In addition, the extrusion process temperature is above 100°C, which limits the water content in extruded textured soy and pea proteins, so the extruded textured proteins need to be pre-hydrated before use. Importantly, after extrusion, the protein is destroyed, so its ability to retain water within the protein structure itself (rather than between protein pockets like a sponge) is destroyed, so other ingredients are needed to retain water to mimic the natural moisture retained within the meat structure.

[0009] Thus, there is a clear need for a ready-to-eat, fresh, texturized, plant-based meat analog product that replicates the texture of meat or fish; that can be easily made at home, if desired, without the need for industrial equipment or chemicals (such as extruders and distiller or calcium chloride); that does not require the use of temperatures above 100°C to texturize the proteins; that does not contain allergenic substances such as soy or wheat gluten; and that does not require multiple ingredients such as starches, gums, and emulsifiers to retain the necessary water and oil to mimic the meaty texture and nutritional content of a variety of meat and fish analog products.

[0010] Hemp seed-based meat or dairy products produced using only hemp seed as a protein source have not been marketed or described in the food industry or food science literature. Hemp seed protein is considered inferior to soy and pea proteins for use in food products, especially in terms of the properties required to produce meat and dairy products. Meat and dairy products, which may also be referred to herein as structured protein foods, require proteins that can form a strong gel matrix, and hemp seed protein has not been found to have strong capabilities in this regard.

[0011] According to Wang, “The emulsification and gel-forming properties of hemp seed protein were generally found to be inferior to those of soy protein.” (Wang et al., 2019). While Malomo demonstrated that salt micellization separation of hemp seed protein improves its gel-forming ability, Shen revealed that this complex, expensive, and time-consuming protein separation method can negatively affect the protein structure and may require chemical cross-linking agents to obtain sufficient gel-forming ability in hemp seed protein. (Shen et al., 2021; Malomo et al., 2014; Wang et al., 2019).

[0012] As Wang points out, soy protein is currently preferred over hemp protein for the production of meat-like products. “Currently, most soy proteins are used to simulate animal proteins because they have good gelling properties and thus produce an interlaced fiber 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, mainly due to health and nutrition-related concerns about soy products, IMPOSSIBLE FOODS’s largest competitor, BEYOND MEAT, uses yellow pea protein in its BEYOND BURGER. However, yellow pea “has a much lower gelling ability than soy protein” and “the heat-induced gel of soy protein isolate (SPI) is stronger than that of pea protein isolate (PPI).” (Schreuders et al., 2019).

[0013] While soy and pea proteins have known taste, texture, and phytochemical limitations as protein sources for meat and dairy analogs production, no plant-based alternative protein substitutes have been found to be successful for use in meat analogs. However, hemp protein has been investigated as a potential replacement for soy protein in meat analogs. Recently, Zahari reported that while hemp protein has been recognized for its excellent nutritional and functional properties, it has not yet been used in meat analogs production. "Previous work has shown that hemp protein, in particular, has high protein quality and functionality. However, no studies have used hemp protein as a raw material for meat analogs production." (Zahari et al., 2020).

[0014] Zahari goes on to demonstrate that hemp protein concentrate (HPC) can be used in combination with soy protein isolate (SPI) to produce meat analogs via conventional extrusion, but not as the sole protein source. The study concluded, "HPC may therefore be a promising new material to include in extruded products, and this study demonstrates that the resulting meat analogs have a texture comparable to SPI alone, and that soy protein can be replaced by up to 60% with hemp protein." (Zahari et al., 2020). Regarding the use of higher concentrations of hemp protein in the formulation, the study showed that this resulted in an unacceptable reduction in the hardness and chewiness of the meat analogs. Therefore,

[0015] Zahari, Wang, and Shen teach not to use hemp protein as the sole protein source in the production of meat analogs.

[0016] Despite the need for new and improved plant protein sources to meet the growing demands of the plant-based food industry, hemp protein has yet to gain significant market share in food production. Despite the nutritional and environmental advantages of hemp, soy and pea proteins continue to dominate the plant-based food market. Soy and pea proteins have benefited from decades of research and widespread commercial availability and use in meat analog production and other foods, which has greatly improved ingredient and product quality as well as scale cost-effectiveness.

[0017] Improvements in soy and pea-based imitation meat products have been achieved through extensive research and development at all stages of the production of imitation meat products and imitation dairy products. The production of imitation meat products generally involves four steps. The first step involves isolating the protein from the selected plant material. 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 thermal gelation or extrusion of the raw materials to coagulate and organize the protein. The last step is to use adhesives and water-binding agents, such as carrageenan, cellulose fibers, starch, gluten or flour, to form imitation meat products, which can then be cooked to simulate products such as hamburgers, cutlets, chicken nuggets and pulled pork.

[0018] The first step in the production of meat-like products involves the isolation of proteins from plant materials. Typically, soy and pea proteins are used as plant materials for protein isolation. However, hemp seed protein has excellent digestibility and an ideal essential amino acid composition and is considered as a possible protein source for meat-like products (Tang, Ten, Wang, and Yang, 2006; Wang, Tang, Yang, and 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 seed concluded that hemp seed is an underexploited protein-rich non-legume seed (Aiello et al., 2016).

[0019] Although the nutritional potential of hemp seed protein is high, the nutritional quality of plant protein (measured by its amino acid composition and digestibility) is affected by many factors. Amino acid composition may be affected by genotypic variation or agronomic conditions, such as soil fertility and post-harvest processing (e.g., dehulling) that changes the ratio of kernel components. Protein digestibility may be affected by protein structure and the presence of anti-nutritional compounds in the plant material or by anti-nutritional compounds formed during alkaline or high temperature processing (Sarwar, 1997). However, Aiello found low concentrations of anti-nutritional factors in hemp seeds, including condensed tannins, phytic acid, and trypsin inhibitors (Aiello et al., 2016).

[0020] The functional characteristics of hemp seed protein have hindered its use as a protein source in food. Hemp seed protein concentrate is commercially available due to the production of hemp seed oil. After the hemp seeds are milled and pressed to obtain the lucrative oil, a protein-rich seed cake is produced. The seed cake is green, high in fiber, and the protein concentrate is about 40%. Unfortunately, it has a very green and earthy taste that is unacceptable in most foods. Grinding this cake and dry sieving can increase the protein content to about 50%. Many researchers who recognize the nutritional value of this protein-rich source have used it as a starting material for isolating and improving the taste and functional qualities of hemp seed protein. Tang found that hemp seed protein isolate (HPI) from seed cake is inferior to SPI in making plant-based foods (Tang et al., 2006). Tang showed that for HPI, the poor water solubility of hemp seed globulin is believed to lead to its poor emulsification and water retention properties when compared with soy protein isolate (Tang et al., 2006; Hadnadev 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 functional properties are inferior compared to SPI. The inferior functional properties of HPI are mainly attributed to the formation of covalent disulfide bonds between individual proteins and subsequent aggregation at neutral or acidic pH due to the high content of free sulfhydryl groups in its sulfur-containing amino acids." (Tang et al., 2006). In addition, "Differential scanning calorimetry (DSC) analysis showed that HPI had only one endothermic peak with a denaturation temperature (T(d)) of approximately 95.0 degrees Celsius, which was attributed to the edema protein component." (Tang et al., 2006).

[0021] Despite the apparent inferior functional aspects of hemp seed protein, its excellent nutritional qualities have led to continued interest in its use in food production. To this end, individual proteins have been isolated from hemp seed and further investigated for their potential functional properties. In addition, researchers have investigated whether different methods of extracting and isolating hemp seed protein can improve functionality. "The value and application of hemp seed protein in food is closely related to the protein's structural and functional properties." (Wang et al., 2019).

[0022] To investigate the nutritional and functional properties of individual hemp kernel proteins, the researchers used methods to extract and separate the two main proteins present in hemp kernel. Hemp kernel proteins are primarily composed of the proteins edema and albumin. Eedema is a globulin that accounts for approximately 60% to 80% of the total protein content (Odani and Odani 1998; Tang et al., 2006), while albumin, a globular protein but not a globulin, makes up the difference. Eedema and albumin have different amino acid compositions and functional characteristics.

[0023] Malamo studied the nutritional differences between edema proteins and albumin from hemp seeds and concluded that the edema proteins fraction of hemp seed protein was nutritionally superior, containing higher levels of sulfur (methionine and cysteine), aromatic (AAA), branched-chain, and hydrophobic amino acids (Malomo and Aluko 2015). Malamo separated edema proteins from albumin and measured the nutritional value and functionality of each characteristic. These characteristics included water solubility, amino acid content, and digestibility.

[0024] Malomo reported that albumin was partially soluble in water, while edestin was partially soluble in saline solutions. The solubility of extracted edestin in water at neutral or acidic pH was extremely low, and it was only soluble at high ionic strength or alkaline pH (Malomo and Aluko, 2015). "Many protein functions, such as surface active properties, are related to protein solubility." (Jackman and Yada, 1989; Malomo and Aluko, 2015). In hemp seeds, edestin was found to have better emulsion forming ability, while albumin had higher solubility and foaming ability than edestin (Malomo and Aluko, 2015).

[0025] Studies have shown that edema proteins may only be present in hemp seeds, although edema-like proteins may be present in seeds from the family that includes pumpkin and squash (Vickery, 1940). Therefore, the present disclosure and its applications may relate to edema proteins and edema-like proteins, which may have similar or identical properties to edema proteins. Vickery revealed that potential substitutes for edema proteins may be found in plants of the Cucurbitaceae family, including squash seeds. Hirohata has examined globulins from 38 varieties and species of eight genera in the family and noted close similarities between globulins from closely related species (Vickery, 1940; Hirotata, 1932). Vickery believes that globulins from the Cucurbitaceae family may include edema-like proteins that meet the requirements as a nutritional substitute for edema proteins from hemp seeds (Vickery 1940).

[0026] Ephedrin was first isolated and analyzed by Thomas Osborne (Osborne, 1892). In its intact native form, edestrin is composed of six identical subunits, each consisting of an acidic (AS) and a basic (BS) subunit, connected by a disulfide bond (Farinon 2020; Patel, Cudney, and McPherson 1994). Recently, studies have shown that edestrin can exist in multiple forms, even in a single variety of hemp seed (Docimo et al., 2014). For example, in one hemp seed variety, seven genes encode edestrin, and they result in different forms of two edestrin types. In some hemp seed strains, one type of edestrin is virtually identical to each other, while the second type of edestrin is very different from the first type. Ponzoni identified a type 3 edestin gene, CsEde3, which shows approximately 65% ​​and 58% sequence homology compared to the genomic forms of CsEde1 and CsEde2, respectively (Ponzoni, Brambilla, and Galasso, 2018). The amino acid composition between the two types of edestin may differ significantly, with some types having higher nutritional quality (Docimo et al., 2014).

[0027] Ephedrine itself has a large particle size of 309,000, but depolymerizes to 51,000 in concentrated urea solution when denatured [Burk and Greenberg, 1930] and to 17,000 in dilute HCl [Adair and Adair, 1934]. These units are approximately 1 / 6 and 1 / 18 the size of the natural molecule, respectively. In the natural state, they have a specific polypeptide pattern and may be partially integrated by some form of chemical bonding (such as SS bonds), but mainly by lateral attractions between adjacent CO and NH groups and interactions between free acid and basic groups of the side chains. The high number of these latter groups can be seen from the following analytical data: glutamate, 19-2%; aspartate, 10-2% [Jones and Moeller, 1928]; arginine, 17-76% [Vickery, 1940]; lysine, 2-4%, histidine, 2-03% [Tristram, 1939]; amide-N, 1-73% [Bailey, 1937, 2]. Taking into account the amidated COOH groups, they correspond to a total of 670 charged groups per 309,000 molecules. The spatial arrangement of such charges gives rise to a specific charge symmetry, on which the stability of the molecule must ultimately depend, and which is capable of some variation within well-defined limits of pH, as reflected by variations in the dipole moment. Beyond these limits, further inhibition of the ionization of the acidic or basic groups sets up forces of attraction and repulsion within the molecule, which, especially in the absence of small mobile ions, distort and ultimately destroy the unique polypeptide conformation. (Bailey, 1940).

[0028] Thus, edestin as referred to in this disclosure may incorporate all forms of edestin that may be presently known or presently unknown, which have similar or identical properties to edestin disclosed for the purposes of this disclosure.

[0029] Edestan is rapidly degraded to denatured edestan under weak acidic conditions. Denatured edestan is an intermediate derived from edestan that appears during the denaturation of edestan and was first identified by Osborne (Osborne, 1901; 1902). Denatured edestan is formed when edestan comes into contact with dilute acid. Denatured edestan results in the release of SH groups (Bailey, 1942). Bailey demonstrated that edestan can be rapidly converted to denatured edestan in less than 20 minutes under acidic conditions (Bailey, 1942). This study showed that the release of SH groups accompanies the conversion of edestan to denatured edestan. Bailey also reported that the nitrogen content of denatured edestan was reduced compared to edestan, which could be explained by the reduction of tryptophan in edestan. Edema proteins in their non-denatured, native state have different functional properties than denatured or partially denatured edema proteins or denatured edema proteins.

[0030] Conventional techniques for isolating hemp protein or separating edestin from albumin may result in structural changes in the proteins, some of which may be irreversible. Different protein extraction and separation techniques and conditions (pH, presence or absence of monovalent and polyvalent salts, ionic strength of the medium used for protein extraction, time, temperature, etc.) can affect the functional properties of the proteins (Hadnadev et al., 2018). These changes can have negative effects on the functionality of the proteins (Hadnadev et al., 2018; Shen et al., 2021). These negative effects can include changes in digestibility, protein-oil interactions, taste, solubility, and emulsification and gel-forming abilities (Shen et al., 2021). Therefore, it is critical to maintain the native structure of the proteins as much as possible when extracting and isolating edestin, especially for use in food products.

[0031] Many different techniques have been used to separate hemp protein and edema globulin. These techniques include the use of high temperatures, alkaline or acidic conditions, isoelectric precipitation, isoelectric focusing, micellization, ultrafiltration, and mechanical methods in aqueous or solvent slurries, including pressing, grinding or sieving the kernels or shelled kernels, or grinding the kernels and sieving the kernel slurry. Any of these techniques has the potential to change the protein structure and reduce its function.

[0032] High temperatures generated by mechanical methods can negatively affect protein functionality. For example, milling the kernels to produce flour may generate temperatures high enough to alter the structure of the proteins. These temperatures may cause denaturation of edestin and potentially binding or aggregation between edestin, albumin, or fibers, interfering with their independent separation.

[0033] Dry grinding of kernels can generate temperatures of at least 80°C to 100°C, which may denature edible proteins. Mohammad found that the heat and mechanical forces generated during milling can denature globular proteins (Mohommad, 2015). Mohommad showed that the mechanical stress applied during milling can change the volumetric properties of globular proteins.

[0034] Therefore, the high temperatures caused by dry milling to produce powder may change the structure of hemp protein. Farinon calculated the denaturation temperature of hemp protein (edema globulin) to be 92°C (Farinon et al., 2020). In addition, Malamo showed that heat treatment and changes in pH may change the secondary structure of hemp albumin and edema globulin (Malomo and Aluko, 2015). High temperatures may unfold the proteins, exposing their hydrophobic groups and favoring protein-protein interactions rather than protein-water interactions.

[0035] Heating during the extraction process can be avoided or minimized by using chemical methods of protein extraction, however, many chemical extraction methods first require mechanical particle size reduction. Solvent extraction is a common method for isolating proteins from plant materials and involves the use of a liquid solvent to which the protein-containing material has been added. The solvent may be water, alcohol, acetone, hexane, or other liquid solvents. Solvent extraction may be combined with mechanical or other extraction methods that first break down the plant material, thereby releasing the protein. Solvent extraction may involve the use of solvents that break down plant cell walls or fibrous material, thereby releasing the protein.

[0036] Some solvents used for protein extraction have the disadvantage of denaturing proteins. In addition, these solvents may be toxic and unsuitable for ingestion, even in small amounts. In addition, solvents typically require long extraction times, labor-intensive procedures, leave residual solvents in food, and may be difficult to safely dispose of. Hexane is an example of such solvents. According to the organic food labeling guidelines of the U.S. Department of Agriculture (USDA), many solvents cannot be used to produce certified organic foods.

[0037] An alternative protein extraction method that does not require solvents is water extraction, which involves adding the ground or pressed plant material to water and then separating the proteins based on their solubility in the aqueous fraction or in the fat-containing fraction when the fat in the plant material is separated from the water. Water extraction can be followed by isoelectric precipitation or focusing or salt extraction to separate the proteins.

[0038] Alkaline extraction is a commonly used technique in which highly alkaline solvents disrupt the cell structure, thereby releasing proteins from the cells. However, this method can lead to protein damage, including amino acid racemization, lysine alanine formation, reduced digestibility, and loss of essential amino acids (Moure et al., 2006). According to Xu, under alkaline conditions, polyphenols found in many plant materials, including hemp seeds, oxidize and subsequently react with proteins, resulting in a dark green or brown color in the extracted protein solution (Xu and Diosady, 2002).

[0039] When used during hemp protein extraction, the alkaline extraction pH is usually raised to 9 or 10, which is higher than the pH of soy protein extraction (pH 8), because native hemp protein is tightly compacted and may be tightly bound to other components, such as phenolic compounds (Wang and Xiong, 2019). Alkaline extraction usually precipitates the target hemp protein at the isoelectric point, and after several washing steps, the induced color is usually not removed from the protein isolate.

[0040] Isoelectric precipitation or salt extraction is often performed after aqueous or alkaline extraction to isolate proteins. Isoelectric precipitation can be used to extract soluble proteins after alkaline or solvent extraction and involves adjusting the pH until the charge between the target protein and the solvent is balanced, causing the protein to precipitate from solution. Isoelectric precipitation requires a change in pH, which may alter protein structure and negatively affect protein function.

[0041] Regarding the isoelectric precipitation of edema protein, Bailey disclosed that the isoelectric zone of edema protein is pH 5.5 (Bailey, 1942). In this method, albumin can be largely eliminated at its isoelectric point during the precipitation of edema protein (Papalamprou et al., 2009). This result can be attributed to the high solubility of hemp albumin at pH 5.0 (>75%) compared to hemp protein (<10%) (Malomo and Aluko, 2015). One advantage of isoelectric precipitation compared to other protein separation methods is that the water binding capacity of protein isolates obtained by isoelectric precipitation has been found to be higher than that of the same isolates obtained by micellization extraction (Krause et al., 2002). However, one disadvantage of isoelectric precipitation during the separation of edema protein is that the solubility of the protein is lower compared to edema protein isolated by salt extraction, which indicates that the protein is no longer in its native state (Hadnadev, 2018).

[0042] Compared to alkaline extraction and isoelectric precipitation, salt extraction, which may involve micellization, represents a milder extraction method that does not cause polyphenol oxidation, aggregation, and co-extraction with proteins. Salt extraction involves "salting out" a group of proteins and then "salting out" the target protein. "Salting out" refers to an effect in which increasing the ionic strength of a solution increases the solubility of a solute (e.g., protein). This effect tends to be observed at lower ionic strengths. "Salting out" involves further increasing the salt concentration so that the abundance of salt ions reduces the solvating power of the salt ions, resulting in reduced solubility and precipitation of the target protein.

[0043] One salt extraction method described in U.S. Pat. No. 6,005,076 to Murray includes a micellization step. Salt extraction using micellization involves first dissolving the protein with a salt solution having a certain ionic strength. Next, the salt solvent is diluted in the concentrated protein solution to reduce the ionic strength to below a certain level so that discrete protein particles are formed in the aqueous phase at least partially in the form of protein micelles. The protein micelles then settle to form a large amount of the target protein isolate. The protein isolate can then be separated from the supernatant.

[0044] Salt-based micellization extraction, such as that disclosed by Murray, has the advantage of producing protein isolates with higher solubility compared to isolates obtained by isoelectric precipitation (Karaca et al., 2011; Krause et al., 2002; Paredes-López and Ordorica-Falomir 1986). In addition to increased solubility, protein isolates obtained by micellization techniques also have higher interfacial activity compared to isoelectric precipitation. Moreover, according to Krause and Papalamprou, protein isolates produced by micellization extraction have a better preserved native protein structure compared to isoelectrically precipitated proteins (Krause et al., 2002; Papalamprou et al. 2009). Typically, isoelectric precipitation results in a certain degree of denaturation of the extracted proteins, and this can lead to hydrophobic interactions between protein molecules, resulting in the formation of insoluble protein aggregates. Although salt extraction and micellization are probably the least destructive methods known for the isolation of hemp seed proteins, the addition of salt during the separation does have a negative impact on protein structure and function. “The addition of NaCl also had different effects on the gel structure. Specifically, increasing NaCl concentration (up to 300 mM) promoted protein-protein interactions and aggregation, leading to the formation of HMI [hemp seed protein micellar isolate] gel structures with larger pore sizes.” (Shen et al., 2021).

[0045] Salt extraction was the first method used to isolate edema globulins from hemp seeds (Osborne, 1892). This method was further developed by Malomo, who used micellization technology to extract edema globulins (Malomo and Aluko, 2015). As demonstrated by Malomo, during salt-based micellization extraction, after the salt is removed in the dialysis step, the albumins remain in the supernatant, while the globulins precipitate and can be collected by centrifugation. In Malomo, a globulin isolate was produced by salt extraction of hemp seed powder followed by dialysis against water in dialysis tubing.

[0046] Dialysis of the salt extract of hemp seed powder resulted in precipitation of water-insoluble globulins in the form of micelles, while albumin remained in solution (Malomo and Aluko, 2015). The precipitate was then collected and freeze-dried. When comparing the albumin and globulin fractions of hemp seed protein, the protein solubility and foaming ability of albumin were significantly higher than those of globulins, while no difference in emulsion forming ability was observed between the two protein fractions. Salt extraction and micellization have high labor, time, material, equipment and waste disposal costs and are currently not considered commercially viable for protein extraction for use in food.

[0047] Ultrafiltration is another method that can be used to generate protein isolates with improved functional properties compared to other conventional protein extraction techniques. For example, protein isolates obtained by ultrafiltration generally have better emulsifying properties compared to alkaline extraction. However, one disadvantage of ultrafiltration is the clogging of the membrane due to the formation of precipitates in the final product, which can lead to high extraction costs.

[0048] Newer protein extraction methods include ultrasound-assisted extraction, enzyme-assisted protein extraction, and protein electroextraction methods. These methods have disadvantages including high cost, low yield, protein degradation, and protein impurities. Therefore, conventional extraction methods including salt extraction, alkali extraction, and isoelectric precipitation are still the main methods for extracting protein from plant materials such as hemp seeds.

[0049] Regarding the disclosed method of extracting and isolating hemp seeds using the methods described above, an example of aqueous protein extraction of hemp seeds followed by isoelectric precipitation is disclosed in U.S. Pat. No. 10,555,542 to Crank. Crank discloses first grinding the hemp seeds using any suitable method, including grinding using a hammer mill, a roller mill, or a spiral mill. Milling by these methods is a high energy method that generally results in high temperatures, usually around 140°F to 150°F. These high temperatures are required to obtain a paste because forming a paste from a solid does require a certain high temperature, which is known in the field of peanut butter production. In some cases, depending on the final application of the product, these temperatures may cause undesirable interactions between the protein components of the seed material. In Crank, milling produces a paste or meal (meal when the seeds are first pressed to remove oil), wherein water can be added to the milled material at a ratio of about 4 to about 16 parts by weight per part of plant material. Crank discloses adjusting the pH to about 7.5 by adding a base (e.g., calcium hydroxide) to facilitate protein extraction.

