Methods and compositions for consumer products

Abstract

The present invention relates to methods and compositions for consumer products. Provided herein are methods and compositions relating to plant-based meat substitutes having properties similar to those of meat.

Description

[0001] This application is a divisional application based on the unity of invention issue raised in the first office action of the invention patent application filed on July 12, 2012, with application number 201510387835.1 and entitled "Method and Composition for Consumer Products".

[0002] Furthermore, the aforementioned invention patent application with application number 201510387835.1 is a divisional application of the invention patent application filed on July 12, 2012, with application number 201280041713.1 and title "Method and Composition for Consumer Products".

[0003] This application claims priority to U.S. Application No. 61 / 572,205, filed July 12, 2011, the entire contents of which are incorporated herein by reference. Background Technology

[0004] Animal farming has profound negative environmental impacts. Currently, it is estimated that 30% of the Earth's landmass is dedicated to animal farming, and livestock account for 20% of the total biomass of terrestrial animals. As a result, large-scale animal farming accounts for over 18% of net greenhouse gas emissions. Animal farming is likely the largest anthropogenic source of water pollution and is currently the greatest threat to biodiversity worldwide. It is estimated that if the world's human population could shift from a meat-based diet to one free of animal products, 26% of the Earth's landmass would be freed up for other uses. Furthermore, a shift to a vegetarian diet would significantly reduce water and energy consumption.

[0005] Meat consumption has profound negative effects on human health. The health benefits of a vegetarian diet are well-established. If the human population were to transition to a vegetarian diet, the cost savings in healthcare would be significant.

[0006] Hunger is a global problem, and the world’s four major commodity crops (soybeans, corn, wheat and rice) already supply more than 100% of the human population’s needs for calories and protein (including every essential amino acid).

[0007] Most plant-based meat substitutes fail to induce a shift towards vegetarianism. Current levels of meat substitute composition technology involve extruding soy / grain blends, producing products that largely fail to mimic the experience of cooking and consuming meat. A common limitation of these products is their homogeneous texture and mouthfeel compared to equivalent meat products. Furthermore, because these products must be sold pre-cooked with artificial flavors and aromas, they cannot replicate the smells, flavors, and other key characteristics associated with freshly cooked meat. Therefore, these products primarily appeal to a limited consumer base that has transitioned to vegetarianism / veganism, failing to attract the larger segment of consumers accustomed to eating meat.

[0008] This document discloses improved methods and compositions that more accurately mimic the characteristics valued by consumers in the preparation and consumption of meat and overcome the shortcomings and defects of existing meat substitutes. Summary of the Invention

[0009] In some aspects, the present invention provides a meat substitute composition comprising protein contents, wherein one or more isolated and purified proteins constitute 10% or more by weight of the protein contents, wherein the meat substitute composition precisely mimics the taste, texture or color of meat products from animal sources.

[0010] In one embodiment, the meat substitute composition precisely mimics the color of the meat product in its raw state and in its cooked state after cooking.

[0011] In another embodiment, the one or more isolated and purified proteins constitute 25% or more of the protein contents by weight.

[0012] In another embodiment, the protein of the one or more isolated and purified proteins accounts for 50% or more of the protein contents by weight.

[0013] In another embodiment, the one or more isolated and purified proteins constitute 75% or more of the protein contents by weight.

[0014] In another embodiment, the one or more isolated and purified proteins constitute 90% or more of the protein contents by weight.

[0015] In another embodiment, gluten does not constitute 10% or more of the protein contents by weight.

[0016] In another embodiment, each of the separated and purified proteins is separated and purified separately.

[0017] In another embodiment, the meat substitute composition comprises 1 to 7 isolated and purified proteins.

[0018] In another embodiment, the 1 to 7 isolated and purified proteins are each isolated from different plant species.

[0019] In some embodiments, the meat substitute comprises less than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 individual proteins isolated from one or more plant species.

[0020] In another embodiment, the protein contents contain no more than trace amounts of any other proteins from one or more plant species.

[0021] In another embodiment, the one or more isolated and purified proteins are selected from the group consisting of: legume hemoglobin, non-symbiotic hemoglobin, hemoglobin, myoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3 and cytochrome, Hell's gate globin I, bacterial hemoglobin, ciliate myoglobin. Myoglobin), flavin hemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose-1,5-bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), ribulose-1,5-bisphosphate carboxylase oxygenase activator (RuB isCO activator enzyme), albumin, soybean globulin, con-soybean globulin, globulin, pea globulin, con-albumin, gliadin, gluten, glutenin, barley gliadin, gliadin, bean globulin (protein), protein body, rye gliadin, extensin, wheat gluten, corn gliadin, any seed storage protein, oil protein, oil body calcium protein, oil body sterol protein or other oil body protein, nutrient storage protein A, nutrient storage protein B, mung bean seed storage 8S globulin.

[0022] In another embodiment, the one or more isolated and purified proteins are not isolated from animals.

[0023] In another embodiment, the one or more isolated and purified proteins are isolated from a single plant source.

[0024] In another embodiment, the one or more isolated and purified proteins are isolated from a variety of plant sources.

[0025] In another embodiment, the one or more isolated and purified proteins are isolated from genetically modified organisms.

[0026] In some embodiments, the genetically modified organism is a genetically modified bacterium or yeast organism.

[0027] In some embodiments, the separated and purified proteins have been shaped into fibers.

[0028] In a particular embodiment, the fibers are similar to skeletal muscle fibers.

[0029] In other specific embodiments, the fiber is an asymmetric fiber.

[0030] In some embodiments, the meat substitute composition further comprises one or more isolated and purified iron-containing proteins.

