Methods and compositions for consumables

Abstract

To provide methods and compositions for production of non-meat consumable products.SOLUTION: This invention relates to a method of imparting a meat flavor and / or aroma to a food product, comprising combining a solution containing a heme-containing protein with a consumable to obtain the food product, wherein cooking the food product produces at least two volatile compounds having an aroma associated with meat.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Cross-reference of related applications This application is based on U.S. Patent Application No. 13 / 941,211, filed on 12 July 2013. U.S. Patent Application No. 61 / 908,634, filed on November 25, 2013. Claiming priority from U.S. Patent Application No. 61 / 751,816, filed on January 11, the following applies: Simultaneously pending patent applications: Application No. PCT / US12 / 46560; Application No. PCT / US 12 / 46552; Application No. 61,876,676 filed on 11 September 2013; Application No. 61 / 751,818 filed on January 11, 2013 and March 2012 This relates to application No. 61 / 611,999, filed on the 16th, and all of them are, This is incorporated herein by reference.

[0002] The present invention relates to consumable products, and more specifically, in some embodiments, to non-animal materials as their constituent elements. Animal-based products can be manufactured by breaking them down into their basic components and reconstructing those components into consumables. Regarding non-animal-based imitations of food products. [Background technology]

[0003] The livestock industry has serious negative environmental impacts. Currently, 30% of the Earth's land surface is occupied by animals. It is largely allocated to livestock farming, and it is estimated that livestock account for 20% of the total terrestrial animal biomass. Because of its enormous scale, the animal and livestock industry accounts for more than 18% of net greenhouse gas emissions. It accounts for a large portion. The animal and livestock industry is the largest source of human-borne water pollution, and the animal and livestock industry is overwhelmingly... It is the world's greatest threat to biodiversity. The world's human population is consuming a diet that includes meat. If we could transition to a diet that does not include animal products, 26% of the Earth's land surface could be freed up for other uses. It was estimated that this would be opened up for the purpose of [something]. Furthermore, if they switched to a vegetarian diet, water and energy would be [something]. Ghee consumption will likely decrease on a large scale.

[0004] Meat consumption has serious negative effects on human health. The health benefits of vegetarianism are ten It is well established that if the human population shifts to a more vegetarian diet, healthcare costs will decrease. It is likely.

[0005] Hunger is a global problem, but it affects four major crops worldwide (soybeans, corn, Wheat and rice are rich in calories and protein, including all essential amino acids. It already supplies more than 100% of what the human population needs.

[0006] Plant-based meat substitutes have largely failed to drive a shift towards vegetarianism. Current technology for meat substitute compositions involves extruding a soybean / grain mixture, resulting in the production of a product. While these are related to cooking and eating meat, they are mostly about replicating the experience of cooking and eating meat. This is not possible. A common limitation of these products is that they are less uniform than comparable meat products. It is chewiness and texture. Furthermore, these products are mostly pre-cooked and contain artificial flavors. Since the aroma must be incorporated and sold, they have aroma, flavor and meat Other important characteristics related to cooking cannot be reproduced. As a result, these products This largely consists of a limited consumer base that is already leaning towards vegetarianism / strict vegetarianism. It only piques curiosity and is unlikely to attract the interest of a broader consumer segment than those accustomed to eating meat. I couldn't.

[0007] Food is any substance that is eaten or drunk by any animal, including humans, for nutrition or pleasure. It is usually of plant or animal origin and contains essential nutrients such as carbohydrates, fats, proteins, vitamins or minerals. The substance is ingested by an organism and assimilated by the cells of the organism in an attempt to generate energy, maintain life, and stimulate growth.

[0008] Normally, food has its origin in photosynthetic organisms that are usually plants. Some foods are obtained directly from plants, but furthermore, animals used as food sources are raised by feeding them foods that are of plant origin. Edible fungi and bacteria are used to convert materials from plants or animals into

[0009] other foods, such as mushrooms, bread, yogurt, etc. In most cases, plants or animals are divided into different parts depending on the purpose of the food. Certain parts of plants, such as seeds or fruits, are often more highly valued by humans than others, and these are selected for

[0010] human consumption, while other less desirable parts, such as straw, are usually used for feeding animals.

[0011] Many foods can be eaten raw, but many also undergo some form of cooking for safety, palatability, texture or flavor. At the simplest level, this can include washing, cutting, trimming or adding other foods or ingredients. It can also Alternatively, this may involve cooling or fermentation, and individual foods are used to achieve the desired combination of properties. It can be combined with other foods.

[0012] In recent years, under the fields of food science and molecular gastronomy, scientific rigor has been required for food preparation professionals. Attempts have been made to bring it to Seth. Food science encompasses food preparation safety, microbiology, preservation, While broadly studying chemistry, engineering, and physics, molecular gastronomy transforms food into unexpected forms. To do this, liquid nitrogen, emulsifiers such as soy lecithin and calcium alginate, etc. The focus is on the use of scientific tools such as gelling agents.

[0013] However, the raw materials are usually whole organisms (plants or animals) or steaks, and also fungal fruiting bodies. It is either extracted tissue such as plant seeds. In some cases, it involves grinding seeds into powder. The extracted tissue is used to isolate oils and bulk proteins, etc., before cooking food. It will be altered. [Overview of the Initiative] [Problems that the invention aims to solve]

[0014] All of these items are a mixture of protein, carbohydrates, fats, vitamins, and minerals. Despite the fact that it contains compounds, the physical properties of these materials in the original plant or animal The arrangement determines the intended use of the plant or animal tissue. Improvements for the manufacture of consumables. Methods and compositions described herein are disclosed. [Means for solving the problem]

[0015] This specification provides consumable products and methods for manufacturing them. Consumable products are non Animal-based, for example, primarily plant-based or entirely plant-based proteins and / or It may be a consumable product containing fat, and a beverage (for example, an alcoholic beverage such as cream liqueur) Protein supplements (or protein drinks), baked foods (e.g., bread) (For example, cookies), spices (e.g., mayonnaise, mustard), meat products or meat substitutes It can take the form of (for example, ground beef products). For example, protein drinks are meal replacement beverages. Beer or distilled alcoholic beverages supplemented with protein (for example) The spice can be vodka or rum. The spice can be mayonnaise. The meat product may contain tendons. Meat imitations, patties, sausages, or may contain plant-based fats and / or connective tissue. It can be a meat substitute. Coacervates containing one or more proteins are consumable products. It can be used to help bind ingredients together in a product (for example, a ground beef product).

[0016] Therefore, in this specification, consumables containing isolated and purified plant proteins The product is an isolated and purified plant protein, (i) between approximately 2°C and approximately 32°C It has a solubility in a solution of at least 25 g / L at a given temperature, and the solution has a pH between 3 and 8. (ii) It has a sodium chloride content of 0-300 mM, or (ii) 90°C to 11 It has a solubility in solution of at least 1 mg / ml at temperatures between 0°C, and the solution is 5 to 8 A consumable product is provided that has a pH between 0 and 300 mM and a sodium chloride content of 0 to 300 mM. In some embodiments, consumable products include beverages, protein supplements, and baked foods. These include spices, meat products, or meat substitutes. In some embodiments, the beverage is an alcoholic beverage. It is a food or protein drink. In some embodiments, the alcoholic beverage is cream. It is a liqueur. Cream liqueur is a lipid emulsion that does not contain dairy ingredients. The cream liqueur may contain no animal products. In some embodiments, protein The drink is a meal replacement beverage, beer supplemented with the aforementioned protein, or a protein supplement. It is a distilled alcoholic beverage. The spice may be a mayonnaise imitation. Some implementations In some embodiments, the meat product may be pâté, sausage imitation, or meat substitute. Therefore, the isolated and purified plant proteins are at least 10 kDa in size. In some embodiments, the isolated and purified plant proteins are not completely denatured. In some cases, isolated and purified plant proteins are not derived from soybeans. In some embodiments, isolated and purified plant proteins are used with RuBisCo, mo ong 8S globulin, pea globulin, pea albumin, lentil It contains one or more of the following: meprotein, zein, or oleosin.

[0017] In some embodiments, isolated and purified plant proteins are dehydrin, hydroph It can be dissolved after boiling at a pH and salt concentration equivalent to that of filin, intrinsically denatured proteins, or food. It includes proteins identified based on its ability to remain sexual. In some embodiments, The consumable product further comprises plant-derived lipids or microbial-derived lipids. In some embodiments, Consumable products include a second isolated and purified protein and / or seasonings and flavorings. It further contains emulsifiers, gelling agents, sugars, or fiber.

[0018] This disclosure also relates to coacerbates comprising one or more isolated and purified proteins. The present invention provides consumable products including [a specific component]. In some embodiments, the consumable product is a meat imitation. In some embodiments, the consumable product further comprises plant-derived lipids or microbial-derived lipids. Plant-derived or microbial-derived lipids may contain lecithin and / or oils. It may contain up to approximately 1% by weight of lecithin. The product may contain lecithin and oil. In some embodiments, the oil is canola oil, coconut oil, or cocoa butter. It may contain approximately 1% to 9% oil. One or more isolated and purified proteins The protein may contain plant proteins. One or more plant proteins may be one or more. Numerous pea proteins, chickpea proteins, lentil proteins, lupine May contain proteins, other leguminous plant proteins, or mixtures thereof. In terms of application methods, one or more pea proteins are used as legumin, bicillin, combinator, etc. It is syrin or a mixture thereof.

[0019] This disclosure also relates to muscle mimics comprising one or more isolated and purified proteins. The company provides connective tissue imitation products, adipose tissue imitation products, and meat imitation products including coacervates. Acervate may further contain plant-derived lipids or microbial-derived lipids. Microbial lipids may be lecithin and / or oils. Meat imitations are ground beef. It could be a counterfeit.

[0020] Furthermore, if one or more isolated and purified proteins are derived from non-animal sources... Consumable products including a low-temperature solidified gel containing salt are also provided. In some embodiments, isolated The purified plant proteins include RuBisCo, moong 8S globulin, and E. Pea globulin, pea albumin, lentil protein, zein or Contains one or more of the leosin species. In some embodiments, isolated and purified plant Proteins include dehydrins, hydrophyllines, or intrinsically disordered proteins. In the application form, the low-temperature solidified gel further contains plant-derived lipids or microbial-derived lipids. In the embodiment, plant-derived lipids or microbial-derived lipids are lecithin and / or oils. be.

[0021] This disclosure relates to one or more isolated plant proteins and one or more plants The present invention further provides adipose tissue imitation containing algae-derived oil and optionally phospholipids. In some embodiments, the phospholipid is lecithin. In some embodiments, plant-based The oils are corn oil, olive oil, soybean oil, peanut oil, walnut oil, and almond oil. Sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, sunflower oil, flaxseed oil, coconut oil Palm kernel oil, coconut oil, babassu oil, shea butter, mango butter, cocoa butter, Selected from the group consisting of wheat germ oil, rice bran oil, and combinations thereof. (Some embodiments) So, the fat release temperatures for adipose tissue imitation are 23°C to 33°C, 34°C to 44°C, and 45°C to 5°C. 5℃, 56℃~66℃, 67℃~77℃, 78℃~88℃, 89℃~99℃, 100℃~ 110℃, 111℃~121℃, 122℃~132℃, 133℃~143℃, 144℃~ 154℃, 155℃~165℃, 166℃~167℃, 168℃~169℃, 170℃~ 180℃, 181℃~191℃, 192℃~202℃, 203℃~213℃, 214℃~ 224℃, 225℃~235℃, 236℃~246℃, 247℃~257℃, 258℃~ Between 268°C, 269°C-279°C, 280°C-290°C, or 291°C-301°C Yes. In some embodiments, the fat release percentage of the adipose tissue imitation is 0-1 during cooking. 0%, 10%~20%, 20%~30%, 30%~40%, 40%~50%, 50%~6 These are 0%, 60%-70%, 70%-80%, 80%-90%, or 90%-100%. In some embodiments, isolated and purified plant proteins are used with RuBisCo, mo ong 8S globulin, pea globulin, pea albumin, lentil It contains one or more of the following: meprotein, zein, or oleosin.

[0022] In some embodiments, the adipose tissue imitation contains about 40% to about 90% oil. In this embodiment, the adipose tissue imitation contains approximately 1% to 6% isolated and purified plant proteins. It contains quality. In some embodiments, the adipose tissue imitation contains about 0.05 to about 2% phospholipids. In some embodiments, the hardness of the adipose tissue imitation is similar to that of beef adipose tissue.

[0023] It contains heme-containing proteins and (i) carbon monoxide and / or (ii) nitrite compounds. Further provided are consumable products that do not contain meat. In some embodiments In some embodiments, the heme-containing protein accounts for at least 0.01% of the composition. Consumable products contain one or more ammonium, sodium, potassium, or calcium It further contains um salts. In some embodiments, the isolated and purified protein is crosslinked. It is.

[0024] A consumable product containing a gel-like emulsion, wherein the gel-like emulsion is a) Isolated and purified proteins, b) If not present in the consumable product, a first lipid that is solid within the selected temperature range, c) If not present in the consumable product, a second lipid that is liquid within the selected temperature range and The melting temperature of the mixture of the first and second lipids is the same as the melting temperature of the lipids found in meat. Similarly, the first and second lipids are plant-derived or microbial-derived lipids, and are consumed. More products will be offered.

[0025] This disclosure also relates to a method for manufacturing consumable products, a) Prepare a solution containing isolated and purified plant proteins, where isolated and purified The prepared plant protein is (i) dissolved in at least 25 soluble particles at a temperature between approximately 2°C and approximately 32°C. It has solubility in liquid, the solution has a pH between 3 and 8, and contains 0-300 mM sodium chloride. (ii) Having a thorium content, or at a temperature between 90°C and 110°C, at least 1 It has a solubility in mg / ml solution, and the solution has a pH between 5 and 8, and 0-30 Having a sodium chloride content of 0 mM, b) Adding the solution to a beverage This provides a method that includes [something].

[0026] In some embodiments, the solution comprises two or more isolated and purified plant proteins. In some embodiments, the beverage is clear. In some embodiments, isolated and purified The plant protein is present in solution at a concentration of at least 1% by weight. In some embodiments, The isolated and purified plant proteins are RuBisCo, moong globulin, and soybean. Globulin, pea globulin, pea albumin, prolamin, lentil Select from the group consisting of meproteins, dehydrins, hydrophylins, and intrinsically disordered proteins. Selected. In some embodiments, isolated and purified plant proteins are used to prepare the solution. It is freeze-dried before production. In some embodiments, the beverage is made from isolated and purified proteins. It has an improved texture compared to the corresponding beverage that does not contain chlorine.

[0027] Furthermore, the addition of heme-containing protein to consumable products, and the heme-containing protein Under equivalent storage conditions, it oxidizes more slowly than myoglobin, and has a longer shelf life than meat-free consumable products. Methods for extending the interval are also provided. In some embodiments, the heme-containing protein is the sequence number. It has at least 70% homology to any one of the amino acid sequences shown from 1 to 27. It contains the following amino acid sequence.

[0028] A method for producing a meat imitation containing a low-temperature solidified gel, a) Under conditions in which isolated and purified proteins do not precipitate from solution, from non-animal sources Denaturing a solution containing at least one isolated and purified protein, b) Optionally, add any thermally unstable component to the solution of denatured protein, c) By increasing the ionic strength of the solution and forming a low-temperature solidified gel, approximately 4°C to approximately 2°C The gelation of the denatured protein solution at 5°C, d) Incorporating a low-temperature solidification gel into the meat imitation product. Further methods, including those mentioned above, are also provided.

[0029] In some embodiments, gelation is achieved using 5-100 mM sodium chloride or calcium chloride. It is induced using a substance. In some embodiments, the heat-unstable component is a protein or lipid. It is a substance or a mixture thereof. In some embodiments, the protein is a heme-containing protein. It is of quality. In some embodiments, the cold-solidified gel contains freeze-aligned plant proteins. It is formed within the matrix.

[0030] In some embodiments, isolated and purified proteins from non-animal sources are plant-based. It is a protein. In some embodiments, the plant protein is RuBisCo, moon Globulin, soy globulin, pea globulin, pea albumin, pu Loramine, lentil protein, dehydrin, hydrophylin, and intrinsically denatured proteins It is selected from a group consisting of qualities.

[0031] a) Isolated and purified non-animal proteins, b) Non-animal lipids and, c. A three-dimensional matrix containing fibers derived from non-animal sources and It contains lipids and proteins, which are dispersed in a three-dimensional matrix. Further adipose tissue imitation is provided, which stabilizes the structure of the adipose tissue imitation.

[0032] Furthermore, one or more isolated fibers assembled into a fiber structure by the solution spinning process Connective tissue imitation products containing purified proteins are also provided. In some embodiments, fibers The structure is stabilized by a crosslinking agent.

[0033] In this specification, a method for imparting a beef-like flavor to a consumable product, wherein the consumable composition contains The process includes adding protein containing um, and after cooking, the consumable composition is given a beef-like flavor. A method is provided.

[0034] Furthermore, a method for making a poultry or fish composition taste like beef, wherein the poultry or fish composition Methods are also provided for each, including the addition of heme protein.

[0035] In some embodiments, the heme-containing protein is composed of amino acids as shown in SEQ ID NOs: 1-27. It has an amino acid sequence that has at least 70% homology to any one of the columns.

[0036] A method for producing coacervates, a) Acidify a solution of one or more plant proteins to a pH between 3.5 and 5.5. And here, the solution contains sodium chloride at a concentration of 100 mM or less. b) Isolating the coacervate from the solution and Methods including the above are further provided. In some embodiments, the pH is between 4 and 5. In some embodiments, the plant protein is one or more pea proteins. Chickpea protein, lentil protein, lupine protein, and other leguminous plant proteins Contains physical proteins or mixtures thereof. In some embodiments, pea protein The quality is isolated and purified legumin, isolated and purified bicilin, isolated and purified Contains combined combicillins or combinations thereof. In some embodiments, isolation and purification The pea protein was isolated and purified from bicillin, and isolated and purified It contains combicillin. In some embodiments, the acidification step is performed using plant-derived lipids or micro This is carried out in the presence of bio-derived lipids. In some embodiments, plant-derived or microbial-derived lipids are used. Lipids include oils and / or phospholipids.

[0037] In this specification, a method for producing adipose tissue imitation, comprising one or more isolated A mixture of plant proteins, one or more plant or algae-derived oils, and optionally phosphorus. A method is provided which includes forming an emulsion containing lipids. In some embodiments, If phospholipids are present, then lecithin. In some embodiments, plant-based The oils are corn oil, olive oil, soybean oil, peanut oil, walnut oil, and almond oil. Oils, sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, sunflower oil, flaxseed oil, coconut Oils, palm kernel oil, coconut oil, babassu oil, shea butter, mango butter, cocoa butter Selected from the group consisting of wheat germ oil, rice bran oil, and combinations thereof. Some implementations In this state, the fat release temperature of adipose tissue imitation is 23°C to 33°C, 34°C to 44°C, and 45°C to 55℃, 56℃~66℃, 67℃~77℃, 78℃~88℃, 89℃~99℃, 100℃ ~110℃, 111℃~121℃, 122℃~132℃, 133℃~143℃, 144℃ ~154℃, 155℃~165℃, 166℃~167℃, 168℃~169℃, 170℃ ~180℃, 181℃~191℃, 192℃~202℃, 203℃~213℃, 214℃ ~224℃, 225℃~235℃, 236℃~246℃, 247℃~257℃, 258℃ Between ~268°C, 269°C~279°C, 280°C~290°C, or 291°C~301°C In some embodiments, the fat release percentage of the adipose tissue imitation is 0- during cooking. 10%, 10%~20%, 20%~30%, 30%~40%, 40%~50%, 50%~ 60%, 60%~70%, 70%~80%, 80%~90%, or 90%~100% In some embodiments, isolated and purified plant proteins are treated with RuBisCo, m Oong 8S globulin, pea globulin, pea albumin, lens Contains one or more of bean protein, zein, or oleosin. Some embodiments The emulsion then contains approximately 40% to 90% oil. In some embodiments, the emulsion Lujon contains approximately 1% to 4% isolated and purified plant proteins. Morphologically, adipose tissue mimics contain approximately 0.05 to 1% phospholipids. In some embodiments, The emulsion is formed by high-pressure homogenization, sonic processing, or manual homogenization. ru.

[0038] A method for minimizing undesirable odors or flavors in a composition containing plant proteins. The composition contains ligands that have affinity for one or more lipoxygenases. Further methods are provided, including bringing them into contact.

[0039] Furthermore, to minimize undesirable odors or flavors in compositions containing plant proteins. A method comprising contacting a composition with activated carbon and then removing the activated carbon from the composition. Methods are also provided, including those that include this.

[0040] Furthermore, to minimize undesirable odors or flavors in compositions containing plant proteins. A method comprising adding a lipoxygenase inhibitor and / or antioxidant to a composition. Methods including the following are also provided.

[0041] This disclosure is, a) Sugars and, b) Chocolate flavoring, c) Cream fraction obtained from plant-based milk and We also offer a chocolate-flavored spread, which includes [the ingredient mentioned earlier].

[0042] In this specification, a method for changing the texture of consumables during or after cooking is used. This includes incorporating one or more plant proteins with low denaturation temperatures into the consumables. A method is provided. In some embodiments, one or more plant proteins are used. At least one type is isolated and purified. In some embodiments, one or more plant compounds are used. Proteins include Rubisco, pea protein, lentil protein, or other proteins. Selected from the group consisting of membranaceous plant proteins. In some embodiments, pea methane. The protein contains pea albumin protein. In some embodiments, the consumables are It becomes harder during or after cooking.

[0043] Furthermore, tissue mimics containing frozen and aligned non-animal proteins are provided. In this context, non-animal proteins are plant proteins. In some embodiments, non-animal proteins are plant proteins. The protein is isolated and purified. In some embodiments, the tissue imitation is a muscle tissue imitation. That is the case.

[0044] This disclosure also provides meat imitations, including tissue imitations containing frozen and aligned non-animal proteins. To provide.

[0045] Unless otherwise defined, all technical and scientific terms used herein This is the same as that generally understood by those skilled in the art in the field to which the present invention relates. It has meaning. Methods and materials similar to or equivalent to those described herein are part of the present invention. The following methods and materials may be used to carry out this, but suitable methods and materials are described below. All publications, patent applications, patents, and other references listed are cited by reference to their respective sources. The whole is incorporated. In case of conflict, this specification, including definitions, takes precedence. Furthermore, the material The methods and examples provided are illustrative and not intended to be restrictive.

[0046] Details of one or more embodiments of the present invention are shown in the accompanying drawings and the following description. Other features, purposes, and advantages of the invention are described in the description and drawings, and in the claims. It becomes clear. The word "comprising" in the claims is defined in patent law. According to standard practice, by "consisting essentially of", "Ta" can be replaced with "consisting of". [Brief explanation of the drawing]

[0047] [Figure 1-1] This figure shows the amino acid sequence of an exemplary heme-containing protein. [Figure 1-2] This figure shows the amino acid sequence of an exemplary heme-containing protein. [Figure 1-3] This figure shows the amino acid sequence of an exemplary heme-containing protein. [Figure 1-4] This figure shows the amino acid sequence of an exemplary heme-containing protein. [Figure 1-5] This figure shows the amino acid sequence of an exemplary heme-containing protein. [Figure 2A] This is a bar graph showing the percentage of fat release based on the amount of lecithin. [Figure 2B] This is a bar graph showing the fat release temperature based on the amount of lecithin. [Figure 2C] This is a bar graph showing the hardness of fat imitation products based on the amount of lecithin. [Figure 3] This bar graph shows the fat release percentage of fat imitations containing various oils (canola oil, cocoa butter, coconut oil, or rice bran oil). [Figure 4]This bar graph shows the fat release temperatures of fat imitation products containing various oils (canola oil, cocoa butter, coconut oil, or rice bran oil). [Modes for carrying out the invention]

[0048] I.Consumables Methods and compositions for manufacturing consumables are described herein. In some cases, consumables are made by breaking down non-animal materials into their constituent parts and then reassembling those parts into consumables. It is a non-animal-based imitation of animal-based food that can be manufactured by construction. In certain cases, the consumables are not intended to replicate animal-based food, but rather food It is intended to have desirable, unique characteristics of its own. Furthermore, the consumables are, In that case, rather than fulfilling its primary function as food, it serves as a nutritional supplement or pharmaceutical composition. It can function as a body.

[0049] The advantages of the consumables described herein include, for example, compared to similar foods, In the manufacture of consumables, use less energy or water, To avoid using animals, to manufacture healthier products, and to avoid waste otherwise Using certain raw materials or eliminating certain components (e.g., allergens) from consumables One example is enabling output (or the absence of built-in). Consumables are also, It can have a higher degree of manufacturing consistency and enable improved product quality control. Another advantage The consumables are designed to have desirable characteristics for cooking food that is superior to traditional foods. This means it can be designed intentionally.

[0050] Consumables may be for animal consumption, including human consumption. Consumables are for livestock food. (For example, dog food that can be manufactured according to the present invention) or wild animal food (for example, It could be food for undomesticated carnivorous animals.

[0051] Similar to existing human food products, consumables are available at grocery stores, convenience stores, and large retail stores. Sold at Yobi Club Store, fast food restaurants, schools, event venues, hospitals, It may be prepared in restaurants, including those in military facilities, prisons, shelters, or long-term care facilities.

[0052] Consumables may be approved by the appropriate regulatory authority. For example, consumables may be approved by the U.S. Food and Drug Administration. It can be prepared to suit the needs of the bureau. The method of the present invention is necessary to meet the requirements of the regulatory body. It may include a step.

[0053] The consumables of the present invention reproduce conventional food products (hereinafter referred to as "food products"), and It can compete with, complement, or replace it. Food is any currently existing It may be food. The consumables of the present invention are manufactured to replicate food, for example, equivalent meat products. It is possible. Equivalent meat products can be white meat or dark meat. Equivalent meat products The product may originate from any animal. The equivalent meat product used may originate from any animal, not limited to the animal. For example, cows, sheep, pigs, chickens, turkeys, geese, and ducks. , domesticated animals such as horses and dogs, or rabbits, deer, bison, buffalo, wild boars Wild boar, snake, pheasant, quail, bear, moose, antelope, pigeon, dove ve), ptarmigan, fox, wild pig, goat, kangaroo, emu, alligator, ku Locodile, turtle, woodchuck, marmot, opossum, partridge, squirrel, Snorkel bears, whales, sea lions, ostriches, capybaras, nutrias, guinea pigs, rats, ma Hunting animals such as moose, field mice, any kind of insect or other arthropod (wild or (Any of the farmed ones) or for example, fish, crabs, lobsters, oysters, muscles, scallops Examples include marine products such as shellfish, abalone, squid, octopus, sea urchins, and tunicated animals.

[0054] Many meat products are usually derived from the skeletal muscle of animals, but meat can also come from other muscles of animals. It is understood that it may originate from organs. In some embodiments, it is equivalent to meat. The product is a cut of meat derived from skeletal muscle. In other embodiments, an equivalent meat product is, for example, These include organs such as the kidneys, heart, liver, gallbladder, intestines, stomach, bone marrow, brain, thymus, lungs, or tongue. Therefore, in some embodiments, the compositions of the present invention are consumables similar to skeletal muscle or organs. be.

[0055] Consumables (e.g., meat substitutes) include a first composition containing muscle tissue imitation, and adipose tissue imitation. One of the following: a second composition containing and / or a third composition containing a connective tissue imitation. It may include multiple components, where one or more components repeat the essentials of the physical organization of meat. The method is combined. The present invention also relates to muscle tissue imitation (in this specification, "muscle imitation"). (referred to as "adipose tissue imitations"), adipose tissue imitations (hereinafter referred to as "adipose tissue imitations") Fat imitations (referred to as "fat imitations") and connective tissue imitations (referred to herein as "connective tissue imitations") The individual compositions of the woven imitation (called "woven imitation") are provided. In some embodiments, these compositions The substance consists mainly of components derived from non-animal sources (for example, components of (less than 10% is derived from animal sources). In alternative embodiments, muscle, fat and / or Meat substitute products containing connective tissue imitations or one or more imitations are partially animal-based. Although derived from a source, components derived from non-animal sources are supplemented. In some embodiments, As much as 90% of food comes from animal sources. In some embodiments, about 75% of food is animal It originates from the source. In some embodiments, about 50% of the food originates from animal sources. In this embodiment, approximately 10% of the food is derived from animal sources. In yet another alternative embodiment... The present invention relates to one of the following: a muscle tissue imitation, a fat imitation, and / or a connective tissue imitation. This applies to animal sources that are supplied in multiple ways (e.g., beef, chicken, turkey, or pork products). A meat product that is substantially derived from a non-animal source, where the imitation is substantially or entirely derived from a non-animal source. They provide meat products. Examples of such meat products, though not limited to them, include consumables with low animal fat content. Non-animal fat imitation products that improve texture and mouthfeel while maintaining the health benefits. This is a supplemented product of lean ground beef. Such alternative embodiments involve cooking and consuming meat. It more precisely repeats the important features related to this, but at a lower cost and with fewer rings. Consumer health benefits that are only related to environmental impacts, have little impact on animal welfare, and are improved This can result in products with improved characteristics.

[0056] Other food items that can be replicated and replaced by consumables include beverages (e.g., cream). Liqueur or milk), protein drinks (for example, RuBisCo is beer, Distilled alcoholic beverages such as vodka, fruit juices, meal replacement drinks, or protein supplements in water (Can be used as a ment), paste (e.g., Nutella®, cream, Nacho cheese or mayonnaise imitation, patties, blood sausage, meat fillers, eggs, fish Foods such as sausages, tenders, spam, or chilled foods (e.g., ice cream, yogurt) Examples include tart, kefir, sour cream, or butter imitations.

[0057] The consumables may be meat imitations. The consumables are manufactured to mimic the cut or appearance of meat. It is possible. For example, the consumables may be visually similar to ground beef or a specific cut of beef. They may be indistinguishable. In one example embodiment, the imitation is made from natural ground meat (for example). It is combined in a way that approximates the physical structure of ground beef, ground chicken, or ground turkey. In other embodiments, the imitations include, for example, ribeye, filet mignon, and London. It is combined in a way that approximates different cuts of beef, such as broil. Or, consumables. They may be manufactured to have a unique appearance or look. For example, consumables may have a unique structure. It may contain patterns formed from (e.g., lettering or pictures). In some cases, consumables look like traditional foods after being prepared. For example, traditional beef It is larger than a skewer, but after the consumables are sliced ​​and cooked, it is the same as traditional cooked meat. Consumables that look like consumables can be manufactured. In some embodiments, the consumables are two-dimensional and in the shape of traditional food products. It may be similar in appearance but not in three dimensions. For example, consumables may be in two dimensions. It may resemble a cut of meat (for example, when viewed from above), but traditional It can be considerably longer (or thicker) than a cut. In this example, the composition is traditional It can be repeatedly cut into products that are shaped like meat.

[0058] Consumables can be manufactured from locally sourced materials. For example, consumables can be manufactured in a specific area of ​​the end consumer. It can be manufactured from plants grown within the area. The area can be, for example, 1, 10, 100 or 100. It can be 0 miles. Therefore, in some embodiments, the present invention is 1, 10, 100 Or, to provide a method for manufacturing consumables that do not contain products transported more than 1000 miles. ru.

[0059] This invention creates consistent properties when manufactured from consumables and various supply sources. This provides a method for producing plant-based products from local plants in Iowa, USA. The meat imitation is a plant-based meat imitation manufactured from local plants in Lorraine, France. It has substantially the same taste, aroma, and texture. This consistency This makes it possible to promote locally grown food that possesses certain characteristics. Consistency is key. These components may result from the concentration or purification of similar components at different locations. They can be combined in predetermined proportions to demonstrate this. In some embodiments, they originate from the same plant species. Using components (e.g., isolated or concentrated proteins and fats) to achieve high characteristics Consistency is possible. In some embodiments, components derived from different plant species (e.g., single High-level feature consistency is possible using separated or concentrated proteins and fats. In some embodiments, the same protein can be isolated from different plant species (i.e., Homologous proteins). In some embodiments, the present invention provides similar proteins from plant sources in different locations. To isolate plant components and to collect compositions from both locations provided herein. and a method including selling compositions assembled and sold in different geographical locations. The present invention provides a method for producing compositions that have consistent physical and chemical properties. In some embodiments, the isolated components are derived from different plant populations in different locations. In the embodiment, one or more isolated components are transported to separate geographical locations.