[0050] The resulting solution is then centrifuged to separate the fat portion from the aqueous portion or lipid-reducing extract. The lipid-reducing extract can be used as a lipid-reducing vegetable milk or further processed to produce a protein concentrate or protein isolate. In Crank, the protein in the lipid-reducing extract is concentrated and separated by precipitation to produce a vegetable protein concentrate or isolate from partially defatted plant material. Crank discloses that the protein in the lipid-reducing extract can be precipitated by adding an acid (e.g., citric acid) to the isoelectric point of the protein. Crank does not disclose that water extraction can be used alone to separate edema protein and albumin. In addition, although Crank does mention in the application that hemp seed may be a protein source for food separated according to the Crank method, and that hemp seed contains edema protein, Crank does not disclose the purification and separation of edema protein. Crank discloses discarding the fiber and protein-containing portion of the hemp seed protein after centrifugation.

[0051] Czechoslovak Patent No. 33,545 issued to Beran discloses a method for extracting hemp protein from hemp seeds to produce protein for human consumption. In the background section of the patent, Beran discloses that hemp protein is typically produced in the form of spray-dried hemp protein isolate, which typically utilizes high temperatures and may result in protein denaturation. According to Beran, spray drying may require temperatures between 150°C and 250°C; this is a temperature that may denature hemp protein. Beran disclosed that "heat denaturation of the protein can adversely affect solubility and dispersibility, foaming and emulsification properties."

[0052] To avoid thermal denaturation caused by spray drying during the preparation of edema protein, Beran discloses a process comprising first grinding or pressing the hemp seeds to remove oil, followed by water extraction and isoelectric precipitation or salt extraction to purify edema protein. The preferred particle size reduction method used in Beran appears to be dry milling. According to the patent, the ground powder is then added to water at a water to powder ratio of 5:1. Beran then discloses shaking the solution to produce an albumin containing a water portion and a precipitate portion.

[0053] Beran does disclose that the precipitate contains edema proteins, however, Beran discloses additional steps to isolate the edema proteins for use in food products. These steps include extracting the protein by isoelectric precipitation, salt extraction, or ultrafiltration. Beran does not disclose the purity level of the edema proteins in the precipitate prior to the additional steps of isolating the edema proteins, however, the need for such additional steps suggests that the purity of the edema proteins in the precipitate is insufficient to achieve the stated purpose of the Beran patent, which is to use edema proteins to "increase the protein content of high protein foods and smoothie protein beverages." In summary, Beran discloses that "this product can be used in these foods due to its emulsifying properties and beneficial effects on the organoleptic properties of the final product."

[0054] Both Crank and Beran generally disclose the use of ground or pressed hemp seeds (to remove oil) and subsequently ground into hemp seed powder as the starting material for protein extraction. Thus, the hemp seed powder has been subjected to dry grinding or grinding, as well as oil extraction processes that affect the structure and function of the protein. In addition, both Crank and Beran disclose the separation of edema proteins by at least isoelectric precipitation, which can result in changes in the structure of the protein, thereby reducing its function.

[0055] In the process of extracting protein from kernels for use in plant-based meats, it is also possible to extract oil from the kernels. In some cases, extracting the oil may be the primary goal because plant-based oils have value as food and cosmetic products. Common methods for extracting oil from kernels, kernels, and seeds include extraction methods based on pressing, including cold pressing and expeller pressing, and solvent extraction.

[0056] Pressing the seeds to extract the oil involves mechanical compaction of the plant material to squeeze the oil out of the solid. Solvent extraction involves placing the plant material into a liquid to extract the oil. In some cases, pressing and solvent extraction can be combined. Recovering the oil from the extruder process can be relatively inefficient, and a fairly high percentage of fat can remain in the cake. Therefore, the pressed cake can be further extracted using an oil solubilizing solvent. Commercially available cakes and flours produced by pressing methods or pressing and solvent methods are considered to have reduced protein functionality.

[0057] Conventionally produced hemp seed oil may have a green tint due to the rupture of the protoplasts or chloroplasts during the extraction process. Hemp seeds may contain chlorophyll-containing bodies that release chlorophyll when ruptured. Hemp seeds contain more of these substances than other types of seeds, and therefore hemp seed oil tends to have a green tint when it is extracted using conventional methods.

[0058] According to Leonard et al., “Unrefined hemp seed oil has a dark green color due to its chlorophyll content.” (Leonard et al., 2019). Additionally, the presence of chlorophyll in the oil can cause fat oxidation, which can produce off-flavors. U.S. Patent No. 9,493,749 to Soe discloses that “vegetable oils derived from oilseeds such as soybean, palm or rapeseed (canola), cottonseed, and peanut oils typically contain some chlorophyll. However, the presence of high levels of chlorophyll pigment in vegetable oils is generally undesirable. This is because chlorophyll produces an undesirable green color and can cause oxidation of the oil during storage, causing the oil to deteriorate.”

[0059] In order to remove chlorophyll from vegetable oil, various methods have been adopted. These methods include chemical bleaching and ultrasonic bleaching. Chlorophyll can be removed at many stages of the oil production process, including seed pressing, oil extraction, degumming, alkali treatment and bleaching steps. However, the bleaching step is usually the most important step to reduce chlorophyll residues to an acceptable level. In the bleaching process, the oil is typically heated and passed through an adsorbent to remove chlorophyll and other colored compounds that affect the finished oil appearance and / or stability. The adsorbent used in the bleaching step is typically clay.

[0060] Conventional methods of removing chlorophyll from hemp seed oil are costly and can pose problems for waste disposal. In addition, methods of removing chlorophyll from hemp seed oil after chloroplast disruption allow the oil to oxidize due to temporary exposure to chlorophyll. Therefore, there is a need for improved methods of extracting oil from hemp seed.

[0061] In the production of imitation meat products and imitation dairy products, after obtaining the protein isolate and the preferred oil source, the second step involves combining the isolated protein with water and possible oil to form a material for solidification or extrusion. After the protein is isolated from hemp seeds or other plant products, it must be combined with other components of the imitation meat product or imitation dairy product to form the final imitation meat product. The three basic ingredients for the production of imitation meat products are protein, water and fat. These components can be combined in different concentrations and processed in different ways to form imitation meat products and imitation dairy products.

[0062] In view of the need for gelation or structuring of imitation meat products to produce a chewy meat-like texture, proteins are typically combined with water and possible oils to form a gel, which can be solidified by heating to produce texture. Regarding the use of these components or only using protein and water to form a hemp seed protein isolate gel, studies have shown that hemp seed protein does not have good gel-forming properties. As disclosed above, Wang, Shen and Zahari point out that hemp seed does not have good gel-forming ability, which makes it unlikely to be used as a candidate main protein in imitation meat products or imitation dairy products (Wang et al., 2019; Shen et al., 2021; Zahari et al., 2020). For example, Wang shows that when heated alone, the combination of hemp seed protein isolate and water will not form an ideal gel. Wang also shows that even if oil is added to Wang's protein and water mixture and heated, the mixture of hemp seed protein, water and oil will not form an ideal gel when heated.

[0063] In the production of imitation meat products, the third step of heat coagulation of protein, water and oil typically includes extrusion, which texturizes the product to form a more meat-like material. In order to form texturized meat, an extruder can be used to form texturized vegetable protein (TVP). TVP is typically a soy-based product, but other plant proteins (such as peas) can be used alone or in combination with soy. To produce TVP, plant-based ingredients are fed into an extruder for texturization. Conventionally, dry plant protein is fed into an extruder, and then water, starch and occasionally fat are added to the protein through a separate input as the protein is conveyed through the extruder. After extrusion, the extruder output may undergo a marinating, coating and / or cooling step.

[0064] A common problem with conventional plant-based meat substitutes, including TVP and HMMA, is that the texture is not dispersible and the mouthfeel is rubbery compared to meat. This texture and mouthfeel of conventional meat-like products is due in part to the fact that fat, oil, or a combination thereof, is not incorporated into the molecular structure of the protein peptide chains or "fibers." Meat derived from animals incorporates fat molecules between these muscle fibers, which make up the majority of animal meat products. This fat is released during the chewing process, providing the consumer with positive and continuous sensory feedback in terms of taste and mouthfeel as the chewing continues. The sensory feedback provided during the chewing of current conventional meat-like products is not equivalent to meat, in part because there is no fat between the peptide layers of the protein. In conventional meat-like products, fat is added after the protein is fully denatured, and therefore the fat may surround large pieces of cooked protein, but is not incorporated into the peptide layers of the protein itself.

[0065] The soy and pea proteins commonly used to create the fibers in TVP and HMMA may contain only about 10% fat by weight. Typically, muscle fibers in meat contain from 5% to about 50% of their weight in fat in the protein fibers, depending on the source of the meat. Therefore, with conventional soy and pea meat analogs, most of the fat added to the product during extrusion remains outside the fibers, resulting in a greasy, unappealing product that does not release the fat in a controlled and juicy manner when chewed. As a result, conventional meat analogs primarily appeal to a limited number of committed vegetarian or vegan consumers, while failing to appeal to the majority of meat-eating consumers.

[0066] Different extrusion methods may produce different textures of simulated meat products. For decades, extrusion technology has been developing to produce simulated meat products that are more like meat. The extrusion and extrusion preparation of simulated meat products involve complex chemical changes and processes within the protein components of the extrusion mixture. During extrusion, the protein isolate structure changes significantly, whereby the protein may be partially or completely denatured or unfolded, as well as repositioned and cross-linked with other protein molecules and chemically bonded to other components of the extrusion mixture. In addition to changes in temperature and pressure, the extruder also causes these changes through the application of shear forces applied by the screw as the mixture moves through the machine. The final texture, taste and mouthfeel of simulated meat products produced by extrusion depend on the various types of chemical bonds formed between the components of the extruded mixture before, during and after extrusion.

[0067] Regarding early development of an extrusion process for producing a texturized meat analog product, U.S. Pat. No. 6,319,539 issued to Shemer et al. discloses mixing a protein with a large amount of water and possibly a fat, and heating, gelling and forming the resulting paste in an extruder. During transfer to the extrusion apparatus, Shemer discloses that the paste is heated and conveyed at a determined rate, and then extruded through an opening. The resulting food has a fibrous texture comprising substantially aligned axial fibers. However, a problem with this process is that it has a limited flow rate and can only be implemented using certain raw materials, particularly gluten, which results in a limited variety of products. Gluten is a known allergen, which has also limited the use of soy-based TVPs to date.

[0068] Another disadvantage of the Shemer process and other early extruder processes is that the heated product expands as it is conveyed from the extruder due to the release of water vapor as the hot product cools. The water vapor causes disorder in the aligned protein fibers, which is undesirable for an acceptable texture in a meat analog product.

[0069] To address this problem, WO 2003 / 007729 to Bouvier et al. discloses a twin-screw rotor extruder, rather than a single-screw device, having an elongated cooling chamber that allows the raw materials to be mixed and extruded at a controlled temperature so that steam does not disrupt the alignment of the protein fibers in the final product. In addition to addressing the cooling and water vapor issues, the '729 application also recognized a problem in the prior art of incorporating the desired amounts of oils and fats into the extruded product using conventional raw material formulations.

[0070] To achieve the desired fat content, the '729 application discloses a novel extrusion mixture containing a fat component mixed with lecithin or caseinate, protein, fiber, starch and water. This mixture is kneaded to obtain a paste, which is heated and gelled in an extruder. However, the inclusion of large amounts of carbohydrates (e.g., starch) in meat analog products is undesirable for taste and nutritional reasons.

[0071] To address the problem of introducing fat into a meat analog without adding starch, and without other related problems of introducing oil into an extruder, WO 2012 / 158023 to Giezen et al. discloses an extrusion method for converting a vegetable protein composition, such as soy protein, into a fibrous, meat-like structure. Giezen discloses an extrusion exit temperature above the boiling point of water, thereby producing an open product structure that can be injected with oil to achieve the desired fat content. Problems with Giezen include the addition of a processing step after extrusion, and consumers perceiving the final product as being too greasy and fatty.

[0072] A commonly recognized problem in the art of meat analog extrusion is that higher amounts of oil in the extrusion mixture interfere with obtaining a product with the texture of animal meat. In conventional meat analog extrusion, the presence of oil reduces the high mechanical shear forces within the extruder that form the desired meat analog fiber structure. Thus, using conventional methods, adding an optimal amount of oil results in a meat analog with a suboptimal fiber structure and texture.

[0073] To overcome the problem of suboptimal texture of texturized meat analogs resulting from higher oil content, U.S. Patent Application No. 20180064137 to Trottet et al. discloses adding oil separately from other raw materials during extrusion. This method includes feeding 40-70% by weight of water and 15-35% by weight of vegetable protein into an extruder barrel, and then injecting 2-15% by weight of oil into the extruder barrel at a point downstream of the feeder. According to the present 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 high shear forces to be generated in the first half of the extruder to promote fiber formation, while oil can be added downstream without interfering with fiber formation.

[0074] Although Trottet's process improves the product compared to the prior art, Trottet does not result in a significant amount of oil being incorporated into the core of the protein fibers. If the oil is not incorporated into the fibers, the resulting product would be perceived by the consumer as greasy and the release of fat would be uncontrolled during chewing. This unsatisfactory result is because people are accustomed to eating animal meat, which has a large amount of fat incorporated into the muscle fibers. Animal meat protein fibers incorporate up to about 50% of their weight in fat, although this varies depending on the source of the meat. In the Trottet process, the amount of fat incorporated into the fibers is only about 10% of the weight of plant protein fibers. This problem with the Trottet process is caused by the type of protein used as the extrusion feedstock (disclosed as soy and wheat for Trottet) and the method of adding oil to the protein fibers.

[0075] In another patent application addressing the fat content of simulated meat products, WO 2020 / 208104, issued to Pibarot, was filed in 2020. In a document titled "Meat-like products and extrusion apparatus and method for simulated meat products," Pibarot acknowledges the problem of simulating the fat content of animal meat, which contains adipose tissue both inside and outside the protein matrix. Pibarot believes that this complex structure may affect the appearance of the meat as well as the texture and juiciness of the meat.

[0076] To solve this problem, Pibarot discloses injecting fat into the interior of the extruded mixture when the extruded mixture is cooled in a die. In Pibarot, gaps are created between the protein fibers of the extruded mixture during extrusion. When the heated and sheared product is conveyed through a cooling die, fat is injected between these gaps so that the fat is deposited between the protein fibers. Pibarot suggests that this method will produce a marbled appearance similar to red meat and improve the texture and palatability of the product. Pibarot discloses using this method for soybeans, peas, and other conventional plant protein sources. However, Pibarot does not teach a method for incorporating fat into the molecular structure of protein fibers.

[0077] In summary, the Shemer process can only be used with a limited number of ingredients, and the lack of controlled temperature and cooling results in a poor quality product. Although Bouvier solved the cooling problem of Shemer, in order to achieve the desired fat content, Bouvier blended the extruded raw material with a large amount of starch, resulting in unsatisfactory taste and nutritional quality. Giezen solved the starch problem of the Bouvier process by adding fat after extrusion, however, this required an extra step and resulted in a greasy, unpalatable product. Trottet improved on Giezen and Bouvier by introducing oil later in the extrusion process, however, Trottet still suffered from the problem of low incorporation of oil into the protein fibers of the meat-like product. Pibarot disclosed injecting fat into the meat-like product while the meat-like product was cooling, which introduced fat between the protein fibers, but did not produce a final product in which the fat was incorporated into the protein fibers.

[0078] Hemp seed has great value as a source of protein and a source of oil. Although there are multiple methods for producing food from hemp seed, it is obviously necessary to extract protein and oil from hemp seed more effectively, more efficiently, more cleanly and at a lower cost, to produce a clean and light, unoxidized hemp seed protein with significant purity, gelling function, nutritional value, digestibility and flavor, and an oxidatively stable oil with clean flavor and light color, suitable for cooking and cosmetics. In addition, there is a need for methods and raw materials for producing a variety of imitation meat products with the appearance, taste, texture, juiciness and chewiness of a variety of animal meat products and dairy products. More specifically, a kind of imitation meat product is needed, which has the appearance, texture and taste of meat, and contains the oil or saturated fat of the best amount in the final product, wherein the oil or saturated fat is mixed in the protein fiber at a level close to that of animal meat or dairy products. Summary of the invention

[0079] This document discloses a ready-to-eat meat-like product or food-like material based on natural edible globulin isolate (NEPI). According to the method of the present disclosure, when NEPI is combined with water and oil, a ready-to-eat, textured meat-like product can be formed. The present disclosure describes a method comprising using a NEPI concentrate, or mixing NEPI powder with water to form a hydrated protein suspension, and then adding oil to form a protein-fat hydrosol. When using a NEPI concentrate, it is critical to reach a hydration temperature between 60°C and 70°C after any necessary further dilution by adding water to the NEPI concentrate to reach the moisture content required for the formulation, in order to fully hydrate and open the protein structure. It is critical that the NEPI protein must be fully hydrated and opened at the above specified temperature before adding oil to form the protein-fat hydrosol. It should be noted that if it is necessary to heat the NEPI aqueous suspension before adding oil, it should be heated gently directly, or preferably gently heated in a water bath between 60°C and 70°C, and gently stirred to even out the heating rate until the temperature of the hydrated NEPI reaches the temperature of the water bath. When using dry powdered NEPI to prepare the hydrated protein suspension, it is more preferred to use hot water, preferably at a temperature between 60°C and 80°C. Once the hydrated NEPI protein has been opened, reaching a minimum temperature of 60°C, the oil can be added. After the oil has been added and incorporated into the heated NEPI concentrate or the NEPI protein hydrated with hot water, it is then possible to cool the formed protein-fat hydrosol to refrigeration temperature to maintain microbial stability for at least 48 hours. If the protein-fat hydrosol has been cooled for microbial stability purposes, the protein-fat hydrosol should be gently heated again, preferably in a water bath between 60°C and 70°C, and very gently and occasionally stirred to equalize the heating rate, until the temperature of the protein-fat hydrosol reaches the temperature of the water bath, before subjecting the protein-fat hydrosol to microwave coagulation. Most preferably, the protein-fat hydrosol is prepared and reaches a minimum temperature of 60°C as described above, and is immediately heated using microwaves to further heat and solidify the protein-fat hydrosol into a solid hydrogel in a microwave oven before the protein-fat hydrosol has time to cool below 60°C, so as to solidify the protein-fat hydrosol to form a hydrogel or meat analog. The water, NEPI protein, and oil hydrosol can be blended in a microwaveable container, or blended separately and poured into a microwaveable container. The container is preferably constructed of a microwave insulating material (e.g., paper, glass, plastic, or ceramic) that does not absorb microwaves but allows microwaves to pass through the material in the container. In some embodiments, an irregular surface can help the protein-fat hydrosol and protein-fat hydrogel adhere to the side walls of the container.

[0080] The protein hydrosol may be mixed in a microwaveable container, preferably constructed of a material having an irregular or rough surface, such as paper or rough plastic, and of a size that facilitates the formation of a meat analog product from the protein-fat hydrosol upon microwave heating. The materials required to produce a meat analog product from NEPI powder at home may be provided as a convenient kit for producing a ready-to-eat meat analog product.

[0081] The present disclosure solves the problems in the prior art regarding hemp seed protein separation, raw material input preparation and raw material input processing to produce high-quality plant-based imitation meat products and imitation dairy products. The compositions and methods of the present disclosure include methods for hemp seed protein separation, pasteurization, liquid solutions, gel formation, texturization, and the production of imitation meat products and imitation dairy products. The methods of the present disclosure produce structured protein foods or imitation meat products with superior properties compared to existing products or similar products made using known technologies.

[0082] The preparation of a simulated meat product or simulated dairy product according to the method of the present disclosure can be divided into three major steps. The first step involves extracting or isolating protein from hemp seeds. 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 thermal gelation or extrusion of the raw material to solidify or texturize the simulated meat product. The final simulated meat product can then be cooked to simulate meat products or dairy products, such as chicken, fish and cheese.

[0083] With respect to the first step, hemp seed protein separation, the method of the present disclosure incorporates the known seed processing method disclosed in U.S. Pat. No. 7,678,403 to Mitchell ("Mitchell" or "the '403 patent"), with some modifications. The '403 patent is incorporated herein by reference in its entirety. The Mitchell process discloses water wet milling at low temperatures and screening the resulting product. In the present disclosure, water wet milling may preferably be performed while maintaining the temperature of the slurry between preferably 33°F and 38°F. At higher temperatures, particularly at temperatures of 42°F or higher, microbial growth becomes a problem.

[0084] In some embodiments, whole hemp seeds or shelled hemp seeds (also known as dehulled hemp seeds) can be used for milling. Depending on whether whole or dehulled hemp seeds are used, the final meat-like or dairy-like product may have different colors. Using whole hemp seeds will produce a darker, more beef-like color, while using dehulled hemp seeds will produce a whiter, more chicken-like or fish-like color. Using part whole hemp seeds and part dehulled hemp seeds, in one embodiment, wherein the whole hemp seeds are used at a concentration of about 20-30% by weight relative to the amount of dehulled hemp seeds, results in a beef-like color in the final product. In one embodiment, the husk previously removed by dehulling the hemp seeds can be reintroduced into the dehulled hemp seeds to increase the color; wherein, in one embodiment, in order to obtain a beef-like color, the husk can be added to the dehulled hemp seeds in an amount of about 10-15% by weight relative to the dehulled hemp seeds.

[0085] After wet milling, Mitchell in the '403 patent teaches screening with a different mesh size than the present disclosure, wherein screening with a mesh size of 170 to 200 is preferred in the present disclosure. Mitchell in the '403 patent, when discussing the screening of rice grains for dairy production, discloses a mesh size of 150 or less, which is suitable for some grains, but not for chloroplast removal of hemp seeds. The present disclosure surprisingly found that a mesh size of 170 to 200, preferably, or a size between about 160 and 200 mesh prevents chloroplasts or chlorophyll-containing particles from passing through the filter without significantly reducing protein or nutrient yields, allowing sufficient protein particles to pass through the filter.

[0086] When hemp seed is processed according to the modified Mitchell method, a precipitated by-product containing insoluble protein is produced. This protein-containing material was discovered by Mitchell to have unique and valuable properties and is particularly suitable for the production of imitation meat products and imitation dairy products. This material containing hemp seed protein has not been previously disclosed. Upon further investigation, Mitchell determined that the material is mainly composed of edible globulin and, importantly, it is essentially free of albumin, which is another major protein component of hemp seed. Due to the processing parameters of the Mitchell method, edible globulin appears to be essentially maintained in its natural state. Due to its high concentration of essentially natural edible globulin, the material will be referred to as natural edible globulin isolate (NEPI) hereinafter. NEPI consists of approximately 80% protein and also contains oil, fiber, carbohydrates and ash.

[0087] After grinding and sieving, NEPI can be separated from the aqueous oil albumin emulsion (AOAE) by centrifugation and decantation. The aqueous oil albumin emulsion can optionally be further processed to produce hemp seed oil and albumin. NEPI extracted according to the present disclosure can be used in a variety of different plant-based foods that replicate meat products or dairy products. NEPI can optionally be combined with oil to form protein hydrosols and protein-fat hydrosols, and can be processed to produce evaporated or spray-dried products. The hydrosol can be used to produce plant-based simulated meat products.

[0088] The present disclosure is based on methods and materials for making plant-based products that more closely replicate meat products, including the texture, juiciness, fibrousness and texture uniformity of animal meat. A method for producing a simulated meat product is described herein, the method comprising selecting proteins based on their unfolding or denaturation, properties and fat holding capacity. In addition, the methods described herein include a method for preparing an extrusion mixture or input prior to extrusion, the extrusion mixture or input may be a protein-fat hydrosol, the method incorporating water and fat into the selected protein in a manner such that the water and fat form a liquid matrix (also referred to as a protein-fat hydrosol) with the protein. In some embodiments, the liquid matrix may have additional components, and the protein-fat hydrosol may have no more than protein, fat and water. In addition, the methods described herein include a method for extruding or otherwise heating a liquid matrix, the liquid matrix may also be referred to herein as an extrusion input or extrusion mixture, wherein the liquid matrix may be a fat-protein hydrosol. The method of extruding a liquid matrix comprises feeding the liquid matrix into a pump at a first end of an extrusion chamber. The liquid matrix is ​​fed into an extruder, wherein the extruder is set to parameters suitable for the liquid matrix.