[0031] In some embodiments, the one or more isolated and purified iron-containing proteins are selected from the group consisting of: hemoglobin, myoglobin, legume hemoglobin, non-symbiotic hemoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavoglobin.

[0032] In one particular embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 1. SEQ ID NO 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAA KDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA.

[0033] In one particular embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 2. SEQ ID NO 2: MIDQKEKELI KESWKRIEPN KNEIGLLFYA NLFKEEPTVS VLFQNPISSQSRKLMQVLGI LVQGIDNLEG LIPTLQDLGR RHKQYGVVDS HYPLVGDCLL KSIQEYLGQG FTEEAKAAWTKVYGIAAQVM TAE. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 80% homology with SEQ ID NO 2. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 90% homology with SEQ ID NO 2. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 98% homology with SEQ ID NO 2.

[0034] In one particular embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 3. SEQ ID NO 3: MRKQPTVFEK LGGQAAMHAA VPLFYKKVLA DDRVKHYFKN TNMEHQAKQQEDFLTMLLGG PNHYKGKNMA EAHKGMNLQN SHFDAIIENL AATLKELGVS DQIIGEAAKV IEHTRKDCLGK. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 80% homology with SEQ ID NO 3. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 90% homology with SEQ ID NO 3. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 98% homology with SEQ ID NO 3.

[0035] In some embodiments, the isolated and purified proteins are assembled into one or more gels.

[0036] In some embodiments, the meat substitute composition further comprises one or more types of fat.

[0037] In a particular embodiment, one or more of the fats are derived from plant sources.

[0038] In another aspect, the present invention provides a meat substitute product comprising an indicator indicating the cooking progress from a raw state to a cooked state, wherein the meat substitute product is derived from a non-animal source.

[0039] In some embodiments, the indicator is a visual indicator that accurately simulates the color change of the meat product during the cooking process.

[0040] In one embodiment, the color changes from red to brown.

[0041] In one embodiment, the color changes from pink to white or brown.

[0042] In one embodiment, the visual indicator changes from translucent to opaque during the cooking process.

[0043] In some embodiments, the indicator is an olfactory indicator that indicates the progress of cooking.

[0044] In one embodiment, the olfactory indicator is one or more volatile odorants released during cooking.

[0045] In some embodiments, the indicator comprises one or more isolated and purified iron-containing proteins.

[0046] In a specific embodiment, the one or more isolated and purified iron-containing proteins are in a reduced state before cooking.

[0047] In one embodiment, the one or more isolated and purified iron-containing proteins are selected from the group consisting of: hemoglobin, myoglobin, legume hemoglobin, non-symbiotic hemoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and cytochrome, hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavoglobin.

[0048] In one particular embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 1. SEQ ID NO 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAK DLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA.

[0049] In some embodiments, the one or more isolated and purified iron-containing proteins are not isolated from animals. In some embodiments, the compositions of the present invention do not contain any proteins derived from animals.

[0050] In a particular embodiment, the one or more isolated and purified iron-containing proteins are isolated from one or more plant sources.

[0051] In other specific embodiments, the one or more isolated and purified proteins are isolated from root nodules, roots, seeds, leaves or stems of the one or more plant sources.

[0052] In other specific embodiments, the one or more plant sources are soybean or pea plants.

[0053] In one embodiment, the one or more plant sources comprise one or more legumes.

[0054] In some embodiments, the one or more isolated and purified iron-carrying proteins in a reduced or oxidized state have UV-visible characteristics similar to those of myoglobin proteins from animal sources in an equivalent reduced or oxidized state.

[0055] In one particular embodiment, the difference between the peak absorption wavelength of the one or more isolated and purified iron-containing proteins and the peak absorption wavelength of myoglobin from animal sources is less than 5%.

[0056] In some embodiments, the one or more isolated and purified proteins are isolated from genetically modified organisms.

[0057] In one embodiment, the genetically modified organism is a genetically modified bacterium or yeast organism.

[0058] In some embodiments, the meat substitute product is free of methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0059] In a particular embodiment, the meat substitute product further contains less than 1% wheat gluten.

[0060] In another specific embodiment, the meat substitute product is free of wheat gluten.

[0061] In other specific embodiments, the meat substitute product does not contain soy protein isolate.

[0062] In other specific embodiments, the meat substitute product does not contain soy protein concentrate.

[0063] In other specific embodiments, the meat substitute product does not contain soy protein.

[0064] In another specific embodiment, the meat substitute product contains less than 5% carbohydrates.

[0065] In other specific embodiments, the meat substitute product does not contain tofu.

[0066] In some embodiments, the meat substitute product is free of tofu and wheat gluten.

[0067] In some embodiments, the meat substitute product is free of soy protein and wheat gluten.

[0068] In some embodiments, the meat substitute product contains no animal products and contains less than 5% carbohydrates.

[0069] In some embodiments, the meat substitute product contains less than 1% cellulose.

[0070] In some embodiments, the meat substitute product contains less than 5% insoluble carbohydrates.

[0071] In some embodiments, the meat substitute product is free of soy protein and contains less than 1% cellulose.

[0072] In some embodiments, the meat substitute product is free of soy protein and contains less than 5% insoluble carbohydrates.

[0073] In some embodiments, the meat substitute product is gluten-free and contains less than 1% cellulose.

[0074] In some embodiments, the meat substitute product is free of wheat gluten and contains less than 5% insoluble carbohydrates.

[0075] In another aspect, the present invention provides a muscle tissue replica comprising protein contents, wherein the protein contents comprise one or more isolated and purified proteins, wherein the muscle tissue replica has a similar taste, texture or color to equivalent muscle tissue from an animal source.