[0060] Consumables require fewer sources to manufacture than consumables produced from livestock. This may not be the case. Therefore, the present invention requires less water or energy to produce than meat. We provide meat imitation products that require only energy. For example, the consumables described herein. The items are approximately 10, 50, 100, 200, 300, 500 or 10 per pound of consumables. It may require less than 00 gallons of water. For comparison, producing beef requires 1 pound of meat. It may require more than 2,000 gallons of water per unit.

[0061] Consumables require less land to produce than meat products with similar protein content. It may only require the product. For example, the consumables described herein are similar It takes less than 30% of the land area required to produce meat products with that protein content There's a possibility they won't.

[0062] Consumables may offer health benefits compared to animal products that replace them in diet. Therefore, it contains less cholesterol or lower levels of saturated fat than comparable meat products. It is possible. The American Heart Association and the National Cholesterol Education Program recommend that cholesterol from food can be a factor. Reduce your sterol intake to 30 units per day, which is equivalent to consuming 12 ounces of beef or two egg yolks. It is recommended to limit intake to 0 mg. It is indistinguishable from animal products such as ground beef, and cholesterol. Consumption as described herein, with reduced cholesterol content or without cholesterol The product may help maintain a low-cholesterol diet. In another example, as described herein The listed consumables are cholesterol-free or, compared to animal products, do not contain cholesterol. It may also contain high levels of polyunsaturated fatty acids.

[0063] Consumables may have greater benefits for animal welfare compared to animal products that replace them in food. For example, childbirth, forced feeding, premature weaning, disruption of maternal-offspring interaction or its effects It can be manufactured without requiring the slaughter of animals.

[0064] Consumables may have lower "carbon emissions" than the meat products they replace. For example, consumables These are 1%, 5%, 10%, 25%, and 5% of greenhouse gas emissions caused by replacement animal products. This could result in net greenhouse gas emissions of 0% or 75%. For example, the Environmental Working Group... Environmental Working Group (2011) "Guidelines for Meat Consumption in Climate Change and Health" According to "(meat eaters guide to climate change and health)," beef production is a source of consumption. The production of lamb meat causes emissions equivalent to 27 kg of carbon dioxide per kilogram of beef. The production process generates emissions equivalent to 39 kg of carbon dioxide per kilogram of beef consumed.

[0065] Consumables described herein are not for animals whose consumption is prohibited by religious beliefs. It may be possible to provide substitutes for products or combinations of animal products. For example, consumables may be kosher imitations. It could be a pork chop.

[0066] Consumables may also be transported by component, manufactured in different locations, and assembled. Available In such cases, local ingredients may be used in the manufacture of consumables. Local ingredients may be locally available Non-functional components may be supplied. This means that the necessary components for meat during transport... It is possible to manufacture consumables, such as meat imitation products, using less energy than conventional methods. For example, local water can be used in combination with a kit that provides other components of the consumables. It can be used. Using local water reduces transport weight, thereby saving costs and environmental impact. The impact of the border will be reduced.

[0067] The consumables described herein are not used in areas where animal husbandry is practiced or where permission is granted. It may be manufactured or assembled entirely or partially in areas where it has not been manufactured. Consumables may be manufactured or assembled within an urban environment. For example, to manufacture consumables, A water supply may be provided to the user. The user may use local water, or, for example, Shanghai Plants obtained from a rooftop garden can be used, as in the example shown. In another example, consumables are used in a spaceship. It can be manufactured in a space station or lunar base. Therefore, the present invention is a space travel For use in the line, or for the manufacture of meat replicas for training for space travel. The present invention provides methods and systems for training at an Earth base for space travel. It can be used in islands where keeping livestock is difficult or prohibited. Alternatively, they can be manufactured on artificial platforms in the sea.

[0068] II. Characteristics of Consumables The consumables described herein are typically used to replicate the experience of eating food, such as meat. It is designed to be so. The appearance, texture, and taste of the consumables are similar to those of food, such as meat. It may be present, or indistinguishable from it. Consumables are also desirable characteristics of food. They can be manufactured to have and not incorporate other undesirable features. For example, consumables are , a imitation steak that does not contain tendons or other components not normally consumed in the aforementioned food product. It is possible.

[0069] In certain embodiments of this invention, for example, an animal or human consumes a consumable food, For example, by determining whether it can be distinguished from a specific type of meat, it becomes possible to determine whether it is suitable as a food imitation. This provides a method for determining the suitability of consumables to be classified. One way to determine whether it is comparable to ) is to a) define the characteristics of the meat and b) The objective is to determine whether the consumables have similar characteristics.

[0070] Hardness is a property that can be tested or used to compare or describe food or consumables. Mechanical properties include degree of adhesion, brittleness, chewability, gummability, viscosity, elasticity, and bonding properties. The characteristics of the food that can be tested include the size and shape of the particles, as well as the form of the particles. Geometric properties such as shape and orientation can also be considered. The three-dimensional organization of particles can also be tested. Other characteristics include water content and fat content. These characteristics are hard Use terms like "soft," "firm," or "hard" to describe the degree. To describe the adhesion, use words like "brittle," "crunchy," "easily broken," and "chewy." "Tough," "Soft," "Difficult to bite through," "Crispy," "Powdery," "Glue-like" or "sticky"; "sparse" or "viscous" to describe viscosity "; "Plastic" or "elastic" to describe elasticity; to describe adhesion The "chewy," "sticky," or "greasy" texture; the shape and size of the particles. To describe the texture, use words like "sandy," "gritty," or "coarse"; the shape of the particles and To describe orientation, use terms like "fibrous," "cellular," or "crystalline"; to describe water content. To clarify, "dry," "moist," "damp," or "watery"; Alternatively, terms such as "high in fat" or "greasy" can be used to describe the fat content. This can be explained as follows. Therefore, in one embodiment, a group of people consumes certain foods, for example For example, ground beef may be required to be graded according to the characteristics that describe the food. The consumables listed herein may be graded to determine their equivalence by those individuals. ru.

[0071] The flavor of food can also be evaluated. Flavor is judged by its similarity to other foods, for example, "egg-like." "Like fish," "Like butter," "Like chocolate," "Like fruit" "Eel," "like pepper," "like bacon," "like cream," "milk" It can be graded according to "like" or "like beef." Flavor is based on 7 basic foods. Taste, that is, sweet, sour, bitter, salty, umami (a pleasant aroma), spicy (or They can be graded according to their spiciness and metallic properties. Flavor is determined by chemicals, for example, ji Acetyl (butter-like), 3-hydroxy-2-butanone (butter-like), nona- 2E-Enal (containing fat), 1-Octen-3-ol (of mushrooms), Hexanoic acid (of sweat) (like), 4-hydroxy-5-methylfuranone (HMF, meat-like), pyrazine ( (Like nuts), bis(2-methyl-3-furyl) disulfide (roasted meat), decano N (musty / fruit-like), isoamyl acetate (banana), benzaldehyde (bitter) Almonds, cinnamaldehyde (cinnamon), ethyl propionate (fruit-like) ), methyl anthranilate (grape), limonene (orange), ethyl decadienate (Western Pear, allyl hexanoate (pineapple), ethyl maltol (sugar, cotton candy), ethyl Vanillin (vanilla), butanoic acid (foul-smelling), 12-methyltridecanal (beef-like) For experiences caused by eel or methyl salicylate (wintergreen) These classifications can be explained according to their similarities. These classifications are used to indicate the characteristics of the food. It can be used. The consumables of the present invention then determine how similar the consumables are to food. It can be compared to food to determine. In some cases, then the characteristics of the consumables are The consumables are modified to be more similar to food products. Therefore, in some embodiments, Consumables are graded to be similar to food according to human evaluation. Some implementations In terms of form, consumables are indistinguishable from real meat to humans.

[0072] Consumables may be manufactured to exclude characteristics related to the source of the consumable's components. Example For example, consumables can be manufactured from ingredients derived from beans, but they may have a "bean-like" flavor. It can be manufactured to lack texture. One way to do this is to use component source materials, Decomposition into isolated and purified components and the undesirable characteristic properties of the source This can be achieved by not using ingredients that cause this reaction. Furthermore, as described herein In order to remove any flavor or aroma outside the standard in the isolated and / or purified components ( For example, undesirable flavors or aromas can be removed by deodorizing with activated charcoal, or Or they may be present in trace amounts, such as unsaturated triacylglycerides (linoleic acid or linolenic acid). Enzymes such as lipoxygenase (LOX) can convert these into smaller, more volatile molecules. It can be minimized by removing it. LOX is found in peas, soybeans and peanuts. It is naturally present in leguminous plants such as sorghum, as well as in rice, potatoes, and olives. When plant powders are fractionated into separate protein fractions, LOX is present during maturation or storage. Acting as an undesirable "time bomb" that can cause an undesirable flavor or aroma. Obtain. As shown in Example 34, plant protein (e.g., derived from ground plant seeds) A composition containing ( ) is subjected to purification, and for example, it is bound to LOX, and from a protein sample, LOX can be removed using an affinity resin that removes it. The affinity resin is made of bee Linoleic acid, linolenic acid, stearic acid, which are bound to a solid support such as resin or other materials. It may be oleic acid, propyl gallate, or epigallocatechin gallate. For example, W See O2013138793. Furthermore, depending on the protein component, antioxidants Certain combinations of and / or LOX inhibitors are particularly effective in the presence of fats and oils, and can cause tanning. To minimize the generation of flavors or odors outside the standard in the protein solution It can be used as an effective drug. Such compounds include, for example, one or more β- Carotene, α-tocopherol, caffeic acid, propyl gallate or epigallate Catechins can be cited as an example. These are outside the standards for protein-based foods. To reduce the generation of off-flavors or odors that do not meet standards, during protein purification or It can be included in subsequent food processing steps.

[0073] In some compositions, the subject asked to identify the consumables as a form of food. Alternatively, to identify something as a specific food item, for example, to identify a consumable item as meat. For example, in some compositions, humans identify the consumable as having properties equivalent to meat. In some embodiments, one or more properties of the consumables are, according to human perception, the relationship between meat and... It is equivalent to the corresponding characteristics. Examples of such characteristics that can be tested include: (partially) In this embodiment, a human being uses the consumables of the present invention in comparison to any meat substitute found in the art. Identified as being more meat-like than Rimo.

[0074] The experiment may demonstrate that the consumables are acceptable to consumers. The panel will comply with this specification. It can be used to screen various consumables listed below. Nelist uses multiple consumable samples, namely natural meat versus the consumables described herein. The composition or meat substitute can be tested against the consumable composition described herein. Fat content Variables such as using a mixture of lean and fatty meats, for example, 20% fat It can be standardized to fat. Fat content can be measured using the Babcock method for meat (SS Nielson, Intro duction to the Chemical Analysis of Foods (Jones & Bartlett Publishers, Boston, It can be determined using 1994). Prepare ground beef according to the procedure described herein. A mixture of the manufactured consumables of the present invention can be devised.

[0075] Panelists may be supplied with samples under red or white light in an open consumer panel. (For example, in a booth). To prevent bias, samples are assigned a random three-digit number. The voting position can be rotated. Panelists range from 1 = extremely dislike to 9 = extremely like. Then, using a pleasure / displeasure scale with a median of 5 = neither like nor dislike, we consider softness, juiciness, You may be asked to evaluate the sample for its quantity, texture, flavor, and overall acceptability. The panelists were asked to rinse their mouths with water between samples and were given the opportunity to comment on each sample. It is possible.

[0076] The results of this experiment show a substantial difference or similarity between traditional meat and the composition of the present invention. obtain.

[0077] These results indicate that the compositions described herein are acceptable as true meat products. It can be demonstrated that it is judged to be equivalent to the above. Therefore, these results are used in this specification. The composition described herein is preferred by panelists over other commercially available meat substitutes. This can be demonstrated. Therefore, in some embodiments, the present invention is similar to traditional meat. It provides a consumable product that is more meat-like than any meat substitute known to date.

[0078] The consumables of the present invention may also have physical characteristics similar to those of food, such as traditional meat. In the embodiment, a steel rod of a certain diameter is used to make a 1-inch thick consumable made of the present invention. The force required to penetrate the structure (e.g., putty) is, using a steel rod of a similar constant diameter, The force required to penetrate a similar food structure (e.g., a ground beef patty) with a thickness of 1 inch is large. The ingredients are not different. Therefore, the present invention provides a consumable product having physical strength characteristics similar to meat. Provided. In another embodiment, 100 mm 2 The specimen of the present invention having the cross-sectional area of The force required was measured using the same method, 100 mm 2 Animal tissue (muscle, The force required to tear a sample of fat or connective tissue is not significantly different from that required to tear a sample of fat or connective tissue. For example, TA.XT Plus Texture Analyzer(Textrue Te It can be measured using (Chineses Corp.). Therefore, the present invention is We provide consumables that have physical strength characteristics similar to those of meat.

[0079] The consumables described herein have cooking loss characteristics similar to those of food, such as meat. For example, the consumables have a fat and protein content similar to ground beef, and are comparable to true ground beef. It may have a similar reduction in size when cooked. The similarity of the size loss profiles is Regarding the various compositions of the consumables described herein that are applicable to various types of meat, This can be achieved. The cooking loss characteristics of consumables can also be designed to be superior to those of food. Example For example, it has minimal loss during cooking, but retains the same taste and texture as the cooked product. Consumables that achieve quality can be manufactured. One way is to use consumable compositions. This is achieved by changing the proportion of lipids based on the melting temperature. Alternatively, This is achieved by controlling the protein concentration or by the formation of tissue mimics. This is achieved by modifying the protein composition of the consumables through a specific mechanism.

[0080] In some embodiments, consumables are determined based on olfactometer readings, and are used by animals. It is compared to a food product (e.g., meat). In various embodiments, the olfactometer is To determine the odor concentration, odor threshold or subthreshold odor, and evaluation score compared to a reference gas. It can be used to assess the pleasant / unpleasantness scale score or the relative intensity of odors. Some implementations In this context, the Olfactometer enables the training of expert panels and automated evaluation. Therefore, in some embodiments, the consumables are similar or identical olfactometer readings. It is a product that causes a difference. In some embodiments, the difference is below the detection threshold of human perception. Small enough to spin

[0081] Gas chromatography-mass spectrometry (GCMS) separates various substances within a test sample. a method that combines the features of gas-liquid chromatography and mass spectrometry for identification. Yes. In some embodiments, GCMS may be used to evaluate the properties of consumables. For example, volatile chemicals can be isolated from the headspace surrounding the meat. These chemicals The quality can be identified using GCMS. This allows for the detection of volatile particles in the headspace around the meat. A profile of the emitting chemical substances is created. In some cases, each peak in GCMS is Further evaluation is possible. For example, humans smell the odor of a chemical that is causing a particular peak. This allows us to grade the experience. This information is used to further refine the profile. It is possible. Then, GCMS may be used to evaluate the properties of consumables. GCMS Rofles can be used to refine consumables.

[0082] The distinctive flavor and aromatic components are amino acids, fats, and sugars found in plants and meat. These are mostly produced during the cooking process by chemical reaction molecules, including [specific chemical compounds]. Therefore, In some embodiments, the consumables are tested for similarity to meat during or after cooking. In some embodiments, human grading, human evaluation, and olfactometer readings are performed. Using values, GCMS measurements, or a combination thereof, an olfactory map of cooked meat is created. These are manufactured. Similarly, olfactory maps of consumables, such as meat imitations, can be manufactured. The p is compared, and the degree to which the cooked consumables are similar to meat can be evaluated. In some embodiments, the olfactory map of consumables during or after cooking is used to identify cooked meat or It is similar to, or indistinguishable from, that of meat during cooking. In some embodiments, the similarity This is sufficient to fall below the detection threshold of human perception. Consumables can be manufactured in this manner, and Its characteristics are similar to those of cooked food, but uncooked consumables differ from those of cooked food before cooking. It may possess the following characteristics.

[0083] The shelf life is the period before a consumable product is considered unsuitable for sale, use, or consumption. This is the length of time given to the product. Generally, the storage period decreases with exposure to high temperatures. Therefore, it is important to keep meat products at approximately 2°C.

[0084] The shelf life of meat is determined by the sensory cues of the meat product (odor, appearance of the package, color, taste and texture). Studies over time have shown how long a product remains safe, healthy, and enjoyable. This is determined by laboratory analysis under controlled conditions. While ground beef is used, similar conditions apply to steaks made from other types of meat. This would apply to chops and roasts. Beef, in its natural state, is a dark blue color. It is purple. However, oxygen permeates into the meat, and within the meat, myoglobin reacts with a chemical reaction that leads to the red color. This can trigger a reaction. Continuous exposure to oxygen causes oxidation of myoglobin. This causes red meat to turn brown and develops an "off-kilter" flavor. To control this, various methods for storing and displaying meat products to increase the shelf life of meat products Considerable research has been conducted on the law. These include vacuum packing and regulated atmosphere packing. Modified atmosphere packing (high-concentration oxygen), modified atmosphere packaging (modified atmosp (low-concentration oxygen containing carbon monoxide) and / or high-pressure pasteurization (HPP) (packaging) ) is one example of its use.

[0085] The main determinant of meat color is the concentration of iron-sparing protein in the meat. One of the main iron-storing proteins is myoglobin. (Chicken white meat) It has less than 0.05% myoglobin; pork and veal have 0.1-0.3% myoglobin. Beef contains myoglobin; young beef contains 0.4-1.0% myoglobin; old beef contains It is estimated that it contains 1.5-2.0% myoglobin. Normally, myoglobin in meat is , 3 states: oxymyoglobin (Fe 2+ (Oxygenated = bright red); myoglobin (Fe 2+ )(unoxygenated = purplish / purplish-red); and metmyoglobin (Fe 3+ It exists in an oxidized (brown) state. Oxymyoglobin in the presence of oxygen The transition to metmyoglobin is thought to be the cause of the color change of ground meat from red to brown . Meat preservatives have been developed to extend the lifespan of the red color of meat products, and among others include, but are not limited to, carbon monoxide, sulfite compounds, sodium metabisulfite, Bombal, vitamin E, rosemary extract, green tea extract, catechins and other antioxidants .

[0086] However, a hemoglobin-like heme protein that is essentially more stable, isolated from Aquifex aeolicus (SEQ ID NO: 3) or Methylacidiphilum infernorum (SEQ ID NO: 2) oxidizes more slowly than mesophilic hemoglobins such as myoglobin. The heme proteins described herein (see, e.g., FIG. 1) may also have a reduced heme-Fe state lifespan extended by meat preservatives such as carbon monoxide and sodium nitrite . Heme proteins can be selected for their desired color retention properties. For example, for low temperature vacuum cooking , a relatively unstable heme protein such as one derived from Hordeum vulgare can provide a cooked-like brown product that appears to retain the red uncooked appearance of myoglobin under conditions where the myoglobin would retain its red uncooked appearance. In some embodiments, the heme protein can be selected to have increased stability such that, for example, a meat analog can retain an attractive 2+ medium rare appearance even though it is fully cooked for food safety . . . . . .

[0087] The main determinants of rancid odor and the generation of off - flavor or off - odor from the standard are, but not limited to, the oxidation of the components of the consumable, including fats. For example, the oxidation of unsaturated fatty acids is a known cause of the odor that reaches the nose. In some embodiments, the meat analogue has its chemical properties configured such that taste, texture, smell and chemical properties do not react with oxygen to create an off - flavor or an off - odor from the standard, and thus has an extended shelf life. In some embodiments, the meat analogue is less susceptible to oxidation due to the presence of a higher degree of unsaturated fatty acids than present in beef. In some embodiments the meat analogue contains no unsaturated fatty acids. In other embodiments, the meat analogue contains higher levels of glutathione, vitamin C, vitamin A and vitamin E, any antioxidants and enzymes such as catalase, superoxide dismutase and various peroxidases than present in meat. In other embodiments, components such as lipoxygenase that generate an off - flavor or an off - odor from the standard are absent.

[0088] In some embodiments, the consumables described herein exhibit increased stability under commercial packaging conditions. In some embodiments, the improved shelf life is achieved by using components with increased oxidative stability, such as lipids with reduced unsaturated fatty acid levels, and / or by using more stable hemoproteins such as hemoglobin isolated from Aquifex aeolicus (SEQ ID NO: 3) or Methylacidiphilum infernorum (SEQ ID NO: 2 ). This is improved. In some embodiments, the improved shelf life is used in consumables. Depending on the combination of components. In some embodiments, consumables are used for the desired packaging method. It will be specifically designed.

[0089] III. Composition of Consumables The consumables described herein are one or more isolated and purified proteins Contains protein. "Isolated and purified protein" refers to a single monomer or polymer protein. The accumulated amount by mass of protein components other than the specified protein, which may be a type of protein, The specified protein was found in amounts of 2 times, 3 times, 5 times, or 10 times the amount of the isolated source material. The reduction is more than double, more than 20 times, more than 50 times, more than 100 times, or more than 1000 times. It refers to the product. To clarify, isolated and purified proteins are their starting materials (e.g., For example, it may be stated that it was isolated and purified from a plant or other non-animal source. In some embodiments, the term “isolated and purified” means that the protein preparation is less All are 60% pure, for example, 65%, 70%, 75%, 80%, 85%, 90%, 95% It can indicate that it is over 99% pure. Consumables are added to isolated and purified proteins. The fact that it may contain materials means that this definition usually refers to proteins before they are added to the composition. Therefore, it does not alter the isolated and purified properties of the protein.

[0090] In some embodiments, one or more isolated and purified proteins are consumed by weight. At least 1%, at least 5%, at least 10%, of the protein content of the consumables It also accounts for 20%, at least 30%, at least 40%, or at least 50%. In this embodiment, each of one or more isolated proteins is isolated and It is refined.

[0091] The consumables described herein are substantially or entirely from non-animal sources, for example It may consist of components derived from plant, fungal, or microbial sources. Plant sources are It may be an organically grown source. Protein can be extracted from source material. For example, from animal tissue or plant, fungal, algae or bacterial biomass, or secreted sputum (extracted from the culture supernatant of protein) or a combination of source materials (e.g., multiple plants) It can be extracted from seeds. Consumables can also be a combination of plant-based and animal-based sources. It can be manufactured from. For example, the consumable is made from ground beef supplemented with the plant-based product of the present invention. It can be a good product.

[0092] A. Source of ingredients for consumables As described above, isolated and purified proteins can be used in plants, algae, and fungi (e.g., yeast). It may be derived from non-animal sources such as filamentous fungi, bacteria, or archaea. Some embodiments So, the isolated and purified protein is used in genetically modified bacteria or yeast, etc. It can be obtained from a substitute organism. In some embodiments, the isolated and purified protein is chemically processed. It is obtained by chemical synthesis or in vitro synthesis.

[0093] In some embodiments, one or more isolated and purified proteins are used in plant feed It originates from a single source. Isolated and purified proteins can be isolated from a single plant source. Alternatively, multiple plant sources can serve as starting materials for the isolation and purification of proteins. can be obtained. As described herein, the isolated and purified plant protein is soluble in a solution. The solution can contain EDTA (0 - 0.1 M), NaCl (0 - 1 M), KCl (0 - 1 M), NaSO4 (0 - 0.2 M), potassium phosphate (0 - 1 M), sodium citrate (0 - 1 M), sodium carbonate (0 - 1 M), sucrose (0 - 50%), urea (0 - 2 M) or any combination thereof. The solution can have a pH of 3 - 11. In some embodiments, the plant protein has a solubility in a solution at a temperature between about 2°C and about 32°C (e.g., between 3°C and 8°C, 10°C and 25°C or 18°C and 25°C) of > 25 g / L (e.g., at least 25, 30, 35, 40, 45, 50, 75, 100, 125, 1 50, 175, 200 or 225 g / L), the solution has a pH between 3 and 8 (e.g., a pH of 3 - 6, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 or 8) and has a sodium chloride content of 0 - 300 mM (e.g., 50, 100, 150, 200, 2 50 or 300 mM). In some embodiments, the isolated and purified protein is soluble in a solution at greater than 10, 15, 20, 25, 50, 100, 150, 200, or 250 g / L.

[0094] One of ordinary skill in the art will understand that proteins isolated from any organism in the plant kingdom can be used to produce the consumables described herein. Non-limiting examples of plant sources include, for example, corn, rye, rice, wheat, barley, rye, millet, sorghum, buckwheat, amaranth, quinoa, spelt (wheat-rye) Grain crops such as wheat hybrids and teff (Eragrostis tef) Things; cotton bolls, sunflower seeds, safflower seeds, Crambe genus, Camelina genus Oilseed crops including lina, mustard, and rapeseed (Brassica napus). Plants; Acacia genus, or, for example, clover, Stylosanthes genus. s), Sesbania, Vetch (Vicia), Arachis s), Indigofera, Leucaena, Cyamo End peas such as psis, cowpeas, English peas, yellow peas, or green peas. Beans, or for example, soybeans, broad beans, lima beans, kidney beans, chicks Beans, mung beans, quail beans, lentils, lupines, mesquite, carob beans, Soybeans and peanuts (Arachis hypogaea) Plants derived from legumes such as basil; for example, lettuce, spinach, kale, collardock. Lean, turnip, chard, mustard greens, dandelion greens, broccoli or cabbage Any leafy vegetable; or switchgrass (Panicum virgatum) ), Miscanthus genus, Arundo donax, sugarcane for energy By humans, including biomass crops such as sorghum or other grasses Greens that are not normally consumed, alfalfa, corn stalks and leaves, kelp or Other seaweed, green materials that are usually discarded from harvested plants, sugarcane leaves, tree leaves, etc. Root vegetables such as yassava, sweet potato, potato, carrot, beet or turnip; or corn Konatsu is one example.

[0095] Proteins can be isolated from any part of a plant, including roots, stems, leaves, flowers, or seeds. For example, ribulose-1,5-bisphosphate carboxylase / oxygenase (RuB isCo) is a company that produces, for example, alfalfa, carrots (above ground), corn stalks and leaves, and sugar. Millet leaves, soybean leaves, switchgrass, Miscanthus, for energy Isolated from sugarcane, bamboo (Arundo donax), seaweed, kelp, algae, or mustard. It is possible.

[0096] Proteins abundant in plants can be isolated in large quantities from one or more source plants. Therefore, the compositions provided herein (e.g., muscle, fat, or connective tissue) It is an economical option for use in any of the following (counterfeit goods, meat substitutes, etc.). Therefore, in some embodiments, one or more isolated and purified proteins are It contains abundant proteins found at high levels in plants and can be isolated and purified in large quantities. In some embodiments, the abundant protein is approximately the same as the total protein content of the source plant material. 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% It accounts for 70%. In some embodiments, the abundant protein is the total of the source plant material. Approximately 0.5-10%, 5-40%, 10-50%, and 20-60% of the protein content. It accounts for approximately 30-70%. In some embodiments, the abundant protein is supplied by the plant material. Approximately 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8% of the total weight of the dry material. 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% In some embodiments, the abundant protein accounts for the total weight of the dry material of the source plant material. Approximately 0.5-5%, 1-10%, 5-20%, 10-30%, and 15-40% of the total amount. They account for approximately 20-50%.

[0097] One or more isolated and purified proteins are found at high levels in plant leaves. It may contain abundant protein. In some embodiments, the abundant protein is from the source plant. Approximately 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8% of the total protein content of the leaves. %, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% , accounting for 55%, 60%, 65%, 70%, 75%, or 80%. In some embodiments, The abundant protein accounts for approximately 0.5-10% of the total protein content of the source plant leaves, and about 5%. It accounts for ~40%, approximately 10-60%, approximately 20-60%, or approximately 30-70%. In the embodiment, one or more isolated proteins are selected based on their high solubility and human nutritional value. Because of its amino acid composition, which is close to the optimal ratio of essential amino acids for nutrition, it is particularly suitable for meat imitations. Contains the useful protein RuBisCo. In certain embodiments, one or more of these proteins are included. The isolated protein is ribulose-1,5-bisphosphate carboxylase oxygenate. Contains -zeactivase (RuBisCo activase). In some embodiments, one type Alternatively, multiple isolated and purified proteins include plant storage proteins (VSPs). .

[0098] One or more isolated proteins are found in abundance at high levels in plant seeds. It may contain protein. In some embodiments, the abundant protein is from the seeds of the source plant. Approximately 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% of the total protein content. %, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55 %, 60%, 65%, 70%, 75%, 80%, 85%, or 90% or more In some embodiments, the abundant protein is the total protein content of the source plant seeds. Approximately 0.5-10%, approximately 5-40%, approximately 10-60%, approximately 20-60%, or approximately 3 It accounts for 0-70% or >70%. Examples of undefined substances include seed storage proteins, such as albumin, glycinin, and conglycerides. Sinin, Legumin, Globulin, Bicillin, Conalbumin, Gliadin, Glutelin, Gluten, glutenin, hordein, prolamin, phaseolin (protein), prote Inoplast, secarin, wheat (triticeae) gluten or zein or oleost. Oily substances such as caloleosins or steroleosins Protein is one example.

[0099] Dehydrin, hydrophylli, as one or more isolated and purified proteins. N, proteins that are not naturally folded (also called intrinsically disordered proteins) or Other tans in the late-embryogenesis abundant (LEA) family Examples include highly soluble proteins such as protein. LEA proteins are... It has been found in materials, plants, and microorganisms, and acts as an osmotic protective agent and stress response agent. It is thought to act as a protein. For example, Battaglia, et al., Plant Physiol. See 148:6-24 (2008). Such proteins are also thermally stable. UnaLEA protein is produced between 90°C and 110°C (for example, between 95°C and 105°C). At a temperature of 95°C or 100°C, at least 1 g / L (e.g., 2, 4, 6, 8, 10 In a solution of 15, 20, 25, 50, 100, 150, 200 or 250 g / L It may have a solubility such that the solution has a pH between 5 and 8 (e.g., 5, 5.5, 6, 6). It has a pH of 0.5, 7, 7.5 or 8, and a concentration of 0-300 mM (e.g., 50, 100, 1 It has a sodium chloride content of 50, 200, 250, or 300 mM. In this case, LEA protein is protein extract at 90°C to 110°C (for example, 95°C). After heating to 100°C and centrifuging or filtering the insoluble material For example, it can be isolated by concentrating the LEA protein fraction by ultrafiltration. In some cases, isoionic precipitation is used to further remove non-LEA proteins. The steps of trichloroacetic acid precipitation and / or ammonium sulfate precipitation are performed in the heating step. This can be done before or after. By heating the solution to 90°C to 110°C, most tannins can be removed. This process denatures the protein, allowing most of it to be removed from the solution.

[0100] B. Protein I don't want to get bogged down in theory, but non-animal proteins (for example, plant proteins) By separating and refining, the consumables exhibit greater consistency and quality than the consumables themselves. It is thought that it can be manufactured with great control. In some embodiments, the consumable protein Approximately 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7% of the ingredients. 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50 %, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99 % or more consists of one or more isolated and purified proteins. And the purified protein is 60%, 70%, 80%, 85%, 90%, 95%, 9 It may be more than 9% or even 100% pure.

[0101] The isolated and purified proteins are derived from one or more other components of a non-animal source. They can be isolated. For example, the protein fraction can be isolated from plant isolates. In some cases, the protein can be purified, and here, certain types of proteins , it is isolated from other components found in non-animal sources. Proteins are separated by their molecular weight Based on this, for example, size exclusion chromatography, ultrafiltration through a membrane or dense They can be separated by centrifugation. In some embodiments, the protein has a surface charge Based on this, for example, isoelectrolysis, anion exchange chromatography, or cation exchange chromatography Proteins can also be separated by matrixing, for example, based on their solubility. Separation can be performed by ammonium sulfate precipitation, isoelectrolytic precipitation, surfactant, detergent, or solvent extraction. It is possible. Proteins can also, depending on their affinity for other molecules, form, for example, a hydrophobic phase. Separation can be performed using interaction chromatography, reactive dyes, or hydroxyapatite. Affinity chromatography also uses specific binding parent proteins to target proteins. Antibodies with symmetry, nickel NTA against recombinant proteins with His tags, glycota Lectins that bind to sugar moieties on proteins or that specifically bind to the target protein This includes using other molecules.

[0102] By isolating proteins, it becomes possible to eliminate unwanted materials. (Some embodiments) The isolated and purified protein is then found in seeds, leaves, stems, or other parts of the plant. Unnecessary materials (e.g., nucleic acids such as RNA and DNA, lipid membranes, phospholipids, fats, oils, etc.) Carbohydrates such as corn, cellulose, and glucans, phenolic compounds, and polyphenols It is a protein substantially separated from a compound, aromatic compound, or pigment.