[0089] The present disclosure relates to compositions containing edema proteins or edema-like proteins and methods for isolating edema proteins from hemp seeds. As disclosed herein, edema proteins can be isolated from hemp seeds or other grains and seeds containing edema proteins or edema-like proteins. In one embodiment, hemp seeds are wet-milled in an aqueous protein extraction process to produce an edema-containing fraction and an aqueous oil albumin emulsion.

[0090] In one aspect, the present disclosure can utilize water wet grinding method to separate the fat stored in hemp seed without destroying chloroplast and releasing chlorophyll into oil.Once by this method, seed is milled, the gained milled product is sieved by screens of different sizes.Sieving can remove shell, chloroplast and fiber between about 170 meshes, or in some embodiments between 160 and 200 meshes, or in some embodiments between 200-270 meshes.More preferably, the mesh size between 160 and 200 can be used.In a preferred embodiment, the mesh size of 170 can be used.The opening of the mesh size of 150 is too large, and may make unwanted materials enter the filtrate, including fiber and chlorophyll-containing materials.Surprisingly, the chlorophyll-containing particles keep the size greater than 170 mesh pore openings, and most of the protein-containing particles pass through the screen of this size. According to the method of the present invention, chloroplasts, protoplasts or other chlorophyll-containing particles are separated from the hemp seed oil and protein-containing parts by screening with sieves of different sizes to obtain a light yellow final oil product.

[0091] In the methods of the present disclosure, after sieving, an insoluble portion containing NEPI and albumin oil-water emulsion may be present in the filtrate. The AOAE may be decanted after centrifugation. The insoluble portion and the portion containing particles may be washed to remove any residual oil. In some embodiments, two washes may be performed with cold water.

[0092] In some embodiments, the AOAE can be cooled between about 33°F and 38°F, with 35°F being preferred, until the albumin begins to separate and precipitate from the oil in the emulsion, which can be assisted by centrifugation in some embodiments. According to the methods of the present disclosure, albumin binds strongly to the hemp seed oil, thereby improving the separation of the oil and albumin from the insoluble edema protein fraction or NEPI. Albumin can be separated from the hemp seed oil by this method. Gel electrophoresis shows that essentially all of the albumin can be removed from the NEPI by this method, leaving primarily edema protein in the NEPI. The AOAE can be removed from the NEPI by centrifugation and decantation, leaving the NEPI as a solid material, which can be washed to remove residual material.

[0093] In one embodiment, the NEPI may then be heated to a temperature of about 145°F and maintained for about 30 minutes to pasteurize the product. In some jurisdictions, 145°F may be the legal lower limit for pasteurization. Here, the temperature should be maintained at about 145°F, or between 145°F and 155°F to prevent the granulation observed in the present disclosure from occurring at higher temperatures. Granulation may occur in NEPI at temperatures far below the denaturation of edestin, such as at about 158°F; therefore, it is critical to pasteurize at a temperature lower than that typically used for pasteurization by those of ordinary skill in the art. Those of ordinary skill in the art typically pasteurize protein isolates at temperatures that would result in significant granulation in the present disclosure in order to process the product quickly. After pasteurization is complete, the NEPI may be spray dried or stored as a concentrate for use in meat-like and dairy-like products. Spray drying should be performed at a relatively low temperature, preferably around 145°F to 155°F, to prevent protein granulation or aggregation.

[0094] In some embodiments, particularly for commercial applications, the NEPI exits the production line into a tank heated to 145° F., the product is allowed to incubate at this temperature for 30 minutes, and then is sent to a cooling tank to cool to approximately 35° F. After cooling, if necessary, the NEPI can be shipped for spray drying, freezing, freeze drying, or vacuum microwave drying prior to use in the production of meat and dairy analogs or structured protein foods.

[0095] For the production of simulated meat products, pasteurized products can be prepared by first hydrating NEPI (if dried) or otherwise maintaining the appropriate hydration level. In one embodiment, the amount of water can be about 3 parts water to 1 part NEPI. Before being added to NEPI, the water can be preheated, preferably to about 135°F, to form a protein hydrosol before coagulation. Salt should not be added during this process because it may destroy the protein hydrosol structure. Salt can be added just before coagulation or after coagulation, but not before coagulation. In some embodiments, protein hydration and opening can be carried out at 100°F to 135°F, or in some embodiments between 100°F and 155°F; or in other embodiments, protein hydrosol formation can be carried out at a lower temperature, however, the temperature must be above a low temperature that does not allow the protein to open. Preferably, the temperature in the hydration and protein preparation steps should be kept as close to 145°F as possible, which is considered to be the minimum temperature for pasteurization, but will not reach a temperature that causes the product to granulate.

[0096] Once the protein is hydrated, in one embodiment, oil may be added and mixed with the protein hydrosol to form a protein-fat hydrosol. After the NEPI is fully hydrated, oil should not be added so that the protein hydrosol has a smooth appearance. If oil is added before hydration and protein preparation, particles may form. In addition, according to the method of the present disclosure, the oil should be added before the material solidifies, and solidification means forming protein bonds to produce a more solid gel product in which the proteins aggregate, usually at higher temperatures where the proteins denature. In the case of the present disclosure, there is no free oil during the solidification process, and all the oil is incorporated into the emulsion or protein structure before the product solidifies in an extruder or other hot solidification device. In conventional extrusion, free oil will be present in the material that is partially or completely solidified in the extruder. Therefore, for the present disclosure, it is important to add water before adding oil to fully hydrate and open the NEPI so that there is no free oil during extrusion or solidification. This protein-fat hydrosol can then be heated or extruded to form a simulated meat product. In conventional extrusion using, for example, soy or pea protein, oil is added to the protein material after solidification begins at high temperatures in the extruder more for lubrication than for incorporation of oil.

[0097] Once the protein hydrosol is fully hydrated, oil may be added to form a protein-fat hydrosol. The oil may preferably be preheated, wherein the temperature of the oil may preferably be between about 130°F and 135°F. In other embodiments, the oil may be preheated to 100°F to 135°F or 100°F to 155°F, and in other embodiments, the oil may be added at a lower temperature, however, the oil should not be added at a cold temperature, otherwise the structure of the protein hydrosol may be destroyed and the oil may be prevented from being incorporated into the protein hydrosol to form a protein-fat hydrosol. The material may also be coagulated in a cooking system, although cooking may not produce a fibrous product like extrusion.

[0098] The texture of the cooked NEPI meat analog product is surprisingly good and has textural properties, including hardness and chewiness, that are far superior to commercially available hemp seed protein concentrates and isolates under the same conditions. The method of the present disclosure unexpectedly results in the thermal gelation and extrusion of a high quality fibrous meat analog product made using only hemp seed as a protein source. Due to the properties of conventionally used meat analog and dairy analog proteins (including soy and hemp seed), conventional meat analog and dairy analog products are unable to replicate a textured meat slice similar to an animal meat product, such as chicken breast. The unexpectedly advantageous properties and results produced by the use of NEPI and the process for its use described in the present disclosure have produced a structured meat analog product that is far superior to other commercially available products that use only hemp seed protein as a protein source. Until now, it has only been known that hemp seed protein can be used in combination with soy or other plant proteins to produce meat analog products.

[0099] In one aspect, this document provides a meat analog extrusion input or liquid matrix having a protein to fat ratio in the range of about 4:1 to 0.5:1.

[0100] In one aspect, this document provides a method wherein water is added to a protein isolate at a ratio as disclosed below, or wherein water is maintained in a protein isolate at a specific ratio, and wherein after adding or maintaining water and protein isolate, fat is added to the mixture of protein and water at approximately a certain ratio of water to fat and protein to fat.

[0101] In one aspect, this document provides a product wherein the target water content is between 35 wt% and 75 wt%.

[0102] In any of the methods or compositions described herein, the isolated vegetable protein in the liquid matrix may include seed oil proteins, such as edestin, albumin, globulin, or mixtures thereof.

[0103] In any of the methods or compositions described herein, the isolated protein can first be separated from all other plant proteins in the plant.

[0104] In any of the methods or compositions described herein, the isolated protein used may be isolated in a native or non-denatured state; wherein native can mean completely native, substantially native, partially native, or substantially native as identified by conventional methods of examining protein structure, or as understood by one of ordinary skill in the art to be native.

[0105] In any of the methods or compositions described herein, the protein isolated from the seed protein preferably has a higher cysteine ​​content than is typically found in soy or casein.

[0106] In any of the methods or compositions described herein, the liquid base may include flavorings, starch, fiber or other carbohydrate sources.

[0107] In some embodiments, the meat analogs and dairy analogs provided herein may be free of animal products, wheat gluten, soy protein, or pea protein.

[0108] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art to which the present 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 the event of a conflict, this specification (including definitions) shall prevail. In addition, the materials, methods and examples are illustrative only and not restrictive. All percentages mentioned are by weight.

[0109] The details of one or more embodiments of the invention will be set forth in the accompanying drawings and the description that follows. Other features, objects, and advantages of the invention will be apparent from the description, drawings, and examples, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0111] Figure 1 is a flow chart showing a method for producing natural edema protein isolate or NEPI according to the present disclosure;

[0112] Figure 2 is a flow chart showing a method for producing pasteurized NEPI according to the present disclosure;

[0113] Figure 3 is a flow chart showing a method for spray drying NEPI according to the present disclosure;

[0114] Figure 4 is a flow chart showing a method for producing colored NEPI according to the present disclosure;

[0115] Figure 5 is a flow chart showing a method for extracting hemp seed oil from hemp seeds according to the present disclosure;

[0116] Figure 6 is a flow chart showing a method for forming a hydrosol according to the present disclosure;

[0117] Figure 7 is a flow chart showing a method for producing simulated meat products and simulated dairy products by cooking according to the present disclosure;

[0118] Figure 8 is a flow chart showing a method for extruding NEPI according to the present disclosure;

[0119] Fig. 9 is an SDS-PAGE electrophoresis gel of NEPI protein and hemp seed protein from commercially available hemp seed protein concentrates and isolates under non-reducing conditions according to the present disclosure;

[0120] Fig.10is an SDS-PAGE electrophoresis gel of NEPI protein and hemp seed protein from commercially available hemp seed protein concentrates and isolates under reducing conditions according to the present disclosure;

[0121] Figure 11A is an SDS-PAGE electrophoresis gel of hemp seed powder and hemp seed protein isolate under reducing conditions from prior art publications; Figure 11B is an SDS-PAGE electrophoresis gel of hemp seed protein from hemp seed protein isolates from prior art publications under non-reducing and reducing conditions;

[0122] Fig. 12A This is the differential scanning calorimetry thermogram of NEPI 250 shelled hemp seed spray-dried powder; Fig. 12B is a differential scanning calorimetry thermogram of NEPI 250 whole hemp seed concentrate (slurry);

[0123] Fig.13A This is the differential scanning calorimetry thermogram of VICTORY HEMP shelled hemp seed spray-dried powder; Fig. 13B This is the differential scanning calorimetry thermogram of NUTIVA hemp seed powder;

[0124] Fig.14A It is a photo of a cross section of a poached chicken breast; Fig. 14B Is from Fig.14A A magnified photo of a cross section of a boiled chicken breast; Fig. 14C Is from Fig. 14B Photo of a magnified cross section of a boiled chicken breast;

[0125] Fig.15A is a photograph of a cross-section of a cooked meat analog product using NEPI shelled hemp seed concentrate according to the present disclosure; Fig. 15B Is used from Fig.15A A magnified photograph of a cross section of a cooked meat analog product of NEPI's shelled hemp seed concentrate; Fig. 15C Is used from Fig. 15B A magnified photograph of a cross section of a cooked meat analog product of NEPI's shelled hemp seed concentrate;

[0126] Fig.16A is a photograph of a cross-section of a cooked meat analog product using NEPI shelled hemp seed powder according to the present disclosure; Fig. 16B Is used from Fig.16A A magnified photograph of a cross section of a cooked meat-like product of NEPI's shelled hemp seed powder; Fig. 16C Is used from Fig. 16B A magnified photograph of a cross section of a cooked meat-like product of NEPI's shelled hemp seed powder;

[0127] Fig.17Ais a photograph of a cross-section of a cooked simulated meat product using VICTORY HEMP shelled hemp seed powder according to the present disclosure; Fig. 17B Is used from Fig.17A A magnified photograph of a cross section of a cooked meat-like product of VICTORY HEMP shelled hemp seed powder; Fig. 17C Is used from Fig. 17B A magnified photograph of a cross section of a cooked meat-like product of VICTORY HEMP shelled hemp seed powder;

[0128] Fig.18A is a photograph of a cross-section of a cooked simulated meat product using HEMPLAND shelled hemp seed powder according to the present disclosure; Fig.18B Is used from Fig.18A A magnified photograph of a cross section of a cooked meat-like product of HEMPLAND shelled hemp seed powder; Fig.18C Is used from Fig.18B A magnified photograph of a cross section of a cooked meat-like product of HEMPLAND shelled hemp seed powder;

[0129] Fig.19 are photographs of NEPI extruded from shelled powder and a piece of boiled chicken breast according to the present disclosure to show the textural and fibrotic similarities.

[0130] Fig. 20 is a flow chart of a method for producing a ready-to-eat meat analog according to the present disclosure;

[0131] Fig.21A A container according to the present disclosure is shown; Fig. 21B A container containing a protein-fat hydrosol according to the present disclosure is shown; Fig. 21C showing a container containing a fully expanded protein-fat hydrosol according to the present disclosure; and Fig.21D A container containing a protein-fat hydrogel or a structured protein food according to the present disclosure is shown;

[0132] Fig.22A shows a top view of a protein-fat hydrosol in a plastic container; Fig. 22B showing a side view of a protein-fat hydrosol in a container according to the present disclosure;

[0133] Fig.23A shows a side view of a protein-fat hydrogel in a plastic container; Fig. 23B The plastic container according to the present disclosure is shown Fig.23A Side view of the protein-fat hydrogel rotated 180°;

[0134] Fig.24A shows a top view of a protein-fat hydrogel in a plastic container; Fig. 24BShow Fig.24A A top cross-sectional view of a protein-fat hydrogel in which the top layer of the hydrogel has been peeled off according to the present disclosure;

[0135] Fig.25A shows a top view of a protein-fat hydrogel removed from a plastic container; Fig.25B Showing the Fig.25A Side cross-sectional view of the protein-fat hydrogel in;

[0136] Fig.26A A top cross-sectional perspective view of a protein-fat hydrogel is shown removed from a plastic container and folded; Fig.26B Showing the Fig.26A A front cross-sectional perspective view of a protein-fat hydrogel in FIG.

[0137] Fig.27A shows a front perspective view of a protein-fat hydrogel removed from a plastic container, wherein the bottom of the hydrogel is not substantially molded to the shape of the bottom of the container; Fig.27B A front perspective view of a protein-fat hydrogel removed from a plastic container is shown, wherein the bottom of the hydrogel is partially molded to the shape of the bottom of the container; Fig.27C showing a front perspective view of a protein-fat hydrogel removed from a plastic container according to the present disclosure, wherein the bottom of the hydrogel is substantially molded to the shape of the bottom of the container;

[0138] Fig.28 shows a side cross-sectional perspective view of a protein-fat hydrogel in a container according to the present disclosure, showing the hydrogel meniscus;

[0139] Fig.29A A container insert is shown; Fig.29B A container insert is shown placed in a container; Fig.29C A container insert is shown inserted into a container according to the present disclosure;

[0140] Fig.30 An immersion blender according to the present disclosure is shown;

[0141] Fig.31 An indicator cover according to the present disclosure is shown;

[0142] Fig.32 A kit according to the present disclosure is shown. DETAILED DESCRIPTION

[0143] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art to which the present 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 the event of a conflict, this specification (including definitions) shall prevail. In addition, the materials, methods and examples are illustrative only and not restrictive. Unless otherwise stated, all percentages mentioned are by weight. The details of one or more embodiments of the present invention will be described in the accompanying drawings and the following description. From the specification and drawings, and from the claims, other features, objects and advantages of the present invention will be apparent.

[0144] In general, the present disclosure provides methods and materials for producing plant-based imitation meat products or imitation dairy products (also referred to herein as structured protein foods) from hemp seed protein. In any method or composition described herein, and in some embodiments, the extracted protein-containing product can be separated from other hemp seed proteins. In any method or composition described herein, hemp globulin can be substantially separated from some or all other proteins in hemp seed. In any method or composition described herein, the protein separated from the grain protein preferably has a higher cysteine ​​content than typically found in soy or casein.

[0145] The plant protein used according to the present disclosure may be an isolated plant protein. For the purposes of the present disclosure, a "native" protein is a protein that may have the same tertiary and quaternary structures as in living and active cells. In some embodiments, a "native" protein may be substantially natural. In any method or composition described herein, the isolated protein may be isolated in a generally natural, substantially natural, or non-denatured state. In any method or composition described herein, the protein of the isolated protein used may be isolated in a natural or non-denatured state; wherein natural may mean completely natural, substantially natural, partially natural, or otherwise identified as substantially natural by conventional methods for detecting protein structure, or as understood by a person of ordinary skill in the art. Changes and destruction of subunit structure and tertiary structure may occur with temperature changes (typically above 41° C.) or contact with acid or alkaline aqueous solutions, oxidants or reducing agents, or organic solvents. The destruction of the quaternary structure causes or may cause the protein to lose its biological activity in living cells. However, the tertiary structure of the released subunit has a specific shape generated by hydrogen bonds, van der Waals forces, and disulfide bonds, and may have functional activity and exhibit functions similar to those in living cells. An example of this is the lock-and-key function of enzymes due to the tertiary shape of the protein.

[0146] Therefore, if the quaternary or tertiary structure remains essentially the same after extraction as in living cells, these can be considered "native" proteins for the purposes of this disclosure. The present disclosure has discovered that certain oil globular proteins have unique and superior functional properties, and the globular proteins can be considered native in the sense that the tertiary structure is not denatured by temperature changes (typically above 41°C), aqueous acids or bases, oxidizing or reducing agents, or organic solvents.

[0147] Conventional plant protein extraction methods are known to disrupt the quaternary and tertiary structures of proteins. In some cases, this disruption may result in loss or reduction of functionality of the quaternary or tertiary structures. The tertiary structure may be denatured by the disruption of functional bonds and forces, including hydrogen bonds, van der Waals forces, or disulfide bonds, all of which work together to form a specific tertiary protein structure. Changes in the protein environment and the pattern of tertiary structure denaturation may alter the tertiary structure or shape of the protein as well as its bonds, forces, and connections.

[0148] As used herein, the term "isolated plant protein" means a plant protein (which may include proteins such as edema, gluten, albumin, legumin, pea globulin, fava globulin, glycinin and protein isolates such as from any seed or legume, including soybeans, peas, lentils, etc. or a combination thereof), or a plant protein portion (e.g., a 7S portion) that has been separated from other components of the source material (e.g., other animal, plant, fungal, algal or bacterial proteins) such that the protein or protein portion is at least 2% (e.g., at least 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%) free of other components of the source material on a dry weight basis. For example, an isolated native globular protein having a high cysteine ​​content may be used alone or in combination with one or more other proteins such as albumin or any other protein source such as soy, pea, whey, and the like.

[0149] In any method or composition described herein, fat can be non-animal fat, animal fat or a mixture of non-animal fat and animal fat.Fat can be algae oil, fungal oil, corn oil, olive oil, soybean oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed oil, rapeseed oil, mustard oil, safflower oil, sunflower oil, linseed oil, palm oil, palm kernel oil, coconut oil, field comfrey oil (ahi oil), babassu oil, shea butter, mango oil, cocoa butter, wheat germ oil, borage oil, black currant oil, sea buckthorn oil, macadamia oil, saw palmetto oil, conjugated linoleic acid oil, arachidonic acid-rich oil, docosahexaenoic acid (DHA)-rich oil, eicosapentaenoic acid (EPA)-rich oil, palm stearic acid, sea buckthorn fruit oil, macadamia oil, saw palmetto oil or rice bran oil; or margarine or other hydrogenated fats.In some embodiments, for example, fat is algae oil.Fat can contain flavorings and / or isolated vegetable protein (such as conglycinin). The fat or oil composition of the liquid matrix may be made to preferentially match the saturated and unsaturated composition of the target source material of the analog.

[0150] Therefore, in some embodiments, the isolated protein can be substantially a protein isolated from hemp seed or any other grain that may have edema protein or edema protein-like protein, such as native edema protein. In some embodiments, proteins can be separated based on their molecular weight, such as by size exclusion chromatography, ultrafiltration or density centrifugation through a membrane. In some embodiments, proteins can be separated based on their surface charge, such as by isoelectric precipitation, anion exchange chromatography or cation exchange chromatography. Proteins can also be separated based on their solubility, such as by ammonium sulfate precipitation, isoelectric precipitation, surfactant, detergent or solvent extraction, including water extraction. Proteins can also be separated by their affinity to another molecule, such as using hydrophobic interaction chromatography, active dyes or hydroxyapatite. Affinity chromatography can also include the use of antibodies with specific binding affinity to heme-containing proteins, nickel nitroacetate (NTA) with specific binding affinity to recombinant proteins of His tags, lectins bound to sugar moieties on glycoproteins, or other molecules that specifically bind to the protein. In some embodiments, the plant-based meats described herein are composed substantially or entirely of ingredients derived from non-animal sources, such as plant-based, fungal, or microbial sources. In some embodiments, a plant-based meat product or a plant-based dairy product may include one or more animal-based products. For example, a meat replica may be made from a combination of plant and animal sources.

[0151] References:

[0152] The following documents are incorporated herein by reference in their entirety: US Patent Application Serial No. 17 / 551,163 to Mitchell Ellis; US Patent No. 7,678,403 to Mitchell and Mitchell.

[0153] definition:

[0154] Hemp seeds are generally defined herein as live seeds that are typically used for further propagation and planting. HS may or may not be food grade based on cleaning practices and seed agricultural preservation practices.

[0155] Whole hemp seed (WHG) is generally defined herein as hemp seed including live hemp seed and pasteurized hemp seed.

[0156] Live hemp seeds (VHG) are generally defined in this article as live hemp seeds that have been further cleaned of all dust and foreign matter, meet food grade requirements, and have the core and shell intact.

[0157] Pasteurized hemp seed (PHG) is generally defined herein as hemp seed that has been treated with heat or radiation to destroy seed viability.

[0158] Defatted hemp kernel cake (DHGC) is generally defined herein as the dry solid residue resulting from the non-aqueous removal of oil from hemp kernel.

[0159] Hemp seed oil (HGO) is generally defined herein as the virgin green oil produced by the non-aqueous extraction of hemp seeds.

[0160] Hemp seed oil sludge (HGOS) is generally defined herein as a crude oil slurry produced by the non-aqueous extraction of oil from hemp seeds.

[0161] Hulled hemp seeds (HHG) are generally defined herein as equivalent to hemp hearts or hemp seeds; hemp seeds with the hull removed.

[0162] Defatted hulled hemp kernel cake (DHHGC) is generally defined herein as the dry solid residue resulting from the non-aqueous removal of oil from hulled hemp kernel.

[0163] Hulled hemp seed oil (HHGO) is generally defined herein as a yellow oil produced by the non-aqueous extraction of hulled hemp seeds.

[0164] Hemp protein isolate (HPI) is generally defined herein as an isolate of albumin, edestin, or aggregates thereof.

[0165] Aqueous oil albumin emulsions (AOAE) are generally defined herein as water-based emulsions of oil and soluble albumin.

[0166] Native edema protein isolate (NEPI) is generally defined herein as the product of a protein separation process as disclosed herein, and may refer to NEPI in liquid, slurry, and powder forms as understood by one of ordinary skill in the art in the appropriate context of its use.