[0076] In some embodiments, the one or more isolated and purified proteins constitute at least 50% by weight of the protein content. In some embodiments, the one or more isolated and purified proteins constitute at least 40% by weight of the protein content. In some embodiments, the one or more isolated and purified proteins constitute at least 30% by weight of the protein content. In some embodiments, the one or more isolated and purified proteins constitute at least 20% by weight of the protein content. In some embodiments, the one or more isolated and purified proteins constitute at least 10% by weight of the protein content.

[0077] In some embodiments, the one or more isolated and purified proteins constitute at least 50% by weight of the composition contents. In some embodiments, the one or more isolated and purified proteins constitute at least 40% by weight of the composition contents. In some embodiments, the one or more isolated and purified proteins constitute at least 30% by weight of the composition contents. In some embodiments, the one or more isolated and purified proteins constitute at least 20% by weight of the composition contents. In some embodiments, the one or more isolated and purified proteins constitute at least 10% by weight of the composition contents. In some embodiments, the one or more isolated and purified proteins constitute at least 5% by weight of the composition contents. In some embodiments, the one or more isolated and purified proteins constitute at least 1% by weight of the composition contents.

[0078] In some embodiments, the protein contents are derived from one or more non-animal sources.

[0079] In a specific embodiment, one or more non-animal sources are plant-based.

[0080] In other specific embodiments, the one or more non-animal sources are genetically modified yeast or bacteria.

[0081] In some embodiments, each of the one or more isolated proteins is separately isolated and purified.

[0082] In some embodiments, the one or more isolated proteins are selected from the group consisting of: hemoglobin, myoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and cytochromes, hellgammin I, bacterial hemoglobin, ciliate myoglobin, flavin hemoglobin, ribosomal proteins, actin, hexokinase, lactate dehydrogenase, fructose-2-phosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase. Actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), ribulose-1,5-bisphosphate carboxylase oxygenase activator (RuBisCO activator), albumin, soybean globulin, con-soybean globulin, globulin, pea globulin, con-albumin, gliadin, gluten, gluten, barley gliadin, gliadin, bean globulin (protein), protein body, rye gliadin, extensin, wheat gluten, zein, any seed storage protein, oily protein, oily body calciprotein, oily body sterol protein or other oily body protein, nutrient storage protein A, nutrient storage protein B, mung bean seed storage 8S globulin.

[0083] In one embodiment, the seed storage protein is mung bean 8S protein.

[0084] In some embodiments, the protein contents are suspended in a gel.

[0085] In some embodiments, the protein contents are in the form of a gel.

[0086] In one embodiment, the gel contains a separated and purified cross-linking enzyme.

[0087] In some embodiments, the isolated and purified cross-linking enzyme is selected from the group consisting of: transglutaminase, lysyl oxidase, and amine oxidase.

[0088] In one specific embodiment, the separated and purified cross-linking enzyme is glutamine transaminase.

[0089] In some embodiments, the protein contents have been assembled into fibers.

[0090] In a particular embodiment, the fibers are arranged isotropically.

[0091] In one embodiment, the fiber is an asymmetric fiber.

[0092] In some embodiments, the muscle tissue replica further comprises one or more isolated and purified iron-containing proteins.

[0093] In some embodiments, the one or more isolated and purified iron-containing proteins are selected from the group consisting of: hemoglobin, myoglobin, legume hemoglobin, non-symbiotic hemoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and cytochrome, hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavin hemoglobin.

[0094] In one specific embodiment, the one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 70% homology with SEQ ID NO 1. SEQ ID NO 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAKDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA. In one specific embodiment, the one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 80% homology with SEQ ID NO 1. In one specific embodiment, the one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 90% homology with SEQ ID NO 1. In one specific embodiment, the one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 95% homology with SEQ ID NO 1. In one particular embodiment, the one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 98% homology with SEQ ID NO 1.

[0095] In a particular embodiment, the muscle tissue replica comprises protein contents, wherein (i) a separated and purified protein that is not an iron-containing protein comprises 40 to 95% of the protein contents, (ii) one or more separated and purified iron-containing proteins comprises 1 to 20% of the protein contents, and (iii) one or more cross-linking agents comprises 0.1 to 35% of the protein contents.

[0096] In one embodiment, the protein contents comprise 5 to 50% of the replica by weight or by weight / volume.

[0097] In one embodiment, the isolated and purified protein is mung bean 8S protein.

[0098] In one embodiment, the one or more iron-containing proteins that have been separated and purified are soy leghemoglobin.

[0099] In one embodiment, the one or more cross-linking agents are transglutaminase.

[0100] In some embodiments, the muscle tissue replica does not contain methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, or acacia gum.

[0101] In a particular embodiment, the muscle tissue imitation further contains less than 1% wheat gluten. In a particular embodiment, the muscle tissue imitation further contains less than 5% wheat gluten. In a particular embodiment, the muscle tissue imitation further contains less than 10% wheat gluten. In a particular embodiment, the muscle tissue imitation further contains less than 0.1% wheat gluten.

[0102] In another specific embodiment, the muscle tissue replica is free of wheat gluten.

[0103] In other specific embodiments, the muscle tissue replica does not contain soy protein isolate.

[0104] In other specific embodiments, the muscle tissue replica does not contain soy protein concentrate.

[0105] In other specific embodiments, the muscle tissue replica does not contain soy protein.

[0106] In another specific embodiment, the muscle tissue replica contains less than 5% carbohydrates.

[0107] In other specific embodiments, the muscle tissue replica does not contain tofu.