[0103] Isolated and purified proteins can also be expressed using polypeptide expression techniques (e.g., bacterial cells). Heterogeneous expression techniques using insect cells, fungal cells such as yeast cells, plant cells, or mammalian cells. ) can be produced by recombinant DNA. In some cases, the protein is synthesized To manufacture it, standard polypeptide synthesis techniques (for example, liquid-phase polypeptide synthesis techniques) Alternatively, solid-phase polypeptide synthesis techniques may be used. In some cases, proteins are synthesized. Cell-free translation technology may be used for manufacturing.

[0104] Proteins (one or more) incorporated into consumables can perform nutritional functions. In some cases, proteins also contribute to the characteristics of consumables, such as the flavor, color, and odor of the consumables. And / or works to change the texture. For example, meat substitutes work from a raw state to a textured state. It may include a protein indicator showing the progress of cooking to a processed state, where the meat substitute product is non It originates from animal sources.

[0105] Tamper which can be isolated and purified and used in the consumables described herein Examples of proteins include ribosomal proteins, actin, hexokinase, lactate dehydrogenase, Fructose bisphosphate aldolase, phosphofructokinase, iso triose phosphate Melase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pi Rubinate kinase, protease, lipase, amylase, glycoprotein, lectin, m Chin, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase, activator Chin, translation extension factor, histone, ribulose-1,5-bisphosphate carboxylase oxy Sigenase (RuBisCo), ribulose-1,5-bisphosphate carboxylase oxy Shigenase activase (RuBisCo activase), albumin, glycinin, co Glycinin, globulin, bicillin, conalbumin, gliadin, glutelin, gluc Ten, glutenin, hordein, prolamin, phaseolin (protein), protein Plast, secarin, extensin, wheat (triticeae) gluten, collagen, Zein, caffeline, avenin, dehydrin, hydrophylin, late embryonic development accumulation protein Quality, naturally unfolded proteins, any seed storage proteins, oleosins, Caloleosins, steroleosins, or other oils Plant protein, plant storage protein A, plant storage protein B, mung seed storage 8S globulin Examples include brin, globulin, pea globulin, and pea albumin. ru.

[0106] In some embodiments, the isolated and purified protein interacts with lipids and in the structure Proteins that help stabilize lipids, which bind to lipids and help crosslink lipid structures. It binds to proteins or lipids and crosslinks lipid structures and non-lipid-interacting proteins. It may be a protein that helps in this. I don't intend to be bound by any particular theory, but this specification The use of such proteins in the consumables described herein is prohibited for lipids and / or improve the incorporation of fat substitutes containing other ingredients in meat substitute products, and as a result, Improved texture and mouthfeel may be obtained in the final product. Lipid-interacting plant protein A non-exclusive example is proteins in the oleosin family. ¹ is a lipid-interacting protein found in plant oils. It interacts with lipids and Other, not limited to, plant proteins that can stabilize a marion include Great No Seed storage protein derived from peas, albumin derived from peas, peas Globulins derived from mung beans, 8S globulins derived from kidney beans, 8S globulins derived from green beans Examples include prolamins and lipid transport proteins.

[0107] In some embodiments, one or more isolated and purified proteins are heme-containing. This may be an iron-storing protein such as a protein. When used herein, the term "Heme-containing protein" is also known as "heme-containing polypeptide" or "heme protein" or It can be used synonymously with "heme polypeptide," and is covalent or noncovalent to the heme moiety. It includes any polypeptides that can be bound by bonding. In some embodiments, heme-containing polymer The peptide is globin, and contains a series of 7-9 alpha helices. It may contain globin. Globin-type proteins can be any class (e.g., class I, class I It may be Class I or Class III, and in some embodiments, it transports or stores oxygen. For example, heme-containing proteins are non-symbiotic forms of hemoglobin or leghemoglobin. This is possible. Heme-containing polypeptides are monomers, i.e., single polypeptide chains. It can be obtained, or it can be a dimer, trimer, tetramer and / or a higher-order oligomer. Oxygenated Fe of heme-containing proteins 2+ The lifespan of the state is similar to that of myoglobin. To obtain, or to manufacture, store, and handle heme protein-containing consumables, Under conditions prepared for consumption, 10%, 20%, 30%, 50%, 100%, or It can exceed that. Deoxygenated Fe of heme-containing proteins 2+ The lifespan of the condition is, Mioguro It may be similar to that in a bottle, or heme protein-containing consumables may be manufactured and stored. , under the conditions that it is handled or prepared for consumption, 10%, 20%, 30% 5 It can be 0%, 100%, or even higher.

[0108] Examples of heme-containing polypeptides, not limited to these, include androglobin. Cytoglobin, globin E, globin X, globin Y, hemoglobin, leghemoglobin Flavohemoglobin, Hell's Gate Globin I, Myoglobin, Erythrocruolin Beta hemoglobin, alpha hemoglobin, protoglobin, cyanoglobin, cyanoglobin Toglobin, histoglobin, neuroglobin, chlorocruorin, terminally cleaved hemoglobin Robin (e.g., HbN or HbO), terminally cleaved 2 / 2 globin, hemoglobin 3( Examples include Glb3), cytochrome, or peroxidase.

[0109] The heme-containing proteins that may be used in the consumables described herein are milk Animals (for example, domesticated animals such as cows, goats, sheep, horses, pigs, bulls, or rabbits), birds They may originate from plants, algae, fungi (e.g., yeast or filamentous fungi), ciliates, or bacteria. For example, heme-containing proteins are found in livestock (e.g., cows, goats, sheep, pigs, bulls or It can originate from mammals such as rabbits or birds such as turkeys or chickens. The heme-containing protein is from Nicotiana tabacum or Nicotiana. Nicotiana sylvestris (tobacco); Zea mays ( Corn, Arabidopsis thaliana, Glycine mackerel Glycine max (soybean), Cicer arietinum (galvanic acid) Pisum sativa (such as chickpeas, snow peas, or snap peas) Pisum sativum (pea) subspecies, pea, black bean, white kidney bean, northern bean Phaseolus vulgaris, a common bean species such as corn beans or pinto beans. Igaris subspecies, Vigna unguiculata subspecies (cowpea), Vigna • Radiata (Vigna radiata) (mung bean), White-flowered fan bean (Lupinus albus) (Lupinus) or Medicago sativa (Alfalfa), etc. Leguminous plant; Brassica napus (European rapeseed); Triticum (Triticum) genus species (wheat including grains and spelt); Gossypium hirsuts Gossypium hirsutum (cotton); Oryza sativa (rice); Ziza Zizania (a genus of rice species; wild rice); Helianthus annuus (Sunflower); Beta vulgaris (sugar beet); Pennisetum • Glaucum (Pennisetum glaucum) (Echinochloa crustacean); Chenopodium species ( Quinoa; Sesamum species; Linum usitatissimum (Flax) (um); or plants such as Hordeum vulgare (barley) It can be derived from substances. Heme-containing proteins are found in Saccharomyces cerevisiae. Pichia cerevisiae, Pichia pastoris, Magnaporte oryzae Porthe oryzae), Fusarium graminearum or Fusarium It can be isolated from fungi such as Fusarium oxysporum. Heme-containing Proteins include Escherichia coli, Bacillus subtilis, and Bacillus gigantea. Illus megaterium), Synechocystis species, Aquifex eo Aquifex aeolicus, Methylacidiphyllum infernorum Bacteria such as *Hilum infernorum* or thermophilic bacteria such as *Thermophilus*. It can be isolated from (for example, plants that grow at temperatures above 45°C). Heme-containing proteins are found in crystals. It can be isolated from algae such as Chlamydomonas eugametos. Heme-containing proteins are found in Paramecium caudatum or Tetrahymena pili. It can be isolated from protists such as Tetrahymena pyriformis. Some implementations In this state, bacterial hemoglobin is produced by Aquifex aeolicus, - Thermobifida fusca, Methylacidiphyllum inphenolum (Methylacidiphilum infernorum) (Hell's Gate), Synechocystis s) Selected from the group consisting of a genus or species of Bacillus subtilis. Contains a large amount of heme. The protein sequence and structure are publicly known. For example, Reedy, et al., Nuceic Acids Rese arch, 2008, Vol. 36, Database issue D307-D313 and http: / / hemeprotein.info / heme See the heme protein database available on the World Wide Web in .php format.

[0110] For example, non-symbiotic hemoglobin is found in soybeans, germinated soybeans, alfalfa, and golden soybeans. Flax (golden flax), black beans, cowpeas (black-eyed pea), northern n) Chickpeas, mung beans, cowpeas, pinto beans, pod peas, dried Peas, quinoa, sesame, sunflower, wheat grains, spelt, barley, wild rice It may be derived from plants selected from the group consisting of rice.

[0111] Heme-containing proteins described herein that may be used to manufacture consumables Each of them contains the corresponding wild-type heme-containing protein or heme-binding motif. For the amino acid sequence of the fragment, at least 70% (e.g., at least 75%, 80%) It has sequence identity of 85%, 90%, 95%, 97%, 98%, 99%, or 100%. Obtain. For example, heme-containing proteins include Vigna radiata (SEQ ID NO:). 1) Hordeum vulgare (SEQ ID NO: 5), Zea m ays) (Sequence No. 13), Oryza sativa subspecies japonica )(rice)(Sequence ID 14) or Arabidopsis thaliana Non-symbiotic hemoglobin derived from (SEQ ID NO: 15), methyl acydiphyllum-in Hells, such as those derived from Fernorum (Methylacidiphilum infernorum) (SEQ ID NO: 2). Gate (Hell's gate) globin I, Aquifex aeolicus Flavohemoproteins such as those derived from (SEQ ID NO: 3), Glycine Max (Glycine Max) max) (SEQ ID NO: 4), Pisum sativum (SEQ ID NO: 16), or Big Leghemo derived from Vigna unguiculata (SEQ ID NO: 17), etc. Globin, Magnaporthe oryzae (SEQ ID NO: 6), or Fusarius Heme-dependent peroxides such as those derived from Fusarium oxysporum (SEQ ID NO: 7) Xidase, derived from Fusarium graminearum (SEQ ID NO: 8) Cytochrome C peroxidase, Chlamydomonas moewus sii) (Sequence No. 9), Tetrahymena pyriformis (distributed) Row number 10, Group I terminal truncated type), Paramecium caudatum (Sequence number 11, Hemoglobin derived from Group I terminal truncated hemoglobin, and Aspergillus niger mold. )(Sequence ID 12) hemoglobin or bovine (Bos taurus) (Sequence ID 18) myo Globin, wild boar (Sus scrofa) (SEQ ID NO: 19) Myoglobin, horse (Equus caball) (us) (Sequence ID 20) Mammalian myoglobin proteins such as myoglobin, benthamia Nicotiana benthamiana (SEQ ID NO: 21), Bacillus subtilis ( Sequence ID 22), Corynebacterium glutamicum ( SEQ ID NO: 23), Synechocystis genus PCC6803 (SEQ ID NO: 24 ), Synechococcus species PCC7335 (sequence number 25), Nostoc commune (Nostoc commune) (SEQ ID NO: 26) or giant fungus (Bacillus megaterium) (SEQ ID NO: 26) 27) Including the heme protein derived from, at least the amino acid sequence shown in Figure 1 It can have 70% sequence identity. See Figure 1.

[0112] The percentage of identity between two amino acid sequences can be determined as follows. First, amino The acid sequence was obtained from an independent type of BLASTZ containing BLASTP version 2.0.14. Alignment using the BLAST2 Sequences (Bl2seq) program This standalone version of BLASTZ is available on the Fish & Richardson website. (For example, www.fr.com / blast / ) or the U.S. government's National Biotechnology Information Center National Center for Biotechnology Information website (www.n It can be obtained from cbi.nlm.nih.gov. Instructions for using the Bl2seq program. Instructions explaining this can be found in the readme file included with BLASTZ. Yes, it is possible. Bl2seq uses the BLASTP algorithm to analyze between two amino acid sequences. A comparison will be conducted. To compare the two amino acid sequences, the following Bl2seq options will be used: Set as follows: -i is set to the file containing the first amino acid sequence to be compared. (For example, C:\seq1.txt); -j is the second amino acid sequence to be compared. Set to a file containing (e.g., C:\seq2.txt); -p means blas Set to tp; -o is set to any desired filename (for example, C:\outp ut.txt); All other options will remain at their default settings. For example Then, use the following command to create an output file containing a comparison between two amino acid sequences. Can be produced: C:\Bl2seq -ic:\seq1.txt -jc:\seq2 .txt -p blastp -oc:\output.txt. Two types compared If the sequences share homology, the specified output file will be the aligned sequence. This indicates the region of homology between them. If the two compared sequences do not share homology, The specified output file does not show the aligned array. A similar procedure is used for blas The nucleic acid sequence can be traced, except that tn is used.

[0113] Once aligned, the number of positions in which the same amino acid residue is shown in both sequences is counted. The number of matches is determined by the total polypeptide amino acid. The identity percentage is the number of matches. The value is determined by dividing it by the length of the acid sequence, and then multiplying the resulting value by 100. The identity percentage value is rounded to the nearest tenth. For example, 78.11. 78.12, 78.13 and 78.14 are rounded down to 78.1, and 78.15, 7 8.16, 78.17, 78.18, and 78.19 are rounded up to 78.2. Furthermore, the length value is always an integer.

[0114] Some nucleic acids can encode polypeptides that have a specific amino acid sequence. This can be acknowledged. The degeneracy of the genetic code is well known in this field; that is, a large number of A Regarding amino acids, there are nucleotide triplets of two or more types that function as codons for amino acids. For example, the codons in the coding sequence of a given enzyme can be assigned to a suitable codon bias table of that kind. It is used to modify the expression to achieve optimal expression in a specific species (e.g., bacteria or fungi). It is possible.

[0115] Heme-containing proteins are derived from source materials (e.g., animal tissue or plants, fungi, algae). Alternatively, they are extracted from bacterial biomass, or, in the case of secreted proteins, from the culture supernatant. ) or can be extracted from a combination of source materials (e.g., multiple plant species). The bottles contain agricultural leguminous crops (e.g., soybeans, alfalfa, or peas). It is readily available as an unused by-product. In the United States, in the roots of these crops The amount of leghemoglobin in this meat exceeds the myoglobin content of all red meat consumed in the United States. ru.

[0116] In some embodiments, the heme-containing protein extract is obtained from the source material (e.g., other (e.g., derived from animal, plant, fungal, algae, or bacterial proteins, or combinations of source materials) For example, one or more non-heme-containing compounds derived from various animals, plants, fungi, algae, or bacteria. Contains protein.

[0117] In some embodiments, the heme-containing protein is supplied using the above technique from a source material (for example). If isolated from other components of other animals, plants, fungi, algae or bacterial proteins And purified. As used herein, the terms “isolated and purified” mean The heme-containing protein preparation is at least 60% pure, for example, 65%, 70%. Indicates purity above %, 75%, 80%, 85%, 90%, 95%, or 99%. vinegar.

[0118] Heme-containing proteins are also used in polypeptide expression technologies (e.g., bacterial cells, insect cells, algae). Heterogeneous expression techniques using fungal cells such as cell-like cells, yeast cells, plant cells, or mammalian cells. ) can be produced by recombinant DNA. For example, heme-containing proteins can be produced by recombinant DNA using E. coli (E. coli). It can be expressed in coli cells. Heme-containing proteins are used in protein purification. To support this, use FLAG, polyhistidine (e.g., hexahistidine, HIS tag). hemagglutinin (HA), glutathione-S-transferase (GST), or ma Tags can be attached using heterologous amino acid sequences such as luthose-binding proteins (MBPs). In some embodiments, a protective device is used to enable the cutting of the HIS tag and the HIS tag. Recombinant heme-containing proteins that include an ase (e.g., TEV) moiety are found in Escherichia coli (E. coli). It is expressed in His tag affinity chromatography (Talon resin, C It can be purified using loneTech. In some cases, heme-containing proteins. To produce it by synthesis, standard polypeptide synthesis techniques (e.g., liquid-phase polypeptides) are used. Synthetic techniques or solid-phase polypeptide synthesis techniques may be used. In some cases, heme Cell-free translation techniques may be used to synthesize the contained proteins.

[0119] In some embodiments, the isolated and purified protein is substantially its natural fall It is located in the 5 o'clock position and is water-soluble. In some embodiments, the isolated and purified protein is 5 0, 60, 70, 80, or more than 90% are present in its natural fold. In some embodiments, The isolated and purified proteins are water-soluble, with over 50, 60, 70, 80, or 90% water content. That is the case.

[0120] The proteins used in consumables are modified (e.g., hydrolyzed, cleaved) , crosslinked, modified, polymerized, extruded, electrospinned, spray dried or It can be freeze-dried, or derivatized or chemically modified. For example, a protein can be Other substances including sugars, lipids, cofactors, peptides or phosphates, acetates, and methyl compounds. Chemical groups and other natural or unnatural molecules can be modified by covalent bonding. For example, the peptide backbone of a protein is affected by exposure to an acid or protease or Proteins can be cleaved by other means. For example, proteins can be cleaved by exposure to heat or cold, p Changes in H, exposure to denaturing agents such as detergents, urea, or other chaotropic agents. It can be altered by mechanical stress, including shear, that is, by secondary, tertiary or The quaternary structure can be altered. Protein alliance in solution, colloid, or solid aggregates. The coating affects mechanical properties including tensile strength, elasticity, deformability, hardness, or hydrophobicity. It can be controlled to control the boss.

[0121] Proteins are also assembled into fibers that can form a matrix for the structure of a composition. It is possible. The three-dimensional matrix of protein fibers can be, for example, formed by intermolecular disulfide bridges. Chemicals that promote growth (mixed glutathione, dithiothreitol (DTT), ba It may contain ter-mercaptoethanol (BME). In some embodiments, the chemical is These are proteins (thioredoxin, glutaredoxin). In some embodiments, tan The protein is an enzyme (disulfide isomerase). In some embodiments, the fiber is N -Hydroxysuccinimide (NHS) esters, imide esters, aryl fluorides , aldehyde, maleimide, pyridyl dithiol, haloacetyl, aryl azide, dia Two selected from the group consisting of diline, carbodiimide, hydrazide, and isocyanate. It is crosslinked by a chemical crosslinking agent having two reactive groups.

[0122] In some embodiments, a coacervate containing one or more plant proteins is formed. For example, it can be used as a binder in meat or other imitations. In this process, a homogeneous solution of charged polymers undergoes phase separation, resulting in a dense, polymer-rich phase. This process yields a coacervate and a solvent-rich phase (supernatant). Coacervates, which are polysaccharides, have been used in the development of biomaterials. For example, Boral and Bohidar (2010) Journal of Physical Chemistry B. Vol 114 (37): 12027-35; Bi Liu et al., (2010) Journal of Agricultural and Food Chemistry, Vol 58:552-556 See also. The formation of such coacervates is due to the bonding between oppositely charged polymers. Driven by interaction. However, as described herein, core cerve Proteins can be formed using proteins (for example, plant proteins can be one or multiple types). Numerous pea proteins, chickpea proteins, lentil proteins, lupine (Contains proteins, other leguminous plant proteins, or mixtures thereof). Generally, Acervate contains pea melegmine or bicillin (for example, bicillin containing pea melegmine or bicillin). Sylin fraction), a combination of both bisyllin and legumin, or unfractionated pea methane Low ionic strength containing one or more isolated and purified plant proteins such as chlorine Prepare a solution (for example, a buffer solution with a concentration of 100 mM sodium chloride or less) and adjust the pH to 3.5-5.5 (e.g. For example, it can be formed by acidifying the pH to 4-5. Under these conditions, the protein The substance is separated from the solution, and the mixture is centrifuged to completely separate the coacervate. This can be done. Unlike precipitates, this coacervate can be stretched by pulling. It is a viscous material that melts when heated. This process involves oil (up to 70%, for example). This process is carried out in the presence of coconut oil or other oils, and can form a cream-like material. This can be done by changing the composition of the solution (the ratio of bisilin to legumin, the oil used). The type and amount of coacervates, as well as their binding properties, can be adjusted as desired. In the embodiment, one or more types of rubber (e.g., ara) are used to form the coacervate. Coacervates (such as beer gum or xanthan gum) may be used. Coacervates are fat- and muscle- As a binder in beef patty imitations to bind and hold together connective tissue imitations. It can be used.

[0123] In the presence of glycerol (0-30%) or plasticizers such as polyethylene glycol, A combination of wheat gluten (0-20%) and pea protein fraction (0-50%) By combining these materials, binding materials with various adhesive and cooking characteristics can be prepared. Depending on the circumstances, leghemoglobin or other heme-containing proteins may be added to the mixture. There are also combinations. During mixing, the ingredients are incorporated into the beef patty imitation in such a way that any lumps are removed. It is possible.

[0124] In some embodiments, the protein texture is obtained without extrusion. To arrange the materials, they can be subjected to freezing and alignment. This method allows the material to form ice crystals. This includes slow freezing of proteins. When cooled from one side, ice crystals form on the cooled side. Ice is preferentially formed in the direction perpendicular to the surface. After freezing, the ice is removed from the material in a freeze-dryer. Then, a material with several layers may remain. Next, it is heated under pressurized and humid conditions. This stabilizes the structure, and a material that can be used in meat imitation products can be manufactured. Protein freeze alignment was described by Lugay and Kim (1981). ment: A novel method for protein texturization. Page 177-187, Chapter 8 in: DW Stanley, ED Murray and DH Lees eds. 1981. Utilization of Protein Resources. See Westport, CT: Food & Nutrition Press, Inc. Frozen-aligned proteins The quality is subjected to further processing (in a solution containing beef flavor and / or leghemoglobin). (By immersion) to form a beef imitation, a fat- and connective tissue imitation and They can be used in combination. The imitations also have cold set gel (e.g., EN) around them. (containing pea protein and myoglobin) or cross-linked gel (e.g., pea meta) It is used as a structure in which protein and leghemoglobin can be formed, and then, It is combined with fat and connective tissue.

[0125] C. Lipids The consumables described herein may contain lipid components. Lipids are isolated and / or It can be purified into triglycerides, monoglycerides, diglycerides, free fatty acids, and sphi Sphingosides, glycolipids, phospholipids, or oils or aggregates of such lipids The body (for example, a membrane, lecithin, lysolecithin, or a bulk aqueous phase containing small amounts of lipids) It may be in the form of lipid droplets. In some embodiments, the lipid source is also genetically engineered Oils obtained from non-animal sources, including bacteria, algae, archaea, or fungi (e.g., plants, algae) (Oils obtained from fungi such as yeast or filamentous fungi, seaweed, bacteria or archaea) Examples of vegetable oils that are not limited to corn oil include olive oil, soybean oil, and peanut oil. Oils, walnut oil, almond oil, sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, Sunflower oil, flaxseed oil, coconut oil, palm kernel oil, coconut oil, babassu oil, shea butter, man Examples include goat butter, cocoa butter, wheat germ oil or rice bran oil, or margarine. The oil may be hardened (e.g., hardened vegetable oil) or unhardened.

[0126] Some sources indicate that lipids are triglycerides, monoglycerides, diglycerides, and free glycerides. Fatty acids, sphingosides, glycolipids, lecithin, lysolecithin, phospha Tidic acid, lysophosphatidic acid, phosphatidylcholine, phosphatidylinositol , phospholipids such as phosphatidylethanolamine or phosphatidylserine; sulf Sphingolipids such as ingomyelin or ceramide; stigmasterol, sitosterol Lol, Campestrol, Brascastrol, Sitostanol, Campestanol Lu, ergosterol, thymosterol, fecosterol, dinosterol, lanosterol Sterols such as cholesterol or episterol; N-palmitol Lorin, N-steroylglycine, N-palmitoylglycine, N-arachidonoylglycine Syn, N-palmitoyltaurine, N-arachidonoylhistidine, or anandamide Lipid amides such as palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristic acid Stolic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanoic acid, li Nolic acid (C18:2), eicosanoic acid (C22:0), arachidonic acid (C20:4), Eicosapentanoic acid (C20:5), docosapentaenoic acid (C20:5) 22:5), docosahexanoic acid (C22:6), erucic acid (C22:1), conjugated linoleic acid Acids, linolenic acid (C18:3), oleic acid (C18:1), elaidic acid (oleic acid (Trans isomer of), trans-vaccenic acid (C18:1 trans-11) or conjugated o Free fatty acids such as leic acid; or monoacylglyceride esters of such fatty acids, Such lipids, including diacylglyceride esters and triacylglyceride esters. It could be an ester of balsic acid.

[0127] Lipids may include phospholipids, lipid amides, sterols, or neutral lipids. Phospholipids are, Multiple amphiphilic groups including fatty acids (see above, for example), glycerol, and polar groups It may contain molecules. In some embodiments, the polar group is, for example, choline, ethanolamine, Serine, phosphate, glycerol-3-phosphate, inositol or inositol It is an acid. In some embodiments, the lipid is, for example, a sphingolipid, ceramide, sphingolipid. Gongoelin, cerebroside, ganglioside, ether lipids, plasmalogens or pe It is a glycated lipid.

[0128] In some embodiments, the lipids used in consumables are not limited to those mentioned above, but are also known as Maize seeds, safflower seeds, sesame seeds, rapeseed, almonds, macadamia nuts, grapefruit Fruits, lemon, orange, watermelon, pumpkin, cocoa, coconut, mango, Japanese corn Pumpkin, cashew, Brazil nut, chestnut, hazelnut, peanut, pecan, corn Cream painting made from seeds, nuts, and legumes, including lumi and pistachio. It is a fraction. As used herein, the term "cream fraction" refers to lipids, proteins, It can refer to an isolated emulsion containing minerals and water.

[0129] To obtain a cream fraction from seeds, nuts, or legumes, follow these steps: One or more methods may be performed. Seeds, nuts, or legumes may be treated for 1 to 30 minutes. They can be blended. For example, seeds, nuts, or legumes can be blended at a maximum speed of 4 minutes. The blend is achieved by gradually increasing the speed each time, and then blending at maximum speed for one minute. It can be. Seeds, nuts, or legumes can be: EDTA (0-0.1M), NaCl (0~1M), KCl (0~1M), NaSO4 (0~0.2M), potassium phosphate (0 (~1M), sodium citrate (0~1M), sodium carbonate (0~1M) and / or This is sucrose (0-50%), and water containing all or part of a pH range of 3-11. These are blended in solution to obtain a slurry. The slurry is heated to 20°C to 50°C. Then, it can be centrifuged to obtain the cream fraction (the top layer, also called "cream"). Further purification of the cream fraction involves washing the cream fraction with a 0.1M to 2M urea solution. This can then be achieved by further isolating the cream fraction by centrifugation. The remaining liquid (also called the "skim" layer), which is a protein-containing solution, is also used. It is possible.

[0130] The "cream" can be used as is or subjected to further refining steps. For example, washing and heating remove color and flavor molecules (e.g., unwanted molecules) or unwanted It can remove coarse particles and improve texture and creaminess. Especially high pH. Washing with a buffer (pH > 9) can remove bitter compounds and improve texture, and urea Washing can remove storage proteins, and washing at pH below 9 followed by washing at pH above 9 Washing with salt can remove unwanted color molecules, and / or washing with salt can reduce flavor compounds. It is possible. Heating can increase the removal of coarse particles, color, and flavor compounds. For example, cream The fraction may be heated at a temperature of 25°C to 80°C for 0 to 24 hours. In some embodiments, The creamy fraction contains seed storage proteins. In some embodiments, seed storage The stored proteins are substantially removed from the resulting creamy fraction.

[0131] D. Fibers The fibers are isolated and / or purified for inclusion in the consumables described herein. It is possible. The fibers are arabinoxylan, cellulose and, derived from any plant source. Indigestible starch, indigestible dextrin, inulin, lignin, wax, chitin, pe Non-starch polysaccharides such as kuchin, beta-glucan, and other plant components such as oligosaccharides. It could refer to...

[0132] The fibers are extruded proteins and solution-spun fibers as described herein. It can refer to protein.

[0133] E. sugar In some embodiments, the consumable may also contain sugar. For example, the consumable may contain only sugar. It is not limited to glucose (dextrose), fructose (fructose), galactose, ma Contains rinose, arabinose, xylose (D- or L-xylose), and ribose. Metamonosaccharides, and not limited to them, but also sucrose, lactose, melibiose, trehalose Disaccharides including cellulose or maltose, arabitol, mannitol, dulci Sugar alcohols such as sorbitol or sorbitol, galacturonates, glucuronates, It contains sugar acids such as gluconates, polysaccharides such as oligosaccharides and glucans, corn starch, and ja Starches such as potato starch, and pectins such as apple pectin or orange pectin. , raffinose, stachyose or dextran; plant cell wall degrading agents such as salicin It may contain substances and / or sugar derivatives such as N-acetylglucosamine.

[0134] F. Gel formation The components of the composition can be formed into a gel. In some embodiments, the gel is made from a non-animal source ( For example, plant sources or other non-animal sources such as genetically modified yeast or bacteria. Contains derived proteins. The gel can be formed using various methods. Protein concentration The temperature, enzyme concentration, pH, and / or processing temperature affect the rate of gel formation and the final tissue imitation. It will affect the quality.

[0135] The gel can be completely stabilized by physical crosslinking between its components. In some embodiments, The gel can be produced by a heating / cooling cycle, in which case the gel is made of protein molecules. It is stabilized by physical interactions (entanglement, hydrophobic interactions) between them. For example, gels. The protein solution should be heated at least at 40°C, 45°C, 50°C, 60°C, 70°C, 80°C, and 9°C. To heat to a temperature of 0°C or 100°C, then to room temperature or a temperature below 40°C. It can be formed by cooling.

[0136] In some embodiments, the gel is made of protein and any other components (e.g., lipids). It can be formed by subjecting a composition containing to high-pressure treatment.

[0137] In some embodiments, the gel may be produced by adjusting the pH of the solution. For example If the pH of the concentrated protein solution is also determined by hydrochloric acid or other acids or sodium hydroxide, The pH is adjusted to near the isoelectric point of the main protein component by adding other bases. obtain.

[0138] In some embodiments, the gel is produced by immersing protein powder in a solution. To obtain, for example, protein powder contains at least 1%, 5%, 10%, 20% (wt / v) Alternatively, it can be immersed in a concentrated sodium hydroxide solution. In other examples, The protein powder can be immersed in a mixed water / ethanol solution.

[0139] In some embodiments, the low-temperature solidified gel modifies or decomposes any thermally unstable components (e.g., It is formed to avoid oxidation of iron in the heme portion or the generation of undesirable flavors. For a general methodology for forming a warm-solidifying gel, see Ju and Kilara A. (1998) J. F. food Science, Vol 63(2): 288-292; and Maltais et al., (2005) J. Food Science, V See ol 70 (1): C67-C73). Generally, low-temperature solidified gels are protein solutions that solidify to their maximum capacity. It is formed by initial thermal denaturation below a small gel concentration (pH and protein Depending on the type, for spherical plant proteins such as pea protein, pH is usually used. <8% (w / v) at 6-9. The protein solution should not precipitate from the solution (e.g., 0 (Apply to a temperature above the protein denaturation temperature under ~500 mM sodium chloride, pH 6~9) It can be heated. The solution can be cooled to below room temperature and any thermally unstable components (e.g., heme-containing) may be removed. Proteins and / or oils) are present in a solution that has been sufficiently cooled, but before gelation. It can be mixed with sodium chloride or calcium chloride (e.g., 5-100). Gelation occurs with sodium chloride or calcium chloride (e.g., 5-100). The addition of mM can induce gel formation, and the solution should be kept below room temperature to enable gel formation. It can be incubated (usually for several minutes to several hours). The resulting gel can be used as is in the meat imitation. It may be used or further processed (e.g., stabilized) before being incorporated into meat imitation products. )obtain.

[0140] In some embodiments, the gel may contain a crosslinking enzyme or at least partially crosslink the gel. It can be produced (and stabilized, for example) by enzymes. Crosslinking enzymes include, for example, transglucan. Taminase, tyrosinase, lipoxygenase, protein disulfide reductase, Protein disulfide isomerase, sulfhydryl oxidase, peroxidase It may be hexose oxidase, lysyl oxidase, or amine oxidase.

[0141] In some cases, gels promote the formation of intermolecular disulfide crosslinks between proteins. It may contain chemical substances. In some embodiments, the chemical substance is a protein (e.g., thiole Doxin (glutaredoxin). In some embodiments, the protein is an enzyme (dis). It is rufidoisomerase.