[0167] All products depicted in the flow diagrams may exist in various physical forms, including liquid, gel or solid, as understood by one of ordinary skill in the art in the appropriate context of their use.

[0168] The present disclosure may relate to compositions containing natural edema protein isolate (NEPI), which contain edema proteins or edema-like proteins, and methods for extracting and using NEPI to produce meat-like and dairy-like products. The present disclosure solves the problems in the prior art regarding hemp seed protein separation, raw material input preparation, and raw material input processing to produce high-quality plant-based meat-like and dairy-like products. The compositions and methods of the present disclosure include methods for hemp seed protein separation, pasteurization, sol formation, gel formation, texturization, and meat-like and dairy-like product production. The methods of the present disclosure produce meat-like or dairy-like products with superior properties compared to existing products or similar products made using known technologies.

[0169] In addition to protein separation, this document is also based on methods and materials for making plant-based products that more closely replicate meat products, including the texture, juiciness, fibrousness and texture uniformity of animal meat. A method for producing a simulated meat product is described herein, which may include selecting proteins based on the unfolding or denaturation, properties and fat retention capacity of the protein. In addition, the method described herein includes a method for preparing an extrusion mixture before extrusion, wherein the method incorporates water and fat into the selected protein in a manner such that water and fat form a liquid matrix with the protein (which may also be referred to herein as a liquid fat hydrosol, hydrosol, extruder or extrusion input, and input material). Further, the method described herein includes a method for extruding or otherwise heating the liquid matrix. The method for extruding the liquid matrix includes feeding the liquid matrix into a pump at the first end of the extrusion chamber. The liquid matrix is ​​fed into the extrusion chamber of the extruder, wherein the extruder is set to parameters suitable for the liquid matrix.

[0170] As disclosed herein, NEPI can be extracted from hemp seeds or other grains, kernels or seeds containing edema proteins or edema-like proteins; hemp seeds are currently believed to be the only source of edema proteins. In one embodiment, hemp seeds are wet-milled and subjected to water extraction to produce an insoluble edema protein-containing extract, which is referred to herein as NEPI, and an aqueous oil albumin emulsion.

[0171] The method according to the present disclosure can produce pasteurized and functional hemp seed protein concentrate, wherein the concentrate can be a concentrated liquid from a production line or from centrifugation and decantation, or a NEPI powder, which in some embodiments may have a low content of trypsin inhibitors or no trypsin inhibitors and has high nutritional value and functionality. The method may not use isoelectric extraction, alkali or CO2 dissolution methods. The organized protein NEPI concentrate or NEPI powder is believed to be produced by oil extraction and albumin separation, utilizing the natural pH and oil content of hemp seeds in combination with water. The emulsion forming ability of soluble albumin can form an emulsion that can be easily separated from insoluble hemp globulin by centrifugation. No freeze drying, pH readjustment and ultrafiltration separation are required. In addition, fiber and chlorophyll may be removed during the NEPI process. Maintaining a low temperature, preferably between 33°F and 38°F, promotes the insolubility of globulin and the coagulation of albumin.

[0172] One aspect of the present disclosure relates to the isolation of edema proteins and edema-like proteins from plant materials, including hemp seeds. Eedema proteins are present in the hemp plant; in particular, in hemp seeds. Although hemp seeds are considered the most common or only source of edema proteins, other plants may also contain edema proteins.

[0173] The edema protein extract composition or natural edema protein isolate (NEPI) prepared according to the method of the present disclosure can be used to prepare a protein-containing composition. The NEPI may preferably be composed of about 80% dry basis protein; in some embodiments, the NEPI may contain at least 65% dry basis protein, and in some embodiments may contain at least 90% dry basis protein. Therefore, NEPI can be defined as a composition containing edema protein produced according to the method described in the present disclosure, resulting 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, including hemp seed oil or kernel oil and albumin.

[0174] The present disclosure may be practiced using suitable grains, seeds or plant materials containing edestin or edestin-like proteins, wherein such edestin-like proteins may be homologous or have similar structure and function.

[0175] The grain used in the present disclosure can be substantially full-fat vegetable grain, i.e., grain that has not been defatted or pressed prior to milling. In some embodiments, the grain can be partially defatted. Partially defatted grain includes any plant material from which at least a portion of the fat has been removed.

[0176] The fat (or oil) content of substantially full-fat hemp seed can be 10% fat or more by weight, as known to those of ordinary skill in the art. In the present disclosure, the terms fat and oil are used interchangeably. Suitably, the fat content of substantially full-fat seeds is at least about 10%, 15%, 20%, 30%, 40% or even 50% by weight. The fat content of hemp seed is typically at least 30%. The fat content of partially defatted plant material can be greater than about 5%, 10% or 15% fat by weight. After removing the husk, the edible portion of hemp seed contains an average of 46.7% oil and 35.9% protein.

[0177] like Figure 1 As shown in , hemp seeds 102 can be selected for use in the structured protein food method 100. Whole hemp seeds 101 and shelled hemp seeds 105 can be used. Pasteurized whole hemp seeds 103 produced by a pasteurization method 107 can also be used. The hemp seeds 102 used in accordance with the present disclosure can be prepared for processing by a suitable means, including but not limited to drying, conditioning to achieve a balanced moisture level, shelling, crushing and cleaning by countercurrent air suction, screening methods, pasteurization without damaging live seeds, or other methods known in the art. In the present disclosure, the hemp seeds 102 can be selected from the kernels of hemp containing no more than 0.3% THC. The hemp seeds 102 can be whole hemp seeds or shelled (shelled) hemp seeds 102, wherein the hemp seeds 102 can be shelled prior to processing in the structured protein food method 100 to produce shelled hemp seeds 150, such as Figure 4 as shown in .

[0178] View now Figure 1 The hemp seed 102 in the structured protein food method 100 is subjected to a natural hemp globulin separation method 200 (eg Figure 2 ) to extract natural edible protein isolate (NEPI) 250. Natural edible protein isolate (NEPI) slurry 252 or powder 254, or NEPI 250 can be used to produce structured protein food 120, which can be a meat-like product or a dairy-like product. Conventional methods of extracting hemp protein or edible protein from hemp seeds or producing hemp protein isolates may cause edible protein and albumin aggregation or protein denaturation, and may not produce satisfactory structured protein foods or meat-like products. However, NEPI 250, when used as the sole protein source in meat-like products, is able to produce high-quality and novel meat-like products without the need to be combined with soy or other types of plant-based protein isolates, as described by Zahari (Zahari et al., 2020).

[0179] like Figure 1As shown in FIG. 1 , in some embodiments, NEPI 250 may be pasteurized 104 and combined 106 with water to form a protein hydrosol 108. NEPI 250 may be combined 106 with preheated water to form a protein hydrosol 108 (e.g., Figure 6 NEPI 250 should be present with water at least 20% w / w, and with water at up to 80% w / w or higher. The protein is fully hydrated. The hydration time will depend on the conditions. Mixing under high shear is preferred to promote hydration.

[0180] Oil can then be added to the protein hydrosol 110, followed by high shear mixing 112. In some embodiments, after high shear mixing 112, the mixture can optionally be incubated without mixing 113. Adding oil 110 and mixing 112 produces a protein-fat hydrosol 114.

[0181] The protein-fat hydrosol 114 is used as an input for heating the protein-fat hydrosol to solidify the product 116. Solidification may involve heating by means including microwaves, steam tunnels, ovens, retorts, and extrusion (e.g., Figure 7 and Figure 8 ). The heating and coagulation method may include other methods known to those skilled in the art for heating protein-based or starch-based foods to form coagulation. Coagulation of protein-fat hydrosol 114 produces structured protein food 120. Structured protein food 120 may be a meat-like product or a dairy-like product.

[0182] like Figure 2 As shown in , to produce NEPI 250, hemp seeds 102 can be added to cold water 202 to form a hemp seed mixture 204. The extraction temperature during milling and throughout the natural hemp protein isolation method 200 can be more preferably 35°F, or between 33°F and 38°F, or less than about 120°F, which can be added to the hemp seeds 102 to form the hemp seed mixture 204. The hemp seeds can be extracted with an aqueous solution, suitably with water. As used herein, the term "aqueous solution" includes water that is substantially free of solutes (e.g., tap water, distilled water, or deionized water) and water that contains solutes. According to the present disclosure, the aqueous solution can be free of additives such as salts, buffers, acids, bases, and demulsifiers. In some embodiments, the ionic strength of the aqueous solution can be lower than the ionic strength that will change the protein structure. More or less water can be used.

[0183] In the present method, no pH adjustment is required to isolate NEPI. Preferably, the pH is maintained approximately neutral between 6.5 and 7 throughout the structured protein food method 100. In one embodiment, the pH of the solution does not change to any significant extent during the milling of the grain.

[0184] The hemp seed mixture 204 can be wet milled 206 substantially as described in U.S. Pat. No. 7,678,403 to Mitchell. In one embodiment, a Silverson rotor stator type mill can be used to grind the hemp seed 206. Wet milling 206 can be performed as part of a water extraction method. Suitably, the water wet milling 206 can be performed for a suitable period of time, and more suitably, the wet milling 206 is performed for a suitable period of time. As will be appreciated by those skilled in the art, longer extraction periods of time can be used. In some embodiments, enzymes can be used to assist in processing. For example, alpha-amylases with dextrinizing activity can be used to achieve liquefaction to produce a liquefied slurry. Such enzymes can include amylases or other carbohydrases known in the art of food processing. In one aspect, the present disclosure can utilize a water wet milling method to separate fat stored in the hemp seed 102 without destroying chloroplasts and releasing chlorophyll into the oil. Calcium chloride can be added to the NEPI 250 to improve the flavor after centrifugal decantation 222.

[0185] After the wet grinding of hemp seed 206, the extract can be separated from at least a portion of insoluble byproducts or fiber slurry 210 (e.g., insoluble fiber portion) by a sieve. In some embodiments, the hemp seed slurry 208 can be screened in two steps. Screening can remove unwanted impurities that can bring unpleasant color or taste to hemp protein. Insoluble fiber can be removed by the first screening step. Another undesirable product that can be surprisingly removed by screening without significantly affecting protein yield is chlorophyll from chloroplasts in hemp seed and shelled hemp seed, which can produce unwanted color, taste and fat oxidation in the oil part or protein part. In some embodiments, chlorophyll-containing particles can be removed in the second screening step 212. After screening 212, chloroplasts and fiber sludge may be in the retentate, and raw hemp seed milk with a fat to protein ratio of about 1:3:1 on DSB is in the filtrate.

[0186] In the first screening step, in some embodiments, the hemp seed slurry can be sieved through 30 meshes to remove shell. The by-product of the first screening step can be a fiber slurry 210. In the second screening step 212, the hemp seed slurry can be sieved 212 with about 170 meshes to remove chloroplasts, or in some embodiments with 160 to 200 meshes, or in some embodiments with 200 to 220 meshes to remove chloroplasts or chlorophyll-containing material and any remaining fiber. The opening of 150 mesh sizes may be too large usually, and may allow unwanted materials to enter filtrate, including fiber and chlorophyll-containing particles. Surprisingly, the chlorophyll-containing particles keep the size greater than 170 mesh pore openings, and most of the protein-containing particles are through the screen of this size. Screening with different sizes of screens separates chloroplasts, protoplasts or other chlorophyll-containing particles from the part containing hemp seed oil and protein, and obtains a light yellow final oil product.

[0187] In some embodiments, the chloroplasts isolated by the edible protein extraction method 100 can be used as a food supplement. According to the method of the present disclosure, chlorophyll-containing particles 214 are selectively removed from the hemp seed slurry 208 while allowing protein-containing particles to pass through to enter the filtrate. This method is effective for both whole hemp seeds and dehulled hemp seeds that have not been hulled prior to water milling.

[0188] After the hemp seed slurry is screened with 170 mesh to remove chlorophyll-containing particles 212, the resulting product is an aqueous oil albumin emulsion (AOAE) and edestin globulin mixture 220, which may also contain other components of the hemp seed 102 to a greater or lesser extent. The AOAE and edestin globulin mixture 220 can be centrifuged and decanted 222 to produce NEPI 250 and AOAE 230. After separation from NEPI 250, AOAE 230 can be further processed to produce albumin 550 and hemp seed oil 560, such as Figure 5 as shown in .

[0189] In some embodiments, NEPI 250 may be composed of approximately 76% protein, 2% oil, 4% fiber, 1% carbohydrates, and 17% ash. AOAE 220 may be composed of approximately 14% protein, 76% oil, 3% fiber, 4% carbohydrates, and 3% ash. In some embodiments, NEPI may preferably be composed of about 80% dry basis protein; in some embodiments, NEPI may contain at least 65% dry basis protein, and in some embodiments may contain at least 90% dry basis protein. Thus, NEPI may be defined as a composition containing edestin produced according to the methods described in the present disclosure, resulting in a product having the functional characteristics described in the present disclosure. In some embodiments, NEPI may contain at least about 65%, 75%, 85%, or 90% protein by dry weight.

[0190] Table 2 shows the approximate nutritional data of NEPI 250 and commercial hemp protein products. Table 2 shows that the NEPI 250 product has the same high protein content and protein to fat ratio as VICTORY HEMP. The protein content and protein to fat ratio of other commercial products are much lower. This indicates that among the products tested, NEPI 250 and VICTORY HEMP may be far superior to the other products.

[0191] Figure 9-1 1 shows the SDS PAGE gel data of NEPI 250 product and commercial product, indicating the protein composition, structure and integrity (non-reducing conditions are shown in Fig. 9 The reduction conditions are shown in Fig.10 About Fig. 9 , 910 is edestrin dimer, and 920 is albumin. Fig.10 , 930 is the acidic subunit of edestin, 940 is the basic subunit of edestin, and 950 is albumin. FIG. 11 shows prior art SDS PAGE data illustrating the known molecular weights of edestin and edestin products under similar conditions. The lanes are labeled as follows and are applicable to Fig. 9 and Fig.10 :

[0192] M = molecular weight standard

[0193] 1=DP-276HempLife pwd SD HPI

[0194] 2=DP-276HempLife pwd SD HPI

[0195] 3=DC-344 HempLife Liquid Concentrate HPI

[0196] 4=GH-350Good Hemp pwd HPI

[0197] 5=A-560Anthony's pwd HPC

[0198] 6=LP-643Hulled HempLife SD pwd HPI

[0199] 7=VH-794Victory Hemp pwd V70 HPI

[0200] 8=N-950Nutiva pwd HPC

[0201] 9=N-950Nutiva pwd HPC

[0202] Figure 11A shows the prior art SDS PAGE of hemp seed proteins published by Mamone and Wang (Mamone et al., 2019; Wang and Xiong, 2019). Figure 11B shows the prior art SDS PAGE of hemp seed proteins published by Shen (Shen et al., 2020).

[0203] In general, Figure 9-10 The NEPI 250 product is shown to have a different protein composition from other commercial products and is generally more complete in structure, with VICTORY HEMP being closest in terms of native edema protein content and undegraded protein products. Interestingly, as predicted, the NEPI 250 product is substantially free of albumin. It is assumed in the present disclosure that albumin interferes with the ability of hemp seed protein isolate to form a well-structured protein food 120 with excellent texture properties. This theory is supported by the texture analysis data shown in Table 2, where the NEPI product has a higher hardness and chewiness compared to commercially available hemp seed protein products. It is also possible that the excellent natural structural features of edema protein in NEPI 250 contribute to the unexpectedly excellent texture properties of NEPI 250 shown in Table 2. Table 3 and Figures 12 and 13 further show the excellent structural preservation of edema protein in the native state, which show the differential scanning calorimetry data of the product.

[0204] Table 3 shows differential scanning calorimetry thermograms that provide structural information about the edestin proteins contained in NEPI 250 and commercial products. DSC thermograms of two NEPI products (FIG. 12) and two commercial hemp protein powders, VICTORY HEMP and NUTIVA (FIG. 13). Based on the DSC results, the NEPI products are superior in structure to the commercial products, and it is shown that the edestin proteins in NEPI 250 are in a more natural state than the commercial products.

[0205] Compared to the hemp protein isolate previously described in the background of the invention produced by conventional methods, the hemp protein in NEPI 250 is of superior quality. In addition, the protein extraction methods of the prior art have significant disadvantages and limitations compared to the methods of the present disclosure. For example, salt extraction and dialysis in the HMI method do not remove residual phenolic substances from the final product. In addition, the commercial viability of HMI is low.

[0206] The method of the present disclosure has many advantages over the prior art. The method can release phenolic substances and tocopherols from NEPI 250 and AOAE 230. The method of the present disclosure can make the hemp seed oil 560 more oxidatively stable. In the method of the present disclosure, phenolic substances can be separated from the hemp seed oil 560 during the water wet milling process, thereby providing stability.

[0207] The method of the present disclosure is different from conventional methods for extracting protein from hemp seeds, because conventional methods generally involve pressing the seeds to extract oil and produce hemp seed cake, which can then be ground and sieved to produce powder. The resulting cake or powder may contain aggregated edema globulin and albumin, as well as oil, carbohydrates, phenolic substances and minerals. In some cases, the seeds can also be directly dry-milled to produce powder.

[0208] Mechanical methods that result in high temperatures or pressures, such as pressing the kernels, can result in the formation of chemical bonds between edestin and albumin. Pressing whole or dehulled hemp kernels can result in aggregation of edestin and albumin.

[0209] High pressure changes protein structure and causes protein aggregation. According to Yang, high pressure modification of proteins involves changes in the secondary, tertiary, and quaternary structures of proteins, from native to intermediate to fully denatured states (Yang et al., 2016). High pressure changes protein structure primarily by changing non-covalent bonds-electronic interactions, hydrophobic interactions, and hydrogen bonds. High pressure also leads to the formation of new disulfide bonds, which stabilize denatured proteins or produce protein aggregation (Yang et al., 2016).

[0210] It is well known that heat also changes protein structure. The heat generated by friction during grain milling can cause changes in protein structure. Heat can lead to protein denaturation and the formation of protein aggregates. Aggregation between edestrin and albumin is likely to occur during dry milling where temperatures can reach 100°C or higher.

[0211] In one embodiment, the NEPI may then be heated to a temperature of about 145°F and maintained for about 30 minutes to pasteurize the product. In some jurisdictions, 145°F may be the legal lower limit for pasteurization. In one embodiment, the temperature may be maintained at about 145°F, or between 145°F and 155°F, to prevent granulation. It has been observed in the present disclosure that the formation of granules occurs at a temperature of about 158°F. Granulation may occur in the NEPI at a temperature well below the denaturation temperature of edestin (e.g., at about 158°F), which has been demonstrated to be about 95°C. It is critical to pasteurize the NEPI at a temperature lower than that typically used by those skilled in the art to pasteurize plant proteins used in food. Those skilled in the art typically pasteurize protein isolates at a temperature that results in significant granulation in the present disclosure in order to process the product quickly. The pasteurized NEPI 270 is the result of washing with cold water and diluting 232.

[0212] like Figure 3 As shown in , after pasteurization 104 is complete, NEPI 250 can be spray dried via a NEPI spray drying process 300 or refrigerated as a concentrate for use in producing structured protein food products 120. Immediately after separation in the centrifugal decanter, the solids content of the NEPI concentrate is in the range of about 35% to 45% and is a thick, viscous paste that is difficult to pump. Cold water is added at this point to reduce the solids of the NEPI 250 concentrate to preferably about 30% to facilitate rapid pumping of the slurry through heated pipes maintained at a temperature not exceeding 158F. Dilution allows for more turbulence and better heat distribution to heat to 145F and allow for pasteurization without the formation of overheated protein aggregates and particles that are undesirable in the finished dried edestin product. Prior to spray drying, the NEPI concentrate can be maintained in a tank at approximately 145°F or the pasteurization 104 temperature prior to spray drying. Spray drying 306 may then be performed at a higher spray drying 306 temperature, or where the exiting product may reach temperatures of about 158°F and above, which may cause protein aggregation and produce less functional NEPI 250. This protein aggregation can be seen on a non-reducing SDS-PAGE gel at approximately 100 kDa (e.g., Fig. 9), where bands other than the expected edestin or hemp protein are visible. Bands present at high molecular weights above the expected approximately 50 kDa band for edestin dimers under non-reducing conditions may represent aggregation caused by overheating during spray drying 300. Thus, in some embodiments, one potential method of measuring whether the maximum temperature of 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, where NEPI 250 is maintained at a low temperature, such as between 130F and 140F, during microwave drying while removing moisture under vacuum pressure.

[0213] Figure 4 A method of adding color to a structured protein food 120 is shown. White and dark meat analog process 400 can produce white meat NEPI 412 that can replicate chicken or fish and dark meat NEPI 422 that can replicate beef or dark meat chicken. To produce white NEPI 422, shelled hemp seed 105 can be used. In one embodiment, shelled hemp seed 105 can be subjected to a natural edible globulin separation process 200, which produces white meat NEPI 412, which can be used in structured protein food process 100 to produce white meat replicas. To produce dark meat NEPI 412, whole hemp seed 101 can be used. In one embodiment, whole hemp seed 101 can be subjected to a natural edible globulin separation process 200, which produces dark meat NEPI 412, which can be used in structured protein food process 100 to produce dark meat replicas. In one embodiment, dark NEPI 412 or intermediate color NEPI 432 can be produced using part whole hemp seed and part shelled hemp seed, wherein the whole hemp seed is used at a concentration of about 20-30% by weight relative to the amount of shelled hemp seed. In one embodiment, the husk previously removed by hemp seed shelling can be reintroduced into the shelled hemp seed 105 to increase the dark flesh color; wherein, in one embodiment, the husk can be added to the shelled hemp seed in an amount of about 10-15% by weight relative to the shelled hemp seed 105 to produce an intermediate color NEPI 422 in order to obtain a dark flesh color.

[0214] Figure 5A method 500 for oil and albumin extraction is shown. AOAE 230 is the product of the natural hemp protein separation method 200 and can be processed to produce albumin 550 and hemp seed oil 560. In the oil and albumin extraction method 500, AOAE 230 can be evaporated to concentrate 506. The product can be homogenized 504 and heated to pasteurize 530. It may be useful to clarify the AOAE 502. Heating to 180F 520 may break up emulsions. Preferably evaporate until there is more oil than water 506. Cool to near freezing or freezing point 508. Centrifuge with a cream separator 510 to obtain albumin 550 or hemp seed oil 560.

[0215] Figure 6 A hydrosol forming method 600 is shown in which NEPI 250 can be combined with preheated water to form a protein hydrosol 108, substantially as described in Figure 2 In the hydrosol formation method 600, preheated water at about 135°F may be added to the NEPI 250 and mixed 106 under high shear to form the protein hydrosol 108. The protein hydrosol may be pasteurized at 145°F before, during, or after the protein hydrosol is formed. Pasteurization conditions should be maintained or created after the NEPI 250 is produced. The pasteurized 104 product may be prepared as follows: if spray dried 306 to form the NEPI powder 308, by first hydrating the NEPI 250, or otherwise maintaining the appropriate hydration of the NEPI 250 and maintaining pasteurization conditions as much as possible. In one embodiment, the amount of preheated water added to the NEPI 250 may be such that the solution reaches approximately 3 parts water to 1 part NEPI by weight of dry solids. In some embodiments, the NEPI may be frozen 310 in a cooler and freeze dried 312 to produce the NEPI powder 308. It may be useful to heat the hydrosol to 130F 111. It may be useful to heat the oil to 110-115F.