[0108] In some embodiments, the muscle tissue replica is free of tofu and wheat gluten.

[0109] In some embodiments, the muscle tissue replica is free of soy protein and wheat gluten.

[0110] In some embodiments, the muscle tissue replica is free of animal products and contains less than 5% carbohydrates.

[0111] In some embodiments, the muscle tissue replica contains less than 1% cellulose.

[0112] In some embodiments, the muscle tissue replica contains less than 5% insoluble carbohydrates.

[0113] In some embodiments, the muscle tissue replica does not contain soy protein and contains less than 1% cellulose.

[0114] In some embodiments, the muscle tissue replica does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0115] In some embodiments, the muscle tissue replica is free of wheat gluten and contains less than 1% cellulose.

[0116] In some embodiments, the muscle tissue replica is free of wheat gluten and contains less than 5% insoluble carbohydrates.

[0117] In some embodiments, the muscle tissue replica does not contain methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, or acacia gum.

[0118] In a specific embodiment, the meat imitation contains less than 1% wheat gluten. In a specific embodiment, the meat imitation contains less than 5% wheat gluten. In a specific embodiment, the meat imitation contains less than 10% wheat gluten. In a specific embodiment, the meat imitation contains less than 0.1% wheat gluten.

[0119] In another specific embodiment, the meat imitation does not contain wheat gluten.

[0120] In other specific embodiments, the meat imitation does not contain soy protein isolate.

[0121] In other specific embodiments, the meat imitation does not contain soy protein concentrate.

[0122] In other specific embodiments, the meat imitation does not contain soy protein.

[0123] In another specific embodiment, the meat imitation contains less than 5% carbohydrates.

[0124] In other specific embodiments, the meat imitation does not contain tofu.

[0125] In some embodiments, the meat imitation does not contain tofu and does not contain wheat gluten.

[0126] In some embodiments, the meat imitation is free of soy protein and wheat gluten.

[0127] In some embodiments, the meat imitation contains no animal products and contains less than 5% carbohydrates.

[0128] In some embodiments, the meat imitation contains less than 1% cellulose. In some embodiments, the meat imitation contains less than 0.1% cellulose. In some embodiments, the meat imitation contains less than 10% cellulose. In some embodiments, the meat imitation contains less than 5% cellulose.

[0129] In some embodiments, the meat imitation contains less than 5% insoluble carbohydrates.

[0130] In some embodiments, the meat imitation does not contain soy protein and contains less than 1% cellulose.

[0131] In some embodiments, the meat imitation does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0132] In some embodiments, the meat imitation is free of wheat gluten and contains less than 1% cellulose.

[0133] In some embodiments, the meat imitation is free of wheat gluten and contains less than 5% insoluble carbohydrates.

[0134] In another aspect, the present invention provides an adipose tissue replica comprising a gel emulsion, said gel emulsion comprising a protein solution in which fat droplets are suspended.

[0135] In some embodiments, the fat droplets are derived from non-animal sources.

[0136] In some embodiments, the fat droplets comprise one or more vegetable oils.

[0137] In some embodiments, the one or more vegetable oils are 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, algae oil, palm oil, palm kernel oil, coconut oil, and babassu oil. Oils, including butyrate, mango butter, cocoa butter, wheat germ oil, rice bran oil, oils produced by bacteria, algae, archaea, or fungi, or genetically engineered bacteria, algae, archaea, or fungi, triglycerides, monoglycerides, diglycerides, sphingomyelin, glycolipids, lecithin, lysophosphatidylcholine, phosphatidic acid, lysophosphatidic acid, oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidin, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentanic acid, docosahexaenoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid, or esters of the following substances: oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidin, undecanoic acid. The following are triglyceride derivatives of oleic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidic acid, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidic acid, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid.

[0138] In one embodiment, the one or more vegetable oils are rice bran oil or rapeseed oil.

[0139] In some embodiments, the protein solution contains one or more isolated and purified proteins.

[0140] In some embodiments, the one or more separated and purified proteins account for 75% or more of the proteins in the protein solution.

[0141] In some embodiments, the one or more isolated and purified proteins are derived from non-animal sources.

[0142] In some embodiments, the non-animal source is a plant source.

[0143] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0144] In some embodiments, each of the one or more isolated proteins is separately isolated and purified.

[0145] In some embodiments, the one or more isolated proteins are selected from the group consisting of: hemoglobin, myoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and cytochromes, hellgammin I, bacterial hemoglobin, ciliate myoglobin, flavin hemoglobin, ribosomal proteins, actin, hexokinase, lactate dehydrogenase, fructose-2-phosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase. Actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), ribulose-1,5-bisphosphate carboxylase oxygenase activator (RuBisCO activator), albumin, soybean globulin, con-soybean globulin, globulin, pea globulin, con-albumin, gliadin, gluten, gluten, barley gliadin, gliadin, bean globulin (protein), protein body, rye gliadin, extensin, wheat gluten, zein, any seed storage protein, oily protein, oily body calciprotein, oily body sterol protein or other oily body protein, nutrient storage protein A, nutrient storage protein B, mung bean seed storage 8S globulin.

[0146] In some embodiments, the one or more isolated and purified proteins are albumin proteins, seed storage proteins, or pea globulin proteins.

[0147] In a particular embodiment, the albumin protein is isolated pea albumin protein.

[0148] In some embodiments, the seed storage protein is mung bean 8S protein.