[0142] The gel contains N-hydroxysuccinimide (NHS) ester, imide ester, and alley Lufluoride, aldehyde, maleimide, pyridyldithiol, haloacetyl, aryl From the group consisting of azides, diazilines, carbodiimides, hydrazides, and isocyanates Stabilized by chemical crosslinking with a selected chemical crosslinking agent having two reactive groups. obtain.

[0143] In some embodiments, the gel may be stabilized by the addition of starch and rubber.

[0144] In some embodiments, two or more of these approaches are used in combination. For example, transglutaminase-crosslinked gels can be further stabilized by heating / cooling treatment. It can be standardized.

[0145] G. Muscle imitation Many meat products contain a high proportion of skeletal muscle. Therefore, this invention relates to the important aspects of animal skeletal muscle. The present invention provides compositions that can be derived from non-animal sources that reproduce or approximate the characteristics thereof. Compositions derived from non-animal sources that replicate or approximate animal skeletal muscle are consumable. It can be used as an ingredient in products, for example, meat imitations. Such compositions are described herein as having odor It is labeled as "muscle imitation." In some embodiments, muscle imitation and / or muscle model Meat substitute products, including those made from meat, are partially derived from animal sources. In some embodiments, muscle molds are used. Meat substitute products, including artificial and / or muscle imitations, are derived entirely from non-animal sources.

[0146] Muscle tissue imitation contains one or more isolated and purified proteins. The muscle tissue imitation may contain high-quality ingredients and has the same taste and texture as muscle tissue derived from animal sources. It approximates texture or color.

[0147] Many meat products have a striated skeleton in which individual muscle fibers are mainly organized anisotropically. It contains muscle. Therefore, in some embodiments, the muscle imitation is organized to some extent anisotropically. It contains fibers. The fibers may contain protein components. In some embodiments, the fibers are , about 1% (wt / wt), about 2%, about 5%, about 10%, about 15%, about 20%, about 30%, Approximately 40%, approximately 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, approximately 95%, approximately 99% ( Contains protein components of wt / wt or more.

[0148] The connective tissue components of skeletal muscle substantially contribute to the texture, mouthfeel, and cooking behavior of meat products. Connective tissue consists of proteins (collagen, elastin) ranging from 0.1 to 20 microns. ) Composed of fibers. In some embodiments, to replicate the fibrous composition of animal connective tissue. Mixtures of fibers with diameters of <1 to 10 microns and 10 to 300 microns are produced. In some embodiments, the three-dimensional matrix of fibers replicates the tensile strength of animal connective tissue. It is stabilized by protein crosslinking. In some embodiments, the three-dimensional matrix of the fiber The product contains isolated and purified crosslinking enzymes. Crosslinking enzymes include, for example, transglutamyl crosslinking enzymes. Taminase, tyrosinase, lipoxygenase, protein disulfide reductase, Protein disulfide isomerase, sulfhydryl oxidase, peroxidase It may be hexose oxidase, lysyl oxidase, or amine oxidase.

[0149] Some proteins (for example, 8S globulin derived from mung bean seeds or peas) The albumin or globulin fraction of legume seeds is similar to that of animal muscle or adipose tissue. Due to its ability to form a gel with a texture, it is preferred for constructing meat imitations. It possesses certain characteristics. See also the proteins identified in sections IIIA and B. The material can be artificially designed to emulate the physical properties of animal muscle tissue.

[0150] In some embodiments, one or more isolated and purified proteins are used in meat imitation products. Approximately 0.1%, 0.2%, 0.5%, 1%, 2%, 3%, and 4% of the protein content by weight. %, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35% 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% , accounting for 90%, 95%, 99%, or more. In some embodiments, consumables tan The isolated and purified proteins with a protein content of one or more were approximately 0.1% and 0.2%. %, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15 %, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65 %, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more .

[0151] The skeletal muscle of animals such as beef cattle typically accounts for approximately 1% of the total muscle tissue mass at the time of meat processing. It contains a considerable amount of glycogen. After meat processing, this glycogen fraction is metabolized. This continues, resulting in products containing lactic acid, which lowers the pH of muscle tissue, an undesirable outcome in meat. Contributes to superior quality. Glycogen contains branching points with alpha (1→6) glycosidic bonds. Glucos having linear alpha(1→4) glycosidic bonds linked together It is a branched polymer of alpha. Plant-derived starch, in particular amylopectin, is also alpha A linear alpha(1→4) glycoside having a branching point containing a (1→6) glycosidic bond. It is a branched polymer of glucose linked together by bonding, and therefore, meat imitation It can be used as an analog of glycogen in construction. Therefore, in some embodiments Muscle or meat imitations contain starch or pectin.

[0152] Further components of animal muscle tissue include sodium, potassium, calcium, and magnesium. Mu and other metal ions, lactic acid and other organic acids, free amino acids, peptides, Nu Examples include creotides and sulfur compounds. Therefore, in some embodiments, muscle mimicry The ingredients include sodium, potassium, calcium, magnesium, iron, zinc, copper, nickel, and phosphate. Other metal ions such as thium or selenium, and other organic acids such as lactic acid and fatty acids. Free amino acids, peptides, nucleotides and sulfur compounds, glutathione, beta-mercury It may contain putoethanol or dithiothreitol. In some embodiments, muscle imitation or sodium, potassium, calcium, magnesium, and other metal ions in consumables Lactic acid, other organic acids, free amino acids, peptides, nucleotides and / or sulfur The concentration of the compound is within 10% of the concentration found in the reproduced muscle or flesh. .

[0153] The present invention also provides a method for manufacturing muscle imitation products. In some embodiments, the method is The process involves forming the composition into asymmetrical fibers and then incorporating them into consumables. In this configuration, these fibers replicate muscle fibers. In some embodiments, the fibers are spun fibers. In other embodiments, the fibers are extruded fibers. Therefore, the present invention is non-paired. The present invention provides a method for producing crystalline fibers or spun protein fibers. In some embodiments, The fibers are formed by extruding protein components through an extruder. The extrusion method is This is well known in the art, for example, U.S. Patent No. 6,379,738 and No. 3,693,5 This is described in Patent No. 33 and U.S. Patent Publication No. 20120093994, which are references. These methods are incorporated herein by reference. These methods are used to create compositions provided herein. This can be applied to the manufacture of [the product / service].

[0154] Extrusion is performed, for example, by Leistritz Nano-16 twin-screw co-rotating extrusion. Machine (American Leistritz Extruder Corp. USA, This can be done using Sommerville (NJ) to limit protein denaturation. Therefore, active cooling of the barrel portion may be used to prevent expansion and excessive moisture loss of the extruded product. To limit the process, active cooling of the die portion may be used. The protein feed and liquid are Proteins are added separately. The protein is supplied via a volumetric plunger feeder or a continuous auger type. Supplied by a feeder, the liquid can be added to the barrel through a high-pressure liquid injection system. Press For the precise control of the extrusion pressure, cooling rate, and product expansion, nozzles with various inner diameters and channel lengths can be used. In some examples, the extrusion parameters are a screw rotation speed of 100 - 200 rpm, a die diameter of 3 mm, a die length of 15 cm, and the product temperature at the end of the die was 50 °C, a feed rate of 2 g / min, and a water flow rate of 3 g / min. The product temperature at the die during extrusion is measured by a thermocouple.

[0155] Spinning fibers can be produced by preparing a highly viscous protein "dope" in a concentrated protein solution or by adding sodium hydroxide to precipitated protein and pushing the solution through a small steel capillary tube (in some cases, a syringe with a syringe pump) through a small steel capillary tube (in some cases, a 27-gauge hypodermic needle) into a coagulation bath. In some examples, the bath is filled with a concentrated acid solution (e.g., 3 M hydrochloric acid). In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath. (in some cases, a syringe with a syringe pump) through a small steel capillary tube (in some cases, a 27-gauge hypodermic needle) into a coagulation bath. In some examples, the bath is filled with a concentrated acid solution (e.g., 3 M hydrochloric acid). In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath. (in some cases, a 27-gauge hypodermic needle) into a coagulation bath. In some examples, the bath is filled with a concentrated acid solution (e.g., 3 M hydrochloric acid). In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath. In some examples, the bath is filled with a concentrated acid solution (e.g., 3 M hydrochloric acid). In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath. In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath. (in some cases, a 27-gauge hypodermic needle) into a coagulation bath. In some examples, the bath is filled with a concentrated acid solution (e.g., 3 M hydrochloric acid). In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath. (in some cases, a 27-gauge hypodermic needle) into a coagulation bath. In some examples, the bath is filled with a concentrated acid solution (e.g., 3 M hydrochloric acid). In some examples, the bath is filled with a buffer solution having a pH approximately equal to the isoelectric point of the protein. The jet of the coagulating protein solution forms fibers that collect at the bottom of the bath.

[0156] A bundle of spinning fibers can be produced by forcing the protein "dope" through a spinneret having a number of small holes. In some examples, the spinneret is a stainless steel plate with approximately 2 2 5,000 holes per cm and each hole has a diameter of approximately 200 microns. In some embodiments, a muscle tissue construct is a three-dimensional matrix of fibers (connective tissue (in some cases, a 2) and each hole has a diameter of approximately 200 microns. In some embodiments, a muscle tissue construct is a three-dimensional matrix of fibers (connective tissue Immersing the imitation in a protein solution and incorporating a three-dimensional matrix of fibers It is manufactured by creating a protein gel.

[0157] H. Fat imitation Animal fat is important to the experience of eating cooked meat and is part of the nutritional value of meat. This is important for the chemical composition and physical properties of ground beef, for example. By using ingredients that mimic properties, the texture and / or flavor are enhanced. The present invention provides compositions derived from non-animal sources, reiterating the key characteristics of physical fats. Therefore, the present invention relates to a meat substitute product comprising a composition derived from a non-animal source, which is the main point of animal fat. To provide. Such compositions are referred to herein as “adipose imitations” or It is labeled as "fat imitation." In some embodiments, it is labeled as fat imitation and / or Meat substitute products, including fat imitation products, are partially derived from animal sources. Consumables are also Tex Non-animal ingredients including flavor, hardness, fat release percentage and / or fat release temperature This may also include fat imitation products that reiterate the key characteristics of fat. The fat content of consumables is low. 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, It can be 50%, 55%, 60%, 70%, 80%, 90%, or 95% fat.

[0158] Ground beef usually contains adipose (fat) trimmed from steaks. Lean beef is mixed with fatty tissue (Cox 1993) which is added at a ratio of 16-30%. Therefore, it is prepared. Meat that has been ground without using fat is tough, brittle, and quick to chew. It dries out. Fat is added to lean beef, and as a result, the fat released during cooking This helps to provide a liquid surface that generates the important beef flavor, which is largely due to the production of fatty acids. It is an ingredient. It plays a similarly important role in the texture and flavor of plant-based ground beef. Designing adipose tissue imitation is a crucial driver of texture and flavor.

[0159] The adipose tissue imitation described herein has a fatty acid composition in which the amount of saturated fat is reduced. Because it can be controlled to be more beneficial, it offers greater health benefits than beef adipose tissue. Plant-based fat substitutes do not contain cholesterol. Plant-based fat substitutes are It contains a lower percentage of total fat, and furthermore, it has the desired cooking characteristics, flavor and texture. It may have an equal amount of fat released or retained for that purpose.

[0160] As described herein, composition (e.g., fatty acid composition), cooking characteristics (e.g., The fat release temperature or fat release percentage and physical properties (e.g., hardness) are controlled. Therefore, plant-derived lipids can make it possible for plant-based compositions to mimic animal-based fats. Furthermore, a fatty model containing an emulsion of one or more isolated and purified proteins. Products can be manufactured. Adipose tissue imitations may contain (1) triacylglycerides of fatty acids. (1) Vegetable oil, (2) One or more isolated and purified proteins from non-animal sources (For example, plant proteins), as well as (3) phospholipids such as lecithin. The quality is that of plant or microbial proteins such as those mentioned above (e.g., RuBisCo, oleosin). This may include albumin, globulin, or other seed storage proteins. (Section IIIA) See also the proteins described in B. Vegetable oils are described herein. It could be any of the oils used. See, for example, Section IIIC.

[0161] The fat imitation may be a gel-like emulsion. In some embodiments, the gel is protein It is a soft, elastic gel containing carbohydrates, and optionally, a gel-like emulsion. Lujon involves the isolation and purification of multiple proteins, for example, 1 to 5 types or 1 to 3 types. It may contain a protein solution containing the protein, and the protein solution is the volume of the emulsion. It accounts for 1-30% of the total. Gel-like emulsions may contain lipid droplets, and lipid droplets are part of the emulsion. It accounts for 70-99% of the volume. The gel-like emulsion is isolated and purified cross-linked yeast. It may contain elements, and the cross-linking enzyme is 0.0005%~0.5% emulsion weight / volume, 0.5 ~2.5% emulsion weight / volume or 0.001% or less emulsion weight / volume It occupies a large portion. Emulsions of lipid droplets in protein solutions are cross-linked by enzymes such as transg By forming an emulsion into a gel using lutaminase, the protein solution is added. By gelling proteins through heating and cooling, cold set gel By forming a ligament, by forming a coacervate, or in node C These techniques described for coacervates are combined with gel formation in Section F. It can be stabilized by doing so.

[0162] In some embodiments, the fat mimicry undergoes a reaction that leads to covalent crosslinking between proteins. It contains a catalyzing cross-linking enzyme. The cross-linking enzyme modifies the desired structure and texture of the adipose tissue imitation. To create or stabilize an equivalent desired animal fat, to mimic the desired texture. It can be used for the following purposes. In some embodiments, the crosslinking enzyme is isolated and purified from a non-animal source. Examples and embodiments thereof are described herein. In some embodiments, fat Counterfeit products are at least 0.0001%, at least 0.001%, at least 0.01% , containing at least 0.1% or at least 1% (wt / vol) of crosslinking enzyme. Examples of these include transglutaminase, tyrosinase, lipoxygenase, and proteins. Disulfide reductase, protein disulfide isomerase, sulfhydryl ox Sidase, peroxidase, hexose oxidase, lysyl oxidase and amine It can be selected from oxidases. In some embodiments, the crosslinking enzyme is transglutamin Lysyl oxidase (e.g., Pichia pastoris lysyl oxidase) It is an oxidase or other amine oxidase.

[0163] Fat imitation products may contain a gel in which fat droplets are suspended. The fat droplets used are In some embodiments of the invention, the source may be derived from various sources. In some embodiments, the source is , non-animal sources (e.g., plant sources). For example, provided in Section IIIC. See the example. In some embodiments, fat droplets are animal products (e.g., butter, cream). Derived from lard and / or suet. In some embodiments, the fat droplets are from the pulp. In other embodiments, the source is derived from seed oil. In other embodiments, the source is algae, yeast, or Yarowia li It could be an oily yeast or mold such as Yarrowia lipolytica. For example, In this embodiment, triglycerides derived from Mortierella isabellina are used. Celides may be used. In some embodiments, the lipid droplets contain synthetic or partially synthetic lipids. do.

[0164] In some embodiments, lipid droplets are not limited to phospholipids, lecithin and It is stabilized by the addition of surfactants, including lipid membranes. The lipid membrane is formed by algae, fungi, or plants. It may be derived from materials. In some embodiments, surfactants constitute less than 5% of the fat imitation. Lipid droplets can have diameters ranging from 100 nm to 150 μm in some cases. The diameter of these stabilized droplets is determined by homogenization, high-pressure homogenization, extrusion, or ultrasonic treatment. It is possible to obtain it.

[0165] In some embodiments, the vegetable oil is modified to resemble animal fat. The vegetable oil is flavored. Food or heme protein, amino acids, organic acids, lipids, alcohols, aldehydes, ketones, Other agents such as lactones, furans, sugars, or the taste of meat during and after cooking. The essential scents can be modified using other flavor precursors. Therefore, the present invention Some embodiments describe the qualitative similarity between the cooking properties of animal fats and the cooking properties of vegetable oils in consumables. Includes methods for investigation.

[0166] In some embodiments, further polysaccharides, including flaxseed polysaccharides and xanthan gum, are used in lipids. It can be added to fat imitation products.

[0167] The preparation of plant-based fat mimics requires the stabilization of oil-in-water emulsions. Typically, Animal adipose tissue contains approximately 95% fat and is composed of a phospholipid bilayer and associated proteins. It is stabilized. The fat imitations described herein mimic the properties of animal fat. Furthermore, up to 95% fat is used, and in some cases, 80% fat is used under numerous conditions. It can be made using or with a smaller amount of fat (e.g., 50% or less). Achieving a stable emulsion is controlled by stabilizing the emulsion.

[0168] Composition (e.g., fatty acid composition), cooking characteristics (e.g., fat release temperature or fat release temperature) (Cent) and physical properties (e.g., hardness), type and amount of fat, amount of protein By controlling the type and amount of lecithin, the presence of additives, and the gelation method, the process can be controlled. It can be done.

[0169] In some embodiments, the protein component is approximately the same as the fat imitation by dry weight or gross weight. 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15% or 20%, 25% or so This constitutes the above. In some embodiments, the protein component is determined by dry weight or gross weight. It constitutes approximately 0.1-5% or 0.5-10% or more of the fat imitation. In this embodiment, the protein component is 0.5 to 0.5 by dry weight or gross weight of the fat imitation. The percentage is 3.5% or 1-3%. In some embodiments, the protein component is one or multiple. It contains a solution containing several isolated and purified proteins. The type of protein is EMA RuBisCo and pea albumin may affect the stability of the lujean. This allows fat imitation products to be manufactured from over 90% fat. Flaxseed and xanthan The addition of polysaccharides, including gum, helps emulsify the mixture and allows for an increase in fat content.

[0170] The type and amount of fat can be controlled by selecting the fat source and its lipid composition. It is possible. Generally, the more saturated fatty acids an oil has, the lower the protein concentration. Oils that can be emulsified well and have more unsaturated fatty acids have a higher protein concentration. It requires emulsification. Proteins are necessary to stabilize the emulsion. Increasing the protein content enhances stability. The amount of protein added affects the amount of fat. If there is too little to emulsify, the mixture will separate into layers.

[0171] Lecithin is also an emulsion modulator, and the amount of protein present is Depending on the type of oil used, the emulsion may be stabilized or destroyed. For example, lecithin breaks down the protein / fat matrix, making the less stable ema Lujon can be produced, but it can also be added at low levels to modify other physical properties. Emulsions made from oils containing a large amount of unsaturated fats contain a large amount of lecithin (1 It can be destabilized by %), and as a result, the emulsion will not solidify. Large amounts of saturated fat An emulsion made from an oil containing can be solidified with a large amount of lecithin (1%). It is extremely soft.

[0172] As described herein, the range can be from extremely soft to extremely hard. Fat imitation products can be prepared. The composition and amount of fat control the hardness of the imitation product. The harder the oil containing long-chain saturated fats, the harder the gel it produces. The oil produced typically contains more unsaturated or short-chain saturated fatty acids. The hardness of the gel increases as the total fat percentage increases, provided the emulsion is retained and does not separate. It increases as it progresses. The amount of protein also contributes to the hardness of the imitation. Generally, protein Increasing the concentration of the material increases the hardness of the imitation. The amount of lecithin is a modulator of the hardness of the imitation. Therefore, if the gel is formed with a high protein percentage (3%), a large amount of lecithin is needed. N (1%) is considerably softer than a small amount of lecithin (0.05%). Protein is reduced. Then (1.8%) all gels soften, and the emulsion is low when retained. There is almost no difference in hardness between the low level (0.05%) and high level (1%) of lecithin. .

[0173] This applies not only to counterfeit products, but also to xanthan gum and flaxseed paste. The addition of polysaccharides can increase the hardness of the fat-imitation gel.

[0174] When a fat imitation is cooked, as it cooks, the fat leaks out of the imitation that was constructed. There is often fat remaining in cooked products, and this fat is released to aid in the cooking process. It is important to achieve a balance between the fat content and the fat that remains for texture and flavor. The percentage of fat released (per total fat) is the amount of fat released during cooking. This can be determined by measuring the quantity. The percentage of released fat is equal to the total fat of the imitation. This is reported as the weight of fat released per unit. For example, as described herein. The percentage of fat released from adipose tissue imitation materials is 0-10%, 10-20%, and 20% during cooking. %~30%, 30%~40%, 40%~50%, 50%~60%, 60%~70%, 70 It can be %~80%, 80%~90%, or 90%~100%. Fat imitation products are usually, Under standard cooking conditions, it releases 0-90% of its fat. In comparison, beef adipose tissue typically releases the same amount of fat. Under these conditions, 40-55% of fat is released.

[0175] Fat imitation products can be manufactured using vegetable oils that have a specified melting point while releasing fat. The temperature range is wide. The fat release temperature is the temperature at which fat is visibly released from the food on the cooking surface. This is the temperature at which the fat is released. As described herein, the fat release temperature of a fat imitation is The type and amount of fat, the amount of protein, the type and amount of lecithin, the presence of additives, and emulsification. The method and gelation method can be combined. The resulting fat imitation can be stored at 23°C~ 33℃, 34℃~44℃, 45℃~55℃, 56℃~66℃, 67℃~77℃, 78℃~ 88℃, 89℃~99℃, 100℃~110℃, 111℃~121℃, 122℃~132 °C, 133°C~143°C, 144°C~154°C, 155°C~165°C, 166°C~167°C °C, 168°C~169°C, 170°C~180°C, 181°C~191°C, 192°C~202°C °C, 203°C~213°C, 214°C~224°C, 225°C~235°C, 236°C~246°C °C, 247°C~257°C, 258°C~268°C, 269°C~279°C, 280°C~290°C Beef fat may have a fat release temperature of ℃, or between 291℃ and 301℃. It was measured to release fat at 50°C.

[0176] Emulsification is also a factor in controlling the temperature of fat release: fat is converted into protein or When incorporated into the imitation along with protein and lecithin, the temperature at which fat is released is The temperature rises significantly above the melting point of fat on its own.

[0177] Fatty acid composition is also a factor in fat release temperature and fat release percentage. Vegetable oils containing a high proportion of unsaturated fatty acids have a low melting point and are mostly liquid at room temperature. Vegetable oils containing a higher proportion of saturated fatty acids have a higher melting point and are more volatile. It is solid at warm temperatures. Imitations with a higher amount of unsaturated fats are made with more saturated fatty acids. It has a higher temperature of fat leakage than similar imitations. The mixture is handheld homogenized. A larger amount of unsaturated fatty acids, emulsified by Zar and gelled using a heat-cooling method. 75% has a high protein content (3%) and a minimum lecithin content (0.05%) The gel produced from the oil was heated to 200°C with little to no fat release. It is possible. Imitations containing oils with more long-chain saturated fats are usually high in protein. It contains oils that have a higher fat release in terms of quality, but also contain more short-chain fatty acids. Compared to imitations, it releases a lower total fat percentage compared to products with a lower protein percentage. It contains a higher percentage of short-chain saturated fatty acids, high protein content (3%), and minimal lecithin. A gel produced from oil containing a quantity of 0.05% released fat with little to no fat release. It can be heated to 200°C.

[0178] The percentage of fat released when fat imitation products are cooked also depends on the amount of protein and lecithin. It is a function of the amount of protein. Typically, fat imitation products contain 1-3% protein by mass. Increasing protein content leads to a higher temperature for fat release, and the released fat... Reduce the fat content. Increasing the lecithin content to 1% raises the fat release temperature from 60°C to 11°F. The temperature can be reduced to 5℃, and the percentage of fat released can be increased (e.g., 25-30%). The source or composition of lecithin can regulate the amount of fat release and the temperature threshold for fat release. I don't want to get bogged down in any specific mechanism, but lecithin is involved in protein-protein interactions. It is thought that the emulsion is destabilized by destroying its properties. In one embodiment, Increasing the lecithin content to 1% at a high protein concentration of 3% raises the fat release temperature to 55 The temperature was reduced to ~60℃, increasing the percentage of fat leaked to 60-65%.

[0179] The method used to produce the emulsion is also a factor in determining the amount of fat released. This forms a homogeneous mixture of fats that are retained within a matrix of proteins and lecithin. The emulsification method may include high-pressure homogenization, ultrasonic treatment, or manual homogenization. Alternative methods This results in characteristic differences in the size of oil droplets in the emulsion, which in turn affects the resulting emulsion. This affects John's stability and the maximum fat concentration at which a stable emulsion can be formed.

[0180] The method of gelling the imitation is also a factor in determining the amount of fat released. While fat imitation products may be formed without this process, gelation creates a harder, more stable emulsion. It brings about. The gelation method is described above, for example, transglutaminase ( This includes adding cross-linking enzymes such as TG or subjecting the emulsion to a heating / cooling cycle. It is possible. For example, treatment using TG or heating / cooling methods is possible as described above. Lujon can be converted into a gel. Furthermore, it is formed by crosslinking catalyzed by TG. Gel-like emulsions are typically emulsions that have been gelled by heating / cooling techniques. It releases fat at a higher temperature than other methods. The gel formed by cross-linking using TG Furthermore, it releases less fat than gels typically formed by heating and cooling techniques. Release.

[0181] In some embodiments, the fat imitation has a protein content of <1.5% and a minimum lecithin content. It can be manufactured using a quantity (0.05%), with a fat release temperature of 45-65°C and high release temperature. These gels contain a large amount of fat (e.g., 70-90%). It is located at the higher end of the terminal.

[0182] In some embodiments, the fat imitation has a lower protein content (<1.5%) and a higher protein content. It can be manufactured using lecithin (>1%) and has a lower fat release temperature (e.g., 30-50°C). For example, at temperatures of 30-45°C and with an intermediate percentage of leaked fat (45-65%). Therefore, from oils with low protein concentration and short-chain or long-chain fatty acids In the formed gel, lecithin may play a role in stabilizing the emulsion.

[0183] In some embodiments, to produce a fat imitation having more than 70% fat, >2% Bisco or pea malbumin may be used. In some embodiments, >3% single The gel formed using the separated and purified proteins contains more than 70% fat. It brings counterfeit goods.

[0184] In some embodiments, a higher proportion of long-chain saturated fatty acids, 3% protein content and the most Fat imitations made from oil with a small lecithin content (0.05%) are similar to beef fat. It can release fat at temperatures similar to those that do so (50-100°C), and at low to intermediate levels. It can release fat (15-45%).

[0185] In some embodiments, higher protein concentrations (>3%) and lecithin content (>1%) are used. The fat imitation product has a fat release temperature of 50-70°C and releases a larger amount of fat (50-8°C). It may contain 0%). It has a high protein and low lecithin concentration and higher saturated fatty acids. The gel typically contains about 10% more than the corresponding gel formed using unsaturated fats. It leaks fat.

[0186] In some embodiments, protease treatment of protein components prior to gel formation leads to lipid release. This could lead to an increase in [the number of cases].

[0187] In some embodiments, the adipose tissue imitation matrix stabilized by a cross-linking enzyme is More than adipose tissue imitation matrix stabilized by heating / cooling protein denaturation. It releases fat. In one embodiment, mung bean 8S protein and canola oil or Adipose tissue matrix composed of an equal mixture of coconut, cocoa, olive, and palm oil. Crosslinks are formed more easily during heating / cooling denaturation than when crosslinks are formed using enzymes. It holds more mass when combined with Rubisco and Cocoa. By heating / cooling a pre-formed protein-oil emulsion containing teratin, The adipose tissue matrix formed is a similar composition stabilized by cross-linking enzymes. It has a higher melting point than fat imitation materials.

[0188] In some embodiments, 1.4% wt / v mung bean 8S protein and 90% v / v Adipose tissue imitation constructed from canola oil and 0.45% wt / v soy lecithin It can be homogenized in the presence of a variable concentration of sunflower oleosin. The concentration of oleosin is 1 :10~1:10 6 Molar ratio of oleosin:triglycerides can vary. (Referring to adipose tissue imitation) As the concentration of oleosin increases, an increase in mass retention after cooking is observed.

[0189] The hardness of adipose tissue imitation constructed as a stabilized protein-lipid emulsion is It can be modified by altering the protein concentration within the adipose tissue imitation matrix. For example, using 70-80% v / v sunflower oil and different concentrations of Rubisco A series of adipose tissue imitation materials formed by 0% and 0.18% (wt / The adipose tissue imitation containing vol)Rubisco was extremely soft, but 1.6% ( The imitations formed using wt / vol Rubisco are soft, with 1.9% (wt / vol) The imitation made using vol)Rubisco had an intermediate hardness.

[0190] In one embodiment, the protein oil emulsion was formed by stabilizing it. The hardness of the fat imitation material can be altered by varying the amount of protein in the fat imitation material. Obtained. In one embodiment, adipose tissue produced from Rubisco and 70% sunflower oil. The imitation tissue contains 1% R, which is higher than the 3% Rubisco concentration in the adipose tissue imitation. It is softer at lower concentrations, such as uBisCo.

[0191] In another embodiment, the present invention provides a method for producing a fat imitation. The fat is isolated, Homogenization is possible. For example, an organic solvent mixture can help solubilize lipids in a gel. Used, then removed, to provide the final gel. At this point, the lipids are frozen. It can be dried or stored. Therefore, in one embodiment, the present invention has similar characteristics to animal fat. The present invention provides a method for isolating and storing lipids that have been selected for this purpose. The membrane or lipid cake can be hydrated. Hydration can be carried out by stirring or by temperature changes. This can occur in the precursor solution to the gel. After hydration, the lipid suspension exhibits the properties of the lipids in the solution. To change this, it can be processed with sound waves, homogenized, high-pressure homogenized, or extruded. .

[0192] In some embodiments, the fat imitation is assembled to approximate the organization of adipose tissue in meat. In some embodiments, some or all of the components of the fat imitation are gels (e.g., t It is suspended in an protein gel. In other embodiments, the gel is a hydrogel, organogel. It may be a gel or xerogel. In some embodiments, the gel is a polysaccharide or protein. The viscosity can be increased to a desired consistency using a base agent. For example, the structure of a consumable product. Alternatively, to increase the viscosity of the gel that forms the structure, starch (fecula), arrowroot, Corn starch, potato starch, sago, tapioca, arginine Guar gum, locust bean gum, xanthan gum, collagen, egg white, fercere Orchid, gelatin, agar, carrageenan, cellulose, methylcellulose, hydroxymethyl Cellulose, acadia gum, konjac, starch, pectin, amine Lopetin or derived from legumes, grains, nuts, other seeds, leaves, algae, and bacteria, Bacterial proteins can be used individually or in combination.

[0193] In some embodiments, the tensile strength of the fat imitation mimics the tensile strength of adipose tissue. The tensile strength of the emulsion can be increased by incorporating fibers. Not necessarily, but watermelon, jackfruit, pumpkin, coconut, green algae, and corn It may be derived from non-animal sources, including sorghum and / or cotton. In some embodiments, The fibers are derived from the autopolymerization of proteins, such as oleosin and prolamin. In this embodiment, the fibers are electrospinned or extruded into the protein. It originates from a three-dimensional mesh or strand in which each fiber may have a diameter of less than 1 mm. It can form.

[0194] Fat mimics are suspended therein as solutions and droplets of one or more proteins. It may be an emulsion containing one or more types of fats. The oil phase is slowly added to the aqueous phase. This can provide a more robust emulsion and prevent accidental emulsification failures. Adding tin can, in some cases, stabilize emulsions with proteins. This destabilizes the mixture and allows for increased fat leakage when the imitation is cooked. In this state, the emulsion is stabilized into a gel by one or more cross-linking enzymes. In this embodiment, the emulsion is prepared by heating-cooling or cold-set gel technology. The gel is then stabilized by a matrix formed by proteins that have been induced into the gel. Heating an emulsion stabilized by a protein will cause the protein to denature. This can lead to an increase in the hardness of the fat imitation. Heating to a sufficient temperature is also important for natural milk The viability of Cloflora can be reduced by at least 100 times. In some embodiments, emulsion The process involves one or more protein cross-linking enzymes and heating / cooling technology or cold setting. Stabilized by a gel-like protein matrix formed by a combination of Togel technologies. After the emulsion has cooled sufficiently but before gelation is complete, the fat is more... To provide the final product with a natural pink color and / or improved flavor, amino acids and sugars are added. To give one of more flavor compounds such as thiamine or phospholipids, heme One or more optional components, such as contained proteins, may be added (e.g., 0.1 Up to approximately 0.4%, such as 5%, 0.2%, 0.25%, 0.3%, or 0.4%.