[0216] In some embodiments, the preheated water may be tap water, and in some embodiments 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 hydration and protein preparation because it may destroy the protein hydrosol 108 or protein-fat hydrosol 114 structure. Salt may be added after coagulation, but not before coagulation. In some embodiments, protein hydration and opening (so that, without being bound by theory, the protein structure may be slightly changed or opened to allow proper interaction with oil during the formation of the protein-fat hydrosol 114) may be performed at 100°F to 135°F, or in some embodiments between 100°F and 155°F; or in other embodiments, protein hydrosol formation may be performed at a lower temperature, however, the temperature must be higher than a low temperature that does not allow protein hydration and opening. Preferably, the temperature during the hydration and protein preparation steps should be kept as close to 145°F or the pasteurization 104 temperature as possible without reaching a temperature that may cause protein aggregation and granulation. Once the protein hydrosol is formed, the preheated oil 109 may be heated to 110°F to 115°F in some embodiments, and to 100°F to 155°F in other embodiments, or in some cases maintained at a temperature above that which is considered cold so that the protein hydrosol structure is disrupted by the addition of the oil, but below a temperature that produces granulation of the protein-fat hydrosol 114.

[0217] In some embodiments, the protein-fat hydrosol 114 can be produced by combining a fat with a warm hydrated protein suspension (e.g., a protein isolate containing edema globulin) at a pH between 6.5 and 7.8 (e.g., pH 7.5). Rapid stirring, such as in a Waring blender or handheld homogenizer, or homogenizing the mixture results in the formation of an emulsion. The physical properties of the protein-fat hydrosol 114 can be controlled by varying the protein type, protein concentration, pH during homogenization, homogenization speed, and fat to water ratio.

[0218] To form the protein-fat hydrosol 114, a polyunsaturated fatty acid (PUFA) oil or fat (which may be preferably coconut oil or fat) may be heated to just above the melting point of the fat and added to the protein hydrosol 108. Without being bound by theory, the fat may form a layer around the hydrated native edema protein, thereby forming a liquid matrix or protein-fat hydrosol 114 that substantially encapsulates the hydrated protein, forming an oil-in-hydrated protein emulsion that effectively produces a viscous and stable gel. The oil may effectively seal and protect the hydrated protein structure. Compared to dry protein, hydrated protein can accommodate significantly more fat in a gel state. In general, it has been found that, as discussed in this application, native globular proteins that are first hydrated and then gently heated to below their denaturation temperature can accommodate up to twice their weight of fat. In some embodiments, the moisture content of the protein-fat hydrosol may be in the range of about 30% by weight to about 70% by weight. Moisture content refers to the moisture content in a material measured by an analytical method, calculated as the percentage change in mass of the sample after water evaporation.

[0219] In any method or composition described herein, the protein-fat hydrosol 114 may include flavorings or other additional ingredients. Based on the final protein-fat hydrosol 114, the following ingredients may be optionally added in an amount typically less than 2% by weight: fat-soluble or other flavor systems, salts including sodium chloride, plant-based albumin sources, plant-based insoluble or soluble fibers. Starch may be added alone or in combination with other soluble carbohydrates, including complex carbohydrates or sugars, where necessary, in an amount of up to about 10% by weight, but more preferably less than 5% by weight. Auxiliary ingredients may be added to the protein-fat hydrosol 114 before solidification to improve and change flavor or texture. Fiber may be added to reduce the "squeak" of the structured protein food 120.

[0220] In one embodiment, protein-fat hydrosol 114 can include about 15 wt % to about 25 wt %, or more preferably about 18 wt % to about 22 wt % protein on the one hand, wherein the protein can be natural oil seed protein; wherein in one embodiment, about 75 wt % to about 85 wt % protein isolate comprises globular protein, and preferably, protein isolate comprises albumin less than 15 wt %, and more preferably less than 5 wt % albumin. More importantly, globular protein can be in its natural state and preferably has the amino acid cysteine ​​of significant content, and its amount is greater than casein or soy protein isolate. In some embodiments, the remainder of protein composition can be mainly minerals, such as calcium and phosphorus. Natural oil seed globulin preferably can have a large amount of cysteine.

[0221] In one aspect, the protein-fat hydrosol 114 can include from about 40% to about 70%, or more preferably 40%-60% water by weight.

[0222] In one aspect, the protein-fat hydrosol 114 can include about 0% to about 35% fat by weight; the ratio of saturated fatty acids to polyunsaturated fatty acids (PUFA) is between 100% saturated fat by weight and 100% PUFA by weight. Depending on the amount of protein used in conjunction with the fat, the combination of these two amounts of fat provides a variety of unique textures that have not been reported to date.

[0223] In some embodiments, the protein-fat hydrosol 114 may optionally include from about 0% to about 5% by weight starch. The amount of starch added may depend on the amount of water added, in excess of the amount of water required for protein hydration added to the protein.

[0224] The protein-fat hydrosol 114 can be formed by manually or mechanically mixing the ingredients for forming the protein-fat hydrosol 114. Preferably, the hydrated protein is first warmed to just below the granulation temperature of the protein, oil and / or melted fat is added, and the mixture is preferably gently homogenized.

[0225] In one aspect, the protein-fat hydrosol 114 can be combined at a temperature between 120°F and 150°F. It has been found that the temperature range for placing the protein in a heated environment without destroying the formed gel or matrix is ​​between 70°C and 100°C. These temperatures are significantly lower than the extrusion temperatures typically required to extrude conventional meat-like proteins (e.g., soy). Under the conditions typically used in an extruder, the denaturation and fiberization temperature of soy protein is in the range of about 130°C to 140°C. According to the present disclosure, good texturization can be obtained by heating the protein-fat hydrosol 114 in an oven and / or by pressure cooking (retorting) the protein-fat hydrosol 114 to actively coagulate the protein.

[0226] The physical properties of the protein-fat hydrosol 114 are those of the hydrosol. Viscosity depends on the oil, fat and water as well as the protein content. Higher water content changes and can significantly reduce viscosity, even if the protein to fat ratio is low. Likewise, very low protein to fat ratios and low water content can result in very high viscosities. The quality and selection of the fat system and protein system can also significantly affect viscosity.

[0227] The formation of the protein-fat hydrosol 114 can be carried out below the denaturation point of the native protein. However, according to the present disclosure, it is not desirable to store the protein-fat hydrosol 114 at said temperature because it is microbiologically unstable. It is preferably immediately treated by heating to set the protein shape. The liquid matrix can be additionally cooled to below 6° C. by a heat exchanger or other method for storage before further processing.

[0228] Figure 7 A NEPI cooking process 700 is shown to produce a structured protein food 120. The NEPI protein-fat hydrosol 114 is dispensed into formed TETRAPAK 200 mL containers 702, in one embodiment, each container is filled with 180 g. The top can be sealed using a TETRA RECART machine 704. The packaged NEPI protein-fat hydrosol can be placed into a cooking machine 706. The NEPI protein-fat hydrosol can then be heated 708 under cooking conditions to solidify 710. In some embodiments, this process produces a structured protein food 120.

[0229] With respect to cooking according to the present disclosure, Figures 14-18 show photographs of the cooking results of various NEPI products and commercially available hemp seed protein powders. Each figure contains an enlarged view of the cooked product. Boiled chicken was used as a standard. Table 6 below shows the results of the texture analysis of the cooked structured protein food. Tables 7 and 8 show the colorimetric data for each product produced and tested by cooking, using boiled chicken breast as a standard.

[0230] Figures 14-18 show photographs of cooked NEPI hulled powder 250, where the solids are approximately 2: 1 protein to fat (NEPI 250 to coconut oil) and the solids to liquid (water) ratio is approximately 2: 3. After preparation of the protein-fat hydrosol, the cooked product is produced as known to those of ordinary skill in the art.

[0231] Fig.14A It is a photo of a cross section of a poached chicken breast; Fig. 14B Is from Fig.14A A magnified photo of a cross section of a boiled chicken breast; Fig. 14C Is from Fig. 14B Photo of a magnified cross-section of a boiled chicken breast.

[0232] Fig.15A is a photograph of a cross-section of a cooked meat analog product using NEPI shelled hemp seed concentrate according to the present disclosure; Fig. 15B Is used from Fig.15A A magnified photograph of a cross section of a cooked meat analog product of NEPI's shelled hemp seed concentrate; Fig. 15C Is used from Fig. 15BA magnified photo of a cross section of a cooked meat analogue made from NEPI's shelled hemp seed concentrate.

[0233] Fig.16A is a photograph of a cross-section of a cooked meat analog product using NEPI shelled hemp seed powder according to the present disclosure; Fig. 16B Is used from Fig.16A A magnified photograph of a cross section of a cooked meat-like product of NEPI's shelled hemp seed powder; Fig. 16C Is used from Fig. 16B A magnified photo of a cross section of a cooked meat-like product of NEPI's shelled hemp seed powder.

[0234] Fig.17A is a photograph of a cross-section of a cooked simulated meat product using VICTORY HEMP shelled hemp seed powder according to the present disclosure; Fig. 17B Is used from Fig.17A A magnified photograph of a cross section of a cooked meat-like product of VICTORY HEMP shelled hemp seed powder; Fig. 17C Is used from Fig. 17B A magnified photograph of a cross section of a cooked meat analog product of VICTORY HEMP shelled hemp seed powder.

[0235] Fig.18A is a photograph of a cross-section of a cooked simulated meat product using HEMPLAND shelled hemp seed powder according to the present disclosure; Fig.18B Is used from Fig.18A A magnified photograph of a cross section of a cooked meat-like product of HEMPLAND shelled hemp seed powder; Fig.18C Is used from Fig.18B A magnified photograph of a cross section of a cooked meat-like product of HEMPLAND shelled hemp seed powder;

[0236] Figure 8 A method 800 of extruding a NEPI 250 to produce a texturized structured protein food 120 is shown. Figure 8The steps of providing an extruder with a heated auger are shown, preferably, in one embodiment, a hollow steam heated auger 801, or other types of heated auger extruders. In one embodiment, the extruder can be a POWERHEATER PH 100 provided by SOURCETECHNOLOGY. The technology used in this machine that may be used in the present disclosure may be described in the following U.S. Patents and U.S. Patent Applications: No. 10,893,688, No. 10,624,382, No. 10,149,484, No. 10,092,013, No. 10,028,516, No. 9,931,603, No. 2010 / 0062093, No. 2011 / 0091627, No. 2019 / 0299179, No. 2020 / 0113222, No. 2020 / 012095, and No. 2020 / 02680205, which are incorporated herein by reference in their entirety. The POWERHEATER PH 100 can allow for better control of the temperature of the auger and the inner wall of the extrusion tube or chamber because the hollow auger design allows steam to be introduced into the auger to heat the auger and provide a more uniformly heated protein-fat hydrosol in the present disclosure, which is critical for proper coagulation of the present disclosure. Conventional extruders, such as those developed by CLEXTRAL or WENGER were tested in the present disclosure and did not provide a satisfactory final product. Conventional extruders cause the protein-fat hydrosol of the present disclosure to adhere to the inner wall of the extruder tube.

[0237] POWERHEATER PH 100, although known for use with fiberized input materials, is generally known for coagulating starch, rather than protein, in its input materials. Protein coagulation extrusion is generally performed at temperatures well above 100°C, and therefore protein coagulation input materials are not considered for use with POWERHEATER PH 100. However, the protein-fat hydrosol of the present disclosure is effectively texturized and fiberized by POWERHEATER PH 100 at 75°C when fiberizing the protein-fat hydrosol of the present disclosure (which is accomplished at relatively low temperatures between about 75°C-85°C), and wherein the auger and extruder may be preheated to 75°C-85°C 802, and extrusion may occur in the range of about 70°C-95°C. In one embodiment, the protein-fat hydrosol extruder uses an 8 mm screw size, rather than a 3 mm screw size, and uses POWERHEATER PH 100 at 75°C. The protein-fat hydrosol may be introduced into the POWERHEATER PH 100 using a suction pump or a filler pump, wherein the starting temperature may be about 85° C. 802. After the protein-fat hydrosol is pumped into the extruder 804, extrusion may be performed at about 75° C.-85° C., wherein the protein-fat hydrosol does not adhere to the inner wall of the extrusion tube 806. This process produces a texturized structured protein food 120. The texturized structured protein food 120 extruded according to the present disclosure has been demonstrated in testing to have a texture, fibrosis, and color similar to cooked chicken breast, which is superior and unexpected when considering the prior art and the knowledge of those of ordinary skill in the art.

[0238] Fig.19 Photos of NEPI extruded from shelled powder and a piece of boiled chicken breast according to the present disclosure are shown to show the texture and fibrosis similarities, as extruded on the POWERHEATER PH 100 described above. Boiled chicken breast 1910 is shown next to an extruded NEPI 250 chicken product 1920, produced from spray-dried shelled hemp seed NEPI and processed according to the present disclosure. This result is unexpected for hemp seed, which uses only three ingredients, NEPI 250, coconut oil and water, in ratios of 2:1:3, respectively.

[0239] In most extrudates, including soy-based meat analogs, it has been seen that the ratio of protein to fat is typically greater than 10: 1. Thus, extruded, denatured, and fiberized soy can hold very little fat. However, according to the present disclosure, hydrated gels of natural globular proteins such as edestin can hold up to twice their weight in fat, even after forming the gel in a coagulum or solid form produced by application of radiation, microwaves, or other forms of heating, including direct heating or extrusion.

[0240] According to the method of the present disclosure, the protein-fat hydrosol 114 can be coagulated into a solid state at a temperature between about 70° C. and 100° C., depending on the concentration of protein in the system. The lower coagulation temperature is consistent with the denaturation of the native protein in NEPI 250.

[0241] According to the present disclosure, the solid structure formed during extrusion can be cooled and represents a coagulum, but with incomplete denaturation, similar to uncooked protein or "raw" meat. Further heating of the "uncooked" protein enhances shape, elasticity, texture, etc. by further denaturing the protein, a process that will eventually release some water. According to the method of the present disclosure, it is not desirable to heat the product to the extent that a large amount of water is released from the coagulum in the extruder, but rather it is desirable to only solidify the shape or texture of the gel and protein. In one embodiment, the present disclosure describes a method for preparing a raw imitation meat product or imitation dairy product or structured protein food 120 similar to raw animal meat in an extruder. Further cooking of this raw imitation meat product by traditional or commercial means can enhance the strength and toughness of the meat.

[0242] The process according to the present disclosure is in contrast to the prior art, in which simulated meat textures are created by using fully denatured proteins and then blending with other binders including fats, starches and other proteins to create the appearance of a hamburger-type material. According to the prior art, this type of coagulation is primarily achieved during the cooking process by gelatinization of starch or added crude proteins (e.g., gluten).

[0243] The final texture of the structured protein food 120 may depend on the characteristics of the liquid matrix, including the ratio of protein, fat and water, and the extrusion conditions. As described herein, the extruded mixture of isolated vegetable protein may be referred to as a structured protein food 120, which may be a meat-like product, and the fibrousness and tensile strength of the meat-like product may be controlled by co-variation of extrusion parameters such as temperature, pressure, output, and die size. For example, a combination of lower extrusion temperature, medium / low output, and smaller die is favorable for producing a high fibrous tissue with low tensile strength, while a higher extrusion temperature, higher output, and larger die is favorable for producing a low fibrous tissue replica with very high tensile strength.

[0244] The fiber size and tensile strength of the meat analog product can also be adjusted by changing the composition of the extrusion mixture. For example, by increasing the ratio of isolated vegetable protein to fat and water, or by reducing the water content in the extrusion mixture, a meat analog product with finer fibers and greater tensile strength can be produced.

[0245] Extruding the liquid matrix includes feeding the liquid matrix into an extruder. In some embodiments, the extruder can be a SOURCE TECHNOLOGY POWERHEATER PH 100. CLEXTRAL and WENGER twin screw extruders were tested, but the results were not satisfactory. In extrusion, according to the method of the present disclosure, cooling is important to achieve a temperature below 21° C. so that the saturated fat can easily solidify in the structure and the product can be more effectively cooled to refrigeration or freezing temperatures.

[0246] For each product, the wet ingredient blend will be transferred to a feeder, which can meter the liquid matrix through the feed port of an extruder at a specific input rate. In conventional extrusion, a dry protein product is fed to the input end of the machine. When the dry product moves through the machine, water and fat are introduced from a separate input end. By contrast, in the method according to the present disclosure, firstly hydrated protein and oil are mixed as described above to closely regulate the chemical reaction that occurs during the formation of protein-fat hydrosol 114. Therefore, in some embodiments, additional water, starch or fat may be added or not added to the extruder during extrusion. In some embodiments, fiber may also be added.

[0247] In conventional extrusion of plant-based imitation meat products, adding water and fat before starting extrusion may result in unwanted steam release because water escapes from the product as the temperature rises. Therefore, for the present disclosure, the process of adding water and fat is strictly controlled during extrusion. In the method according to the present disclosure, the liquid matrix extrusion mixture is specifically designed to prevent water from being released from the product by forming a gel. During the preparation of the liquid matrix according to the present disclosure, oil is added to the hydrated protein to form an emulsion gel, which prevents the release of water from the product during extrusion, otherwise the water would be released from the machine as steam. The formation of the gel also allows for maintaining high moisture in the liquid matrix during extrusion and in the final product, which is desirable for the excellent texture of the structured protein food 120.

[0248] The temperature during extrusion is very important for the resulting product. The temperature should be gradually increased and maintained between about 70°C and 100°C, or between 100°C and 110°C. In conventional extrusion, the temperature in the extruder is usually higher than 130°C. In the method of the present disclosure, low temperature prevents the destruction of the protein-fat hydrosol 114, thereby allowing the molecular structure of the compound to remain intact substantially or partially. The temperature of the protein-fat hydrosol 114 can be maintained between about 75°C and 85°C, preferably, to solidify the protein-fat hydrosol 114, and then cool to reduce the temperature to less than 21°C during extrusion. For the method of the present disclosure, it is important to maintain a lower temperature than the temperature used during conventional extrusion. Here, the temperature is only increased to a point that allows disulfide bonds to solidify, so that fat is fully incorporated between all peptide layers of the protein. The residence time in the extruder or any heating environment should be sufficient to enable the input temperature of the liquid matrix to reach 70°C to 110°C, or preferably 75°C to 85°C.

[0249] Preferably, the extruder rotates the protein-fat hydrosol 114 at a relatively low screw speed (measured in revolutions per minute (rpm)) during extrusion to form a meat analog that maintains a gel structure and maintains a high moisture content in the product. The screw speed can be closely monitored to prevent temperature increases and prevent chemical structural damage to the liquid matrix.

[0250] In order to prevent the structure of the loose protein-fat hydrosol 114 formed by the hydration of protein and fat encapsulation from being destroyed, the gel may have to be slowly moved through the heating system to keep the initial gel solidification (partial protein denaturation), while forming shape and some fibrosis. Fermentation (occurring in cheesemaking) or cooking and denaturation completely will eventually occur in the later use process of the product. In some embodiments, the final extruded product with a moisture content between 35 % by weight and 75 % by weight can then be fermented, refrigerated or frozen to keep microbial stability, until if necessary, it will be cooked completely at higher temperatures by common or commercial cooking methods, to obtain required final texture before eating. Additional related extrusion parameters may include product temperature and feed rate of die diameter, die length, die end.

[0251] After extrusion, the structure of the final product may be more similar to animal meat than conventional or known structured protein foods, such as imitation meat products and imitation dairy products. Without being bound by theory, according to the present disclosure, the extrusion of the protein-fat hydrosol 114 can form proteins into substantially arranged protein fibers, wherein the protein fibers can be defined as continuous filaments of discrete lengths consisting of proteins held together by intermolecular forces, such as disulfide bonds, hydrogen bonds, electrostatic bonds, hydrophobic interactions, peptide chain entanglements and Maillard reaction chemistry, to produce covalent crosslinks between protein side chains. The coagulation strength after the initial extruder is not complete or as strong as possible. In fact, it may be necessary to remove the heat-coagulated finished product and further heat it by direct or indirect heating, common cooking methods such as boiling, baking, frying, baking, microwaves, fermentation and pressing (such as in cheese making, which may include salting and adding acid) and the like to complete the coagulation of the strength or form of the initial coagulated product.

[0252] According to the method of the present disclosure, the preparation and extrusion conditions of the protein-fat hydrosol 114 can allow the substantially aligned protein fibers to retain up to about 50% by weight of fat in the protein in some embodiments. Therefore, the final product is not greasy, and the mouthfeel and fat release when chewed are closer to animal meat than existing simulated meat products. Mouthfeel can refer to a combination of characteristics including moistness, chewiness, bite force, degradation and fatness, which together provide a satisfactory sensory experience.

[0253] The desired final structure of the structured protein food 120 may vary based on the composition of the protein-fat hydrosol 114. In one embodiment of the present disclosure, the desired final composition of the structured protein food 120 by weight of protein, weight of carbohydrates (if any), weight of lipids, and weight of water along with any other possible components is shown in Table 4. 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. The extruded product can be subjected to post-processing steps.

[0254] Meat analog products, which may also be referred to herein as structured protein foods 120, may be produced from protein-fat hydrosol 114 by methods other than extrusion. Other methods of producing meat analog products from protein-fat hydrosol 114 include application of mechanical energy (e.g., shear, pressure, friction), radiant energy (e.g., microwave, electromagnetic), thermal energy (e.g., heating, steam texturization).

[0255] The present invention will be further illustrated by the following examples, which do not limit the scope of the invention described in the claims.

[0256] Example

[0257] Example 1

[0258] Preparation of Natural Enoki Protein Isolate (NEPI)

[0259] Hemp seeds were obtained from Hemp Oil Canada, Manitoba Canada, and River Valley Specialty Farms, Manitoba Canada. Shelled hemp seeds were obtained from River Valley Specialty Farms, and whole hemp seeds were obtained from Hemp Oil Canada.

[0260] HHG contains 5.5% moisture by weight, 46% protein on a dry basis Kjeldahl, 35% fat on a dry basis, and a protein to fat ratio of 1.3 to 1. WHG contains 8.8% moisture by weight, 22% protein on a dry basis Kjeldahl, 30% fat on a dry basis, and a protein to fat ratio of 0.7 to 1.

[0261] In an 800-gallon agitator tank, 1000 pounds of HHG were mixed with 5000 pounds of water at 34°F. The HHG was wet-milled, keeping the temperature between 34°F and 38°F. The hemp seed slurry was milled at a rate of 56 gallons per minute in a SILVERSON rotor stator slot for 30 minutes to wet-mill the HHG. The diluted slurry was kept for an average of 30 minutes. The extract was separated from the insoluble byproducts using a 120-mesh SWECO 60-inch screen to remove most of the solids. The 120-mesh screen was then passed through a 200-mesh screen on another SWECO vibrating screen to obtain a slurry, which was then transferred to a 500-gallon jacketed tank to keep the temperature of the slurry between 34°F and 38°F. The slurry was then fed to a DELAVAL centrifugal decanter at a rate of 13gpm to achieve separation of the edible globulin solids from the AOAE emulsion. The AOAE emulsion was then pasteurized for 10 minutes through a tubular heat exchanger system at a maximum temperature of 185F. The AOAE was then stored in a 900 gallon tank for processing. The edestin solids at 40% solids were diluted to 30% solids with cold water and pumped through a preheated tubular system set at below 150F and exited the system at 146F into a jacketed storage tank at 145F in the jacket. After 30 minutes, the material was cooled to 35F through a heat exchanger and placed in totes in a refrigerator for further processing and drying through a spray dryer.