[0149] In some embodiments, the gel emulsion comprises: a protein solution containing 1 to 3 isolated and purified proteins, wherein the solution comprises 30 to 70% of the volume of the emulsion; a vegetable oil, wherein the vegetable oil comprises 30 to 70% of the volume of the emulsion; and an isolated and purified cross-linking enzyme, wherein the cross-linking enzyme comprises 0.5 to 5% of the emulsion by weight / volume; wherein the vegetable oil is emulsified in the protein solution, and wherein the emulsion forms a gel by the cross-linking enzyme.

[0150] In other embodiments, the gel emulsion comprises: a protein solution containing 1 to 3 isolated and purified proteins, wherein the solution comprises 1 to 30% of the volume of the emulsion; a vegetable oil, wherein the vegetable oil comprises 70 to 99% of the volume of the emulsion; and an isolated and purified cross-linking enzyme, wherein the cross-linking enzyme comprises 0.5 to 5% of the emulsion by weight / volume; wherein the vegetable oil is emulsified in the protein solution, and wherein the emulsion forms a gel by the cross-linking enzyme.

[0151] In some embodiments, the fat imitation further comprises a cross-linking enzyme.

[0152] In some embodiments, the cross-linking enzyme is glutamine transaminase.

[0153] In some embodiments, one of the 1 to 3 isolated and purified proteins is mung bean 8S protein, pea albumin protein, or pea globulin protein.

[0154] In a particular embodiment, the vegetable oil is rice bran oil or rapeseed oil.

[0155] In some embodiments, the adipose tissue imitation does not contain methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, or acacia gum.

[0156] In a particular embodiment, the adipose tissue imitation further contains less than 1% wheat gluten.

[0157] In another specific embodiment, the adipose tissue imitation does not contain wheat gluten.

[0158] In other specific embodiments, the adipose tissue imitation does not contain soy protein isolate.

[0159] In other specific embodiments, the adipose tissue imitation does not contain soy protein concentrate.

[0160] In other specific embodiments, the adipose tissue imitation does not contain soy protein.

[0161] In another specific embodiment, the adipose tissue replica contains less than 5% carbohydrates.

[0162] In other specific embodiments, the adipose tissue imitation does not contain tofu.

[0163] In some embodiments, the adipose tissue imitation does not contain tofu and does not contain wheat gluten.

[0164] In some embodiments, the adipose tissue imitation is free of soy protein and wheat gluten.

[0165] In some embodiments, the adipose tissue replica is free of animal products and contains less than 5% carbohydrates.

[0166] In some embodiments, the adipose tissue replica contains less than 1% cellulose.

[0167] In some embodiments, the adipose tissue replica contains less than 5% insoluble carbohydrates.

[0168] In some embodiments, the adipose tissue imitation does not contain soy protein and contains less than 1% cellulose.

[0169] In some embodiments, the adipose tissue imitation does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0170] In some embodiments, the adipose tissue imitation is free of wheat gluten and contains less than 1% cellulose.

[0171] In some embodiments, the adipose tissue imitation is free of wheat gluten and contains less than 5% insoluble carbohydrates.

[0172] In another aspect, the present invention provides a connective tissue replica comprising protein contents containing one or more isolated and purified proteins, wherein the protein contents have been assembled into a structure that approximates the texture and visual appearance of connective tissue or skin.

[0173] In some embodiments, the protein contents are derived from non-animal sources.

[0174] In some embodiments, the non-animal source is a plant source.

[0175] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0176] In some embodiments, the one or more isolated proteins constitute 50% or more of the protein contents by weight.

[0177] In some embodiments, the isolated and purified protein comprises 90% or more of the protein contents by weight.

[0178] In some embodiments, each of the one or more isolated proteins is separately isolated and purified.

[0179] In some embodiments, the one or more isolated proteins are selected from the group consisting of: hemoglobin, myoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and cytochromes, hellgammin I, bacterial hemoglobin, ciliate myoglobin, flavin hemoglobin, ribosomal proteins, actin, hexokinase, lactate dehydrogenase, fructose-2-phosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase. Actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), ribulose-1,5-bisphosphate carboxylase oxygenase activator (RuBisCO activator), albumin, soybean globulin, con-soybean globulin, globulin, pea globulin, con-albumin, gliadin, gluten, gluten, barley gliadin, gliadin, bean globulin (protein), protein body, rye gliadin, extensin, wheat gluten, zein, any seed storage protein, oily protein, oily body calciprotein, oily body sterol protein or other oily body protein, nutrient storage protein A, nutrient storage protein B, mung bean seed storage 8S globulin.

[0180] In some embodiments, the one or more isolated and purified proteins are protonin family proteins.

[0181] In some embodiments, the one or more isolated and purified proteins are zein.

[0182] In some embodiments, the protein contents are suspended in a gel.

[0183] In some embodiments, the gel contains a separated and purified cross-linking enzyme.

[0184] In some embodiments, the isolated and purified cross-linking enzyme is selected from the group consisting of: transglutaminase, lysyl oxidase, and amine oxidase.

[0185] In some embodiments, the separated and purified cross-linking enzyme is glutamine transaminase.

[0186] In some embodiments, the protein contents are shaped into fibers.

[0187] In some embodiments, the fibers are produced by an extrusion process.

[0188] In some embodiments, the fibers are stabilized by protein cross-linking.

[0189] In some embodiments, the fiber contains a separated and purified cross-linking enzyme.

[0190] In some embodiments, the isolated and purified cross-linking enzyme is selected from the group consisting of: transglutaminase, lysyl oxidase, and amine oxidase.

[0191] In some embodiments, the separated and purified cross-linking enzyme is glutamine transaminase.

[0192] In another aspect, the present invention provides a meat substitute product comprising: a muscle imitation; an adipose tissue imitation; and a connective tissue imitation; wherein the muscle imitation, adipose tissue imitation, and / or connective tissue imitation are assembled in a manner that closely approximates the physical structure of meat.