[0195] One or more proteins in the solution are isolated and purified proteins, for example, A fraction rich in purified pea albumin, purified pea meglobulin Rich fraction, purified mung bean 8S globulin rich fraction and / or Rubi It may contain a fraction rich in SCO. In other embodiments, one or more fats are of plant origin. It is derived from oil (rice bran oil or canola oil). See, for example, Section IIIC. In some cases, the composition is transglutaminase, lysyl oxidase or It contains other cross-linking enzymes such as amine oxidases. Therefore, in some embodiments, fats Tissue mimicry involves isolating and purifying one or more proteins; one or more proteins. To prepare a solution containing proteins; to emulsify one or more fats in a solution. ; and by stabilizing the solution into a gel-like emulsion using one or more crosslinking reagents. It can be manufactured.

[0196] In some embodiments, the fat imitation is emulsified with 40-80% rice bran oil, and 0.5 Purified enzymes stabilized on a gel using ~5% (wt / vol) transglutaminase. This is a high-fat emulsion containing a protein solution of daisy bean albumin.

[0197] In some embodiments, the fat imitation is 40-80% rice bran oil or 40-80% ka It is emulsified with nora oil and processed using 0.5-5% (wt / vol) transglutaminase. Contains a protein solution of isolated mung bean 8S globulin stabilized in a gel. It is a high-fat emulsion.

[0198] Fat can be isolated from plant tissue and emulsified. Emulsification involves rapid blending and homogenization. High-pressure homogenization, ultrasonic treatment, shearing, stirring, or temperature changes may be used. Lipid suspensions are solutions To further modify the properties of the lipids inside, they can be subjected to ultrasonic treatment or extruded. In some embodiments, other components of the consumables are added to the solution, followed by a gelling agent. It is added. In some embodiments, a crosslinking agent (e.g., to) is used to bind the components of the consumable. Lance glutaminase or lysyl oxidase is added. In other embodiments, A gelling agent is added, and the lipid / gel suspension is subsequently combined with further components of the consumable.

[0199] Controlling the melting point by controlling the fat composition The process of cooking meat is essential to the experience of using and enjoying it. An important characteristic is that as the meat is heated, fat is released from the meat, which lubricates the cooking surface. This increases heat transfer and is a component of the visual, auditory, and olfactory experience of cooking meat. The amount of fat released during cooking compared to the amount retained varies depending on the cooking temperature, and the amount of fat released from the meat varies. It contributes to the visual, auditory, and olfactory experience of cooking.

[0200] The composition and proportion of fatty acids in triglycerides and phospholipids are determined by the proportion of phospholipid heads and Together, they contribute to the creation of individual flavor profiles in cooked meat. For example, the increase in fat Significant levels of phosphatidylcholine and phosphatidylethanolamine are stronger It provides a beef flavor. As discussed above, the flavor of meat imitations comes from various oils and meat The imitation can be modified by changing the proportion and type of phospholipids that make up the imitation. For example. Furthermore, the flavor of cooked meat imitations can be altered by changing the amount of phospholipids, sterols, and lipids. It can be controlled by (for example, 0.2-1% wt / wt). In one embodiment, it is cooked The flavor of meat imitations can be controlled by changing the proportions of various phospholipid heads.

[0201] In some embodiments, the phospholipid comprises a plurality of fatty acids, glycerol, and polar groups. It contains amphibolecular molecules. For example, fatty acids, phospholipids, polar groups and phospholipids associated with phospholipids. For examples of terols, see Section IIIC. For examples of useful vegetable oils, see Section III. See C.

[0202] In various cuts of meat, fat has structurally important properties in bacon, and is also important in Wagyu beef. Beef marbled fat exhibits different characteristics in terms of its softening and melting behavior.

[0203] By controlling the melting point of the adipose tissue imitation in consumables, the cooking experience for various types of meat can be recreated. It is possible to express this. For example, fat made from fats having a melting point of 23°C to 27°C. Fat tissue imitation products may have a melting point similar to that of fat tissue obtained from Wagyu beef; 35°C to 40°C. Adipose tissue imitation made from fat with a melting point is similar to adipose tissue obtained from ordinary ground beef. It may have a melting point; adipose tissue imitation made from fat having a melting point of 36°C to 45°C, It may have a melting point similar to the adipose tissue obtained from bacon. The adipose tissue imitation may melt during cooking. Fat tissue imitation allows for the proportion of fat released and the proportion of fat retained, similar to meat, for example. It can be manufactured to have similar fat properties to ground beef and incorporated into consumables.

[0204] In some embodiments, the fat release temperature of the fat imitation is determined by triacylglycerides and phosphorus. Controlled by mixing vegetable oils in various proportions that contain lipids (e.g., lecithin). It is possible. The melting point of fat is governed by the chemical composition of fatty acids. Generally, saturated fatty acids (e.g.) For example, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0, C Fat containing 22:0) can be stored at refrigeration temperatures (e.g., about 1°C to about 5°C) and room temperature (e.g., about 2°C). It is solid at temperatures between 0°C and 25°C. In adipose tissue imitation products, the temperature at which fat is released during cooking is controlled. By doing so, during refrigeration (for example, at approximately 1.5°C to approximately 4°C) and at ambient temperature (for example, approximately The hardness of adipose tissue imitation can be controlled at 20°C to 25°C. Monounsaturated fatty acids (e.g., Fats containing C16:1 or C18:1 are generally solid at refrigeration temperatures and liquid at room temperature. It is a body. Polyunsaturated fatty acids (e.g., C18:2, C18:3, C20:5 or C2 Fat containing 2:6) is generally liquid at refrigeration and room temperature. For example, virgin fat. Coconut oil melts at approximately 24°C, while hydrogenated coconut oil melts at 36-40°C.

[0205] For example, it contains triglycerides and phospholipids that are liquid at room temperature (approximately 20°C to 25°C). The adipose tissue imitation contains triglycerides and phospholipids that are solid at refrigerated temperatures. It becomes softer than adipose tissue imitation.

[0206] Adipose tissue imitation materials are liquid at both refrigerated and ambient room temperature, and are supplied from one or more sources. The oils obtained from them (for example, canola oil, sunflower oil and / or hazelnut oil) ) may contain. In one embodiment, the adipose tissue imitation is solid at refrigeration temperature, but at room temperature Oils obtained from one or more liquid sources (e.g., olive oil, coconut oil) It contains (and / or rice bran oil). In one embodiment, the adipose tissue imitation is solid at room temperature. However, oil obtained from one or more sources that is liquid at oral body temperature (approximately 37°C) (for example For example, it contains palm kernel oil, coconut oil and / or cocoa butter. In one embodiment, Fat tissue imitation materials are solid at oral temperature (approximately 37°C) and are derived from one or more sources. It contains the obtained oil (for example, oil obtained from mango butter).

[0207] In one embodiment, the adipose tissue imitation has a high proportion of saturated fatty acids, triglycerides It contains phospholipids and higher proportions of monounsaturated and polyunsaturated triglycerides and lipids. It is harder than adipose tissue imitation containing certain substances. For example, adipose tissue imitation containing sunflower oil. It is softer than adipose tissue imitation containing cocoa butter. Adipose tissue imitation is 70% , along with 80% or 90% v / v sunflower or cocoa butter, 0%, 0.18% Cocoa butter can be formed using 1.6% or 2.4% wt / v Rubisco. Each adipose tissue imitation containing - was harder than the imitation formed using sunflower oil. That's it.

[0208] In one embodiment, a stable emulsion of mung bean 8S protein with sunflower oil is used. The adipose tissue imitation produced is made from stable mung bean 8S protein and cocoa butter. Softer than adipose tissue imitation manufactured as an emulsion. 70%, 80%, or 9% 0% v / v sunflower or cocoa butter along with 2%, 1%, or 0.5% wt / v moon A mimic of adipose tissue formed using Gumame 8S protein. It contained cocoa butter. Each adipose tissue imitation was harder than the imitation formed using sunflower oil.

[0209] In one embodiment, a stable emulsion of mung bean 8S protein with canola oil and The adipose tissue imitation produced is made from mung bean 8S protein, coconut, cocoa, Adipose tissue model prepared as a stable emulsion with an equal mixture of olive and coconut oil. Softer than artificial tissue. Fatty tissue imitation is 50%, 70%, or 90% v / v sunflower. Alternatively, it is formed using 1.4% wt / v mung bean 8S protein along with an oil mixture. It is possible. Each adipose tissue imitation containing an oil mixture was formed using sunflower oil. It was harder than the counterfeit versions.

[0210] In one embodiment, it is produced as a stable emulsion of soy protein with sunflower oil. The prepared adipose tissue imitation is a stable emulsion of soy protein and cocoa butter. It is softer than adipose tissue imitation manufactured by [unspecified method]. Adipose tissue imitation is available in 50%, 70%, and 80% concentrations. Along with a mixture of % or 90% v / v sunflower or oil, 0.6%, 1.6%, or It was formed using 2.6% wt / v soybeans. Each fatty acid compound contained an oil mixture. The woven imitation was harder than the imitation formed using sunflower oil.

[0211] In some embodiments, 0% cocoa butter is used along with 70%, 80%, and 90% v / v cocoa butter. Adipose tissue imitation products containing 0.18%, 1.6%, and 2.4% wt / v Rubisco are It is solid at room temperature but melts at approximately oral temperature. In some embodiments, 50%, 70% , along with 0.6%, 1.6%, and 2.6% w / v cocoa butter, 80% and 90% v / v cocoa butter. The t / v soybean-containing adipose tissue imitation is solid at room temperature but melts at approximately oral temperature. In some embodiments, coconut, cocoa, and o Along with an equal mixture of leaf and coconut oil, 1.4% wt / v mung bean 8S protein. Adipose tissue imitation containing is solid at room temperature but melts at approximately oral temperature. In one embodiment, In some embodiments, the melting temperature of the adipose tissue imitation is the same as that of beef fat. The product contains an oil having a 1:1 ratio of saturated to unsaturated fatty acids. In some embodiments, The adipose tissue imitation contains equal amounts of cocoa and mango butter. In some embodiments, The adipose tissue imitation is made from equal parts coconut oil, cocoa butter, olive oil, and palm oil. It contains oil.

[0212] In one embodiment, a fatty tissue imitation containing triglycerides and phospholipids is used to mimic the smell of beef. The proportion of fatty acids is similar to that seen in (C14:0 0-5% wt / wt, C16:0 0~25%, C18:0 0~20%, C18:1 0~60%, C18:2 0~25 Contains %, C18:3, 0-5%, C20:4, 0-2%, and C20:6, 0-2%. For example, adipose tissue imitation is made from equal proportions of olive oil, cocoa butter, and coconut oil. It may contain oil and mango butter. In another example, the fatty tissue imitation is made from equal proportions of olives. May contain oils and rice bran oil.

[0213] In one embodiment, the melting temperature of the adipose tissue imitation is the same as that of Japanese Wagyu beef fat. In application, the fat imitation is an oil having saturated to unsaturated fatty acids in a 1:2 ratio (for example) For example, it includes (1 part coconut oil to 2 parts sunflower oil). In some embodiments, fat The tissue imitation contains equal parts olive oil, rice bran oil, cocoa butter, and mango butter. To possess.

[0214] I. Connective Tissue Imitations Animal connective tissue provides important texture characteristics, which are a key component of the experience of eating meat. Therefore, the present invention is a non-animal source that reiterates the essential features of animal connective tissue. The present invention provides compositions derived from important texture characteristics and visual properties of animal connective tissue. To reiterate the key points of the sensory characteristics, we further provide meat substitute products containing compositions derived from non-animal sources. Such compositions are referred to herein as “connective tissue imitations.” In terms of application, connective tissue imitations and / or meat substitute products containing connective tissue imitations are partially It originates from animal sources.

[0215] Animal connective tissue can generally be divided into fascial and cartilaginous tissues. Fascial tissues are, It is highly fibrous, resistant to stretching (has a high elastic modulus), and has a high protein content. It has a high fat content, a moderate water content (about 50%), and low to no fat. It has a polysaccharide content. Therefore, the present invention reiterates the key features of fascial tissue. We provide connective tissue imitation products. In some embodiments, the connective tissue imitation product is approximately 50% by total weight. It contains approximately 50% protein by liquid weight and has low levels of fat and polysaccharide components.

[0216] The fibrous structure of fascial connective tissue is largely composed of collagen fibers. The fibers have been observed to be in the form of cords or tapes, with a width of 1 to 20 microns. These fibers are tightly packed thin collagen microfibers, 30-100 nanometers in diameter. It consists of thicknesses. These fine fibers are also individual fibers that can be 200 nanometers thick. It is associated with an elastic, reticulated fibrous network having [certain properties].

[0217] In one embodiment, the fascial connective tissue imitation may be fibrous or fiber-based, consisting of protein. It consists of various structures. In some embodiments, the protein content is derived from non-animal sources (e.g., plants). Sources originating from algae, bacteria, or fungi (see, for example, sections IIIA and B). In some embodiments, the isolated protein is 50% of the protein content by weight, 60% It accounts for %, 70%, 80%, or 90% or more. In some embodiments, multiple The isolated proteins are separately isolated and purified to constitute the total protein content.

[0218] In fascial connective tissue, the prolamin family of proteins functions individually or in combination. The combination is highly abundant, and the overall amino acid composition is similar to that of collagen (proli (High fraction of nitrate and alanine), suitable for processing on film, and for protein components To demonstrate eligibility. In some embodiments, prolamin family proteins are used. In (found in corn), hordein derived from barley, gliadin derived from wheat , sekarin and extensin derived from rye, and cafilin or charasum derived from sorghum Selected from the group consisting of avenin derived from ginseng. In some embodiments, one or more mono The separated and purified protein is zein. In some embodiments, physicochemical and To complement prolamins in order to achieve the target specifications of nutritional properties, other proteins Quality may be used. Any major seed storage protein, animal-derived or recombinant collagen. Or extensin (cell wall, for example, Arabidopsis thal iana) contains abundant hydroxyproline-rich glycoproteins, and its monomers are "collagen" See the list in sections IIIA and B, which includes "rod-like" flexible molecules. That.

[0219] The protein is freeze-dried, ground, and mixed with one or more other ingredients (e.g., co It can be combined with wheat gluten, bamboo fiber or other fibers, or soy protein isolates. .

[0220] Fibrous or fibrous structures can be formed by extrusion. In some embodiments, The extrusion process is carried out using a Leistritz Nano-16 twin-screw co-rotating extruder (Amer ican Leistritz Extruder Corp. USA, Sommer This is carried out using Ville (NJ). Mechanical properties, degree of swelling, and moisture content of the fibers are assessed. To optimize the volume, active heating and cooling of the barrel section are used. For example, hard knots To produce synthetic tissue imitation products, the moisture content can be adjusted to approximately 50%. Protein feed The liquid is added separately. The protein is fed by a volumetric plunger feeder. The supplied liquid is added to the barrel through a high-pressure liquid injection system. In some examples, The output parameters were a screw speed of 200 rpm, a product temperature of 120°C on the die, and a feed rate of 120°C. The die velocity was 2.3 g / min and the water flow velocity was 0.7 g / min. The product temperature is measured using a thermocouple.

[0221] A fibrous or fibrous structure is a combination of fine fibers and multiple fine fibers used to manufacture a fibrous structure. It can be formed by extrusion through a die. In some embodiments, the dimensions and composition of the fibers To produce mixed fibrous tissue imitation with precise control over a wide range, 10-300 mm A die incorporating multiple different orifice sizes within the range of the chromium may be used. Fibers of different sizes may be incorporated into the composition to control its properties.

[0222] Electrospinning is used to produce fibers in the range of <1 to 10 microns. To obtain. In some embodiments, to produce fibers with a diameter range of <1 to 10 microns, Lectrospinning is used. For example, mung containing 400 mM sodium chloride. Mameglobulin concentrated solution (140 mg / ml) is poly(vinyl alcohol) or poly (Ethylene oxide) (9% w / v) solution mixed with 22.5 mg / ml mungma A mixed solution containing meglobulin and 6.75% w / v of each polymer can be obtained. The obtained solution is then transferred from a 5 ml syringe to a Teflon tube using a syringe pump. And slowly (for example, 3 μl / min) through a 21 gauge needle with a blunt tip. It is pumped by a pump. The needle is supplied by a high-voltage power supply (e.g., Spellman CZE). It is connected to a positive electrode (30kV) and fixed 20-30cm from the collection electrode. The collection electrode is A It is an aluminum drum (approximately 12 cm long and 5 cm in diameter) wrapped in aluminum foil. The drum is a spindle rotated by an IKA RW20 motor at approximately 600 rpm. It is mounted on the ground terminal of the high-voltage power supply. Portein / polymer fibers are deposited onto the foil, and electrospinning is completed. Afterward, it is recovered from the foil and added to the tissue imitation.

[0223] The dimensions and composition of the fibers produced by the method of the present invention are such that the texture and taste of the tissue imitation are as follows: Effective against charring and mechanical properties. 1 to 50% in the range of <1 to 10 microns A tissue containing fibers between and between 10 and 50% of the fibers in the range of 10 to 300 microns. In terms of taste, texture, and mechanical properties, it most closely resembles animal connective tissue.

[0224] Cartilage-type tissue is macroscopically homogeneous, resistant to compression, and has a high water content (up to 80%). %), low protein (collagen) content and high polysaccharide (proteoglycan) content (each It contains approximately 10%. Compositionally, cartilage-type connective tissue imitation materials more closely resemble "meat" connective tissue. It is similar to a fascial tissue imitation, each with its own relative proportions adjusted to mimic. During this time, the moisture content can be adjusted to approximately 60% to produce a soft connective tissue imitation. .

[0225] The method of forming cartilaginous connective tissue is similar to that of fascial connective tissue, but isotropic nonfibrillary. A method for producing a fibrous gel is preferred.

[0226] Connective tissue imitation involves isolating and purifying one or more proteins and one Alternatively, it can be produced by precipitating multiple proteins, and precipitation is a binding process. It possesses one or more proteins that form a physical structure that approximates the physical organization of tissue. Precipitation is the process of solubilizing one or more proteins in the first solution. This may include pushing the first solution into the second solution, where one or more tans Proteins are insoluble in the second solution and can be extruded by one or more proteins. It induces precipitation.

[0227] In some embodiments, some or all of the components of the consumables are gels (e.g., proteins). It is suspended in a hydrogel. In various embodiments, the gel is a hydrogel, an organogel, or It may be a xerogel. The gel uses a polysaccharide or protein-based agent. Viscosity can be increased. For example, the viscosity of a consumable structure or the gel forming the structure can be increased. Therefore, starch (fecula), arrowroot, cornstarch, potato starch, ja Potato starch, sago, tapioca, arginine, guar gum, locust bean gum, ki Santan gum, collagen, egg white, fercerelan, gelatin, agar, carrageenan, ce Lurose, methylcellulose, hydroxymethylcellulose, acadia gum um), konjac, starch, pectin, amylopectin or legumes, grains, nuts Fungal proteins, derived from seeds, leaves, algae, bacteria, etc., either alone or in combination. It can be used in conjunction with other materials. To form the structure or structure of consumables, covalent bonds between proteins Enzymes that catalyze reactions leading to crosslinking can also be used alone or in combination. For example, forming the structure or form of a consumable by crosslinking component proteins. Therefore, transglutaminase, tyrosinase, lysyl oxidase or other Minoxidase (for example, Pichia pastoris lysyloxidase) Ze (PPLO)) can be used alone or in combination. In some embodiments, Multiple gels having various components are combined to form consumables. For example, plant Gels containing derived proteins can be associated with gels containing plant-derived fats. In one embodiment, the protein fibers or strings are oriented parallel to each other, and then It is then held in place by the application of a gel containing plant-based fats.

[0228] The composition of the present invention can be used in the following ways in the art: stir-frying, baking, microwave heating. Heating, such as in a forced air system or in an air tunnel, causes expansion. It can expand.

[0229] In some embodiments, multiple gels having different components are combined to form a consumable product. For example, a gel containing plant-derived protein is related to a gel containing plant-derived fat. They can be linked together. In some embodiments, protein fibers or strings are flat against each other. Oriented in rows, and then held in place by the application of a gel containing plant-based fats. ru.

[0230] J. Omission from composition Consumables can be assembled from specific components that can be isolated and purified, so specific It is possible to manufacture consumables that do not contain the ingredients. This makes it possible in some cases This may contain ingredients that are undesirable to consumers (for example, ingredients that may be omitted if they cause allergies in some people). This makes it possible to manufacture consumables that lack the proteins or additives that cause [the condition]. In this configuration, the consumables contain no animal products whatsoever. In some embodiments, the consumables are wheat It contains no gluten or less than 1% gluten. In some embodiments, the consumables are methyl It contains no cellulose at all. In some embodiments, the consumables contain no carrageenan at all. No. In some embodiments, the consumables do not contain any caramel color. Therefore, the consumables do not contain any konjac powder. In some embodiments, the consumables are made from ara It contains absolutely no gum arabic (also known as acacia gum). No. In some embodiments, the consumables do not contain any wheat gluten. In this state, the consumables do not contain any soy protein isolate. In some embodiments, The consumables contain no tofu at all. In some embodiments, the consumables contain less than 5% carbohydrates. It contains. In some embodiments, the consumables contain less than 1% cellulose. In some embodiments, the consumables contain less than 5% cellulose. The product contains less than 5% insoluble carbohydrates. In some embodiments, the consumables contain less than 1%. It contains insoluble carbohydrates. In some embodiments, the consumables do not contain any artificial colors. In some embodiments, the consumables contain no artificial flavors at all.

[0231] In some embodiments, the consumables include one or more of the following characteristics: animal-made No ingredients; no methylcellulose; no carrageenan; no konjac powder; no gum arabic. wheat gluten less than 1%; wheat gluten-free; tofu-free; approximately 5% carbohydrates; Less than 5% cellulose; less than 5% insoluble carbohydrates; less than 1% insoluble carbohydrates; caramel Red, paprika, cinnamon, beet color, carrot oil, tomato lycopene extract, raspberry Lee powder, carmine, cochineal extract, annatto, turmeric, saffron, FD& C Red #3, Yellow #5, Yellow #6, Green #3, Blue #2, Blue #1, Purple #1, FD&C Food coloring such as Red 40 - Allura Red AC and / or E129 (red) No artificial flavors; and / or no artificial flavors. In some embodiments, consumables are soybean tannins. It contains no protein isolates at all. In other embodiments, the consumables are soy protein or It contains absolutely no protein concentrates.

[0232] In some embodiments, the muscle tissue imitation is less than 10%, less than 5%, less than 1%, or 0%. It further contains less than 1% wheat gluten. In some embodiments, the muscle tissue imitation is It contains absolutely no wheat gluten.

[0233] IV. Combinations of ingredients A. Meat imitations Meat substitutes (or meat imitations) may include the compositions described herein. For example, meat imitations include muscle imitations, adipose tissue imitations and connective tissue imitations (or May include (partial combinations): muscle imitations, adipose tissue imitations and / or connective tissue imitations. It can be assembled in a manner that approximates the physical organization of meat. In some embodiments, imitations Binding agents such as coacervates are used to help them bond to each other.

[0234] The percentages of various components can also be controlled. For example, muscle, adipose tissue, connective tissue. And non-animal-based substitutes for blood components are used to best approximate the appearance and impression of meat. They can be combined in various proportions and physical arrangements. Various components can be combined between pieces of consumables. The components may be arranged to ensure consistency. The components ensure that no waste is generated from the consumables. It can be arranged in such a way. For example, traditional cuts of meat may have parts that are not usually eaten. However, meat imitations do not contain these inedible parts (e.g., bone, cartilage, connective tissue or general malaise). The meat can be improved by not including other ingredients (such as tendons). Through good, all products that are manufactured or transported become available for consumption, and this Therefore, waste and transportation costs are reduced. Alternatively, meat imitations mimic the experience of meat consumption. In order to mimic the original, it may include parts that are not edible. These parts include bone, cartilage, and knots. Synthetic tissue or other materials commonly referred to as streaks or components thereof are included to imitate them. Materials can be listed. In some embodiments, consumables are designed to serve a secondary function. It may contain imitation inedible parts of meat products. For example, imitation bones The design distributes heat during cooking, allowing consumables to be cooked more quickly or more evenly than meat. It can be measured. In other embodiments, the imitation bone is also maintained at a constant temperature during transport of the consumables. It may help to maintain it. In other embodiments, the imitation inedible part is biodegradable ( For example, it could be biodegradable plastic.

[0235] In some embodiments, the meat substitute composition contains between 10-30% protein and 5-80% The composition contains water between 5% and fat between 5% and 70%, and consists of one or more isolated and purified components. Contains processed protein. Such meat substitutes contain no animal protein at all. In some embodiments, the meat substitute composition contains transglutaminase.

[0236] In some embodiments, the meat substitute product is a muscle imitation, adipose tissue imitation, and connective tissue imitation. Including the materials, muscle imitation accounts for 40-90% of the product by weight, and adipose tissue imitation accounts for... Connective tissue imitation accounts for 1-60% of the product, and by weight, it accounts for 1-30% of the product.

[0237] In some embodiments, the meat substitute product contains 60-90% water and 5-30% protein. It also contains 1-20% fat, and the protein content is one or more isolated and purified proteins. Contains plant protein.

[0238] In some embodiments, the consumables contain ingredients that replicate the components of meat. Main components of meat It is usually skeletal muscle. Skeletal muscle is usually about 75 percent water and 19 percent Protein, 2.5 percent intramuscular fat, 1.2 percent carbohydrates and 2 It consists of 0.3 percent other soluble non-protein substances. These are organic acids, sulfurized It contains nitrogen compounds such as compounds, amino acids and nucleotides, and inorganic substances such as minerals. Therefore, some embodiments of the present invention reproduce an approximation of this composition of the consumables. Provides, for example, in some embodiments, the consumables consist of approximately 75% water and 19% tan. Protein, 2.5% fat, 1.2% carbohydrates, and 2.3% other soluble substances. It is a plant-based meat imitation containing non-protein substances. In some embodiments, the consumables are Water between 60-90%, protein between 10-30%, fat between 1-20%, and 0.1-5% Plant-based, containing carbohydrates and 1-10 percent of other soluble non-protein substances. It is a meat imitation. In some embodiments, the consumables are water between 60 and 90%, and 5 to 10% Protein, 1-20% fat, 0.1-5% carbohydrates, and 1-10% It is a plant-based meat substitute containing other soluble non-protein substances. In some embodiments, The consumables consist of 0-50% water, 5-30% protein, and 20-80% fat. 0.1-5% carbohydrates and 1-10% other soluble non-protein substances It contains plant-based meat substitutes.

[0239] In some embodiments, the meat imitation contains heme-containing protein between 0.01% and 5% by weight. It contains nitrite. In some embodiments, the imitation contains nitrite between 0.01% and 5% by weight. It contains guhemoglobin. Some meats also contain myoglobin, a heme-containing protein. And this is the main cause of the red color and iron content in some meats. The quality of meat can vary, and meat imitations are made to approximate the natural variations in meat. It is understood that it can be produced. In embodiments including heme-containing protein and optional flavors, k-carrageenan is used to absorb some of the liquid and heme solution that contribute to the flavor. This can be done, and the ground tissue will not become excessively moist. When adding the flavor heme mixture solution, k-carrageenan powder is used for tissue mixing to ensure homogeneity in the final ground product. It is uniformly dispersed within the material.

[0240] If the protein is supplied as a solution, freeze-drying is used to concentrate the protein. It is acknowledged that water removal techniques such as drying or spray drying may be used at the discretion of the user. Next, the protein is reconstituted with a certain amount of liquid to prevent the ground tissue from becoming too wet. It is possible.

[0241] Furthermore, in some cases, the present invention does not contain these components in unnatural percentages. To provide an improved meat imitation. The concentration of heme-containing protein enhances the flavor and aroma of meat. It is an important determinant. Therefore, for example, meat imitations have a higher hematin content than regular beef. It may have a protein content. For example, a meat imitation product that has a higher fat content than the normal average may be manufactured. It can be manufactured. The percentage of these components also increases other desirable properties. It may be changed.

[0242] In some cases, when meat imitations are cooked, the percentage of ingredients is the same as when cooked. It is designed to be similar to cooked meat. In this way, in some embodiments, uncooked meat Consumables have different percentages of components than uncooked meat, but when cooked, Consumables are similar to cooked meat. For example, raw meat has a higher moisture content than usual. A meat imitation product having the following characteristics can be manufactured, but when cooked in a microwave, the resulting product , non-starch polysaccharides such as arabinoxylan and cellulose, and resistant starch, resistant Dextrin, inulin, lignin, wax, chitin, pectin, beta-glucan And numerous other plants, such as oligosaccharide percentages with similar components to meat cooked over fire. It contains physical components.

[0243] In some embodiments, the consumable is a meat imitation having a lower moisture content than usual for meat. In some embodiments, the present invention hydrates the meat imitation so that the meat imitation has the same water content as real meat. The present invention provides a method for having a certain moisture content. For example, a low moisture content in meat, for example, Meat imitations containing 1%, 10%, 20%, 30%, 40%, or 50% water are approximately It can be hydrated with 75% water. Once hydrated, in some embodiments, it is then used to create a meat mold. The products are prepared for human consumption.

[0244] The consumables may contain protein components. In some embodiments, the protein content of the consumables This is 10%, 20%, 30%, or 40%. In some embodiments, the tamper of the consumables The protein content is the same as that of meat. In some embodiments, the protein content in the consumables is the same as that of meat. It is more expensive than other things. In some embodiments, the consumables have less protein than meat.

[0245] The proteins in consumables can come from various combinations of sources. Non-animal sources are consumables. May provide some or all of the protein in the product. Non-animal sources include vegetables and carrots. Aerial parts and the genus Miscanthus, seaweed, fruits, nuts, grains, algae, bacteria Alternatively, non-food biomass such as fungi can be cited. For example, see Sections IIIA and B. See below. Proteins are isolated or concentrated from one or more of these sources. It is possible. In some embodiments, the consumables are proteins obtained solely from non-animal sources. It contains meat imitation products.

[0246] In some embodiments, the protein is formed into asymmetrical fibers for incorporation into the consumables. In some embodiments, these fibers replicate muscle fibers. In some embodiments, Proteins are spun fibers. Therefore, the present invention relates to asymmetrical fibers or spun fibers. The present invention provides a method for producing protein fiber. In some embodiments, the consumables are used for human nutrition. This refers to a protein that contains all the amino acids found in essential proteins (singular or It contains (multiple). In some embodiments, the protein added to the consumables contains amino acids It will be resupplied.

[0247] Physical organization can be a determinant of how meat substitutes respond to cooking. For example, the wind of meat The taste is modified by the size of the particles. Minced meat that has been finely ground into a paste is finely ground during cooking. It offers a different flavor from coarsely ground beef. The relative size of the individual tissue imitations and The ability to control orientation modifies the flavor and aroma profile of consumables during cooking. This makes it possible. For example, muscle tissue imitations and adipose tissue imitations are cooked independently. It provides different flavor profiles when used individually or when mixed. Different tissue imitations are mixed. Further changes in the flavor profile can be observed based on the mixing method.

[0248] The physical organization of meat substitute products is muscle, fat and / or as described herein. Manipulated by controlling the localization, organization, assembly, or orientation of connective tissue imitation materials. To obtain. In some embodiments, the product is such that the imitation described herein is like meat. They are designed in such a way that they are related to each other. In some embodiments, consumables are used in cooking Later, the counterfeits described herein may be associated with one another, such as in cooked meat. It is designed to be so.

[0249] The characteristic flavor and aromatic components of meat are largely derived from substrates found in both plants and meat. It is produced during the cooking process by chemical reactions involving amino acids, fats, and sugars. Therefore, in some embodiments, the consumables are similar to the meat during or after cooking. It is then tested. In some embodiments, in order to create an olfactory map of cooked meat, Grade, human evaluation, olfactometer reading or GCMS measurement or These combinations are used. Similarly, olfactory maps of consumables, such as meat imitations, are created. These maps assess how similar cooked consumables are to meat. It can be compared for the purpose of olfactory management of consumables during or after cooking. In some embodiments, the olfactory management of consumables during or after cooking The topping is similar to or indistinguishable from that of cooked meat or meat being cooked. In some embodiments, the difference is small enough to fall below the detection threshold of human perception.

[0250] In some embodiments, individual tissue imitation materials are layers, sheets, blocks in a specified position and orientation. It is assembled into a rack and strings.