[0262] In an 800-gallon agitated tank, 1000 pounds of WHG was mixed with 5000 pounds of water at 34°F. The HHG was wet milled to maintain the temperature between 34°F and 38°F. The hemp seed slurry was milled at a rate of 48 gallons per minute in a SILVERSON rotor stator slot for 30 minutes to wet mill the WHG. The diluted slurry was kept for an average of 30 minutes. The extract was separated from the insoluble byproducts using a 60-mesh screen on a double-stage Sweco 60-inch screen to remove the husk. The second stage of the SWECO was equipped with a 200-mesh screen so that the slurry from the SILVERSON first passed through the 60-mesh screen to remove the husk and immediately fell on the top of the 200-mesh screen to remove the chloroplasts and fine fibers. The rate through the SWECO was about 6 gallons per minute, and the screened slurry went directly into a jacketed 500-gallon jacketed tank to maintain the temperature of the slurry between 34°F and 38°F. When the tank was full, the slurry without husks, fibers or chloroplasts was fed to a DELAVAL centrifugal decanter at a rate of 13 gpm to achieve separation of edema protein solids from the AOAE emulsion. The AOAE emulsion was then pasteurized for 10 minutes at a maximum temperature of 185F through a tubular heat exchanger system. The AOAE was then stored in a 900 gallon tank for processing. The light brown edema protein solids with a solid content of 40% coming out of the decanter were diluted to a solid content of 30% with cold water and pumped through a preheated tubular system set below 150F and left the system at 146F and entered a jacketed storage tank with a temperature of 145F in the jacket. After 30 minutes, the material was cooled to 35F through a heat exchanger and placed in a tote bag in a refrigerator for further processing and dried by a spray dryer. The NEPI dry matter basis yield based on the WGH weight starting material was 15% or 79% of the theoretical value. The AOAE yield was 25.3% DSB, and the shell, fiber and chloroplast fraction was 46.9%, with an overall DSB recovery of 92%. The NEPI yield of HHG was 30% or 86% of theoretical. The AOAE yield was 40.9% DSB, and the shell, fiber and chloroplast fraction was 22.5%, with an overall DSB recovery of 98%. The analysis of the NEPI products obtained from WGH and HHG is shown in Tables 1 and 2 below.

[0263] Table 1 Composition of natural hemp globulin isolate from non-pasteurized shelled and dehulled hemp seeds

[0264]

[0265]

[0266] The NEPI concentrate prepared by the process of the present disclosure exhibits high microbial activity prior to pasteurization and spray drying to powder even while maintaining the process temperature below 38F. (See Table 1). The total plate count (TPC) of incoming raw materials from hemp seeds or dehulled hemp seeds is typically in the range of 2,000 TPC to 250,000 TPC. In a protein-rich aqueous medium, the temperature must be maintained well below 42F, and preferably below 38F. Despite the low temperature, if pasteurization is not performed immediately after the start of water milling, the TPC will continue to increase and cause protein deterioration. The short duration of the process and the ability to pasteurize both the AOAE and edema protein slurries immediately after separation by a centrifugal decanter are important factors in the process. The edema protein product obtained by pasteurization at a low temperature of 145F retains the gelling function mentioned above. The AOAE can be heated at much higher temperatures, in excess of 145F and more preferably 195F for a short time, which facilitates further processing to remove the remaining insoluble solids by centrifugation and then breaking the emulsion to separate the aqueous albumin and oil phases. The success of pasteurization of the NEPI product in final powder form is reflected in the TPC of the product in Table 1.

[0267] Table 2

[0268] Composition of natural edible protein isolate (NEPI) and commercial hemp protein products

[0269]

[0270]

[0271] Table 3 shows the DSC thermograph. The structure of NEPI measured by DSC thermograph (e.g. Fig. 12A -B and Fig.13A -Part B) can be compared with the following commercially available products.

[0272] Table 3 Differential Scanning Calorimetry

[0273]

[0274] Further structural and compositional analysis of NEPI and commercially available hemp seed protein products as measured by SDS-PAGE gel electrophoresis is shown in Fig. 9 and Fig.10 middle.

[0275] Example 2

[0276] Spray Drying NEPI

[0277] The frozen slurry of NEPI obtained from Example 1 was sent to a commercial spray dryer for drying. An ALFA LAVAL model spray dryer with a nozzle capable of removing 1200 pounds per hour of water was used to dry the powder. The refrigerated product was pumped into a jacketed 250 gallon tank that maintained the jacket at 155F using a set water temperature. The tank had a slow agitator and the product required several hours to heat approximately 200 gallons of 30% concentrated edema globulin slurry. Once the product reached temperature, it was sent to another tank that fed the dryer. It should be noted that NEPI dries very easily and does not stick to the walls of the dryer. The final outlet temperature of the dried product was 85F. For each NEPI (WG and HHG) product obtained from Example 1, the composition of the dried product is given in Table 2 below.

[0278] Example 3

[0279] Production of protein-fat hydrosols from NEPI and commercial hemp seed powders

[0280] The protein hydrogel is easily prepared in a 5 gallon plastic bucket by adding 14 pounds of water that has been preheated to 140F. Slowly add 14 pounds of NEPI dry powder to the water while stirring with a 1 / 4 horsepower handheld industrial homogenizer wand. Keep homogenizing until all the powder is added. After holding the temperature, now at 130F, for about 15 minutes, add 7 pounds of canola oil all at once and briefly blend the mixture with the homogenizer wand for about 1 minute or until the slurry appears well blended and the oil is incorporated as a uniform emulsion.

[0281] Example 4

[0282] Protein-fat hydrosol formulations and properties of different types of meat and dairy analogs

[0283] Example 4 discloses formulations comprising a liquid matrix for producing various types of meat analog products. According to the present disclosure, depending on the ratio of protein, fat and water, different types of meat analog products can be produced, including plant-based meat analog product targets that replicate seafood, white meat, dark meat, eggs and cheese.

[0284] Table 4

[0285] Different types of protein-fat hydrosol formulations for meat-like and dairy-like products

[0286]

[0287] With respect to Table 4, the moisture content target is between 35 wt% and 75 wt%. A minimum of 70 wt% of the globular natural vegetable protein has an albumin content of less than 15 wt%, preferably less than 5 wt%. The liquid matrix temperature should be maintained at 140°F from blending to processing. Due to the ability of natural seed oil protein (which may be natural edible globulin in Table 4), the amount of fat may vary to obtain different types of simulated meat products. The structural characteristics of the resulting products are similar to those of the materials they replicate. For example, the seafood texture is white with a very elastic structure, similar to raw shrimp or scallops. The white meat is white and has a texture similar to that expected from a partially cooked chicken steak. The dark meat is slightly light brown and has a texture similar to chicken legs, with more fat and moisture than the white meat. The eggs are similar to what is expected from scrambled eggs and are also white. The cheese is similar to cheese curds and actually squeaks when bitten into a piece similar to fresh cheese curds.

[0288] Table 5 Protein-fat hydrogel formulations and physical properties

[0289]

[0290] Example 5

[0291] Producing structured protein foods by cooking

[0292] The cooking conditions were a temperature increase from 77F to a peak of 270F in 15 minutes and a decrease to 95F at 15 minutes. The pressure was 0.20 bar at 1 minute and increased to 3.0 bar at 4 minutes and decreased to 0.8 bar at 15 minutes. The machine used was a SUNDRY RETORT TYPE: AP-95, Serial No.: 705.

[0293] Table 6

[0294] Texture analysis

[0295] Cooked structured protein foods

[0296]

[0297]

[0298] Table 7

[0299] Colorimetric comparison

[0300] Steamed products

[0301] Whiteboard Standards

[0302]

[0303] Colorimeter-Chroma Meter CR-400-Konica Minolta 2021-12-03Table 8

[0304] Colorimetric comparison

[0305] Steamed products

[0306] Boiled chicken standard

[0307]

[0308] Colorimeter - Chroma Meter CR-400 - Konica Minolta 2021-12-03 Table 9 Texture Analyzer Cutting Test

[0309]

[0310] Example 6

[0311] Producing structured protein foods through extrusion

[0312] The protein-fat hydrosol of Example 3 was used in a Power 100 Source Technology extruder at 185F set at a flow rate of 6 pounds per minute and a 3MM auger diameter to produce a structured gel with the appearance and texture of white meat chicken. For a pictorial comparison of white meat chicken and extruded hydrogel structured protein foods, see Fig.19 .

[0313] The present disclosure unexpectedly demonstrates that surprisingly superior hemp seed-based structured protein products can be produced using only three ingredients: hemp seed, oil, and water. Hemp seed meat analogs produced according to the present disclosure are shown herein to replicate chicken to a surprising degree in color, texture, and taste. When used for this purpose, commercially available protein products, some of which claim to produce superior meat analogs, cannot compare to native hemp protein isolate in terms of taste, color, or texture.

[0314] No commercial products have been found that use only hemp seed protein to produce meat-like products. In addition, the prior art teaches that hemp seed protein alone is not a viable protein for producing structured protein foods, such as meat-like products and dairy-like products. The present disclosure proves that this is not the case.

[0315] definition:

[0316] As used herein, the articles "a" and "an" when used herein, for example "an anionic surfactant" or "a fiber," should be understood to mean one or more of the materials claimed or described.

[0317] "Basic weight" is the weight of a sample of web material (on one side) per unit surface area (in the machine / cross plane) expressed in grams per square meter (gsm). Basis weight may be specified in the manufacturing specification or may be measured and reflects the weight of the material before any liquid composition is added.

[0318] As used herein, "network structure" means a network- or sheet-like hydrogel comprising units of at least a line, a sheet, and a portion adjacent to the side wall of a container or adjacent to the bottom.

[0319] As used herein, "container" means an object that is capable of containing a liquid protein-fat hydrosol and can be used in a microwave oven.

[0320] The "container material" used herein may include materials capable of holding liquids, and may be composed of preferably 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, paper blended with materials such as plastics that allow heat treatment or can hold boiling water, and paper containers including polylactic acid (PLA) instead of conventional plastics; and ceramic materials, including glazed and unglazed ceramics; glass, including microwave glass and other materials known to those of ordinary skill in the art.

[0321] As used herein, "expansion ratio" refers to the V of a protein hydrosol or a protein-fat hydrosol after microwave heating. max Divide by the V of the protein hydrosol or protein-fat hydrosol i As used herein, volume measurements of an expanded product will include voids 2124 formed by gas bubbles, unless otherwise specified.

[0322] The final volume (V f ) means the temperature from the final height (H f ) is the volume of the protein-fat hydrogel in the container measured by , and the final height used in this article refers to the highest point on the container where the materials are combined after collapse.

[0323] The final meniscus center volume (V mc ) means the volume of the protein-fat hydrogel in the container after microwave heating, as measured after the top surface of the collapsed material has collapsed from the container after microwave heating has terminated. This calculation may be an estimate and is not always an accurate measurement of volume, as the shape of each run may vary significantly even if all conditions are the same.

[0324] As used herein, "hydrogel meniscus" means a complete or partial meniscus or concave surface formed by a hydrogel material that may exist after microwave heating according to the present disclosure. The hydrogel meniscus may be formed by a top layer of the hydrogel material. Without being bound by theory, this top layer may be referred to as a protein film or a protein-oil film. This definition may include a complete or partial meniscus that may be formed when bubbles and hydrogel material collapse when microwave heating stops. At this point, a certain amount of hydrogel material may be bonded to the side walls of the container, and a portion of the hydrogel in the center of the container may drop below the maximum height of the material bonded to the side walls of the container, thereby forming a meniscus or partial meniscus having a crescent or concave shape. In general, the center of this meniscus may be approximately located at the center of the container, which may be the bottom of the meniscus or partial meniscus. The meniscus may have a final height (H) from the bottom of the concave portion of the meniscus to the hydrogel. f ) is measured as the meniscus depth, wherein the final height is measured at the highest point where the hydrogel is bonded to the side wall of the container after microwave heating.

[0325] As used herein, "inclusions" means edible materials that may be included in a hydrogel formulation.

[0326] As used herein, "layer" refers to a thickness, ply or fold of protein-containing material disposed on or beneath another layer.

[0327] As used herein, "meniscus ratio" means the meniscus depth divided by the final height (H) of the protein-fat hydrogel in the container. f ).

[0328] As used herein, "microwave oven" means a form of electromagnetic radiation having a wavelength ranging from about one meter to one millimeter, corresponding to frequencies between 300 MHz and 300 GHz, respectively. In all cases, microwaves may include at least the entire SHF band (3 to 30 GHz, or 10 to 1 cm). Typically, consumer-grade microwave ovens operate at a nominal frequency of about 2.45 gigahertz (GHz), or 12.2 centimeters (4.80 inches) in the 2.4 GHz to 2.5 GHz ISM band, while large industrial or commercial microwave ovens typically use 915 megahertz (MHz), or 32.8 centimeters (12.9 inches). For purposes of this disclosure, all wavelengths of microwave radiation are contemplated, but preferably, conventional microwave radiation for cooking food in a home, commercial or industrial setting may be utilized.

[0329] As used herein, "particulate" means a granular material or a powder.

[0330] "Predominantly" or its forms, with respect to the proportion of a component of a structure or composition, means that the component constitutes the majority by weight of the structure or composition.

[0331] As used herein, "visually discernible" means visible to the naked eye.

[0332] Herein, when the amount of a component of a fibrous web 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 accounts for X percent of the total weight of the material it comprises.

[0333] With respect to a web or fibrous web structure, "z-direction" means the direction normal to the general plane of the extent of the web structure.

[0334] Fig. 20 A flow chart describing a method for producing a ready-to-eat meat analog is disclosed. In one embodiment, heating water to boiling 110 or a sufficiently high temperature can be the first step of the microwave texturization method 2000 of the present disclosure. The water can be heated to boiling 2010, as known to those of ordinary skill in the art. The heating method can include a stove, a microwave oven, a hot plate, etc. In one embodiment, hot water 2002 (which may not be higher than 90° C. in some embodiments) is poured into a measuring container, and then the hot water 2002 is added to a container 2012 (shown in Figure 21). In one embodiment, after the water is added to the container 2100, the water temperature preferably does not exceed 85° C. In some embodiments, the amount of NEPI 250, hot water 2002 or similar liquid, and oil 110 added to the container 2100 may be 25 grams of NEPI 250 or similar material, 110 grams of hot water 2002 (which may be homogenized prior to addition), and 12.5 grams of oil 110 to produce a simulated chicken breast product or other simulated meat product 120 (which in some embodiments may resemble chicken breast; wherein the container 2100 may have a 16 oz. paper Comfort In one embodiment, heating the protein-fat hydrosol 114 to produce a protein-fat hydrogel 120 (which is also interchangeably referred to herein as a structured protein food product 120) may be performed A microwave oven (model HMC54151UC / 05, manufactured in May 2018, input 1700W, output 1000W) is used for about 1.5 minutes at 50% power to provide a preferred protein-fat hydrogel 120; wherein the cycle time of the microwave oven is 30 seconds. In one embodiment, only 20% of the water 200 is lost during microwave heating, wherein the remaining water 200 can be incorporated into the protein-fat hydrogel 120.

[0335] The container 2100 should be able to adequately hold the hot water 2002. In some embodiments, the container 2100 may be similar to a food grade, microwave-safe paper. Comfort In other embodiments, the paper container 2100 may contain polylactic acid (PLA), and may be a Basics Compostable 20 oz. Thermo Paper Cup. The container 2100 can be disposable, recyclable, or compostable. In some embodiments, the container 2100 can be constructed from various types of paper, as known to those skilled in the art. In other embodiments, the container 2100 can be constructed from plastic. In some embodiments, this plastic can be a lidded container for use in a dip stick blender. 24 oz plastic polycarbonate measuring cup jar, The interior walls of the container may become sufficiently roughened due to regular use of the blender over a period of time that the protein-fat hydrogel may adhere to the container side walls 2102. The 24 oz plastic pitcher can also adhere to other microwaveable materials, which may include porcelain, pottery, glass, Pyrex, glass ceramics, paper, silicone, and thermoplastics. In some embodiments, the container 2100 may have a rough interior so that the protein-fat hydrogel 120 can bind or adhere to its surface.

[0336] In some embodiments, the container 2100 may be constructed of a material suitable for heating food products in a microwave oven while insulating the container so that when the cooked interior material reaches 150°F to 212°F, the exterior temperature of the container 2100 remains below 140°F, or cool enough to handle comfortably. Preferred materials may include ceramic, HDPE, polypropylene, double or triple layer paper containers, or similar materials. The materials may preferably be recyclable and environmentally sustainable. In some embodiments, the preferred container 2100 may be a double or triple layer paper container 2100. An example of a ceramic container 2100 for use in accordance with the present disclosure is a W&P® 1000® with a protective silicone sleeve. Ceramic mug. An example of a microwaveable but non-insulated polypropylene container for use in the present disclosure may be the CHOICE 32 oz. Microwavable Translucent Round Deli Container. An example of a microwaveable and insulated three-layer paper container is the Chinet Comfort

[0337] In some embodiments, the container 2100 may preferably be composed of a rough or coarse surface material (e.g., paper). In some embodiments, the fibrous or organized or roughened nature of the container 2100 may allow the expanded protein-fat hydrosol 2110 and protein-fat hydrogel 120 to bond with the container sidewall 2102 of the container 2100 and cool rapidly (e.g., Fig. 21C), which can promote the formation of a more organized final product, which in some embodiments can resemble chicken breast or partially separated or chopped chicken breast. Using smooth materials (such as glass or coated ceramics, which are also good thermal insulators) may be less effective in allowing the protein-fat hydrogel 120 to not only bond to the container sidewall 2102, but also maintain the heat of melting and slow the cooling and solidification of the hydrogel, which may produce a different final product that may be more similar to different types of meat and may be inferior for certain desired products (such as chicken breast).

[0338] In some embodiments, the container 2100 can be transparent so that during microwave heating the protein-fat hydrosol 114 can be observed to expand and rise within the container 2100. In some embodiments, visually observing the complete expansion and rise of the expanded protein-fat hydrosol 2110 during microwave heating and stopping heating when a visual cue (e.g., a visual peak of rise or rise to a desired level) occurs can be part of the microwave texturization method 2000.

[0339] In some embodiments, the container 2100 may have different shapes for producing certain types of imitation meat products or imitation dairy products, such as chicken breast shapes or chicken nuggets. In some embodiments, the container 2100 may have specific dimensions required for certain types of meat products. For example, in some embodiments, when the amount of protein-fat hydrosol 114 added to the container 2100 is 110 grams of water, the container 2100 may preferably have a bottom diameter to height ratio of about 1 to 2.5 and a bottom diameter to top diameter ratio of about 1 to 1.5; and wherein the container volume may preferably be about 16 ounces. In some embodiments, the container 2100 may be generally cylindrical, with an open top, and a diameter of approximately 2 inches to 3.5 inches.

[0340] Other embodiments can produce different types of simulated meat products, including seafood, white meat, and dark meat, as shown and described in U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis, which is incorporated herein by reference in its entirety; more specifically, as shown and described in Example 4 of U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis, which is incorporated herein by reference in its entirety.

[0341] In some embodiments, the preferred size of the container 2100 may be a 16 ounce or 32 ounce container 2100 having a diameter of 2 to 4 inches to support the use of a conventional sized immersion blender, and a height of approximately 5 inches to 8 inches. The size and shape of the container 2100 may vary to accommodate the amount of protein-fat hydrosol 114 used in the microwave texturization method 2000. The preferred amount of protein-fat hydrosol 114 used may vary and is related to the microwave time, settings, and size of the container 2100. Preferably, a minimum amount of microwave time is required to solidify or coagulate the protein-fat hydrogel 120, which may also be referred to herein as a structured protein food or simulated meat product. Uniform heating or uniform heat distribution within the protein-fat hydrosol 114 is desired. The expansion or gas rise of the molten protein-fat hydrosol 114 is very important to achieve elongation of the hydrogel, which gives the protein-fat hydrogel 120 a fibrous appearance when solidified. Without being bound by theory, in the present microwave texturization method 2000, expansion and solidification can occur simultaneously, and the expanded product can be solidified or further solidified by the rapid addition of cold water 128, thereby cooling the protein-fat hydrogel 120 to a more rigid, more ductile state. After the protein-fat hydrogel 120 solidifies, preferably no liquid or material remains in the container, indicating that the heat distribution during the heating process is complete and uniform.

[0342] After adding 112 hot water or hot water 2002 to container 2100, NEPI 250 or a similar base material (which may be flavored NEPI 250) may be added to the hot water 2002 or similar liquid in container 2100. NEPI 250 may be produced as described in U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis, entitled “Native Edestin Protein Isolate And Use As A Texturizing Ingredient,” which is incorporated herein by reference in its entirety; and wherein the NEPI 250 extraction method is more particularly described in paragraphs

[0071] to

[0080] ,

[00151] to

[00162] , and

[00163] of U.S. Patent Application No. 17 / 551,163. Figure 2 , which patent application is incorporated herein by reference in its entirety.

[0343] When NEPI 250 is added to hot water 2002, in some embodiments, the temperature of hot water 2002 is preferably between about 60°C and 80°C, or between 75°C and 85°C; generally, the temperature of hot water 2002 should allow for rapid hydration of proteins and interaction with hot water 2002 when NEPI 250 is added, but the temperature is insufficient to cause granulation of NEPI 250. NEPI 250 may not function properly in microwave texturization process 2000 if heated to a temperature above about 70°C, at which granulation may occur, and therefore it is desirable that the temperature of hot water 2002 in container 2100 is at or below about 80°C prior to adding NEPI 250. If hot water 2002 is at 80°C when NEPI 250 is added, the temperature of the mixture can be quickly reduced to about 70°C when NEPI 250 is added, thereby preventing interference with the function of NEPI 250 in microwave texturization process 2000.

[0344] In some embodiments, it may be important to add NEPI 250 to hot water 2002 after hot water 2002 is already in container 2100. The ratio of hot water 2002 to NEPI 250 can vary depending on the desired texture and type of meat analog product, as described in U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis, entitled "Natural Edema Protein Isolate and Its Use as a Texturizing Ingredient," which is incorporated herein by reference in its entirety.

[0345] In one embodiment, water is first added to the container 2100 and heated separately in a microwave oven. Additional water is added to the NEPI 250 concentrate, and the mixture is then slowly heated to a target temperature of 60°C. Oil 110 may then be added to the mixture. Optionally, the protein-fat hydrosol 114 mixture may then be cooled, and then gently preheated to 60°C prior to microwave heating.

[0346] The present disclosure utilizes a protein-containing solution, a protein hydrosol 108, or more preferably a protein-fat hydrosol 114, to produce a protein-fat hydrogel 120 in a microwave oven. Under the conditions described in the present disclosure, the protein-fat hydrosol 114 will form a unique protein-fat hydrogel 120 when heated in a microwave oven. In some embodiments, oil 110 may not be added to the protein hydrosol 108, and the method will be performed without the addition of oil 110. Important conditions for producing the claimed protein-fat hydrogel 120 may include the content of the protein-fat hydrosol 114, the amount or volume of the protein-fat hydrosol 114 in the container 2100, the size and shape of the container, the material constituting the container, the power setting of the microwave oven, and the type or structure of the microwave oven. Other conditions that may be important for the production of protein-fat hydrogel 120 (which may also be referred to as structured protein food product 120) include the addition of additives (including particulates or inclusions) to protein-fat hydrosol 114, the presence or absence of a lid on the container during heating, the temperature of protein-fat hydrosol 114 prior to microwave heating, and different types of homogenizers used to prepare protein-fat hydrosol 114. In some embodiments, the addition of salt to protein-fat hydrosol 114 may also affect the final product by increasing the expansion ratio. These factors that may be important for the production of protein-fat hydrogel 120 are not exclusive, and additional factors may be included or considered, as will be understood by one of ordinary skill in the art.

[0347] An important element of the products and methods of the present disclosure is that when the protein-fat hydrosol 114 is heated in a microwave oven, voids 2124 are formed within the protein-fat hydrosol 114 as the volume of the protein-fat hydrosol 114 expands. These voids may have a diameter or width of at least 1 mm, or at least 2 mm, or at least 5 mm. For the purposes of the present disclosure, the volume expansion of the protein-fat hydrosol 114 (which may also be simply referred to herein as expansion) is defined as at least a portion of the protein-fat hydrosol 114 rising vertically within the chamber of the container 2100. Without being bound by theory, the expansion of the protein-fat hydrosol 114 during microwave heating may be related to the formation of steam pockets within the molten hydrosol protein film, which form bubbles within the protein-fat hydrosol 114. Under certain conditions, according to the present disclosure, the protein-fat hydrosol 114 will expand or rise 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.