[0193] In some embodiments, the meat substitute product comprises two or more of the muscle imitation, adipose tissue imitation, and connective tissue imitation.

[0194] In some embodiments of the meat substitute product, the muscle imitation comprises 40 to 90% of the product by weight, the adipose tissue imitation comprises 1 to 60% of the product by weight, and the connective tissue imitation comprises 1 to 30% of the product by weight.

[0195] In some embodiments, the meat substitute product comprises: 60 to 90% water; 5 to 30% protein content; and 1 to 20% fat or fat imitation; wherein the protein content comprises one or more isolated and purified plant proteins.

[0196] In some embodiments, the protein contents are derived from non-animal sources.

[0197] In some embodiments, the non-animal source is a plant source.

[0198] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0199] The protein contents comprise 50% or more by weight of one or more isolated and purified proteins.

[0200] In some embodiments, each of the one or more isolated proteins is separately isolated and purified from different plant species.

[0201] In some embodiments, one or more of the isolated proteins are selected from the group consisting of: hemoglobin, myoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, phycoglobin, HbO, Glb3, and cytochrome, hellgammin I, bacterial hemoglobin, ciliate myoglobin, flavin hemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose diphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, and pyruvate decarboxylase. Actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), ribulose-1,5-bisphosphate carboxylase oxygenase activator (RuBisCO activator), albumin, soybean globulin, con-soybean globulin, globulin, pea globulin, con-albumin, gliadin, gluten, glutenin, barley gliadin, gliadin, bean globulin (protein), protein body, rye gliadin, extensin, wheat gluten, zein, any seed storage protein, oily protein, oily body calciprotein, oily body sterol protein or other oily body protein, nutrient storage protein A, nutrient storage protein B, mung bean seed storage 8S globulin.

[0202] In some embodiments, the meat substitute product further comprises one or more isolated and purified iron-containing proteins.

[0203] In some embodiments, the one or more isolated and purified iron-containing proteins are selected from the group consisting of: hemoglobin, myoglobin, legume hemoglobin, non-symbiotic hemoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavin hemoglobin. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 1. [SEQ ID NO 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAK DLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA]

[0204] In some embodiments, the meat substitute product is free of methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0205] In a particular embodiment, the meat substitute product further contains less than 1% wheat gluten.

[0206] In another specific embodiment, the meat substitute product is free of wheat gluten.

[0207] In other specific embodiments, the meat substitute product does not contain soy protein isolate.

[0208] In other specific embodiments, the meat substitute product does not contain soy protein concentrate.

[0209] In other specific embodiments, the meat substitute product does not contain soy protein.

[0210] In another specific embodiment, the meat substitute product contains less than 5% carbohydrates.

[0211] In other specific embodiments, the meat substitute product does not contain tofu.

[0212] In some embodiments, the meat substitute product is free of tofu and wheat gluten.

[0213] In some embodiments, the meat substitute product is free of soy protein and wheat gluten.

[0214] In some embodiments, the meat substitute product contains no animal products and contains less than 5% carbohydrates.

[0215] In some embodiments, the meat substitute product contains less than 1% cellulose.

[0216] In some embodiments, the meat substitute product contains less than 5% insoluble carbohydrates.

[0217] In some embodiments, the meat substitute product is free of soy protein and contains less than 1% cellulose.

[0218] In some embodiments, the meat substitute product is free of soy protein and contains less than 5% insoluble carbohydrates.

[0219] In some embodiments, the meat substitute product is gluten-free and contains less than 1% cellulose.

[0220] In some embodiments, the meat substitute product is free of wheat gluten and contains less than 5% insoluble carbohydrates.

[0221] In another aspect, the present invention provides a food product comprising one or more isolated and purified iron-containing proteins, wherein the food product is configured for animal consumption.

[0222] In some embodiments, the one or more isolated and purified iron-containing proteins are derived from non-animal sources.

[0223] In some embodiments, the non-animal source is a plant source.

[0224] In some embodiments, the plant source comprises one or more legumes.

[0225] In some embodiments, the one or more legumes are soybeans or peas.

[0226] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0227] In some embodiments, the iron-containing protein is selected from the group consisting of: hemoglobin, myoglobin, legume hemoglobin, non-symbiotic hemoglobin, hemochloroglobin, invertebrate hemoglobin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanobacterial globin, HbO, Glb3, and cytochrome, hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavoglobin.

[0228] In one embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 1. [SEQ ID NO 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAK DLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA]

[0229] In some embodiments, the food product is free of methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0230] In a particular embodiment, the food product further contains less than 1% wheat gluten.

[0231] In another specific embodiment, the food product is free of wheat gluten.

[0232] In other specific embodiments, the food product does not contain soy protein isolate.

[0233] In other specific embodiments, the food product does not contain soy protein concentrate.

[0234] In other specific embodiments, the food product does not contain soy protein.

[0235] In another specific embodiment, the food product contains less than 5% carbohydrates.

[0236] In other specific embodiments, the food product does not contain tofu.

[0237] In some embodiments, the food product is free of tofu and wheat gluten.

[0238] In some embodiments, the food product is free of soy protein and wheat gluten.

[0239] In some embodiments, the food product does not contain animal products and contains less than 5% carbohydrates.

[0240] In some embodiments, the food product contains less than 1% cellulose.

[0241] In some embodiments, the food product contains less than 5% insoluble carbohydrates.

[0242] In some embodiments, the food product is free of soy protein and contains less than 1% cellulose.

[0243] In some embodiments, the food product is free of soy protein and contains less than 5% insoluble carbohydrates.