[0251] In some embodiments, the counterfeit is set to less than 1 / 2 inch (for example, 1 / 4 inch). The process involves passing the meat through a plate of a meat grinder, which has holes. Dadder offers several functions to reduce particle size, which is similar to what is usually done with ground meat. Then, the material is further mixed or worked and formed and provided to the cylindrical part. Assembly, During grinding and shaping, the imitation material structure is kept at a low temperature (e.g., 4-15°C) to promote microbial growth. It controls the flavor response, maintains the fat in a solid state, and as a result, the grinding process It is important that individual pieces of fat are maintained throughout.

[0252] Before grinding, the imitation material is usually broken down to a predetermined particle size in several ways. For example, In some embodiments, individual tissue imitation specimens have a diameter of less than 1 cm or less than 5 mm. It can be formed into small pieces and then combined with other tissue imitation materials. Adipose tissue is approximately 3- It can be crushed into particles as small as 7mm. This affects the appearance of the final ingredient and the leakage of fat during cooking. It is important for both behaviors. This range of size is the natural distribution of fat spots in the final raw product. To enable appearance. If the fatty tissue is too small (e.g., less than 2 mm), cooked In some cases, the amount of fat leaked from the product may be insufficient.

[0253] The soft connective tissue imitation was broken down into small pieces about 1-3 mm long with irregular ends. Obtain. If the small pieces are too large (for example, more than about 4mm), the texture of the final product will be... Marimo can be something like a bead.

[0254] Each consists of small pieces of amorphous or long noodle-like texture imitation, with a chewy texture. The tissue or noodle tissue imitation and raw tissue imitation are made by hand into small pieces with a diameter of approximately 1-3 cm. It can be destroyed. By achieving particles in this size range, the final ground material This allows for proper mixing and suitable homogeneity.

[0255] In some embodiments, the hard connective tissue imitation is available in three levels (e.g., coarse, medium, and It can be engraved in fine detail. Engraving in three levels results in more unevenness than a single-level engraving process. It provides uniformity and makes the texture of the final product more similar to ground beef.

[0256] In gluten-containing formulations, a further function of the food grinder is to produce gluten. The goal is to develop a gluten network of aligned gluten molecules. Regarding formulations containing gluten, it is possible to minimize the interaction between fat and gluten network. This is important. This involves pre-cooling the fat imitation and the ground tissue imitation, and then combining them. This is done by minimizing the amount of processing required after the fat has been added. If too much gluten is produced in fat substitutes, it can break down or shorten the gluten network. "

[0257] Finally, regarding gluten-containing ingredients, the patties should be left at room temperature for 30 minutes before cooking. It is permitted to keep it at 4°C overnight. This allows the gluten network to loosen, and the whole This allows for time to provide a good texture to the target.

[0258] In some embodiments, connective tissue imitation materials are incorporated into a protein solution, and then muscle A tissue imitation is formed.

[0259] In some embodiments, the connective tissue imitation is directly incorporated into the emulsion, and then the lipid A fat tissue imitation is formed.

[0260] In some embodiments, fat is used to replicate the effect of marbled or striped bacon. Fat tissue imitation is added to muscle tissue imitation in strands and sheets.

[0261] Mixed meat tissue imitations can enhance the sense of flavor, and such flavors are not limited to that alone. It smells like fruit / green bean / metal, like nuts / green, Peanut butter / moldy, raw potato / roasted / earthy, vinegary, spaghetti Sweet / caramel / almond, creamy, sweet, fruity / light Beer with a musty smell / nutty / coumarin / licorice / walnuts / bread, coconut / Woody / sweet, piercing / nauseating, minty or toasty It contains several aromatic compounds associated with caramel aroma.

[0262] In some embodiments, the mixed meat imitation is 2-pentyl-furan;4-methylthiazole Ethylpyrazine; 2,3-dimethylpyrazine, acetate; 5-methyl-2-furancarboic acid Cialdehyde; Butyrolactone; 2,5-Dimethyl-3-(3-methylbutyl)pyrazine ;2-cyclopenten-1-one, 2-hydroxy-3-methyl;3-acetyl-1h- Pyrroline; Pantolactone; 1-Methyl-1(H)-pyrrole-2-2-carboxaldehyde do;caprolactam;2,3-dihydro-3,5-dihydroxy-6-methyl-4(H) -Increases the presence of volatile odor substances such as pyran-4-one. In some embodiments, It's not limited to just that, but it's like gasoline, petroleum, sour / rotten / fishy, ​​tasteless No / woody / yogurt, fat / honey / citrus, tingling / sweet / caramel Undesirable flavors, including nutty / roasted green aromas, are present in individual combinations. It forms only in woven imitation products, but does not accumulate in mixed meat imitation products. In some embodiments, These tissue imitations are not limited to these, but include nonane, 2,6-dimethyl, 3-methyl3 -Hexene; Pyridine; Acetoin; Octanal; 1-Hydroxy-2-Propanone; and / or increase the presence of volatile odor substances, including ethenylpyrazine. Some implementations Morphologically, the level at which all of the above compounds accumulate during cooking is large in tissue imitation units. It varies depending on the size and how they are mixed (coarse, fine, or blended). ru.

[0263] In some embodiments, the mixed meat tissue imitation enhances the sense of flavor, and is not limited to that. It's like fruit / green bean / metallic, like nuts / green , peanut butter / moldy, raw potato / roasted / earthy, vinegary, s Spicy / caramel / almond, creamy, sweet, fruity / flavorful Flat beer, musty / nutty / coumarin / licorice / walnuts / bread, coconut Woody / Sweet, piercing / Nauseating, minty or toasty It contains several aromatic compounds associated with a caramel aroma. In some embodiments, it contains mixed meat Counterfeit products increase the presence of volatile odor substances, and not only that, but also phenylacetal Dehyde, 1-octen-3-one, 2-n-heptylfuran, 2-thiophenecarboxy Aldehydes, 3-thiophenecarboxyaldehyde, butyrolactone, 2-undecene L, methylpyrazine, furfural, 2-decanone, pyrrole, 1-octen-3-ol (E)-2-octenal, decanal, benzaldehyde Hydro, (E)-2-nonenal, pyrazine, 1-hexanol, 1-heptanol, dimethicone Tiltrisulfide, 2-nonanone, 2-pentanone, 2-heptanone, 2,3-butane Dione, heptanal, nonanal, 2-octanone, 1-octanol, 3-ethyl syl Clopentanone, 3-octen-2-one, (E,E)-2,4-heptadienal, ( Z)-2-heptenal, 2-heptanone, 6-methyl-, (Z)-4-heptenal, (E,Z)-2,6-nonadienal, 3-methyl-2-butenal, 2-pentyl- Lan, thiazole, (E,E)-2,4-decadienal, 1-ethyl-5 hexanoate -Methylcyclopentene, (E,E)-2,4-nonadienal, (Z)-2-decena Lu, dihydro-5-pentyl-2(3H)-furanone, trans-3-nonene-2-o (E,E)-3,5-octadiene-2-on, (Z)-2-octen-1-all , 5-ethyldihydro-2(3H)-furanone, 2-butenal, 1-penten-3-ol (E)-2-hexenal, formic acid, heptyl ester, 2-pentyl thiophene (Z)-2-nonenal, 2-hexylthiophene, (E)-2-decenal, 2- Ethyl-5-methylpyrazine, 3-ethyl-2,5-dimethylpyrazine, 2-ethyl -1-Hexanol, Thiofen, 2-Methylfuran, Pyridine, Butanal, 2-E Chil-furan, 3-methyl-butanal, trichloromethane, 2-methyl-butanal, Metacrolein, 2-methyl-propanal, propanal, acetaldehyde, 2-p Ropil-furan, dihydro-5-propyl-2(3H)-furanone, 1,3-hexadie N, 4-decine, pentanal, 1-propanol, heptanoic acid, trimethyl-ethane All, 1-butanol, 1-penten-3-one, dimethyl sulfide, 2-ethyl phosphate Lan, 2-pentylthiophene, 2-propenal, 2-tridecene-1-ol, 4 -Octene, 2-methylthiazole, methylpyrazine, 2-butanone, 2-pentyl- Furan, 2-methyl-propanal, butyrolactone, 3-methyl-butanal, methyl - Thiirane, 2-hexyl-furan, butanal, 2-methyl-butanal, 2-methyl -Fran, furan, octanal, 2-heptenal, 1-octene, heptyl-S formate The tel, 3-pentyl-furan, and 4-penten-2-one are included in some embodiments. The level at which all of the above compounds accumulate during cooking depends on the size of the tissue units and so It depends on how they are mixed (coarse, fine, or blended).

[0264] The production of volatile odor substances occurs when fat, muscle, and connective tissue imitations are in contact with each other. It can be enhanced to have an average size of 5 mm for each individual tissue imitation. When fat, muscle, and connective tissue are closely mixed, the production of volatile odor substances is enhanced. In some embodiments, the fat has an average size of 2 mm for each individual tissue imitation. Furthermore, the production of volatile odor substances is enhanced when muscle and connective tissue are closely mixed. In some embodiments, the fat, muscle, etc., have an average size of 1 mm for each individual tissue imitation. When the connective tissue imitation is closely mixed, the production of volatile odor substances is enhanced. .

[0265] In some embodiments, the meat substitute is optimized for a specific cooking method (microwave). Optimized for cooking in the oven, or optimized for cooking in a stew. (ru).

[0266] In some embodiments, the meat substitute is optimized for dehydration.

[0267] In some embodiments, the meat substitute is rapidly exposed to water after dehydration. It is optimized for rapid rehydration.

[0268] In some embodiments, the meat substitute is used as emergency, camping, or astronaut food. It is optimized for use in this way.

[0269] The methods described herein are meat imitation methods optimized for specific cooking techniques. It can be used to provide meat imitations that have specified cooking characteristics, enabling the manufacture of such products. For example, stews require slow cooking to gelatinize the connective tissue in the meat, The connective tissue imitation gelatinizes more easily, and therefore the stew can be prepared more quickly. A meat imitation that makes this possible can be designed.

[0270] B. Guidelines for cooking meat The consumables may include a composition that indicates the consumables are being cooked or have been cooked. The release of odor-causing substances during processing is an important aspect of meat consumption. In some embodiments, consumables When cooked, it produces an aroma that is recognizable to humans as being characteristic of beef during cooking. It is a meat imitation composed entirely of non-animal products. In some embodiments, the consumables are When cooked, it develops a characteristic flavor in pork, bacon, chicken, lamb, fish, or turkey during cooking. It produces an aroma that is recognizable to humans. In some embodiments, the consumables are used in cooking. It contains odorous substances that are sometimes released or produced by chemical reactions that occur during cooking. These are meat imitations composed primarily or entirely of ingredients derived from non-animal sources. In this embodiment, the consumables are proteins, peptides, amino acids, nucleotides, sugars and When cooking, a mixture of polysaccharides and fats is used to produce odor-causing substances and flavor-producing compounds. It contains these compounds in combinations and spatial arrangements that allow them to undergo chemical reactions. These are meat imitations composed primarily or entirely of ingredients derived from non-animal sources.

[0271] In some embodiments, the consumables absorb volatile or unstable odor substances released during cooking. A meat imitation that has, has, has had, has had, has had, has had, has had, has had, a meat imitation that ru.

[0272] In some embodiments, the indicator is a visual representation that accurately mimics the color transition of the meat product during the cooking process. It is an indicator. The color transition is, for example, from red to brown, pink to white, during the cooking process. It can be yellowish-brown or transparent to opaque in color.

[0273] In some embodiments, the indicator is an olfactory indicator that shows the progress of cooking. In one embodiment, The sensory indicator is one or more volatile odor substances released during cooking.

[0274] In some embodiments, the indicator is one or more isolated, purified iron-containing proteins. Includes. In some embodiments, one or more isolated, purified iron-containing proteins (e.g.) Heme-containing proteins (see Section IIIB) are in a reduced state before cooking. In one embodiment, one or more isolated and purified iron-retaining cells in a reduced or oxidized state are used. Proteins, when in equivalent reduced or oxidized states, are myoglos derived from animal sources. It has a UV-VIS profile similar to that of vin protein. Aquifex eolic acid Aquifex aeolicus hemoglobin has a peak absorbance wavelength of 413 nm; methyl Methylacidiphilum infernorum hemoglobin is It has a peak absorbance wavelength of 412 nm; Glycine max (reghemoglobin) Robin has a peak absorbance wavelength of 415 nm; Hordeum vulgare are and Vigna radiata nonsymbiotic hemoglobin are, respectively, 412n It has a peak absorbance wavelength at m. Bovine (Bos taurus) myoglobin has a peak absorbance wavelength at 415 nm. It has an absorbance wavelength.

[0275] In some embodiments, one or more isolated and purified iron-containing proteins are used. The difference between the absorbance wavelength and the peak absorbance wavelength of myoglobin derived from animal sources is, It is less than 5%.

[0276] The odor-causing substances released during meat cooking are reactants, including fats, proteins, amino acids, Reagents may include peptides, nucleotides, organic acids, sulfur compounds, sugars and other carbohydrates. It is produced by the response. In some embodiments, odor substances that combine during the cooking of meat are consumed. They are identified within the product, located close to each other, and as a result, odor-causing substances combine during the cooking of consumables. They combine. Therefore, in some embodiments, the characteristic flavor and aroma components are derived from plants and This is produced during the cooking process by chemical reactions involving amino acids, fats, and sugars found in meat. It is produced. Therefore, in some embodiments, the characteristic flavor and aroma components are derived from plants and One or more amino acids, fats, peptides, nucleotides, organic acids found in meat, During the cooking process, a chemical reaction involving sulfur compounds, sugars, and other carbohydrates occurs. Almost all of them are manufactured.

[0277] Several reactions that produce odor-causing substances released during meat cooking involve iron, particularly myoglo. It can be catalyzed by heme iron in the bottle. Therefore, in some embodiments, a characteristic flavor is And some of the flavoring components are released during the cooking process through a chemical reaction catalyzed by iron. It is manufactured. In some embodiments, some of the characteristic flavor and aroma components are derived from heme. It is produced during the cooking process by a catalyzed chemical reaction. In some embodiments, Some of the characteristic flavor and aroma components are catalyzed by heme iron in leghemoglobin. It is produced during the cooking process by a chemical reaction. In some embodiments, a characteristic wind Some of the flavor and aroma components are produced through chemical reactions catalyzed by heme iron in heme proteins. It is produced during the cooking process. For example, heme protein (e.g., acifex) Aquifex aeolicusm, Methylasydiphyllum infernorum (M Ethylacidiphilum infernorum), Glycine max, Hordeum u Lugare (derived from Hordeum vulgare or Vigna radiata) is GC- When analyzed by MS, any subset of the three components is cysteine ​​and This study presents significantly different profiles of volatile odorants when heated in the presence of glucose. Provided. The volatile flavor components that are increased under these conditions are not limited to these, Furan, acetone, thiazole, furfural, benzaldehyde, 2-pyridinecarbone Cialdehyde, 5-methyl-2-thiophenecarboxyaldehyde, 3-methyl-2-thiophenecarboxyaldehyde Examples include offencarboxyaldehyde, 3-thiophenmethanol, and decanol. Under these conditions, cysteine ​​and glucose, individually or ferrous glucanate, In the presence of iron salts such as these, a sulfurous odor was produced, but the addition of heme protein produced a sulfurous odor. It reduces, and is not limited to, chicken broth, roasted mushrooms, molasses and bread. It was replaced by a different flavor.

[0278] Furthermore, heme proteins (for example, Aquifex aeolicus) Methylacidiphilum infernorum, glycy Glycine max, Hordeum vulgare, or Vigna • Radiata (derived from Vigna radiata) is detected by GC-MS when heated in the presence of ground chicken. Therefore, analysis reveals that certain volatile odor substances are elevated in beef compared to chicken. That's impressive. While not the only volatile flavor components that increase under these conditions, Propanal, butanal, 2-ethyl-furan, heptanal, octanal, tran S-2-(2-pentenyl)furan, (Z)-2-heptenal, (E)-2-octenor Lupirol, 2,4-dodecadienal, 1-octanal, or (Z)-2-decena One example is Ru-2-Undecenaar.

[0279] C. Color index The color of meat is an important part of the experience of cooking and eating it. For example, the cut of beef In its raw state, it is a distinctive red color, which gradually changes to a brown color during cooking. For example, white meats such as chicken or pork have a characteristic pink color when raw. It has a color that gradually changes to a white or brownish color during cooking. The amount of color change depends on the cooking of the beef. The process is shown, and the amount of cooking time and temperature is determined to achieve the desired degree of doneness. Used for the purpose of providing a visual indicator of the cooking progress of non-meat dishes. In some embodiments, the present invention provides a visual indicator of the cooking progress of non-meat dishes. It provides a base meat substitute product. In some embodiments, a visual indicator is color transition during cooking. This is a color indicator that receives the signal. In some embodiments, the color indicator is used to indicate the progression of the meat from raw to cooked. The key points of color transitions as the meat is cut are repeated. In more embodiments, the color indicators The meat substitute product is colored red to indicate its raw state before cooking, and the meat substitute is colored red during the cooking process. The substitute product is transferred to a brown color. In other embodiments, the color indicator is used to indicate the meat substitute product during cooking. Previously, the meat substitute was colored pink to indicate its raw state, and then white or brown during the cooking process. Transition to the desired color.

[0280] The main determinant of the nutritional definition of meat color is the concentration of iron-sparing protein in the meat. In the skeletal muscle components, one of the main iron-storing proteins is myoglobin. Sea urchin, myoglobin content is less than 0.05% in white meat of chicken, compared to old cattle. It changes to 1.5-2.0% in meat. Therefore, in some embodiments, consumables are iron storage It is a meat imitation containing a protein (e.g., heme-containing protein). In some embodiments, Meat imitation products are approximately 0.05%, 0.1%, and 0.2% of the dry weight or gross weight. Approximately 0.3%, approximately 0.4%, approximately 0.5%, approximately 0.6%, approximately 0.7%, approximately 0.8%, approximately 0.9% %, approximately 1%, approximately 1.1%, approximately 1.2%, approximately 1.3%, approximately 1.4%, approximately 1.5%, approximately 1.6% %, approximately 1.7%, approximately 1.8%, approximately 1.9%, approximately 2%, or more than approximately 2% iron-storing protein ( For example, it contains heme-containing proteins. In some cases, iron-sparing proteins are supplied. It is isolated and purified from the source. In other cases, iron-sparing proteins are isolated and purified. It is not purified. In some cases, the source of iron-sparing protein is an animal source. This includes plants, fungi, or genetically modified organisms such as plants, algae, bacteria, or fungi. Which non-animal source is it? In some cases, the iron-sparing protein is myoglobin. In some embodiments, the consumable is a plant-based consumable having added animal myoglobin. It is a meat imitation. For example, a young beef imitation contains about 0.4-1% myoglobin. It may have a consumable. In some embodiments, the consumable is an added leghemoglobin or cyto It is a plant-based meat imitation containing chromium. Therefore, for example, a young beef imitation is, It may contain approximately 0.4-1% leghemoglobin or cytochrome.

[0281] Another example of an iron-storing protein is hemoglobin in the red blood cells of vertebrates, and iron-containing oxygen bonds. There are complementary proteins. Hemoglobin is similar in color to myoglobin. Some implementations In this context, the present invention secures blood from the animal and livestock industries and recycles it to replenish the color of consumables. The present invention provides a method for obtaining blood from a slaughterhouse and obtaining hemoglobin from the blood. It is used to enhance the color of the consumables. In some embodiments, the consumables are hemoglobin It is a plant-based meat imitation containing .

[0282] Further iron-containing proteins are naturally present. In some embodiments, consumables are myog Contains iron-containing protein other than myoglobin. In some embodiments, the consumable is myoglobin. It does not contain. In some embodiments, the consumables do not contain hemoglobin. In terms of form, consumables are meat containing iron-containing proteins other than myoglobin or hemoglobin. It is a counterfeit. For examples of heme-containing proteins, see, for example, Section IIIB and Figure 3. See reference. For example, in some embodiments, the consumables are hemoproteins (e.g., hemog Robin, myoglobin, neuroglobin, cytoglobin, leghemoglobin, non-symbiotic Hemoglobin type I, Hell's gate globin I, bacterial hemoglobin, ciliate Contains myoglobin-like substances or flavohemoglobin.

[0283] Leghemoglobin is similar to myoglobin in terms of structure and physical properties, and agricultural As unused by-products of leguminous plant crops (e.g., soybeans or peas) It is readily available. In the United States, leghemoglobin in the roots of these crops is It exceeds the myoglobin content of all red meat consumed.

[0284] In some embodiments, consumables are made from components derived primarily or entirely from non-animal sources. It is composed of heme proteins (e.g., leghemoglobin or globin protein family). It is a meat imitation containing (members of Lee). For example, meat imitation is mainly or completely non Composed of components derived from animal sources, including muscle tissue imitation, adipose tissue imitation, and connective tissue. Contains imitation and heme proteins. In some embodiments, the consumables are mainly or completely Meat composed of components derived from non-animal sources, with a high iron content derived from heme protein. It is an imitation. In some embodiments, the iron content is the same as that of meat. In some embodiments, The consumables have a distinctive meaty red color, and this color is provided by leghemoglobin. Ta.

[0285] Heme proteins (for example, heme-containing proteins described in Section IIIB) are consumables. It can be used as an indicator that cooking is complete. Therefore, one embodiment of the present invention is that the product This includes detecting leghemoglobin that has moved from the inside to the surface of a consumable item when it is cooked. This is a method for preparing consumables. Another embodiment of the present invention is a method for preparing products. This is a method for cooking consumables, which includes detecting a color change from red to brown.

[0286] In some embodiments, the increased shelf life is due to the location of the food (e.g., non-meat-based meat substitutes). Provided by extending the lifespan of the desired red color.

[0287] In one embodiment, the present invention provides a heprotein that provides a desired color to a non-meat substitute. In some embodiments, the hemoprotein is derived from plants, fungi, or, for example, plants, algae. It originates from non-animal sources such as genetically modified organisms like bacteria or fungi. For example, see Section I. See IIB. In some embodiments, the lifespan of hemoproteins is extended by the meat preservation agent. It is extended by a procedure using [this method].

[0288] In some embodiments, the meat preservation extender is carbon monoxide, sulfite compounds, or metabisulfite. Sodium, Bombal, rosemary extract, green tea extract, catechins, and others Selected from a group consisting of antioxidants.

[0289] In one embodiment, the present invention provides a desired flavor profile to a food (e.g., a non-meat substitute). Provides a hemoprotein to provide to a desired hemoprotein. In some embodiments, the hemoprotein is desired Its ability to generate flavor profiles is similar to that of myoglobin.

[0290] In some embodiments, the lifetime of the ability to produce a desired flavor profile of hemoproteins This is 10%, 20%, 30%, 50%, or 100% more than myoglobin. It's bigger than this.

[0291] D. Foods containing isolated and purified heme protein In some embodiments, the heme protein described herein is used in meat or consumables. To enhance the properties of the substance, it is added to meat or consumables described herein. For example, to enhance the sensory characteristics of the meat during cooking by adding a "beef-like" flavor, hemta The protein-containing solution is injected into raw (for example, raw white meat) or cooked meat. It can be (for example, white meat such as chicken).

[0292] In another example, to enhance appearance, a heme protein solution is used in meat or the consumables of the present invention. It can be dripped upwards. In one embodiment, it is used for advertising and photographing food products such as meat or meat substitutes. Alternatively, video recording can be enhanced using heme proteins.

[0293] In another embodiment, heme protein is added to consumables as an iron supplement.

[0294] In one application of the present invention, heme protein can be used as a food coloring. In one embodiment Heme protein is used in various applications, including FD&C Red 40 - Allura Red A It can be used as a safe, digestible substitute for C, E129 (red meat). The list of uses, not limited to, includes, in particular, forms such as body painting, or This includes depicting blood in an exaggerated or artificial way.

[0295] In some embodiments, the present invention relates to heme proteins (e.g., leghemoglobin) derived from plants. This provides a method for obtaining leghemoglobin. Leghemoglobin can be obtained from various plants. Leguminous plant species and their varieties (e.g., soybeans, broad beans, lima beans, cowpeas, ginseng) Red peas, yellow peas, lupine, green beans, chickpeas, peanuts, almonds Farfa, vetch, clover (Lespedeza or quail bean) is a type of grass. Guhemoglobin plays an important role in regulating oxygen concentration in nitrogen-fixing root nodules (for example, Contains root nodules derived from the pea plant. In one embodiment, leghemoglobin protein The quality is achieved using ion exchange chromatography on leguminous plants (e.g., soybeans, sorella). It is purified from the root nodules of the pea plant (or pea). In one embodiment, leghemoglobin is, It is refined from soybeans, broad beans, or sweet pea root nodules.

[0296] In some cases, plants do not receive fertilizer, and the soil is naturally rhizobium (Rhizobium Except for being rich in nitrogen-fixing bacteria of the genus m, it is cultivated using standard farming methods. It is possible. Either the whole root or the root nodules are harvested and a grinder-blender is used. For example, 20 mM potassium phosphate pH 7.4, 100 mM potassium chloride and 5 mM It can be dissolved in M ​​EDTA. During this process, leghemoglobin is buffered. It is released into the rhizome. The nodule lysate containing leghemoglobin is filtered through a 5 μm filter. Cellular debris can be removed by filtration. In some embodiments, filtration is performed using centrifugation (7000). The process continues for 20 minutes. Next, the clarified solution containing leghemoglobin is administered. Filtered through a 200nm filter, and then rapidly processed using a protein liquid chromatography instrument (G Anion exchange chromatography column (High P) on E Healthcare) rep Q; High Prep DEAE, GE Healthcare) on The leghemoglobin is collected in the flow-through fraction and filtered on a 3 kDa filtration membrane. It is concentrated to the desired concentration. The purity (partial abundance) of the purified leghemoglobin is SD Analyzed by S-PAGE gel: In the lysate, leghemoglobin was present at 20-40% It is present, and after anion exchange purification, it is present at 70-80%. In another embodiment, anions The soybean leghemoglobin flowthrough obtained from exchange chromatography is size Exclusion chromatography (Sephacryl S-100 HR, GE Health) It is applied to care. Soybean leghemoglobin corresponds to dimeric and monomeric forms. It is eluted as two fractions. The purity (partial abundance) of leghemoglobin is measured using SDS. -Analyzed by PAGE, it was determined to be approximately 90-100%.

[0297] Proteins in lysates of leguminous plant root nodules are found in 10 mM sodium carbonate at pH 9.5, 50 It is transferred to a mM sodium chloride buffer and filtered through a 200 nm filter. Anisotropic ion on a rapid protein liquid chromatography instrument (GE Healthcare) It can be applied to an exchange chromatography column. Leghemoglobin is an anion. It is bound to an exchange chromatography matrix and eluted using a sodium chloride gradient. It is possible. The purity (partial abundance) of leghemoglobin is analyzed by SDS-PAGE. Therefore, it can be determined that the percentage is approximately 60-80%.

[0298] Undesirable small molecules derived from leguminous plant roots undergo anion exchange with leghemoglobin in solution. These can be removed from purified leghemoglobin by passing it through a resin. The offspring casts a fluctuating brown shadow on the root-nodule lysate, and therefore the leghemoglobin solution It degrades the quality of the color. In one embodiment, the anion exchange resin is FFQ, DEAE, Am Berlite IRA900, Dowex 22, or Dowex 1x4. Sulfur By fractionation of ammonium sulfate (60% wt / v and 90% wt / v ammonium sulfate) or by anion exchange chromatography, the reghemoglobin is purified. Robin buffered 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride. The solution was exchanged and passed through one of the anion exchange resins mentioned above. The flow-through was collected. The resulting coloration can be compared to the color of the solution before it passed through the anion exchange resin. Therefore, an improvement in the color of the purified leghemoglobin solution, as evaluated, can be observed. (From yellow / brown to a more pronounced red), but the removal of the yellow-brown tint varies in degree.

[0299] Alternatively, heme-containing proteins can be produced by recombinant DNA, as described in Section IIIB. It is possible. For example, non-symbiotic hemoglobin derived from mung beans can be detected by E. coli. It is expressed by recombinant expression and can be processed by anion exchange chromatography or cation exchange chromatography. The cell lysates can be purified using chromatography. It can be loaded onto FF-Q resin on GE Healthcare equipment. Non-symbiotic hemoglobin from the mung bean eluted in the flow-through fraction. The purity (partial abundance) of moglobin was analyzed by SDS-PAGE, and the total protein As a fraction of quality: 12% in E. coli lysate, and 31% after purification with FFQ. It was determined that... UV-Vis analysis of the purified protein revealed that heme-binding proteins are characteristic... It showed a typical spectrum.

[0300] Alternatively, the cell lysate can be processed using a rapid protein liquid chromatography (GE Hea) instrument. It can be loaded onto FF-S resin on (thcare). Mung bean non-symbiotic hemoglobin The FF-S column is coupled to a sodium chloride gradient (50 mM to 1000 mM). It can be eluted using [a specific method]. The purity (partial abundance) of mung bean non-symbiotic hemoglobin is SD Analysis was performed using S-PAGE, and 13% of E. coli lysate was obtained, followed by 35% purification using FFQ. It can be determined to be %. UV-Vis analysis of purified protein indicates heme-binding protein It may exhibit a characteristic spectrum.

[0301] In some embodiments, heme protein is used as an ingredient in foods where a blood flavor is desired. The heme-containing protein of the present invention was tasted by a volunteer panel, and each In some cases, it was described as having a blood-like taste.

[0302] Heme proteins, such as leghemoglobin, are found in other plant-based meat substitutes. They can be combined. In some embodiments, heme proteins are other components, such as lipids. They are captured in a gel containing proteins and / or other proteins. In some embodiments, multiple Several gels are combined with non-gel-based heme proteins. In some embodiments, To ensure that heme proteins can diffuse into consumables, heme proteins and consumables Other compound combinations of consumables are made. In some embodiments, the consumables are hemeproteins A solution containing a substance, for example, a leghemoglobin solution, containing, for example, 1, 5, 10, 15, 30 Alternatively, it can be immersed for 45 minutes or for 1, 5, 10, 15, 20, or 30 hours.

[0303] Considering the usefulness of heme proteins for coloring consumables, if a product contains a specific heme protein... Detecting whether or not a substance is present is useful. Therefore, the present invention is useful in some practical applications. The method includes a method for determining whether a product contains heme protein. For example, Guhemoglobin or other heme-containing proteins in food such as meat or meat imitations. To determine whether it is present, ELISA, proximity ligation assay, and luminescence assays are used. A smear assay or Western blot analysis may be performed. In one embodiment, the meat is leg We will examine whether it has been altered using hemoglobin or other heme-containing proteins. A detection method is then implemented.

[0304] E. Counterfeit Mayonnaise Spread Mayonnaise is a thick, creamy sauce. Traditional mayonnaise is It is a stable emulsion of oil and egg yolk. Lecithin and egg yolk-derived proteins are used in the process. It is thought to stabilize the marron. Traditional commercially available mayonnaise is usually 70-80 Contains % (wt / wt) fat and 5% (wt / wt) egg yolk. Low-fat commercial product. It may contain approximately 20% wt / wt of fat. The consumable has properties comparable to mayonnaise. It may include the following composition.

[0305] In one embodiment, the visual impression and texture are similar to traditional mayonnaise, and the appearance is stable. To produce a creamy protein-fat emulsion, purified plants Protein can be used as a substitute for egg protein. Fat (approximately 20-80% wt / wt) refers to a single or multiple source of supply, as described herein. It is possible. Non-traditional mayonnaise products can be used in all dishes where traditional mayonnaise is used. It can be used for application. In one embodiment, vinegar and / or lemon are used as flavoring additives. and / or lime juice is added. In one embodiment, purified plant protein The quality is not soy protein. In one embodiment, the flavor is mustard, spices, and It may be modified by the addition of ve and / or pickles.

[0306] Mayonnaise imitations may contain a mixture of non-animal proteins. In one embodiment, mayonnaise The imitation of Neese contains 50% (wt / v) rice bran oil and 7% (wt / v) mung bean 8S tahini. It is a mixture of proteins. In one embodiment, the mayonnaise imitation is 70% (wt / v) Sunflower oil or cocoa butter, 2.4% (wt / v) RuBisCo, 0.29% (w A mixture of t / wt soy lecithin and, optionally, 8 μM oleosin.