[0348] During microwave heating, the protein-fat hydrosol 114 may first begin to "melt", forming a film structure, and then trapping water molecules. When the water molecules reach a temperature of 100°C and form a gas, the protein film begins to expand until it reaches a temperature at which it can solidify, or a temperature at which substantially all of the protein-fat hydrosol 114 can solidify. Solidification can be defined as a transition from a liquid state to a solid state, wherein the solid state initially remains moldable when hot or warm. Once solidified, according to the present disclosure, the product is referred to as a protein-fat hydrogel 120 in a moldable or non-moldable solid state, or as a protein hydrogel 108 if no fat is added. Further heating beyond the solidification point of the protein-fat hydrogel 120 may cause the quality of the protein-fat hydrogel 120 or the structured protein food 120 to deteriorate.

[0349] Reference now Fig. 20 After adding 116 NEPI 250 to the hot water in container 2100, NEPI 250 and hot water 2002 may be blended 2018 to form a smooth and uniform open protein hydrosol 108. Blending may be performed by an immersion blender 2600, such as Fig.30 As shown. 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 container bottom 2104 or base (e.g. Fig.30 2600). The handle 2602 can be used to rotate the blade end 2604 to facilitate thorough mixing of the ingredients. An immersion blender 2600 suitable for use in the present disclosure can be Immersion Blender. In some embodiments, the blender can be a hand blender, an immersion blender or a stick blender, a single-serving blender, a portable blender, a benchtop blender, a stand blender, or a commercial blender.

[0350] In some embodiments, flavoring agents may be added 114 to NEPI 250 before or after NEPI 250 is added to hot water 2002. In some embodiments, batch preparation of protein-fat hydrosol 114 may be performed in any blender ( However, in order to solidify the protein-fat hydrosol 114 to form the protein-fat hydrogel 120, a preferred amount of the protein-fat hydrosol 114 should be added to a container 2100 of a preferred size and shape, and a preferred microwave power setting and time should be used to achieve the desired results described herein.

[0351] After a minimum blending time of 2018, in order to fully hydrate and open the protein, as evidenced by a smooth and uniform emulsion with no observable visible particles or particles, thereby forming a suitable protein hydrosol 108, oil 110 may be added 120 to the protein hydrosol 108 (2020). Depending on the desired product type, different amounts of oil 110 may be added, as described in U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis and in the following examples. A variety of different types of oil 160 may be used, including sunflower oil, coconut oil, olive oil, and vegetable or animal oils. The type of oil 110 may depend on the type of simulated meat product desired. For example, if a simulated pork product is desired, bacon grease may be added to the protein hydrosol 108.

[0352] After adding the oil 110, the mixture of the protein hydrosol 108 and the oil 110 may be blended 2022 in the manner previously described to form the protein-fat hydrosol 114. In some embodiments, the protein-fat hydrosol 114 may have the appearance of a thick pudding before the pudding is refrigerated and solidified. Excessive blending of the protein-fat hydrosol 114 may reduce the viscosity of the protein-fat hydrosol 114 to the viscosity of a loose milkshake. In some embodiments, after blending, the protein-fat hydrosol 114 may preferably be maintained at a temperature between about 65°C and 70°C or between 0.5°C and 23°C. Maintaining the protein-fat hydrosol 114 at a temperature between 0.5°C and 23°C or between 65°C and 70°C may retard microbial growth.

[0353] The protein-fat hydrosol 114 can then be heated in a microwave oven 2026, and optionally, the container 2100 can be covered by an indicator cover 2024. The microwave oven can be a standard household microwave oven. In some embodiments, the microwave oven can be set to 50% power; wherein, in some embodiments, the operating time in the microwave oven can be approximately 1 to 2 minutes, or more preferably 1 minute and 30 seconds.

[0354] In conventional microwave ovens, variable power levels increase the flexibility of microwave cooking. Each power level provides microwave energy a certain percentage of the time. For example, power level 7 provides microwave energy 70% of the time. Power level 3 is energy 30% of the time.

[0355] In one embodiment, complete cooking is desired so that no residual material remains in the container 2100 after heating. In some embodiments, a double-walled container 2100 may be desired to prevent a user from burning their hands when removing the container 2100 after heating. In some embodiments, a coozie or container sleeve, such as that used with beer cans, may be used to contain the container 2100 after heating.

[0356] Figures 21A-21D The protein-fat hydrosol 114 in the container 2100, the container side wall 2102, the container bottom 2104, the container base 214, Fig. 21C Swelling protein-fat hydrosol 2110, Fig.21D The protein-fat and protein-fat hydrogel 120 in the present invention. Heating the protein-fat hydrosol 114 can make the protein-fat hydrosol 114 rise and form an expanded protein-fat hydrosol 2110, such as Fig. 21C The expanded protein-fat hydrosol 2110 may rise vertically to a certain height and then collapse or degas as the microwave heating stops, wherein the degassed protein-fat hydrogel 120 is as shown in FIG. Fig.21D shown.

[0357] In some embodiments, the protein-fat hydrogel 120 or the protein hydrogel in the case where fat is not included in the protein-fat hydrogel 120 may also be referred to herein as a meat analog 120. Heating in the microwave oven may preferably be stopped when the expanded protein-fat hydrosol 2110 reaches a certain height, wherein a certain amount of the hydrosol material is in an expanded state before collapsing when heating is stopped. The material in the expanded state may be defined as material that is not in the container bottom adjacent portion 2400, wherein the container bottom adjacent portion 2400 may be defined as material that is substantially uniform and in contact with the container bottom and is not part of the container sidewall adjacent portion, wherein the container sidewall adjacent portion may be defined as material that is generally bonded or adhered to the sidewall, but is not part of the line and sheet located more centrally in the container 2100. When the major amount of protein-fat hydrosol 114 solidifies, in some embodiments at least about 99% of the material is in a swollen 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%, and in other embodiments 86%. 3%, 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 approximately at least 40% protein-fat hydrosol 114.

[0358] Further heating may cause the protein-fat hydrogel 120 to dehydrate and deteriorate the final product quality. Having the proper material parameters and container 2100 shape may allow for a preferred product at the maximum height of the expanded protein-fat hydrosol 2110. Adhesion of the expanded protein-fat hydrosol 2110 to the container sidewall 2102 may be important for producing a properly structured or texturized protein-fat hydrogel 120 or structured protein food 120. Too much oil 110 in the protein-fat hydrosol 114 may result in the expanded protein-fat hydrosol 2110 not adhering to the container sidewall 2102, resulting in a denser, solid, unstructured mass of the protein-fat hydrogel 120, like a stopper, lacking the desired texture.

[0359] In one embodiment, the preferred microwave texturization method 2000 may include the shortest heating time to produce a fully cooked protein-fat hydrogel 120, wherein there is no residual material after the protein-fat hydrogel 120 is removed, and this can be indicated by the maximum volume increase or vertical rise of the expanded protein-fat hydrosol 2110. When used with the microwave texturization method 2000, other protein materials, including soy or pea protein isolates and other hemp seed protein isolates tested (such as disclosed in U.S. Patent Application No. 17 / 551,163 to Mitchell-Ellis) failed to produce comparable expanded protein-fat hydrosols 2110 or significant vertical rise, and did not produce acceptable protein-fat hydrogels 120. Other hemp seed protein isolates tested, such as those produced by VICTORY HEMP and HEMPLAND, formed products with loose textures and low elasticity and could not be considered functional meat-like or dairy-like products.

[0360] The examples included herein are for illustrative purposes only, and as will be understood by those of ordinary skill in the art, the methods may vary. In Example 8, 100 mL of protein-fat hydrosol 114 was heated in a microwave oven at a power setting of 5. As observed by a transparent plastic container during heating at power 5 in a Bosch microwave oven, after the first 30 second cycle, 10 seconds into the second cycle, the protein-fat hydrosol 114 began to expand and rise significantly in the container 2100, and then collapsed at the 45 second point when the magnetron was turned off. In the third 30 second cycle, about 5 seconds later, at 1 minute 5 seconds, the protein-fat hydrosol began to expand again. At about 1 minute 20 seconds, the protein-fat hydrosol solidified and collapsed.

[0361] For the present disclosure, the maximum protein-fat hydrosol 114 volume (V max) is defined as the approximate maximum volume to which the protein-fat hydrosol 114 expands before collapsing, which typically occurs when microwave heating is stopped. In some cases, by additional heating, V max It is possible to increase, however, for the present disclosure, V max Defined as the maximum volume achieved for a particular method, not the absolute maximum volume achievable during heating and expansion. V max The approximate maximum protein-fat hydrosol 114 height (H) to which the protein-fat hydrosol 114 rises in the container 2100 during heating in the microwave oven can be used. max ) to measure.

[0362] Generally, the protein-fat hydrosol 114 can expand and the top surface maintains a somewhat uniform height within the container. The bubbles make the top surface partially non-uniform; however, when the top surface rises and reaches a peak height, the height of the top surface can be observed and estimated. The final height used herein has a different meaning from the maximum height, where the final height (H f ) is the highest level of binding of the protein-fat hydrogel 120 to the container sidewall 2102 of the container. The initial protein-fat hydrosol 114 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 .

[0363] When the protein-fat hydrosol 114 reaches V max When solidified later, it has an uneven network structure 2160, such as Fig.24A The fibrous web 2160 may include additives such as softeners or fortifiers, flavorings, inclusions, nutritional additives, fibers, and other additives in addition to the NEPI 250, oil 110, and hot water 2002, which may constitute the core material of the product of the present disclosure.

[0364] like Fig.24A As shown, the network structure 2160 of the protein-fat hydrogel 120 can have a line 2130 that can span the width of the container 2100. The line 2130 can be joined to a sheet of material 2128, wherein the sheet 2128 can be significantly thinner than the line 2130. There can also be a container adjacent sidewall portion 2300, such as Fig.26A and 26B As shown, it is composed of protein-fat hydrogel 120, and in some cases, may be thicker and larger than thread 2130 and sheet 2128. Thread 2130, sheet 2128 and adjacent side wall portion 2300 of the container can be interconnected and form a semi-continuous structure similar to chicken breast fillet or other simulated meat products. These structures are shown in Figures 23-26 and are composed of Fig.22A and 22B The protein-fat hydrosol 114 shown is produced. Figures 22-26 are composed of the same protein-fat hydrosol 114 processed through the steps described in this disclosure.

[0365] Fig.23A and 23B The protein-fat hydrogel 120 is shown in a container 2100. The container sidewalls 2102 and the container bottom 2104 are also shown. The protein-fat hydrogel 120 is shown at its final height (H f ) 2122. The voids 2124 in the protein-fat hydrogel 120 are shown in Fig.23A and 23B middle.

[0366] In one embodiment of the present disclosure, when the protein-fat hydrosol 114 solidifies, substantially all of the protein-fat hydrosol 114 has expanded from the bottom surface of the container and is present in the container adjacent sidewall portion 2300, line 2130 sheet 2128, so that the lower portion of the protein-fat hydrogel 120 does not significantly take the shape of the bottom of the container 2100. In some embodiments, when solidified, the percentage of protein-fat hydrosol 114 in the container adjacent sidewall portion 2300, line 2130 or sheet 2128 (as opposed to pooling at the bottom of the container) is about 5%, or more preferably 10%, or more preferably 15%. For most purposes of the present disclosure, a large amount of protein-fat hydrogel 120 at the bottom of the container 2100 is generally considered undesirable.

[0367] In some embodiments, there may be voids 2124 between portions of the hydrogel 120. These voids 2124 may be caused by the presence of bubbles within the protein-fat hydrosol 114 during solidification. Without being bound by theory, these bubbles may be formed by steam generated by microwave heating of the protein-fat hydrosol 114. In some embodiments, the shape and size of the voids 2124 may be heterogeneous and may provide a desired non-uniform structure to the hydrogel 120. The voids 2124 may form a layer 2200 in the hydrogel 120, such as Fig.25A and 25B In one aspect, layer 2200 can be formed from wires 2130 and sheets 2128 on either side of voids 2124 .

[0368] like Fig.24A and 24B As shown, when V max After the protein-fat hydrosol 114 solidifies, the protein-fat hydrogel 120 may collapse and form a hydrogel meniscus 2121 (e.g. Fig.28 ), the meniscus has a centerline 2126 along the meniscus (as shown in FIG. Fig.28 The center of the meniscus 2119 is as shown in FIG. Fig.24A and 24B As shown. Fig.24A and 24B A plurality of voids 2124 can be seen in the sheet. These voids 2124 can be heterogeneous and range in size from about at least 2 mm or 5 mm to 1 cm or more. The voids 2124 can be formed by bubbles formed during the microwave heating process. The sheet 2128 can be formed in Fig.24A and 24B 21. The mesh 2160 is shown between the threads 2130. The mesh 2160 can include threads 2130 and sheets 2128 throughout the collapsed protein-fat hydrogel 120. In some embodiments, the hydrogel meniscus 2121 can be formed by a protein-oil film 2131 that forms the top of the protein-fat hydrosol 120 during the solidification process. Bubbles can escape through the protein-oil film 2131 to allow the protein-fat hydrogel 120 to collapse and form the hydrogel meniscus 2121.

[0369] like Fig.25A and 25B As shown, as the lines 2130 and sheets 2128 collapse within the protein-fat hydrogel 120, a layer 2200 in the protein-fat hydrogel 120 may form in the hydrogel 120. Immediately after the collapse, and after the microwave heating is stopped, the protein-fat hydrogel 120 may still be malleable and hot, and in some embodiments, may be formed into a different shape before being cooled 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 portions, lines and sheets, and not in the container bottom adjacent portion 2400. This may correspond to, in an Oyster 24 ounce pitcher having a height of about 7 inches and a bottom diameter of about 2.75 inches and a top diameter of about 3.75 inches, where 100 ml of the starting protein-fat hydrosol is about optimal, about 6 mm to 10 mm of material in the container bottom adjacent portion 2400 is unacceptable for a meat analog product.

[0370] Fig.26A and 26BSheet 2128 and layer 2200 of protein-fat hydrogel 120 are shown, along with adjacent portions 2300 of a container sidewall and container sidewall contacting surface 2302. These portions of protein-fat hydrogel 120 are formed by adhesion to the sidewall of container 2100 during expansion and solidification of the product due to microwave heating. Container sidewall contacting surface 2302 can be generally flat, like container sidewall 2102, which can be an aesthetic advantage for simulated meat products 120 prepared according to the present disclosure, as certain portions and surfaces of conventional meat (e.g., chicken breast) can be naturally smooth and flat.

[0371] Figures 27A-27C A container-adjacent sidewall portion 2300 and a container-sidewall-adjacent surface 2302 are shown, as well as a container-adjacent bottom portion 2400 and a container-adjacent bottom surface 2402. In some embodiments, it is desirable to prevent the protein-fat hydrogel 120 from taking on the shape of the bottom of the container 2100. The present disclosure describes conditions that produce a bottom portion of the protein-fat hydrogel 120 that is not molded to the shape of the bottom of the container 2100. Fig.27A shows a front perspective view of a protein-fat hydrogel removed from a plastic container, wherein the bottom of the hydrogel is not substantially molded to the shape of the bottom of the container; Fig.27B A front perspective view of a protein-fat hydrogel removed from a plastic container is shown, wherein the bottom of the hydrogel is partially molded to the shape of the bottom of the container; Fig.27C A front perspective view of a protein-fat hydrogel removed from a plastic container is shown, wherein the bottom of the hydrogel is substantially molded to the shape of the bottom of the container, in accordance with the present disclosure.

[0372] Fig.28 A cross-sectional view of the protein-fat hydrogel 120 after collapse in the container 2100 is shown. It can be observed that a hydrogel meniscus 2121 can be formed in the protein-fat hydrogel 120. This hydrogel meniscus 2121 may be due to degassing of the protein-fat hydrogel 120 after the heating is stopped, resulting in a significant decrease in the height of the hydrogel 120 in the center portion of the material due to the outer portion of the material being fibrillated and bonded to the sidewalls of the container 2100. In some embodiments, the depth of the hydrogel meniscus 2121 can be related to the quality of the protein-fat hydrogel 120 as a product.

[0373] The hydrogel meniscus ratio may be a useful tool for evaluating the quality of a protein-fat hydrogel for a particular application, such as for producing an acceptable meat analog. The hydrogel meniscus ratio may range between 0 and 1. It is also possible that 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.

[0374] A meniscus may not form when at least a portion of the bottom of the meniscus lies flat against the bottom surface of the container, disrupting the shape of the bottom meniscus curve.

[0375] 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 protein-oil film 2131 is flat and has a height equal to H f When the hydrogel meniscus ratio is 0, the meniscus depth is 0, resulting in a hydrogel meniscus ratio of 0 / 1, where 1 is H f .

[0376] According to the present disclosure, for the production of simulated meat products, a hydrogel meniscus ratio between about 0.3 and 0.7 may be preferred. This ratio is obtained using the method of the present disclosure using The ratios are calculated for a plastic 24 ounce jug and a substantially optimal protein-fat hydrosol 114 composition, and as discussed elsewhere in this disclosure, some variation may occur under different conditions. A hydrogel meniscus ratio above 0.7 generally indicates an undercooked product for a meat analog, while a hydrogel meniscus ratio below 0.3 generally indicates an overcooked product for a meat analog. However, in some cases, an overcooked product may be desired.

[0377] The formation of a hydrogel meniscus appears to be unique to the method of the present disclosure, as other protein isolates tested, including soy protein isolate, potato protein isolate, and other commercially available hemp seed protein isolates, were unable to form a hydrogel meniscus under the conditions tested and described in the present disclosure.

[0378] In some embodiments, a protein-fat hydrogel meniscus range of between about 0.3-0.7 may be preferred; in some embodiments, a range of between about 0.4-0.6 may be preferred, in some embodiments, a range of between about 0.45 to 0.55 may be preferred, in some embodiments, a hydrogel meniscus ratio of about 0.5 may be preferred. In some embodiments, a hydrogel meniscus ratio between 0.2 and 0.8 may be preferred. In some embodiments, a hydrogel meniscus ratio between 0.3 and 0. In some embodiments, a hydrogel meniscus ratio between 0-1 may be preferred. In some embodiments, a hydrogel meniscus may not be present. In some embodiments

[0379] Additionally, certain chemical compounds, including carbonate compounds and salts, may increase swelling in the present disclosure and may alter the hydrogel meniscus ratio. Even though such changes may alter the scope of what is claimed, they are considered to be within the scope of the present invention. In some embodiments of the present disclosure, where spray-dried NEPI powder is used, for example, 1% calcium carbonate is added to NEPI 250 to a concentration of 1%. The addition of calcium carbonate, with increasing concentration, may enhance swelling of the protein-fat hydrosol. This may be a result of the formation of carbon dioxide gas in the material during microwave heating, resulting in increased swelling. In some embodiments, calcium carbonate may be added for flavoring purposes.

[0380] For example, under the conditions of the present disclosure, without being bound by theory, protein isolates such as soy and pea may not form a hydrogel meniscus 2121 due to the lack of expansion and the accompanying fibrosis and organization along the sidewalls of the container. The formation of a hydrogel meniscus 2121 or a significant and substantial hydrogel meniscus 2121 requires significant bonding with the container sidewall 2102 and a high degree of bubble formation within the protein-fat hydrogel 120. Prior art materials may not be able to achieve this. As used herein, the meniscus ratio refers to the meniscus depth divided by the final height (H) of the protein-fat hydrogel 120. f ) ratio, as evidenced by the height of the protein-fat hydrogel 120 associated with the container sidewall 2102.

[0381] After heating in the microwave oven is stopped, the protein-fat hydrogel 120 can be quickly immersed in cold water (typical temperatures of cold tap water for household use are between 50°F and 70°F, which is sufficient to cool the product by quickly reducing the product temperature to below 100°F). Cold tap water typically found in a household kitchen sink is generally sufficient in terms of temperature. In some embodiments, the simulated meat product 120 can then be separated from the container 2100 by scraping around the interior portion of the container sidewall 2102 using a spatula. The simulated meat product 120 can then be removed from the container 2100 by a spatula or by hand.

[0382] In some embodiments, the simulated meat product 120 can have a texture similar to boiled chicken breast. An advantage of the present disclosure is that, in some embodiments, the void 2124 can exist at or immediately adjacent to the bottom surface of the container so that the finished product does not have the appearance of being molded to the shape of the bottom of the container. Avoiding a molded appearance significantly improves the aesthetic appeal of the protein-fat hydrogel 120.

[0383] Conditions that may produce a microwave texturized protein-fat hydrogel 120 that is not fully molded to the shape of the bottom of the container may vary depending on a variety of variables, including but not limited to the amount of starting material, the size and shape of the container, the material comprising the container, the type of microwave oven, the power of the microwave oven, the temperature of the starting material, and other variables that one of ordinary skill in the art will understand, and routine optimization may produce a product that does not appear to be fully molded to the bottom of the container. In some embodiments, it may be acceptable to partially mold the material to the bottom of the container.

[0384] Use of the methods described in the present disclosure and protein isolates other than those effective for the present disclosure generally results in a final product that is molded to the shape of the bottom of the container and, unlike the protein-fat hydrogel 120 of the present disclosure, cannot be formed after microwave heating. Using protein isolates prepared according to the present disclosure, but using ineffective protein isolate starting materials, soy protein isolate, potato protein isolate, commercial hemp protein isolate, and other protein isolates have been observed to completely mold other protein isolate materials to the bottom of the container. In some embodiments, there may be a thin or insubstantial layer of protein-fat hydrogel 120 material at the bottom of the material that can be molded to the shape of the bottom of the container, but may be so thin or fragile that at least a portion of its shape is lost when removed from the container.

[0385] In some embodiments, the simulated meat product 120 can be sliced ​​and eaten without further cooking, for example, in a salad. The simulated meat product 120 can also be pan-fried and browned, just as chicken breast can be browned and pan-fried.

[0386] like Figures 29A-29C As shown, the container 2100 may have a different shape or include an insert 2500. In some embodiments, the insert 2500 may have a central cylindrical protrusion 2510a, such as Fig.29B 25. The insert 2500 may be formed from a solid microwave material or may be formed from a hollow material so that the hot water 2002 cannot enter the insert 2500. In some embodiments, the insert 2500 may be foldable and may be folded outward from the side wall of the container or folded upward from the base of the container.

[0387] In some embodiments, the insert protrusion 2510 may have a peninsular shape relative to the container sidewall and form a sidewall protrusion 2510b so that the protrusion 2510 can extend from the sidewall to the center of the container 2100 to 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 may be constructed of a microwaveable material, as previously disclosed herein. The insert 2500 may have an insert base 320 that is shaped to correspond to the container base. The insert base 2520 may provide stability to the insert 2500. The insert 2500 may be removable, or foldable, and connected to the container sidewall 2102 or the container base 2104 to allow access to an immersion blender 2600 (such as Fig.30 ). In some embodiments, the insert base 2520 may have holes or perforations to allow liquid to flow. In some embodiments, the container 2100 may have ridges, chambers, sidewalls, or grooves in the container sidewall 2102 and container bottom 2104, or may have paper inserts of various shapes and sizes.

[0388] In some embodiments, the indicator cover 2700 (eg, Fig.31 2700) is placed on top of the protein-fat hydrosol 114 in the container 2100 and can be used to more easily observe the point at which the expanded protein-fat hydrosol 2110 reaches a maximum height in the container 2100. In some embodiments, the indicator cap 2700 or another cap can be used to prevent steam from escaping from the container 2100 during heating, thereby providing a more hydrated product, or to provide some pressure to the expanded protein-fat hydrosol 2110 as it expands and rises.