[0244] In some embodiments, the food product is free of wheat gluten and contains less than 1% cellulose.

[0245] In some embodiments, the food product is free of wheat gluten and contains less than 5% insoluble carbohydrates.

[0246] In another aspect, the present invention provides a method for preparing a meat substitute composition, comprising isolating and purifying one or more proteins; and assembling said one or more proteins into a physical structure that approximates the physical structure of meat.

[0247] In another aspect, the present invention provides a method for producing a muscle tissue replica, comprising isolating and purifying one or more proteins; and assembling said one or more proteins into a physical tissue that closely resembles the physical tissue of skeletal muscle.

[0248] In another aspect, the present invention provides a method for producing adipose tissue replicas, comprising isolating and purifying one or more proteins; preparing a solution containing one or more proteins; emulsifying one or more fats in the solution; and stabilizing the solution into a gel emulsion using one or more crosslinking agents.

[0249] In another aspect, the present invention provides a method for producing a connective tissue replica, comprising isolating and purifying one or more proteins; and precipitating the one or more proteins, wherein the precipitation causes the one or more proteins to form a physical structure that closely resembles the physical structure of connective tissue.

[0250] In some embodiments, the precipitation comprises dissolving one or more proteins in a first solution; and squeezing the first solution into a second solution, wherein the one or more proteins are insoluble in the second solution, wherein the squeezing induces the one or more proteins to precipitate.

[0251] In another aspect, the present invention provides a food product comprising one or more isolated and purified iron-containing proteins, wherein the food product is configured for animal consumption.

[0252] In another aspect, the present invention provides a food product comprising one or more isolated and purified iron-containing proteins, wherein the food product is configured for human consumption.

[0253] In another aspect, the present invention provides a food product comprising one or more isolated and purified iron-containing proteins, wherein the food product is configured for animal consumption. In yet another aspect, the present invention provides a food product comprising one or more isolated and purified iron-containing proteins, wherein the food product is configured for human consumption.

[0254] All publications and patent applications mentioned in this specification are incorporated herein by reference to the same extent that each individual publication or patent application is specifically and individually indicated to be incorporated in general by way of introduction. Attached Figure Description

[0255] This patent or application document contains at least one color drawing. A copy of this patent or patent application publication containing the color drawing will be provided by the Patent Office upon request and payment of the necessary fees.

[0256] The novel features of the invention are given by the characteristics described in the appended claims. The features and advantages of the invention will be better understood with reference to the following detailed description of illustrative embodiments, in which the principles of the invention are used, and wherein the accompanying drawings are provided.

[0257] Figure 1 A comparison of aligned GCMS distributions (mirror-oriented) of otherwise identical muscle sculptors cooked in the presence of either leghemoglobin (LHb, right-hand feature) or ferric ions (left-hand feature) is depicted. In this figure, the vertical axis represents the residence time during the gas chromatographic separation step, and the horizontal axis represents the m / z ratio of the ions produced by fragmenting the corresponding separated volatile compounds. Selected compounds that differ in their representation in the two samples are marked on the right side of the figure.

[0258] Figure 2 The image depicts a portion of the root of a pea plant (Pisum sativum), with the root nodule cut open to reveal the red color imparted by the leghemoglobin contained therein. The cut nodule appears red.

[0259] Figure 3 Depicting leghemoglobin isolated from 1 ounce of pea root. The red color, typically characteristic of meat, is clearly visible in the color photograph.

[0260] Figure 4 The results show that leghemoglobin from different species are homologous and have similar color properties. Figure 4 In Group A, SDS-PAGE gels of dissolved root nodules from the following three leguminous species are shown: (1) broad bean, (2) English pea, and (3) soybean. Arrows indicate the corresponding leghemoglobin. Group B shows the similarity of the UV-Vis spectral distribution of leghemoglobin from two different plant species (broad bean and soybean).

[0261] Figure 5 Showing reduced (heme iron 2+) and oxidized (heme iron 3+) soybean leghemoglobin ( Figure 5 Group A) and equine cardiac myoglobin ( Figure 5 The comparison of Group B shows the similarity of the UV-Vis absorption distribution of the two proteins. We purified soybean leghemoglobin from soybean root nodules using the protocol described here. Purified horse myoglobin was purchased from Sigma-Aldrich. Soybean leghemoglobin was reduced with 1 mm sodium bisulfite ( Figure 5 Group A) and horse myoglobin ( Figure 5 Group B). Showing soybean leghemoglobin ( Figure 5 Group A) and horse myoglobin ( Figure 5 Group B) heme Fe3+ (blue line) Figure 5 and 6 The UV-Vis absorption spectra of the higher peaks in the spectrum and heme Fe2+ (red line). The inset shows a magnified view of the UV-Vis spectrum in the 450 nm to 700 nm region. Figure 5 Group C), the image of a 10 μl droplet (left droplet) of a 40 mg / ml solution of soybean leghemoglobin in the Fe3+ state shows a characteristic rust-red color, and the image of a 10 μl droplet (right droplet) of a 40 mg / ml solution of soybean leghemoglobin in the Fe2+ state shows a characteristic red color. The characteristic rust-red color and the characteristic red color are present in and correspond to the sample of horse myoglobin (right image).