[0307] The mixture can be emulsified, and the stability of the emulsion can be improved by high-pressure homogenization or ultrasonic treatment. This can be controlled by modifying the size of oil-water-protein particles. The oil is It can be added as a liquid. Proteins can be added as a solution in a buffer. Zurecithin can be resuspended in water and subjected to ultrasonic treatment, and then in an oil and protein solution. It is mixed with the oil, protein and lecithin solution, for example, first 50 It can be homogenized at 00 psi, then at 8000 psi, or at 40% duty cycle. The ultrasonic processing can be performed for 2 minutes at the maximum setting in a cycle. The resulting product's density, texture, and clarity are all affected. - The texture and appearance are similar to those of traditional mayonnaise. In some cases ( For example, using mung bean 8S protein and rice bran oil, the product is light, grayish-white. It is the color of.

[0308] F. Cream Liqueur Imitation Traditionally, cream liqueurs contain fresh cream and liqueur as their base. To possess. Examples of liqueurs include whiskey, Irish whiskey, and Scotch whiskey. Key, rum, vodka, grappa, or fermented fruit (e.g., cherry liqueur), Examples include plum brandy, tequila, or herbal bitters. Cream liqueur The product contains fresh cream in the cream liqueur, and does not contain dairy components obtained from plant sources. It can be produced by substitution with the cream fraction. In one embodiment, the cream fraction The cream in the bottle contains vegetable fat and isolated or purified cream of a similar consistency to cream. It can be replaced with a stable emulsion of the protein. In one embodiment, purified plant Proteins and / or vegetable fats are single or multiple as described herein. It may originate from the source of the sunflower cream fraction, RuB. siCo and whiskey, and one or more optional flavorings (e.g., vanilla) May contain (chocolate and / or coffee).

[0309] G. Alcoholic beverages rich in protein Traditionally, alcoholic beverages contain negligible to small amounts of protein. The addition of plant proteins to alcoholic beverages positively modifies their flavor, texture, and physical state. This increases the nutritional protein content. Furthermore, various alcohols used in cocktails The presence of protein in a beverage positively modifies the flavor, texture, and physical state of the cocktail. To increase the nutritional protein content. Different classes of alcoholic beverages contain different amounts of alcohol. It contains alcohol. For example, a wine cooler contains about 4-7% alcohol, and beer contains alcohol. It contains approximately 3-10% alcohol, while wine contains approximately 8-14% v / v alcohol. Dessert wine contains approximately 17-20% alcohol, while whiskey contains approximately 40%. It contains % alcohol, and vodka contains approximately 35-50% alcohol. Some traditional alcoholic beverages contain sugar (for example, 10% wt / v Bacardi razor) include.

[0310] Therefore, alcoholic beverages should contain, for example, 0.1-5% wt / v of purified alcohol. Plant proteins and, optionally, sugars (1-15% wt / v) are added. Sugars can be, for example, cane sugar, brown sugar, sucrose, or glucose. For example, 180 mg / m³ in 20 mM potassium phosphate, pH 7.0, and 150 mM NaCl. 1 liter of refined Rubisco can be added to whiskey. 5% wt / v Rubs Jameson whiskey refilled with ICO has a consistency similar to a traditional Jell-O shot. A soft gel was formed.

[0311] For example, Corona beer, Pineau Grigio wine, and Jameson whiskey, Purified Rubsico, mung bean 8S, and pea globulin proteins are used. By adding them at final protein concentrations of 0.5%, 1%, and 5% wt / v, respectively. Alcoholic beverages rich in rubsico, mung bean 8S, and PA globulin are manufactured. It was done. In a solution of 60% ethanol and 5% sucrose in water, zein was added at concentrations of 0.5% and 1%. It was added at a concentration of 5% wt / v.

[0312] Pea protein powder is mixed with 5%, 20%, or 40% ethanol in water, 5% The enzyme is then resuspended in sucrose solution and incubated at room temperature for 1 hour. Extracted from pea flour. Any undissolved solids were centrifuged at 5000g for 10 minutes. It was removed by separation. The resulting supernatant solution appeared clear. 5% ethanol The solution was particularly useful.

[0313] The sensory panel compared all protein-rich alcoholic beverages with those that were not protein-rich. It was evaluated as having a different aroma and flavor from the beverage. In some cases, the resulting aroma And the flavor is neither one nor the other, and in some cases it is more appealing than the contrast. Then, in some cases, it was determined that the appeal was not very strong. In a specific case, James Mung bean 8S protein in both 0.5% wt / v and 1% wt / v to Son Whiskey The addition of quality softened the aroma and texture of Jameson. (0) The addition of 0.5% wt / v mung beans gives Jameson a slightly creamy flavor. In addition, it had an aroma similar to that of a traditional White Russian cocktail. The addition of 5% wt / v zein to skies is characterized as moldy beans and raw potatoes. It produced added aromas and flavors.

[0314] In another example, Corona beer was enhanced using 0.5% wt / v pea meglobulin. The aroma changes to a hoppy taste, similar to that of an Indian Pale Ale. The flavor has changed to retain the pea note. 0.5% wt / v and 5% wt / The addition of mung bean 8S protein to the V variety enhances the corona aroma and the hop aroma, resulting in a sweeter taste. It was transformed into a peony flower. The flavor is, in the case of 0.5% wt / v mung bean 8S, either It's not that simple, but in the case of 5% wt / v mung bean 8S, it's plant-like - nut-like He had the notebook.

[0315] Pineau Grigio wine is enhanced using 1% wt / v mung bean 8S protein. In another example, sweet, further citrusy aromatic notes were detected, along with a peanut-like flavor. It has changed to retain the buttery notes. It contains 1% wt / v pea meglobulin. The addition modified the aroma to that of strong moldy oak and wet leaves. The flavor was muddy. Modified to retain notes. Addition of 5% wt / v Rubisco, wet and dry. It produced a grassy aroma and flavor.

[0316] A 60% ethanol, 5% sucrose solution rich in zein is equivalent to a solution without zein. It retained the aromatic notes of toasted tortilla chips compared to the solution. There was no difference in flavor. It was.

[0317] 5%, 20%, and 40% ethanol and 5% sucrose, rich in pea protein. All of the solutions produced an earthy aroma and flavor compared to the protein-free control. Furthermore, a pea flavor was detected, and the bitter flavor increased with higher alcohol content. did.

[0318] H. Chocolate Spread Chocolate spread traditionally has cocoa powder, dairy milk, and vegetable oil as its main ingredients. It is a chocolate-flavored spread, which is also sugar. Toss spread is either a hard or soft solid at ambient temperature, and contains cocoa butter. It melts at a lower temperature than other substances. The product is a spread for bread, crepes, pancakes, and more. Cookie and cookie icing, chocolate confectionery filling, or dairy-free ingredients It can be used as a chocolate cake filling.

[0319] In one embodiment, dairy milk and ice cream, whey, cream, yogurt, and sour cream are used. - Dairy products such as cream or butterfat are manufactured as described herein. It is replaced with a cream fraction that does not contain dairy components. In one embodiment, it does not contain dairy components. The non-cream fraction is derived from one or more sources as described herein. In one embodiment, dairy milk and dairy products are any milk described herein. It is replaced with milk that does not contain the component. In one embodiment, dairy milk and dairy products are described in this specification. It is replaced with purified plant protein as described in the book. In one embodiment, Dairy milk and dairy products are made from one or more vegetable oils and one or more refined vegetables. It is replaced by a stable emulsion of a soft solid made from protein.

[0320] I. Other applications: In one embodiment, a milk chocolate bar that does not contain dairy ingredients or a milk chocolate bar that does not contain dairy ingredients is used. To manufacture Le Creuset chocolate confectionery, a dairy-free vegetable cream fraction is used instead of dairy milk. It can also be used as a substitute for dairy products.

[0321] In one embodiment, a milk chocolate bar that does not contain dairy ingredients or a milk chocolate bar that does not contain dairy ingredients is used. To manufacture Le Creuset chocolate confectionery, a dairy-free vegetable cream fraction and refined The processed plant protein can be used as a substitute for dairy milk and dairy products.

[0322] In one embodiment, to produce chocolate mousse, a dairy-free vegetable cream is used. Chocolate fractions and plant proteins may be used. The main ingredients of traditional chocolate mousse are: One embodiment consists of bittersweet or semisweet chocolate, dairy butter, and eggs. Therefore, dairy butter can be replaced with a vegetable cream fraction that does not contain dairy ingredients. In terms of form, dairy butter and eggs are made from a vegetable cream fraction and peas that do not contain dairy ingredients. This can be replaced by foam-stabilizing plant seed storage proteins such as mealbumin.

[0323] In one embodiment, vegan consumables such as putty analogues can be manufactured. The imitation putty is made by finely chopping 10g of fat imitation and finely chopped shallots It can be produced by heating it in a frying pan for 2-3 minutes with a bat. Connective tissue imitation The muscle imitation (20g), manufactured without muscle fibers, is cut into 1 / 2-inch squares and fat-free. The shallots can be further caramelized in the mixture for 3-5 minutes until golden brown. The mixture is homogeneous. It can be pushed through the sieve until it is dry. The bread is rinsed with a spoonful of Madeira while it is still warm. However, do not let it evaporate completely. The liquid obtained from the bread is added to the homogenized mixture. Spices (salt, pepper) are added to taste, and the mixture is then sifted again and pressed out. After being chilled in the refrigerator (for example, for 15 minutes), the pâté is ready to serve.

[0324] In some embodiments, other ingredients are used to produce a redder or fattier putty. A fat-to-muscle ratio is used. For example, the putty ratio is 0.5-10%, approximately 5-40%. It may contain approximately 10% to 60%, or approximately 30% to 70%, or >70% of adipose tissue imitation.

[0325] In some embodiments, to make the pâté a closer imitation of pork or chicken liver pâté... Therefore, muscle tissue imitation materials with a higher iron content can be used in the putty. For example, muscle tissue Counterfeit products may contain approximately 1%, 1.5%, 2%, or >2% heme protein.

[0326] In some embodiments, to make the pâté a closer imitation of chicken or fish pâté In addition, muscle tissue imitation materials with a lower iron content may be used in the putty. For example, muscle tissue imitation The product may contain approximately 1%, approximately 0.5%, approximately 0.2%, or <0.2% heme protein. ru.

[0327] In one embodiment, vegan consumables such as blood sausage analogues can be manufactured. Blood sausage for vegans is made from a solution of heme protein and refined vegetables. It is produced from blood analogues created by mixing proteins. For example, blood A 35 ml solution of leghemoglobin (120 mg / ml) and peas, which approximates the composition of the above. A mixed solution of mamealbumin (100 mg / ml) is added to a slurry of corn flour in brine. (6:5 w / v flour to water ratio) can be carefully mixed. 1 spoonful of chopped t The onions are sautéed with 10g of chopped imitation adipose tissue and mixed with 2-3 raisins. It can be combined, cooled to room temperature, and then mixed with the blood / powder mixture. The mixture can be adjusted to your preference. Seasoned together (for example, using salt, pepper, parsley and / or cinnamon) It can be stuffed into a vegetarian sausage casing and boiled in water just below boiling point for about 45 minutes. After cooling, the sausages can be consumed as is or further cooked, for example, smoked, or They can be crisped up in the bun or baked.

[0328] In some embodiments, muscle imitation products may be included in recipes that mimic meat / blood sausage. In some embodiments, barley, buckwheat, oats, rice, rye, sorghum, and other grains are used. Wheat or other grains may be used in blood sausage. In some embodiments, Bread, chestnuts, potatoes, sweet potatoes, starch or other fillers, It can be added to sage or substituted for grains in blood sausage. Examples [Examples]

[0329] Protein isolation All steps were performed at 4°C or room temperature. The centrifugation step was performed at 8000g. The temperature was 4°C or room temperature for 20 minutes. The powder was suspended in a specific buffer solution, and the suspension was centrifuged. The supernatant is microfiltered through a 0.2 micron PES membrane, and then Spectrum La bs KrosFlo hollow fiber tangential flow filtration system at 3kDa, 5kDa, or 10kD The concentration was achieved by ultrafiltration using a molecular weight cutoff PES membrane.

[0330] After fractionation, store all desired ammonium sulfate precipitate fractions at -20°C until further use. Before using them in the experiment, the precipitate was put into 10 volumes of 50 mM potassium phosphate buffer, pH 7.4 was resuspended in 0.5M NaCl. The suspension was centrifuged, and the supernatant was collected in 0.2 micrometers. Microfiltration is performed through a PES membrane, followed by Spectrum Labs KrosFl o Molecular weight cutoff of 3kDa, 5kDa, or 10kDa in hollow fiber tangential flow filtration systems The protein composition at each fractionation step was concentrated by ultrafiltration through a PES membrane. The protein concentration was monitored by DS-PAGE and measured using the standard UV-Vis method. It was measured.

[0331] (i) Pea Mealbumin: Dried green or yellow pea flour is used as pea mealbumin. Lubumin was used as a source. The powder was added to 10 volumes of 50 mM sodium acetate buffer, pH 5. The soluble protein was suspended and stirred for 1 hour. The unextracted protein and pea seeds were used to extract the soluble protein. Centrifugation (8000g, 20 minutes) or filtration through a 5-micron filter from the small debris. Separation was performed by either the supernatant or the filtrate. This crude protein Solid ammonium sulfate was added to the extract until saturated at 50% wt / v. The solution was stirred for 1 hour. Next, the mixture was centrifuged. Ammonium sulfate was added to the supernatant from this step at 90% wt / v. The solution was added until saturated. The solution was stirred for 1 hour, and then the pea methylalbumin protein in the precipitate was extracted. The precipitate was centrifuged to recover the protein. The precipitate was stored at -20°C until further use. Collect the precipitate and ensure the final buffer solution contains 0-500 mM sodium chloride. It was prepared for use as described above, except for the following:

[0332] In some embodiments, the powder is suspended in 10 volumes of 50 mM NaCl, pH 3.8, and 1 The mixture was stirred for a certain amount of time. Soluble proteins were extracted from unextracted proteins and pea seed debris. The components were separated by centrifugation (8000g, 20 minutes). The supernatant was collected and a 0.2 micron membrane was prepared. The solution was filtered through a filter and concentrated using a 10 kDa cutoff PES membrane.

[0333] (ii) Pea globulin: Dried green pea flour is used to make pea globulin Used to extract protein. The powder was mixed with 10 volumes of 50 mM potassium phosphate buffer (pH 100%). The soluble proteins were suspended in 8 and 0.4 M sodium chloride and stirred for 1 hour. The seeds were separated from the edible bean seed debris by centrifugation. The supernatant was divided into two solutions at 50% and 80% saturation. The ammonium sulfate was subjected to fractionation in the second step. The 80% precipitate containing the target globulin was then obtained. Store at -20°C until use. Collect the protein from the precipitate and use as described above. Prepared for use.

[0334] iii) Soy 7S and 11S globulins: Globulins from soy flour, low fat / Prepare defatted soy flour in 4-15 volumes of 10 (or 20) mM potassium phosphate solution at pH 7.4. It was isolated by suspension. The slurry was centrifuged at 8000 rcf for 20 minutes. The solution was then clarified by 5 micron filtration, and the supernatant was collected. The crude protein extract was 7S It contained both and 11S globulin. Then, before using the solution in the experiment, 0.2 ml Filtered with a cloning filter and using a 10kDa molecular weight cutoff PES membrane with Spectrum L Concentration is performed using abs KrosFlo hollow fiber tangential flow filtration systems, or through anion exchange resin. It shrunk. 11S globulin was separated from 7S protein by isoelectric focusing precipitation. Crude protein The pH of the extract was adjusted to 6.4 with dilute HCl, stirred for 30 minutes to 1 hour, and then precipitated with 11S. The 7S protein was recovered from the supernatant and centrifugation. The 11S fraction was separated before use. The solution was resuspended in 10 mM potassium phosphate pH 7.4, and the protein fraction was microfiltered to concentrate it. It shrunk.

[0335] Soy protein is used to reduce off-flavors in purified protein. Defatted soy flour in 4-15 volumes (for example, 5 volumes) of 20 mM sodium carbonate, pH 9 (also (Water adjusted to pH 9 after adding the powder) or 20 mM potassium phosphate buffer pH 7.4 It can also be extracted by suspending it in 100 mM sodium chloride. The slurry is 1 The mixture was stirred and centrifuged at 8000 × g for 20 minutes. The extracted protein was ultrafiltered. Next, the supernatant is collected and passed through a 0.2 micron membrane, either as described above or alternatively. The sample was filtered and concentrated using a 10 kDa cutoff PES membrane.

[0336] (iv) Moon bean 8S globulin: Moon bean powder, 4 volumes of powder 50mM In potassium phosphate buffer pH 7 (with 0.5 M NaCl added for laboratory-scale purification), Initially, 8S globulin was extracted by suspension. After centrifugation, the supernatant was used. The proteins inside were saturated with ammonium sulfate in two steps, 50% and 90%, respectively. The mixture was fractionated by addition. The precipitate from the 90% fraction contained 8S globulin and was further used. Stored at -20°C until use. The protein was collected from the precipitate and used as described above. It was prepared as follows.

[0337] Moong bean globulin reduces off-flavors in purified protein fractions. To do this, put the powder in 4 volumes of 20 mM sodium carbonate buffer, pH 9 (or Moon Powder) Extraction can also be achieved by suspending the slurry in water (with the pH adjusted to 9 after addition). To remove the particles, centrifuge (or filter), ultrafilter, and then process as described above. did.

[0338] (v) Abundant protein for late embryonic development: powder (not limited to this, but also moon beans and (Containing soy flour) is mixed with 20 mM Tris-HCl, pH 8.0, and 10 mM NaCl. The mixture was suspended, stirred at room temperature for 1 hour, and then centrifuged. The supernatant was concentrated with acid (HCl or acetic acid). The solution was added up to 5% (v / v), stirred at room temperature, and then centrifuged. The supernatant was heated to 95°C for 15 minutes. The mixture was heated for minutes, then centrifuged. Trichloroacetic acid was added to the supernatant until it reached 25%. The mixture was precipitated, centrifuged, and then washed with acetone. The heating and acid washing steps were reversed. The same can be done in direction.

[0339] (vi) Pea prolamin: Dried green pea flour 5 × (w / v) 60% ethanol Suspend in gauze, stir at room temperature for 1 hour, then centrifuge (7000g, 20 minutes), and above The supernatant was collected. The ethanol in the supernatant was evaporated by heating the solution to 85°C, and then... It was then cooled to room temperature. Ice-cold acetone (1:4 v / v) was added to precipitate the protein. Next, the solution was centrifuged (4000g, 20 minutes) to separate the proteins into a light beige color. It was collected as a color precipitate.

[0340] (vii) Zein Prolamin: Corn protein concentrate or powder 5 × (w / v) Suspended in 60% ethanol, stirred at room temperature for 1 hour, then centrifuged. The supernatant was... The tanol was evaporated by heat, and then the solution was centrifuged to collect the protein as a precipitate.

[0341] (viii) Blend RuBisCO leaves in 4 volumes of cold 50 mM potassium phosphate. Buffer pH7.4 buffer (0.5M NaCl+2mM DTT+1mM EDTA) First, the alfalfa green was fractionated by crushing. The resulting slurry was then used as debris. To remove the contaminants, the mixture was centrifuged, and the supernatant (crude solubilized material) was used in further purification steps. The protein in the crude solubilized product is reduced by adding ammonium sulfate until it reaches a 30% (wt / v) saturation. The solution was then fractionated. The solution was stirred for 1 hour, and then centrifuged. The precipitate from this step was discarded. Discard the supernatant and add additional ammonium sulfate until saturated with 50% (wt / v) ammonium sulfate. The solution was stirred for 1 hour and then centrifuged again. The precipitate from this step was RuBisC It contained O and was stored at -20°C until use. Protein was collected from the precipitate and as described above. Prepared for use.

[0342] RuBisCO prepares the crude solubilizer to 0.1M NaCl and uses it in an anion exchange resin. Purification can also be achieved by applying a solution. Weakly bound protein contaminants can be removed using 50 mM potassium phosphate. The solution was washed with buffer solution pH 7.4 + 0.1M NaCl. Then, RuBisCO was applied in high heat. Elution was performed using an on-intensity buffer (0.5 M NaCl).

[0343] The RuBisCO solution was decolorized by passing it through a column packed with activated carbon (pH 7~). 9) The dye was bound to the column, while Rubisco was isolated in the filtrate.

[0344] The RuBisCO solution is packed into the column (or in batch form) using FPX66 (Dow Chemicals) Desorption can also be achieved by incubating the resin and solution. The slurry was colored. The slurry was incubated for 30 minutes, then the liquid was separated from the resin. The element was bonded to the resin, and RuBisCO was recovered in the column-pass fraction.

[0345] In some embodiments, RuBisCO blends spinach leaves in a blender. Volume 20 mM potassium phosphate buffer pH 7.4 buffer + 150 mM NaCl + 0.5 It was initially isolated by grinding with mM EDTA. The resulting slurry was then removed from the debris. To remove it, centrifuge is used, and the supernatant (crude solubilized material) is filtered through a 0.2 micron membrane. The material was concentrated using a 0 kDa cutoff PES membrane.

[0346] In some embodiments, RuBisCO is derived from alfalfa or wheat sap powder. In the renderer, the powder is mixed with 4 volumes of 20 mM potassium phosphate buffer, pH 7.4 buffer + 150 ml. Extraction was performed by mixing with M NaCl (0.5 mM EDTA). The obtained slurry The lee was centrifuged to remove debris, and the supernatant (crude solubilized material) was passed through a 0.2 micron membrane. The sample was filtered and concentrated using a 10 kDa cutoff PES membrane.

[0347] (ix) Leghemoglobin. Soybean root nodules using a grinder-blender. In 0 mM potassium phosphate pH 7.4, 100 mM potassium chloride, and 5 mM EDTA It was suspended and solubilized. During this process, leghemoglobin is released into the buffer. Solubilized root nodules containing moglobin are filtered through a 5-micron filter. The cellular debris was cleared. In some embodiments, filtration was performed using centrifugation (7000g, 20 This continued for several minutes. A clarified solubilized substance containing leghemoglobin was then added to 0.2 microns. Filtered through a filter, and then processed using a fast protein liquid chromatography (GE H) apparatus. Anion exchange chromatography column (High Prep) in Healthcare Applied to Q, High Prep DEAE, GE Healthcare. Ghemoglobin was collected in the pass-through fraction, and Spectrum Labs KrosFlo Concentration to the desired concentration by a 3kDa molecular weight cutoff PES membrane in a hollow fiber tangential flow filtration system. The purity (partial abundance) of the purified leghemoglobin was determined by SDS-PAGE gel. Analysis revealed that leghemoglobin was present in the solubilized product at a concentration of 20-40%, after anion exchange purification. In another embodiment, from anion exchange chromatography Soybean leg hemoglobin pass-through fraction is analyzed using size exclusion chromatography (Sephacr Applied to yl S-100 HR (GE Healthcare). Soybean hemoglobin Robin was eluted as two fractions corresponding to the dimer and monomer molecular species. The purity (partial abundance) of globin was analyzed by SDS-PAGE, and it was found to be approximately 90-100%. It was determined that it was present. Analysis of the UV-VIS spectrum (250-700 nm) showed that heme was low Spectroscopic characteristics consistent with those of the treated leghemoglobin were revealed.

[0348] (x) non-symbiotic hemoglobin from moon beans into the pJexpress401 vector The cells were cloned into (DNA2.0) and transformed into E. coli BL21. Replace tryptone with soyton, kanamycin, 0.1 mM ferric chloride and 10 μg The cells were grown in LB medium containing 5-aminolevulinic acid at a concentration of 0.2 mM I. The cells were induced by PTG and grown at 30°C for 20 hours. moong bean nonsymbiotic hemoglobin E. coli cells expressing globin were collected and treated with 20 mM MES buffer at pH 6. The sample was resuspended in 0.5, 50 mM NaCl, 1 mM MgCl2, and 1 mM CaCl2. Small amounts of DNAase I and a protease inhibitor were added. Cells were then sonicated. It was solubilized. 16,000 g of the solubilized material was separated from the cell debris by centrifugation for 20 minutes. Next, filtration with a 200nm filter was performed. Then, the cell soluble material was processed using fast protein... The FF-S resin was subjected to a liquid chromatography system (GE Healthcare). It worked. Moong bean nonsymbiotic hemoglobin bound to the FF-S column, and sodium chloride Elution was performed using a gradient (50 mM to 1000 mM). moong bean non-symbiotic. Hemoglobin purity (partial abundance) was analyzed by SDS-PAGE: Escherichia coli (E. coli) The solubilized content was determined to be 13%, and after purification with FFQ, it was determined to be 35%. UV-Vis of the purified protein The analysis revealed a spectrum characteristic of heme-binding proteins.

[0349] (xi) Heme protein is associated with the N-terminal His6 epitope tag and TEV cleavage site. Synthesized and cloned into the pJexpress401 vector (DNA2.0) in the large intestine. The transformed cells were converted to the bacterium (E. coli) BL21. Soyton was used instead of tryptone in the transformed cells. Kanamycin, 0.1 mM ferric chloride and 10 μg / ml 5-aminolevulinic acid The cells were grown in the LB medium containing the cells. Expression was induced by 0.2 mM IPTG. The mixture was grown at 30°C for 20 hours. E. coli expressing heme protein was collected. 50 mM potassium phosphate pH 8, 150 mM NaCl, 10 mM imidazole, 1 mM MgCl2, 1 mM CaCl2, DNAase I, and protease inhibitors are reconstituted. The cells were suspended. They were solubilized by sonication and clarified by centrifugation at 9000 × g. The solubilized material was incubated with NiNTA resin (MCLAB) and 5 columns were used. CV) 50 mM potassium phosphate pH 8, 150 mM NaCl, 10 mM imidazole Wash, then add 50 mM potassium phosphate (pH 8), 150 mM NaCl, and 500 mM imidazo. Elution occurred with elution. SDS-PAGE and UV-vis spectra showed the predicted molecular weight. We confirmed complete hemloading in each case.

[0350] In some embodiments, transformed cells were treated with 10 g / L glucose monohydrate and 8 g / L Monopotassium nitrate, 2.5 g / L Sensient Amberferm 6400, 2 0.5g / L Sensient Tastone 154, 2g / L Dyammonium Phosphate 1mL / L Trace Metals Mixture (Teknova 100 0× Trace Metals Mixture (Catalog No. T1001), 1g / L magnesium sulfate, 0.25 mL of 0.1 M ferric chloride solution, 0.5 mL / L Sigm a Anti-foam 204, 1 mL / L kanamycin sulfate 1000× solution The plants were propagated in a seed medium consisting of the following: 250 mL of medium was placed in (4)-1 L baffled shaker. For use in the SCO, take 0.25 mL from each vial of glycerol-preserved culture. Inoculation was performed. The cells were grown in a shaking flask for 5.5 hours at 37°C with 250 RPM agitation. The seed culture medium was steam-sterilized in a 100L bioreactor, cooled to 37°C, and pH was adjusted. Adjust to 7.0, and once the shaking flask OD of 2.5 is achieved, shake the flask culture Inoculation was performed with 800 mL. Air was supplied to the bioreactor at 40 L / min, and agitation was performed. The growth rate was 250 RPM. After 2.2 hours of growth, an OD of 2.20 was achieved, and the culture was 2 2L to the end 4m 3 It was transferred to the bioreactor. The starting medium for the final bioreactor was The following components, steam-heated in place: 1775 L of deionized water, 21.75 L of monopotassium phosphate. kg, diammonium phosphate 2.175 kg, ferric ammonium citrate 4.35 kg , 8.7 kg ammonium sulfate, Sensient Amberferm 6400 1 Composed of 0.875kg of Sensient Tastone 154 and 10.875kg of Sensient Tastone 154. The culture medium components were heated by steam for 30 minutes, then cooled to 37°C and added after sterilization: 0. 2.145 L of 1 M ferric chloride solution, 59.32 kg of 55% w / w glucose monohydrate, T race Metals Mixture(Teknova 1000× Trace Metals Mixture (Catalog No. T1001) 3.9L, 200g / L Phosphorus 10.88 L of diammonium acid, 36.14 L of 1 M magnesium sulfate and Sigma Anti-foam 204 2.175L, Kanamycin sulfate 1000× solution 2.1 75L. pH was controlled to 7.0 by adding 30% ammonium hydroxide. Aeration was 2. 175m 3 Supply at / min and vary the agitation of dissolved oxygen between 60 and 150 RPM. By modifying it, it was controlled to 25%. At two time points (EFT=4 and EFT=8), Rapid, large-scale addition of additional nutrients was supplied. Each addition was Sensient Amberfe rm 6400 5.5kg, Sensient Tastone 154 5.5kg and 4.4 kg of diammonium phosphate in an autoclaved solution (Amberfer 100 g / L for m and Tastetone, and for diammonium phosphate It was added to a 200g / L solution. Sterile glucose solution of 55% w / w glucose monohydrate. This was supplied to the bioreactor to maintain a residual glucose level of 2-5 g / L. Once OD25 is achieved, reduce the temperature to 25°C and add 1M isopropyl β-D Induction was performed with 0.648 L of -1-thiogalactopyranoside. The culture was grown for a total of 25 hours. Then, the culture is diluted 1:1 with deionized water, and then centrifuged, and the centrifugal is obtained. The cells were concentrated to a 50% v / v solid content. The centrifugated cells were frozen at -20°C. Thaw the material at 4°C, then add 20 mM potassium phosphate (pH 7.8), 100 mM NaCl, and 10 ml The solution was diluted in M-imidazole and homogenized at 15,000 PSI. Cells were filtered by tangential flow filtration (TFF) to 0.2 μm, and the filtered solubilized material was charged with zinc I. The MAC column (GE) was loaded directly. The binding protein was loaded into 10 column volumes (CV) of 2 Wash with 0 mM potassium phosphate pH 7.4, 100 mM NaCl, and 5 mM histidine. Elution was performed using 10 CV of 500 mM potassium dihydrogen phosphate and 100 mM NaCl. Ghemoglobin was concentrated and used with a 3kDa molecular weight cutoff PES membrane and TFF. The sample was filtered. The concentrated sample was reduced with 20 mM sodium dithionite and G-20. Desalting was performed using gin (GE). The desalted leghemoglobin sample was frozen in liquid nitrogen, - Stored at 20°C. Leghemoglobin concentration and purity were measured by SDS-PAGE and UV-V. This was determined by an IS analysis.

[0351] (xi) Oleosin. Sunflower oil was refined from sunflower seeds. Sunflower seeds were 10 0 mM sodium phosphate buffer pH 7.4, 50 mM sodium chloride, 1 mM EDTA The oil was mixed in a 1:3 wt / v ratio. The oil was then centrifuged (5000g, 20 minutes). Collect the solution, resuspend it in 50 mM sodium chloride and 2 M urea at a ratio of 1:5 (wt / v) for 30 minutes. The mixture was stirred at 4°C. The 2M urea washing and centrifugation steps were repeated. By centrifugation... The recovered oil was treated with 100 mM sodium phosphate buffer pH 7.4 and 50 mM sodium chloride. The oil was resuspended in the solution. The centrifugal separation and washing steps were repeated once more to obtain the final washed oil. The fraction was obtained from the final centrifugation step. The oil was dissolved in 100 mM sodium phosphate buffer (pH 100 mM). 7.4, 50 mM sodium chloride, 2% wt / v vegetable oil fatty acid salts, rehydrated at 10% wt / v The solution was suspended, homogenized at 5000 psi, and incubated at 4°C for 12 hours. The mixture was centrifuged (8000g, 30 minutes), the top layer was taken, and the soluble fraction was recovered. SDS- PAGE analysis suggests that oleosin is the major protein present in the soluble fraction. The oleosin concentration was 2.8 mg / ml.

[0352] (xii) Pea whole protein: Dried green or yellow pea flour whole pea Used to extract horse protein. The powder was mixed with 10 volumes of 20 mM potassium phosphate solution. The sample was suspended in a saturation solution at pH 8 and 100 mM sodium chloride and stirred for 1 hour. Soluble protein The substance was separated from the pea seed debris by centrifugation. The supernatant was collected and 0.2 ml The solution was filtered through a ron membrane and concentrated using a 10 kDa cutoff PES membrane.