[0389] about Fig.32 , the materials needed to produce a ready-to-eat meat analog product 120 according to the present microwave texturization method 2000 can be provided as part of a kit 2800. The kit can include a container 2100, a NEPI bag 2802 (which includes protein isolate and, in some embodiments, can also include chicken seasoning or other meat seasonings as desired), a bottled oil 2804 and optionally a container sleeve 2806, an indicator cap 2700, an optional insert 2810 or a seasoning packet 2812 (which, in some embodiments, can contain additional seasonings or spices).

[0390] Regarding the source of NEPI 250 material for producing microwave texturized protein-fat hydrogel 120, spray dried powder or concentrate can be used, as previously described herein. In some embodiments, NEPI 250 powder may be more effective than NEPI 250 concentrate in texturizing protein-fat hydrogel 120 in a microwave oven.

[0391] Without being bound by theory, shorter microwave heating times produce superior final products. Therefore, it may be desirable to produce sufficient protein-fat hydrogel 120 in as short a heating time as possible. The superior organization may be related to the degree of expansion of the material during microwave heating.

[0392] In some embodiments, such as Fig.26A and 26B As shown, it may be desirable to fold or roll the protein-fat hydrogel 120 after removing it from the container. In some embodiments, the protein-fat hydrogel 120 is foldable. Folding can produce an appearance that is more similar to chicken breast or other types of simulated meat products. In one embodiment, the protein-fat hydrogel 120 can be rolled up so that the smooth container sidewall contact surface is located on the outer surface of the folded or rolled simulated meat product 120, and the more textured portion of the simulated meat product 120 is located on the inner portion of the folded or rolled simulated meat product 120, as shown. Fig.26A and 26B shown.

[0393] In some embodiments, the present disclosure contemplates the use of protein isolates made from seeds or grains that may have properties similar to edible globulins, as described in this application. This may include globulins from pumpkin and squash (Cucurbita moschata and Cucurbita maxima), watermelon (Citrullus vulgaris), cucumber (Cucumis sativus), tobacco, and cottonseed, among others.

[0394] In some embodiments, after microwave heating and before cooling with water, protein-fat hydrogel 120 can be shaped 2034, such as Fig. 20 As shown. A tool such as a spatula can be used for shaping. The minimum temperature for shaping can be about 150F, or 160F, and more preferably 170F, and the maximum is about 212F. The temperature in the microwave oven generally does not reach a temperature higher than the boiling temperature of the protein-fat hydrosol 114 because it is an aqueous liquid and does not exceed the boiling temperature. However, in general, the product can be shaped between about 150F and 212F, wherein 150F may be more difficult to shape because the layers or parts of the protein-fat hydrogel 120 cannot stick together when pressure is applied.

[0395] The shaping may be performed in the container 2100. In some embodiments, shaping may allow the user to eliminate the molded appearance of the hydrogel 120 that initially assumes the shape of the bottom of the container 2100. The protein-fat hydrogel 120 may be shaped to have a structure similar to a chicken breast, or other shapes that may be more aesthetically pleasing or more convenient to eat.

[0396] In some embodiments, calcium carbonate may be added to NEPI 250, which in some cases may increase the expansion rate of the protein-fat hydrosol 114. 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 fibrosis when combined with NEPI 250. Calcium chloride does not appear to have an effect on the expansion rate. In some embodiments, the addition of about 0.5% calcium carbonate to the protein-fat hydrosol 114 may significantly increase the expansion rate. More or less of certain carbonate compounds or other expansion increasing compounds may result in a relative increase related to the amount of compound added to the protein-fat hydrosol 114.

[0397] Using NEPI 250 concentrate or dry NEPI 250 (which has been rehydrated with hot water and opened) in combination with Oil 110 forms a thickened hydrosol that, when placed in a microwaveable container, dish or tube and exposed to microwaves, is sufficient to first heat the water sufficient to "melt" or transform the hydrosol-gel and form a membrane sufficient to hold the water within the protein-oil film so that when the water becomes a gas it can be trapped within the membrane, forcing the membrane to expand and subsequently cool, resulting in a solid setting of the hydrogel. Thus, heating the NEPI 250, water and Oil 110 hydrosol blend to a temperature above the boiling point of water may produce a fibrous structure similar to cooked meat.

[0398] As disclosed in prior application U.S. Patent Application No. 17 / 551,163, specifically, NEPI, when hydrated, opened and blended with oil to form a hydrosol, solidifies the hydrosol into a hydrogel under the action of a direct or indirect external heat source (e.g., from a stove, oven, frying pan, steam, pressure extrusion, or infrared heating). Surprisingly, we have discovered that a protein-fat hydrosol 114 suspension forms a unique and unexpected fibrous structure when heated with microwaves (e.g., in a microwave oven). It is hypothesized that upon microwave activation, the water encapsulated in the hydrated protein-fat hydrosol 114 suspension initially causes the water molecules to heat the protein-fat hydrosol 114 to a transition point where the protein-fat hydrosol 114 begins to solidify or "melt."

[0399] As the water molecules continue to convert to vapor (gas), they may now be trapped in a unique way within the solidifying hydrogel. The gas may cause the solidifying or molten hydrosol 114 to expand until the protein-fat hydrogel 120 is completely solidified. A similar example may be in the glass blowing industry, where air is blown into a ball of liquid molten glass to expand the glass before the liquid glass cools and becomes solid. A myriad of shapes and forms may then be created. However, just like glass blowing, the temperature of the glass and the amount and rate of addition of the gas may be important to achieve the desired result.

[0400] Heating the protein-fat hydrosol 114 too quickly (between the temperature at which the transition melt is formed and the temperature at which steam is formed) may cause the hydrosol 114 to transition to the hydrogel 120 too quickly without forming a hydrosol 114 "melt", which may then capture or entrain existing water during solidification into the hydrogel 120. Agitation may exacerbate this effect. The slower this process is, the more water may be entrained and cause a greater initial expansion. The maximum amount of water can be entrained when the internal water temperature is maintained at slightly below 100°C or the boiling point of water. When the water temperature exceeds the boiling point of water, the generated gas begins to expand the transition protein-fat hydrosol 114 "melt", which may now be partially composed of protein-fat hydrogel 120, until the protein-fat hydrogel 120 is completely solidified. Cooling the protein-fat hydrogel 120 can complete and stabilize the solidification of the melt.

[0401] Described herein are conditions using a microwave oven that allow water to be entrained as it transitions from the protein-fat hydrosol 114 to the protein-fat hydrogel, while allowing the gases that form to force the molten protein-fat hydrosol 114 to expand, thereby creating a unique structure that simulates the texture and lines typically associated with meat.

[0402] NEPI hydrosols are unique in that the melting temperature of the protein-oil hydrosol is below the boiling point of water. Without being bound by theory, the lower melting temperature allows the water to be completely entrained within the protein-oil film 2131 for the first time. High temperature extruded soy or pea isolate products are typically run between 130°C and 140°C to "melt" the protein and cause fiberization, and leave the extruder with a moisture content of less than 10%. In this case, water cannot be retained in the hydrosol 114 type structure, which is why soy or pea type textured vegetable protein (TVP) initially has very low entrapped moisture and must be rehydrated, and multiple ingredients including starch and gums must be added to suspend and retain moisture.

[0403] Microwaves may not destroy proteins (proteins and fats are zero or unaffected by microwaves). The increase in heat of the water molecules may initially cause the protein-oil film 2131 to begin to solidify into a hydrogel, but almost simultaneously, before the gel is fully solidified and solidified, as the activated water molecules are converted to gas and are now trapped in the solidifying gel being formed, the hydrosol expands as it transforms and solidifies into a hydrogel 120. Eventually some steam is released, where the escaping steam may break a portion of the protein film, causing the structure to collapse, and subsequent condensation may cool the hydrogel, causing it to completely and irreversibly solidify after being stretched into a unique shape. Importantly and critically, water continues to be entrained within the hydrogel. Water activity measurements using a water activity meter capable of measuring the water activity of solid or semi-solid materials (such as hydrosols or meat chunks) can be used to quantify the unique water activity properties of the protein-fat hydrogel 120.

[0404] Presumably, a similar phenomenon may occur when NEPI hydrosols are heated in a microwave oven. Unlike many other proteins (including soy or pea isolates), which require temperatures in excess of 140°C to melt and stretch, NEPI 250 protein-fat hydrosols 114 are able to "melt and stretch" at temperatures below the boiling point of water, thereby trapping the water as it converts to gas and expanding the formed network 2160.

[0405] In this case, we found that the melting and solidification temperatures of proteins are below the temperature at which water forms a gas, 100 °C, allowing the protein-oil film to expand and structure the hydrogel by more or less simultaneous melting and expansion of the entrapped water as the water becomes a gas, followed by cooling to final solidification by either releasing the escaping water or condensing the vapor into water, thereby entrapping the water within the protein-oil hydrogel structure.

[0406] Additional testing can be performed to identify additional effective conditions for producing protein-fat hydrogels 120 according to the present disclosure. For different hemp seed protein isolates, commercially available hemp seed protein isolates (e.g., VictoryHemp) and hemp seed protein isolates described in U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis can be tested to further characterize the differences in capabilities. Testing can be performed under the same conditions as described in the present disclosure, and the resulting texture can be observed and compared to the product obtained using NEPI 250.

[0407] Further testing can be done on changing oils from saturated to unsaturated (vegetable and animal oils) under the same conditions as described in this disclosure, and the texture of the resulting product can be observed and compared to the product obtained using NEPI.

[0408] Further testing of the effect of container size, including from 65 ml to 550 ml, testing the effect on texture at intervals within this range can identify different effects, wherein the container shape is preferably consistent, but only the diameter or width of the container is changed. Conducting such testing according to the methods described in the present disclosure can identify additional benefits and advantages of the present disclosure. The ratio of the container 2100 size to the starting liquid volume of the protein-fat hydrosol 114 is important for the structure of the protein-fat hydrosol 120. The microwave power can also be varied as the container size changes to optimize the results.

[0409] In some embodiments of the present disclosure, it may be important to maximize the amount of water entrapped in the hydrogel in order to obtain better results. In some embodiments of the present disclosure, the water content of the protein-fat hydrogel 120 may correspond to the water content in conventional meat.

[0410] Additional testing of container materials may also produce different effects depending on the container material. In some embodiments of the present disclosure, adhesion or bonding of the protein-fat hydrosol 114 or protein-containing material to the inner sidewall of the container is important for texturization. Too strong a bond may prevent the hydrogel from being effectively removed from the sidewall of the container and may also make cleaning difficult. Ceramic materials, particularly unglazed ceramic materials, can allow for effective microwave heating, expansion, and bonding of the protein-fat hydrosol 114 to the sidewall of the container, however, due to the strength of the bonding of the protein-fat hydrosol 114 to the sidewall and the presence of pores in the ceramic material, the container is difficult to clean after use. Using unglazed ceramic materials that have pores filled by known or unknown methods, such as by soaking the container 2100 in milk and then heating to caramelize the milk and fill the pores, can produce more effective containers for use in the present disclosure.

[0411] Additional testing of the indicator cap 2700 may be performed to potentially identify different effects on the texture of the protein-fat hydrogel 120. Without being bound by theory, the cap 2700 may enable and affect the entrainment of water in the protein-fat hydrogel 120 and the heating rate in the microwave oven. According to the present disclosure, the weight, size, position and material of the cap may be changed to potentially change the texture and potentially entrain different amounts of water in the material by affecting the heating rate. In some embodiments, the cap 2700 may be constructed of any microwaveable material, including but not limited to plastic, paper, glass and ceramic. In some embodiments, the cap 2700 may be located within the container 2100, while in other embodiments, the cap 2700 may be located on top of the container 2100. The cap 2700 may be adapted to rise as the protein-fat hydrosol 114 expands and rises. In some embodiments, the cap 2700 may be adapted to increase the pressure within the container 2100.

[0412] Further testing may be performed to achieve different effects on the addition of flavorings to the protein-containing material. In some embodiments, flavorings may be added to the water during the process, before NEPI 250 is added to the water. In other embodiments, flavorings may be blended with NEPI 250 powder or concentrate. In some embodiments, flavorings may be blended in oil 110. In some embodiments, flavorings may be blended after the protein-fat hydrosol 114 is formed.

[0413] The presence of albumin or albumin-containing complexes in hemp grain protein isolate may interfere with the ability of hemp grain protein to form a protein-fat hydrogel 120 with 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 NEPI 250, acceptable protein-fat hydrogels are not produced using the methods described in the present disclosure. When coagulated in a microwave oven at 25% albumin 75% edema, the protein-fat hydrogel 120 into which albumin had been reintroduced became softer and less elastic. In some embodiments, very low levels of albumin that may be present after NEPI is produced according to the present disclosure can improve texture; however, large or excessive amounts of albumin or albumin-containing complexes can produce a product with a poor texture.

[0414] Example

[0415] Example 7

[0416] With respect to Example 1, a plant-based imitation chicken product was produced according to the method of the present disclosure. As shown in Table 1, for Test Sample 1, boiling water was first added to a 1 / 3 measuring container, and then added to a plastic container having a size of 2.5 inches by 8 inches and a capacity of 20 ounces. For Test Sample 2, 65.0 grams of boiling water as shown in Table 1 was first added to a measuring container, and then added to a straight-walled Pyrex glass beaker having a size of 2.5 inches in diameter, 10 inches in height, and a capacity of 400 ml. For Test Samples 1 and 2, the protein hydrosol was then mixed with an immersion blender (Braun MultiquickMQ7025x) for approximately 1 minute.

[0417] Vegetable oil (Crisco) was then added to the protein hydrosols of both samples, as shown in Table 1, to produce protein-fat hydrosols. The protein to fat ratio of Test Sample 1 was 1.96:1. The protein to fat ratio of Test Sample 2 was 2.04:1. The samples were then mixed using an immersion blender (Braun Multiquick MQ7025x), mixing the protein and water for approximately 1 minute and mixing for approximately 30 seconds when adding the oil to the protein hydrosol. Each sample was blended at 40°C. Heating in a microwave oven (model HMC54151UC / 05, manufactured in May 2018). The heating time for test sample 1 was 1 minute 25 seconds. The heating time for test sample 2 was 1 minute 58 seconds.

[0418] Table 10A

[0419]

[0420] According to visual observation during microwave heating, Test Sample 1 fully expanded and rose during microwave heating. When heating was stopped and the product was immediately rinsed with cold water, the product adhered to the side of the container. After cooling by general cold water immersion and scraping off the hydrogel, the hydrogel exhibited fibrous tendrils, and the hydrogel depth exceeded 1 inch and was approximately 3 inches in length. After slicing, the hydrogel resembled chicken strips of approximately 1 / 4 inch by 2 inches, with good tensile strength and bite characteristics similar to boiled chicken.

[0421] According to visual observation during microwave heating, Test Sample 2 expanded and rose sufficiently during microwave heating. When heating was stopped, the product adhered to the sides of the container. After cooling by soaking in cold water and scraping off the hydrogel, the hydrogel solidified firmly without any further shape changes. The shape had some external tendrils and spikes, similar to the shape expected for chicken shreds.

[0422] Other test samples did not provide satisfactory results when appropriate process parameters were not used. For example, in contrast to container materials with inner side walls of paper and plastic containers or microwave materials that preferably have rough or irregular surfaces, glazed ceramic containers with very smooth inner side walls were used to cook the hydrogel, and it is clear that the protein-fat hydrosol does expand, but it does not bind to the container side walls as expected. However, when parchment paper was used to line the walls after the protein-fat hydrosol was added, the protein-fat-hydrosol expanded very quickly, even expanding to a height above the edge of the ceramic container before collapsing. The imitation meat products made using parchment paper are very thin but have a high fiber content and excellent tensile strength. However, in this embodiment, the product does not have a "filet" type interior, but rather a fibrous, extremely thin meat slice network with irregular shapes.

[0423] Example A

[0424] In Example A, protein-fat hydrosol test samples A-1 to A-10 were prepared according to U.S. Patent Application No. 17 / 551,163 to Mitchell Ellis. This method can be used to prepare simulated chicken products. The composition of the test materials is shown in Table A.

[0425] Example 8

[0426] Table 10 was prepared using the same method as Table 10A above.

[0427] Table 10B

[0428]

[0429] Table 11 Plastic Oster polycarbonate at different microwave power settings

[0430]

[0431] *UC=Unripe; PC=Partially cooked; FC=Well cooked; OC=Over cooked Table 12 Container material composition

[0432]

[0433]

[0434]

[0435] Table 13 Sensory properties of protein fat hydrogel

[0436]

[0437]

[0438] Example 9

[0439] Example 9 shows that about 17% to 38% NEPI in a solution prepared generally as described in Example 1 is effective for producing an acceptable meat analog product according to the present disclosure. Example 9 shows that about 0% to 50% oil in a solution prepared generally as described in Example 1 is effective for producing an acceptable meat analog product according to the present disclosure. There must be at least enough water present in the protein-fat hydrosol to fully hydrate and open the NEPI. The amount of water required to fully hydrate and open the NEPI may be about 30% of the protein hydrosol. The maximum concentration of water in the protein-fat hydrosol is about 80%.

[0440] Table 14 Protein-Fat Hydrosol Formulations for Effective Protein Range Evaluation

[0441]

[0442] Table 15 Analysis results of effective protein range evaluation

[0443]

[0444]

[0445] Table 16 Sensory results of effective protein range evaluation

[0446]

[0447] Table 17

[0448] Protein-Fat Hydrosol Formulations for Effective Oil Range Evaluation

[0449]

[0450] Table 18 Analysis results of effective oil range evaluation

[0451]

[0452]

[0453] Table 19 Sensory results of effective oil range evaluation

[0454]

[0455] Example 10

[0456] The effect of albumin complexes on protein-fat hydrogels was tested.

[0457] The formulations used in Table 20 used products made according to the procedure described in Example 7. A total of 5 simulated meat product samples were used in this experiment, and each point was repeated six times. The prepared simulated meat product blocks were cut into a size of 50×15×15 mm. The values ​​were measured using a texture meter (TA.XTplus, Stable Microsystems) with a 30 kg load cell equipped with a Warner-Bratzler blade, and adjusted to a descent and penetration speed of 2.00 mm / s, a penetration depth of 30 mm, and a contact force of 10 g.

[0458] equipment

[0459] Texture Analyzer - TA.XTPlus Connect Texture Analyzer 650H s / n 2-P6_Z11140-01-V003C98CB

[0460] Texture Analyzer Probe–TA-007

[0461] These results indicate that increasing concentrations of albumin-containing complexes lead to a decrease in the strength of the texture analysis. This result suggests that as the concentration of albumin complexes increases, the strength and toughness of the meat analogs decreases, causing the material to become soft and may no longer have an acceptable texture for meat analogs.

[0462] Table 20 Protein-Fat Hydrosol Formulations for Effective Albumin Complex Range Evaluation

[0463]

[0464] Table 21 Analytical results of effective albumin complex range assessment

[0465]

[0466] Table 22 Texture analysis for evaluation of effective albumin complex range

[0467]

[0468] Embodiment 11

[0469] Evaluation of the effects of chemical additives on protein-fat hydrosols

[0470] The chemical additives shown below affect the expansion rate of the protein-fat hydrosol, increasing it in some cases and decreasing it in other cases.

[0471] Table 23

[0472]

[0473]

[0474] Embodiment 11

[0475] The effects of container material composition and container structure, including the size and shape of the container, were evaluated. Certain container materials were effective in allowing the protein-fat hydrogel to bond or adhere to the container sidewalls during and after expansion of the protein-fat hydrosol and hydrogel. These materials were certain plastic containers, including polycarbonate plastic, and bonding was enhanced by abrading the inner surface of the container sidewalls. Paper containers were also effective in bonding the protein-fat hydrosol and hydrogel.

[0476] The container shape is important in producing the desired expansion. An increase in container diameter has a negative synergistic effect on the expansion rate of the protein fat hydrosol, while a decrease in the diameter of the plastic container has a positive synergistic effect on the expansion rate. In one test example, a 3.25-inch diameter was ineffective in producing expansion, while a 2.75-inch diameter was effective in producing acceptable expansion and fibrosis.

[0477] Table 24

[0478]

[0479]

[0480] Table 25

[0481]

[0482] legend:

[0483] 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 Power 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 = Unacceptable, A = Acceptable, P = Preferred, FA / TF = Acceptable Functionality / Unacceptable Taste, VH = Victory Hemp Competitor, VH = Victory Hemp Competitor, HL = Hemp Land Competitor, PLA = Polylactic Acid, PMP = Polymethylpentene, PP = Polypropylene, PE = Polyethylene, UKR = Ukraine, NEPI = Natural Edema Isolate, E = Edema, AC = Albumin Complex

[0484] E1=100%NEPI

[0485] EA2 = 90% NEPI + 10% AC (albumin complex)

[0486] EA3 = 80% NEPI + 20% AC (albumin complex)

[0487] EA4 = 70% NEPI + 30% AC (albumin complex)

[0488] EA5 = 60% NEPI + 40% AC (albumin complex

[0489] TS = Total Solids

[0490] NEPI in Tables 10 to 25 = NEPI with shell

[0491] NEPI = Industrial production for Tables 10 to 25, except for Example 10, where the NEPI was produced on the benchtop. Higher microwave power produced finer fibers.

[0492] In Tables 10 through 25, the expansion description using "x" means that the height of a 24 ounce Oster container increases by "x" times. This means that a 1x in an Oster container would have a height of 1 inch, while a 3x would have a height of about 3 inches. In an Oster container, 4 inches in height in the liquid holding chamber would be equivalent to about 500 mL. 2 inches would be equivalent to about 225 mL. 3 inches would be equivalent to 350 mL. 4 inches would be equivalent to 500 mL. 5 inches would be equivalent to 675 mL.

[0493] Unless otherwise noted, Examples 7-11 generally used the same method of formulating NEPI as Tables 10A and B.

[0494] Other Implementations

[0495] It should be understood that although the invention has been described in conjunction with the detailed description of the invention, the foregoing description is intended to illustrate rather than limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the following claims.

Claims

1. A method, comprising: adding a protein-fat hydrosol to a container, wherein the protein-fat hydrosol contains natural edema protein isolate, and wherein the container has a bottom and at least one sidewall; placing the container into a microwave oven; heating the protein-fat hydrosol by microwave; forming a plurality of bubbles in the protein-fat hydrosol; causing the protein-fat hydrosol to swell; solidifying the protein-fat hydrosol to form a network protein-fat hydrogel; wherein the networked 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; Cooling the network protein-fat hydrogel with an aqueous solution; as well as The reticulated protein-fat hydrogel is separated from the container.

2. The method of claim 1, further comprising shaping the protein-fat hydrogel after solidifying the protein-fat hydrogel and before cooling the reticulated protein-fat hydrogel.

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

4. The method of claim 1, wherein the power setting of the microwave oven is between 4 and 7 for a conventional household microwave oven.

5. The method of claim 1, wherein a hydrogel meniscus is formed in the reticulated protein fat hydrogel.

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

7.

7. The method of claim 1, wherein the at least one sidewall has at least one continuous sidewall.

8. The method of claim 1, wherein the at least one side wall allows for substantial expansion of the liquid.

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

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

11. A method comprising: adding a protein hydrosol to a container, wherein the protein-fat hydrosol contains protein isolate, and wherein the container has a bottom and at least one sidewall; Allowing the liquid to expand significantly; placing the container into a microwave oven; adjusting the time and power of the microwaves to solidify the protein-fat hydrosol to form a network protein hydrogel; heating the protein hydrosol; forming gas bubbles in the protein-fat hydrosol; causing the protein hydrosol to swell; solidifying the protein-fat hydrosol to form a network protein hydrogel; wherein the network protein hydrogel is heterogeneous and comprises at least one thread, at least one sheet, at least one container-forming sidewall portion, and a plurality of voids; Cooling the network protein hydrogel with water; as well as The network protein hydrogel is separated from the container.

12. The method of claim 12, wherein fat has been added so that the protein hydrosol is a protein-fat hydrosol.

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