[0353] (xiii) Pea mebicillin and pea melegmin: dried green or yellow Pea flour was used as described above to extract all pea protein. The crude pea mixture obtained therefrom was then analyzed using ion exchange chromatography. The material was fractionated into pea mebicillin and pea melegmin. The material was treated with Q Sepharose Fa Load into st Flow resin and change the salt concentration from 100 mM to 500 mM NaCl. The fraction was recovered after the solution was converted. Pea pulp silin was recovered with 350 mM sodium chloride. Meanwhile, pea melegmin was recovered using 460 mM sodium chloride. The recovered fraction was divided into 10 Concentration was performed using a KDa cutoff PES membrane.

[0354] (xv) Lentil total protein: air-classified lentil flour Used to extract the crude mixture of lentil protein. Powder was mixed with 5 volumes of 20 mM phosphate. The solution was suspended in potassium buffer pH 7.4 and 0.5 M sodium chloride and stirred for 1 hour. Soluble proteins are separated by centrifugation (8000g, 20 minutes) to remove unextracted proteins and lecithin proteins. It was isolated from dwarf bean seed debris. The supernatant was collected and filtered through a 0.2 micron membrane. The material was concentrated using a 0 kDa cutoff PES membrane.

[0355] (xvi) Lentil albumin: 5 volumes of air-classified lentil powder in 50 mM chloride The soluble protein was suspended in sodium, pH 3.8, and stirred for 1 hour. The soluble protein was centrifuged (80°C). Unextracted proteins and lentil seed debris were isolated by (00g, 20 minutes). The supernatant is collected and filtered through a 0.2 micron membrane, and a 10 kDa cutoff PES membrane is used. It was concentrated using [a specific method / tool].

[0356] (xvii) Chickpea (Garbanzo bean) Total Protein: Add 5 volumes of chickpea (garbanzo bean) flour to 20 mM potassium phosphate buffer pH 7.4. The soluble protein was suspended in 0.5M sodium chloride and stirred for 1 hour. The soluble protein was centrifuged. Unextracted protein and chickpea seed saturation (8000g, 20 minutes) It was separated from the yellowtail. The supernatant was collected and filtered through a 0.2 micron membrane, and 10 kDa cut-off was performed. Concentration was performed using an off-PES membrane.

[0357] (xviii) Chickpea / Garbanzo bean albumin: Suspend chickpea (garbanzo bean) flour in 5 volumes of 50 mM sodium chloride, pH 3.8. The mixture was then stirred for 1 hour. The soluble protein was separated by centrifugation (8000g, 20 minutes). The extracted protein was separated from lentil seed debris. The supernatant was collected and measured to 0.2 micrometers. The solution was filtered through a membrane and concentrated using a 10 kDa cutoff PES membrane.

[0358] (xix) Amaranth powder dehydrin: Amaranth powder in 5 volumes of 0.5M sodium chloride The soluble protein was suspended in pH 4.0 and stirred for 1 hour. The soluble protein was centrifuged (8000g, 2 Unextracted proteins and lentil seed debris were separated by (0 minutes). The supernatant was then rotated. The sample is collected, filtered through a 0.2 micron membrane, and concentrated using a 3 kDa cutoff PES membrane. Further concentrate of dehydrin from this fraction was boiled in concentrated protein material for 80°C. It was obtained by rotating it at 00g for 10 minutes and collecting the supernatant. [Examples]

[0359] Construction of muscle tissue analogues To prepare muscle tissue imitation materials, use moon bean protein solution (20 mM phosphate solution). 8 ml of saturates (pH 7.4) and 114 mg / ml sodium chloride in 400 mM solution is added to the leg. Hemoglobin solution (6m in 20mM potassium phosphate, 400mM NaCl, pH 7.3) The mixture was mixed with 16 ml of (g / ml leghemoglobin). The resulting mixture was then mixed with Amicon sp Using in concentrators (10kDa cutoff), the final concentration mo ong bean 8S globulin 61 mg / ml and leghemoglobin 6.5 mg / ml It was concentrated. Approximately 400 mg of transglutaminase powder was added to the solution and mixed thoroughly. The mixture was then divided into two 50ml Falcon tubes and incubated overnight at room temperature. The final total protein concentration was 67.5 mg / ml total protein. (Muscle tissue imitation) The product contains a small amount (<1 ml) of dark red inclusions, a reddish-brown opaque gel containing a venous blood-colored liquid. It was formed. [Examples]

[0360] Increased tensile strength of adipose tissue imitation 40ml aliquot of rice bran oil and moong bean protein (114mg / ml) A 40ml aliquot was mixed in a 250ml Pyrex beaker. The beaker was placed in a water bath. The Branson Sonifer 450 sonicat features a 12mm tip. Alternatively, emulsification was performed for 6 minutes using a 60% load cycle (duty cycle) and power level 5.

[0361] In the 18cm x 18cm x 2.5cm synthetic rubber Ikea plastic ice tray, 48 mg of Ctrospan fiber (derived from connective tissue in Example 14) is placed in one 13.97 cm × 1.2 They were placed as evenly as possible vertically on the bottom of a 7cm x 1.5875cm triangular mold. Pour approximately 20ml of rice bran oil / moong bean protein emulsion onto the fibers. Next Then, use an additional 20 ml of emulsion as a control in the same dish of similar size. It was poured into an empty mold.

[0362] The ice tray was floated in boiling water for 15 minutes, then removed and allowed to cool to room temperature.

[0363] Each gel obtained using a razor blade was measured to a length of 4.66 cm and a cross-sectional area of ​​1 cm². 2 It was cut into three segments with the attached TA-96B probe. e Micro Systems TA XTExpress Enhanced te An Xture analyzer was used to evaluate tensile strength. Contains fat imitation. The fiber has a tensile strength of 23 kPa, while the fat imitation without fiber has a tensile strength of 20 kPa. It had a tensile strength of [value missing]. [Examples]

[0364] High-percent fat imitation 3.3% wt / v pea meglobulin, as well as coconut, cocoa, and olive oil Formed from an equal mixture of 70% v / v oil and 0.5% wt / v lecithin. Adipose tissue imitation containing the resulting protein oil emulsion was treated with 2% transglutaminase (Aji It was cross-linked with nomoto Activa(registered trademark) TI). After discharge and dehydration, The resulting gel was moderately soft, and its fat content was confirmed to be 75% (wt / wt).

[0365] Adipose tissue matrix containing protein-oil emulsification at 1.6% wt / v Rubisco It was formed with 80% v / v cocoa butter. The resulting gel was soft. [Examples]

[0366] Method for preparing adipose tissue imitation If necessary, melt the oil by warming it to room temperature or heating it gently. If they are solid at room temperature, keep them near their melting point during the rest of the procedure. Proteins are specific Obtain according to each protocol (see Example 1). Weigh out the lecithin and resuspend it in water. Next, the solution is ultrasonically treated to create a uniform solution. The components are adjusted to specific ratios, and as needed... The solution is a buffer solution (20 mM sodium phosphate containing 50 mM sodium chloride, pH 7.4). Volume is adjusted, and then homogenization is performed to control the particle size in the emulsion. Alternatively, subject to ultrasonic treatment. Afterwards, the emulsion is subjected to (a) heating / cooling, (b) translation. (c) Cross-linking with suglutaminase enzyme or transglutaminase following heating / cooling Gel formation occurs by either the addition of a ze enzyme or by transferring. Control sample (without heating / cooling, transferring For comparison, a sample (without glutaminase crosslinking treatment) was prepared. Heating / cooling treatment was performed. The emulsion to be stabilized is to place the emulsion in a water bath at 90-100°C for 5 minutes, then The sample is prepared by slowly cooling it to room temperature. Transglutaminase crosslinking The emulsion stabilized by the compounding process is obtained by adding transglutaminase at 2% wt / v. Prepare by incubating at 37°C for 12-18 hours. Continue heating / cooling. Emulsions stabilized by the addition of transglutaminase enzymes are heated / cooled. First, the protocol is followed, and then, once the sample has cooled to room temperature, the enzyme is added to prepare it. Prepare the mixture. Incubate all emulsions at 37°C for 8-12 hours. [Examples]

[0367] Methods for analyzing adipose tissue imitation After various gelation treatments, the gel emulsion is brought to room temperature for evaluation. The total volume of the emulsion, and the volume of water and / or oil separated into phases (gel emulsion) Record the hardness of the adipose tissue imitation (if John is not a single phase). It is evaluated by gently poking it. The cooking experiment involves transferring the mass to a heated surface, This is done by measuring the temperature of the liquid immediately after cooking. [Examples]

[0368] Beef fat imitation Adipose tissue imitation is mixed with equal parts cocoa butter, coconut butter, olive oil, and palm oil. It was prepared by gelling a solution of emulsified purified moon bean 8S protein. g soybean 8S protein was purified as described in Example 1, and the result was 20 mM potassium phosphate at pH 7.4. The concentration was adjusted to 140 mg / ml in 400 mM NaCl. The fat mixture was solidified into individual fats. It was prepared by melting the substance from its body into a liquid state at 45°C for 30 minutes. The fats (cocoa butter, coconut butter, olive oil and palm oil) in a 1:1:1:1 ratio. The protein-fat emulsion was mixed in a (v / v) ratio. The protein-fat emulsion was mixed with a 70% v / v liquid fat mixture. Contains 4.2% wt / v Moong Bean 8S Protein and 0.4% wt / v Soy Lecithin. By mixing, vortexing for 30 seconds, and then sonicating for 1 minute, the mixture is emulsified. Formed. After homogenization, the lipid protein emulsion was observed visually. It was determined to be a single liquid phase.

[0369] One adipose tissue imitation emulsion contains 0.2% wt / v transglutaminase enzyme, 3 It was stabilized by cross-linking at 7°C for 12 hours. Another adipose tissue imitation was subjected to 10°C in a water bath. The protein was stabilized by gelation through heating to 0°C followed by cooling to ambient temperature. The adipose tissue imitation was monophase. It was formed by transglutaminase. The adipose tissue imitation matrix is ​​formed by heat / cooling-induced gelation. It was a softer solid than the product matrix. [Examples]

[0370] Wagyu beef fat imitation Adipose tissue imitation is made from purified pea meglobulin protein and an equal amount of cocoa butter. It was made by gelling an emulsion of coconut butter, olive oil, and palm oil. The gentian meglobulin protein was purified as described in Example 1, and 20 mM potassium phosphate was used. The solution was prepared at pH 8, in 400 mM NaCl, at a concentration of 100 mg / ml. The fatty mixture was then divided into individual portions. The fat was prepared by melting it from a solid state to a liquid state at 45°C for 30 minutes. The individual fats (cocoa butter, mango butter, olive oil) are in a 2:1:1 ratio (olive oil). A mixture of oil, cocoa butter, and mango butter in a v / v ratio. Protein-fat emulsion. Mix the liquid fat mixture with a 5% wt / v solution of pea meglobulin in a 1:1 ratio. The mixture was formed by emulsifying it for 30 seconds at the maximum setting using a handheld homogenizer. After homogenization, the lipid-protein emulsion was determined by visual observation. It was a single liquid phase. The emulsion was mixed with 0.2% wt / v transglutaminase enzyme. The tissue was stabilized by cross-linking at 37°C for 12 hours. The resulting adipose tissue imitation was monophase. It was a soft, solid, and had a salty flavor. [Examples]

[0371] Adipose tissue imitation with fatty acid dispersion of beef: Adipose tissue imitation was prepared using purified pea meglobulin protein and an equal amount of cocoa butter. —Made by gelling an emulsion of mango butter, olive oil, and rice bran oil. Pea meglobulin protein was purified as described in Example 1, and 20 mM potassium phosphate was used. The solution was prepared at pH 8, with a concentration of 100 mg / ml in 400 mM NaCl. The fat mixture was then divided into individual portions. The fat was prepared by melting it from a solid state to a liquid state at 45°C for 30 minutes. The individual fats (cocoa butter, mango butter, olive oil and rice bran oil) in a 1:1 ratio: Mixed in a 1:1 v / v ratio. Protein-fat emulsion mixed with 50% v / v liquid fat. Mix the substance with 5% wt / v pea meglobulin and use a handheld homogenizer. It was formed by emulsifying at the maximum setting for 30 seconds. After homogenization, the fat was The protein emulsion was determined to be a single liquid phase by visual observation. The cross-linking was performed using 0.2% wt / v transglutaminase enzyme at 37°C for 12 hours. It stabilized. The resulting adipose tissue imitation was a single phase, a soft solid, and had no flavor. It was salty. [Examples]

[0372] The hardness of adipose tissue at refrigeration and ambient temperatures depends on the melting temperature of the fat in the adipose tissue imitation. Controlled adipose tissue imitation The adipose tissue imitation made as a stable emulsion of RuBisCo containing sunflower oil is Adipose tissue imitation made as a stable emulsion of RuBisCo and cocoa butter Softer than regular adipose tissue. Adipose tissue imitation is 70%, 80%, and 90% v / v sunflower or Contains cocoa butter in 0.18%, 1.6%, and 2.4% wt / v forms in Rubisco. It was done. Each fatty tissue imitation containing cocoa butter is made from the corresponding imitation made from sunflower oil. It was harder than expected. Contains 0.18% v / v cocoa butter, 70%, 80%, and 90%. Adipose tissue imitation products containing 1.6% and 2.4% wt / v Rubisco can be stored at room temperature. It was solid, but melted at near mouth temperature. Rubbed with 70-80% v / v sunflower oil. Adipose tissue formed by varying the concentration of ISCO (0.18, 1.6, 1.9% wt / v) In imitation tissue, the amount of protein increases as the amount of protein in the adipose tissue imitation matrix increases. It became quite hard. Adipose tissue imitation containing 0.18% wt / v Rubisco was very soft. Soft, adipose tissue imitation containing 1.6% wt / v Rubisco is soft, 1.9% Adipose tissue imitation containing wt / v RuBisco exhibited moderate hardness.

[0373] Made as a stable emulsion of Moon bean 8S protein, containing sunflower oil. Adipose tissue mimicry is a stable emulsion of Moon bean 8S protein and cocoa butter. It was softer than the adipose tissue imitation made for testing. and containing 90% v / v sunflower or cocoa butter, 2%, 1%, and 0.5% wt Each adipose tissue model was formed using Moong bean 8S protein and cocoa butter. The replica was harder than the corresponding imitation made from sunflower oil.

[0374] Made with canola oil as a stable emulsion of Moong bean 8S protein. The adipose tissue imitation contains equal parts coconut, cocoa, olive, and palm oil. Softer than the corresponding adipose tissue imitation made as a stable emulsion of g bean 8S protein. It was soft. Adipose tissue imitation was mixed with 50%, 70%, and 90% v / v sunflower or oil. Formed with 1.4% wt / v Moong bean 8S protein, including the oil mixture. Each adipose tissue imitation containing [the specified ingredient] was harder than the corresponding imitation formed from sunflower oil. Equal amounts of coconut, cocoa, olive, and palm oil at 50%, 70%, and 90% v / v The adipose tissue imitation containing 1.4% wt / v Moong bean 8S protein is room It was solid at warm temperatures but melted at near mouth temperature.

[0375] Adipose tissue imitation made as a stable emulsion of soy protein containing sunflower oil. This is a diabetic tissue model made as a stable emulsion of soy protein and cocoa butter. It was softer than the artificial material. Fatty tissue imitation was 50%, 70%, 80%, and 90% v / v. In addition to sunflower or oil mixtures, 0.6%, 1.6%, and 2.6% wt / v soy Each adipose tissue imitation containing an oil mixture was formed using the corresponding imitation made with sunflower oil. It was harder than the manufactured product. Contains 50%, 70%, 80%, and 90% v / v cocoa butter. Adipose tissue imitation products containing 0.6%, 1.6%, and 2.6% wt / v soy protein are available. It was solid at room temperature but melted at near mouth temperature. [Examples]

[0376] Cooking of adipose tissue imitation: The structure of the adipose tissue matrix controls the melting point during cooking. Stabilized protein oil emulsion constructed as described in Examples 5 and 6 above Adipose tissue containing 2% w / v Rubisco as well as 50%, 70%, and 90% Formed with %v / v cocoa butter, and if formed by heating / cooling denaturation, at higher temperatures If melted and formed by cross-linking with transglutaminase, at a lower temperature It melted. [Examples]

[0377] Cooking of adipose tissue: The arrangement and structure of proteins and fats within the adipose tissue matrix. The composition is the amount of fat released during cooking and the amount of fat retained by the adipose tissue imitation. To control. With 2% w / v Rubisco and 50%, 70%, or 90% v / v cocoa butter During the preparation of the adipose tissue imitation matrix containing the formed protein oil emulsion, When adipose tissue imitation is formed by heating / cooling denaturation, cross-linking with transglutaminase occurs. A larger mass of adipose tissue imitation was retained after cooking than when formed by the combined method. The resulting mass appeared to be liquid and oily.

[0378] 2.6% and 0.6% w / v soy protein and 50%, 70%, or 90% w / v / v Adipose tissue imitation matrix containing a protein oil emulsion formed with cocoa butter When kousu is prepared by heating / cooling denaturation, cross-linking occurs with transglutaminase. A larger mass of adipose tissue imitation material was retained than when formed by binding. The mass appeared to be liquid and oily. [Examples]

[0379] Cooking of adipose tissue imitation: The concentration of specific proteins in the adipose tissue matrix after cooking Control the remaining clumps of adipose tissue imitation. Contains 90% v / v canola oil and 0.45% wt / v soy lecithin, totaling 1.4% wt / v. A series of adipose tissue mimics constructed from t / v moong bean 8S protein were homogenized. Then, gradually increasing concentrations of sunflower leocinicin were added to the emulsion at varying concentrations. The leosin concentration is oleosin to triglyceride molar ratio of 1:10 to 1:10. 6 Change The higher the oleosin-to-oil ratio in the adipose tissue imitation, the easier it is to maintain the mass after cooking. An increase was observed.

[0380] A series of lipids formed by varying the concentration of Rubisco, containing 70% v / v sunflower oil. The fat tissue imitation held up in larger chunks during cooking as the concentration of Rubisco increased. Adipose tissue imitation containing 0% wt / v of BisCo completely melted, while 1.9 %wt / v Rubisco retains 10% of its mass, and 2.4% wt / v Rubisco The adipose tissue imitation contained retained 20% of its mass during cooking. [Examples]

[0381] connective tissue analog Connective tissue fiber imitation is mixed with 400 mM sodium chloride and 6.75% w / v poly(vinyl) moong Prepared by electrospinning of a mameglobulin solution (22.5 mg / ml). The obtained solution was then dispensed from a 5 ml syringe at a rate of 3 μl / min using a syringe pump. The fluid was pumped through a lon tube and a smoothed 21-gauge needle. The needle was set to 17kV. Connect to the positive terminal of the fixed Spellman CZE 30kV high voltage power supply, and aluminum wheel An aluminum drum (approximately 12 cm long and 5 cm in diameter) wrapped in rubber was fixed at 12 cm. The drum is attached to a spindle that rotates at approximately 220 rpm by an IKA RW20 motor. I attached it. I connected the spindle to the ground terminal of the high-voltage power supply. Protein accumulated on the foil Polymer fibers were scraped off and used as a connective tissue imitation. [Examples]

[0382] Extension of the lifespan of reduced (heme-FE2+) leghemoglobin I purchased horse myoglobin from Sigma. I took 20mg of myoglobin at 10mg / ml. The solution was resuspended in potassium M phosphate, pH 8.0, 100 mM NaCl. (SDS-PAG) E analysis suggested that the protein purity was approximately 90%.

[0383] Soybean leghemoglobin is extracted from soybean (Glycine max) root nodules by ammonium sulfate precipitate (6 The solution was purified as detailed in Example 1 via (0% / 90% fractionation). The resuspended 90% sulfate solution was then used. Monium leghemoglobin in 20 mM potassium phosphate, pH 8.0, 100 mM Na Anion exchange chromatography in Cl (HiTrapQ FF 5mL FPLC) Further purification was performed using ram. Leghemoglobin eluted into the pass-through fraction. SDS-P AGE analysis suggested that the protein purity was approximately 70%. Replace the buffer with 20 mM potassium phosphate, pH 7.4, 100 mM NaCl, and raise to 3.5k The concentration was increased to 10 mg / ml using a Da membrane concentrator.

[0384] Carbon monoxide treatment: 100 mM potassium phosphate in 20 mM NaCl, pH 8.0. Myoglobin in mg / ml, and 20 mM potassium phosphate, pH 7.4, 100 mM Leghemoglobin in 10 mg / ml NaCl was first degassed under vacuum for 1 hour at 4°C. Next, the mixture was perfused with carbon monoxide gas for 2 minutes. Then, the globin was treated with 10 mM dithionite. Heme-Fe is extracted by adding sodium and 0.1 mM sodium hydroxide for 2 minutes. 3+ mosquito Heme-Fe 2+ It was reduced to its original state. Sodium dithionite and sodium hydroxide were added in two parts. 0 mM potassium phosphate, pH 8.0, 100 mM NaCl and 20 mM potassium phosphate Size exclusion chromatography (PD-1) in 100 mM NaCl at pH 7.4. Each was removed from the protein solution using a desalting column. Globin The fraction was collected as the peak red fraction, which was evaluated by visual estimation. The UV-VIS spectrum is Heme-Fe 2+ The presence of the state was confirmed for both proteins. After desalting, the solution was gasmed again. The solution was perfused for another 2 minutes. The color of the solution was measured every 20 minutes using a nanodrop spectrophotometer. Evaluation was performed by taking UV-Vis spectra (250nm~700nm). The samples were not treated with carbon monoxide.

[0385] Sodium nitrite treatment: 20 mM potassium phosphate, pH 8.0, 100 mM NaCl Myoglobin and 20 mM potassium phosphate at 10 mg / ml, pH 7.4, 100 Leghemoglobin in mM NaCl at 10 mg / ml concentration is reduced by 10 mM sodium dithionite. By adding 0.1 mM sodium hydroxide for 2 minutes, heme-Fe 3+ From Hem -Fe 2+ It was reduced to sodium dithionite and sodium hydroxide with 20 mM phosphorus. Potassium acid, pH 8.0, 100 mM NaCl and 20 mM potassium phosphate, pH 7 4. Size exclusion chromatography in 100 mM NaCl (PD-10 desalting chromatography) Each was removed from the protein solution using (M). The globin fraction was estimated by visual inspection. The peak red fraction, as evaluated by [method], was recovered. The UV-VIS spectrum was heme-Fe 2+ The presence of the state was confirmed for both proteins. Next, sodium nitrite was added to the phosphorus A 100 mM nitrite solution was added from an acid buffer solution at pH 7.4 until the final concentration reached 1 mM. Heme-Fe 2+ The duration of the state is determined by using a spectrophotometer to measure the UV-VIS spectrum (25 The wavelength (0-700 nm) was tracked by recording it as a function of time. The control sample was nitrite. Sodium was not used for treatment.

[0386] Myoglobin and leghemoglobin samples treated with carbon monoxide and sodium nitrite. Regarding the material, heme-Fe 2+ Analyze survival time data in Microsoft Excel This was performed by plotting the amplitude of the absorbance peak at a wavelength of 540 nm. 5 The "baseline" absorbance at 40 nm is determined by the addition of any additive, reduction of dithionite, or removal of the additive. The state was determined by the UV-vis spectral state of the globin solution before salting. The line fitting function is used to create an optimally fitting exponential line, and its exponent is half of the peak amplitude. It is directly correlated with the decay period.

[0387] Heme-Fe 2+ The duration of the condition and the attachment of myoglobin and leghemoglobin solutions The accompanying red color was obtained in the absence of carbon monoxide and sodium nitrite for approximately 6 hours and It took about 4 hours. The addition of sodium nitrite was heme-fe 2+ Duration of the state and attached The accompanying red color was extended for more than 7 days. The addition of carbon monoxide was heme-fe 2+ The continuation of the state The duration and associated red flag were extended for more than two weeks. [Examples]

[0388] The particle size of individual tissue mimics is changed to control aroma generation during cooking. Preparation of meat imitation products Muscle tissue imitation and adipose tissue imitation are prepared separately, and then the aroma generation during cooking is performed. To control the size of individual tissue imitation units, the size of the meat tissue imitation units is changed, and they are incorporated into the meat tissue imitation. Combined, individual fat, muscle, and connective tissue imitation materials were constructed in the following manner:

[0389] Muscle tissue imitation was prepared as in Example 2. The...

Claims

1. A meat imitation comprising a heme-containing protein and a nitrite compound, wherein the meat imitation does not contain meat.

2. The meat imitation according to claim 1, wherein the heme-containing protein has an amino acid sequence that has at least 90% sequence identity with the amino acid sequence shown in Sequence ID No.

1.

3. The meat imitation according to claim 1, wherein the heme-containing protein is leghemoglobin or myoglobin.

4. The meat imitation according to claim 1, wherein the heme-containing protein is selected from the group consisting of leghemoglobin, flavohemoglobin, hell's gate globin I, erythrocruorin, protoglobin, cyanoglobin, chlorocruorin, terminally cleaved hemoglobin, terminally cleaved 2 / 2 globin, and hemoglobin 3.

5. The meat imitation according to claim 1, comprising 0.01% to 5% of the heme-containing protein by weight of the meat imitation.

6. The meat imitation according to claim 1, comprising 0.4% to 1% of the heme-containing protein by weight of the meat imitation.

7. The meat imitation according to claim 1, comprising 0.3% of the heme-containing protein by weight of the meat imitation.

8. The meat imitation according to claim 1, comprising 0.2% of the heme-containing protein by weight of the meat imitation.

9. The meat imitation according to claim 1, further comprising one or more types of plant protein.

10. The meat imitation according to claim 9, wherein the one or more types of plant proteins are selected from the group consisting of Rubisco, pea protein, lentil protein, or other leguminous plant proteins.

11. The meat imitation according to claim 10, wherein the pea protein comprises pea albumin protein.

12. The meat imitation according to claim 9, wherein the one or more types of plant proteins include gluten.

13. The meat imitation according to claim 9, wherein one or more types of plant proteins are present in the meat imitation in an amount of 1% to 30% by weight of the meat imitation.

14. The meat imitation according to claim 1, further comprising glucose, ribose, sucrose, fructose, xylose, maltodextrin, and sugars selected from combinations thereof.

15. The meat imitation according to claim 1, further comprising at least one sulfur compound selected from methionine, cysteine, and thiamine.

16. The meat imitation according to claim 1, further comprising one or more types of ammonium salts, sodium salts, potassium salts, or calcium salts.

17. The meat imitation according to claim 1, which is a taco filling, casserole, sauce, topping, soup, stew or loaf.

18. The meat imitation according to claim 1, which is a sausage or pate.

19. The meat imitation according to claim 1, wherein the meat imitation is smoked or dehydrated.

20. The meat imitation according to claim 1, further comprising a fatty tissue imitation.

21. The meat imitation according to claim 20, wherein the adipose tissue imitation comprises an oil selected from the group consisting of corn oil, olive oil, soybean oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, sunflower oil, flaxseed oil, coconut oil, palm kernel oil, coconut oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, rice bran oil, and combinations thereof.

22. The meat imitation according to claim 20, wherein the adipose tissue imitation is emulsified, ultrasonically treated, or homogenized.

23. The meat imitation according to claim 1, wherein cooking generates at least two volatile compounds having an aroma associated with beef.

24. At least two volatile compounds are (E)-2-decenal, (E)-2-heptenal, (E)-2-hexenal, (E)-2-nonenal, (E)-2-octenal, (E)-4-octene, (E,E)-2,4-decadienal, (E,E)-2,4-heptadienal, (E,E)-2,4-nonadienal, (E,E)-3,5-octadien-2-one, (E,Z)-2,6-nonadienal, (Z)-2-decenal Senal, (Z)-2-heptenal, (Z)-2-nonenal, (Z)-2-octen-1-ol, (Z)-4-heptenal, 1-(1H-pyrrole-2-yl)-ethanone, 1-(2-furanyl)-ethanone, 1-(6-methyl-2-pyrazinyl)-1-ethanone, 1-(acetyloxy)-2-propanone, 1,3-dimethylbenzene, 1,3-hexadiene, 1-butanol, 1-ethyl-5-methylcyclopentene n, 1-heptanol, 1-heptene, 1-hexanol, 1H-pyrrole-2-carboxyaldehyde, 1-hydroxy-2-propanone, 1-methyl-1(H)-pyrrole-2-2-carboxyaldehyde, 1-octanal, 1-octanol, 1-octen-3-ol, 1-octen-3-one, 1-octene, 1-pentanol, 1-penten-3-ol, 1-penten-3-one, 1-propanol, 2(5H) -Furanone, 2,2,4,6,6-pentamethylheptane, 2,2-dimethyl-undecane, 2,3-butanedione, 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one, 2,3-dimethyl-5-ethylpyrazine, 2,3-dimethylpyrazine, 2,4-dodecadienal, 2,5-dimethyl-3-(3-methylbutyl)pyrazine, 2,5-dimethylpyrazine, 2,6-dimethylnonane, 2,6-dimethylpyrazine, 2-acetylthiazole, 2-butanone, 2-butenal, 2-decanone, 2-ethenyl-6-methylpyrazine, 2-ethyl-1-hexanol, 2-ethyl-5-methylpyrazine, 2-ethyl-furan, 2-furan methanol, 2-heptanone, 2-heptenal, 2-hexyl-furan, 2-hexyl-thiophene, 2-hydroxy-3-methyl-2-cyclopenten-1-one, 2-methyl Tilthiazole, 2-methyl-1H-pyrrole, 2-methyl-2-heptene, 2-methyl-butanal, 2-methyl-furan, 2-methyl-heptane, 2-methyl-propanal, 2-n-butylacrolein, 2-n-heptylfuran, 2-nonanone, 2-octanone, 2-pentanone, 2-pentyl-furan, 2-pentyl-thiophene, 2-propenal, 2-propyl-furan, 2-pyridinecarboxaldehyde D, 2-pyrrolidinone, 2-thiophenecarboxyaldehyde, 2-tridecen-1-ol, 2-undecenal, 3,5-octadien-2-one, 3,6,6-trimethyl-bicyclo[3.1.1]hepta-2-ene, 3-acetyl-1H-pyrroline, 3-ethyl-2,2-dimethylpentane, 3-ethyl-2,5-dimethylpyrazine, 3-ethyl-2-1,4-dioxin, 3-ethyl-2-methyl-1,3 -Hexadiene, 3-ethylcyclopentanone, 3-methyl-2-butenal, 3-methyl-2-thiophenecarboxaldehyde, 3-methyl-3-hexene, 3-methylbutanal, 3-methylfuran, 3-octen-2-one, 3-pentylfuran, 3-thiophenecarboxaldehyde, 3-thiophenethanol, 4,7-dimethyl-undecane, 4-cyanocyclohexene, 4-cyclopentene-1,3-dione, 4-decine, 4-methyl-5-thiazoleethanol, 4-methylthiazole, 4-octene, 4-penten-2-one, 5-acetyldihydro-2(3H)-furanone, 5-ethyldihydro-2(3H)-furanone, 5-methyl-2-furancarboxaldehyde, 5-methyl-2-thiophenecarboxaldehyde, 6-methyl-2-heptanone, 6-methyl-5-hepten-2-one, 8-methyl-1-undecene, acetaldehyde, acet Amides, acetic acid, ethenyl acetate, acetoin, acetone, acetonitrile, acetophenone, benzaldehyde, benzene, benzyl alcohol, butanal, butanoic acid, butyrolactone, caprolactam, carbon disulfide, decanal, decanol, dihydro-2-methyl-3(2H)-furanone, dihydro-5-pentyl-2(3H)-furanone, dihydro-5-propyl-2(3H)-furanone, dimethyl sulfide, dimethyl trisulfide, d - Limonene, ethenylpyrazine, ethyl acetate, ethylpyrazine, formamide, heptyl formate, furan, furaneol, furfural, heptanal, heptane, heptanoic acid, heptyl ester, hexanoic acid, isopropyl alcohol, isovaleric acid, metachlorein, methional, methyl ethanolate, methylpyrazine, methyl thiirane, vinyl n-caproate, nonanal, octanal, octane, octanoic acid, oxalic acid, butylpropyl alcohol A meat imitation according to claim 23, selected from the group consisting of pyr ester, pantolactone, p-cresol, pentanal, pentanoic acid, phenol, phenylacetaldehyde, propanal, pyrazine, pyridine, pyrrole, styrene, tetramethylpyrazine, thiazole, thiophene, toluene, trans-2-(2-pentenyl)furan, trans-3-nonen-2-one, trichloromethane, trimethylethanethiol, and trimethylpyrazine.

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