[0262] Figure 6 An example is described of the successful reduction of heme iron from soybean leghemoglobin using sodium bisulfite and titanium citrate. Figure 6 In the images, the UV-Vis spectra of purified soybean leghemoglobin in the oxidized (+3) state of heme iron are represented by blue curves in each group (blue curves have higher peaks on the main plot). The red curves in each group represent the UV-Vis spectra of the same leghemoglobin species reduced to the (+2) state (red line) after addition of (Group A) 1 mM sodium bisulfite or (Group B) a 20 mM potassium phosphate buffer (pH 7.3) containing 0.24% (wt / v) titanium citrate and 100 mM sodium chloride. The inset shows a magnified view of the UV-Vis spectrum in the 450 to 700 nm region. For this example, leghemoglobin was purified from soybean root nodules using 60 / 90% ammonium sulfate fractionation, and the leghemoglobin was exchanged into a 20 mM potassium phosphate buffer (pH 7.4) and 100 mM sodium chloride buffer. A sodium bisulfite stock solution was prepared by dissolving 100 mM sodium bisulfite in 1 mM sodium hydroxide in water. A titanium citrate stock solution was prepared from 20% (wt / v) titanium chloride in hydrochloric acid (by mixing it with 0.2 M sodium citrate (1:10 v / v)). The pH was adjusted to 7.0 using sodium carbonate.

[0263] Figure 7Examples of soybean leghemoglobin purification streams from soybean root nodules are depicted. The diagrams show SDS-PAGE fractionation of different soybean leghemoglobin purification steps: (lane 1) soybean root nodule lysate; (lane 2) soybean root nodule lysate purified by fractionation with 60 / 90% (wt / v) ammonium sulfate. The protein contents of protein globules fractionated with 90% ammonium sulfate are shown, the globules being resuspended in 20 mM potassium phosphate pH 7.4, 100 mM sodium chloride, 1 mM EDTA buffer; the protein from the 90% ammonium sulfate globules was further purified by anion exchange chromatography (FFQ GE Healthcare) with 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride. Leghemoglobin collected in the effluent is shown in lane 3. Size exclusion chromatography (Sephacryl S-100 GE Healthcare) was used to fractionate the anion exchange effluent, and the resulting leghemoglobin fraction is shown in lane 4. The contents of leghemoglobin at different purification steps were determined by analyzing the fractions of the leghemoglobin band on the SDS-PAGE gel using ImageDoc analysis software (BioRad). The purity (partial abundance) of leghemoglobin at the corresponding purification steps was as follows: dissolved matter: 32.7% (lane 1), 60 / 90% (wt / v) ammonium sulfate fractionation: 78% (lane 2), anion exchange chromatography: approximately 83% (lane 3), and size exclusion chromatography: approximately 95% (lane 4).

[0264] Figure 8 The following are SDS-PAGE gel analyses of stained substances: (A) soybean leghemoglobin expressed and purified using recombinant protein technology, and (B) soybean leghemoglobin purified from soybean root nodules. (A) Recombinant soybean leghemoglobin A carrying a His tag and a TEV protease His tag removal site was expressed in *E. coli* strain BL21 and purified using His tag affinity chromatography (Talon resin, CloneTech). The left lane contains molecular weight standards, and the right lane contains purified recombinant soybean leghemoglobin A (arrow). The expected molecular weight of recombinant soybean leghemoglobin A is 17.1 kDa. (B) SDS-PAGE gel of soybean leghemoglobin purified from root nodules. The left lane contains molecular weight standards, and the right lane contains purified soybean leghemoglobin A (arrow). Mass spectrometry analysis of the purified material confirmed the presence of all four soybean leukoglobin isoforms, all of which were full-length (data not shown). The expected molecular weight (MW) of the soybean leukoglobin isoforms is in the range of 15.4 to 15.8 kDa.

[0265] Figure 9This image shows six examples of commercially available meat analogues (Quorn chicken analogues), each approximately 1 cm on one side. Four of these (left and bottom right) have been soaked in a solution of approximately 10 mg / ml soybean leghemoglobin in 20 mM potassium phosphate (pH 7.4) and 100 mM NaCl; the remaining two (top right) have been soaked in the same buffer solution without soybean leghemoglobin. The deep pink of the cubes infused with soybean leghemoglobin contrasts sharply with the pale brown of the uninfused cubes in the color photograph.

[0266] Figure 10 Four cubes of plant-based chicken analogue, infused with bean-based leghemoglobin, are shown cooking in a pan at 350°C. The two lower cubes have been flipped to expose the roasted, browned surface. In the two upper cubes, where the heated portion has turned grayish-brown, the cooler top surface remains pink. In some embodiments, a solution containing heme (e.g., bean-based leghemoglobin solution) is injected into the consumer product until it resembles the color of uncooked meat.

[0267] Figure 11 The mixture was prepared by mixing 43 ml of mung bean protein solution (150 mg / ml in dialysis buffer) with 37 ml of soybean leghemoglobin solution (46.5 mg / ml soybean leghemoglobin and 20 mg / ml other soybean root nodule proteins in 20 mM potassium phosphate, 100 mM NaCl, pH 7.3). 20 ml of transglutaminase solution (20% w / w) was added, and the mixture was thoroughly mixed. The solution was then aliquoted into two 50 ml Falcon tubes and incubated overnight at room temperature. The final protein concentrations were 65 mg / ml mung bean protein, 18 mg / ml soybean leghemoglobin, and 91 mg / ml total protein.

[0268] Figure 12 A “white” muscle analogue was prepared by mixing 43 ml of mung bean protein solution (150 mg / ml) with 45 ml of 11.7 mg / ml leghemoglobin solution and 0.8% (wt / v) transglutaminase solution. The final protein concentrations were 63 mg / ml mung bean protein, 5.2 mg / ml leghemoglobin, and 68 mg / ml total protein.

[0269] Figure 13Describe a mung bean-based adipose tissue analog prepared in an eppendorf tube, which forms an opaque gel with a grayish-white, smooth, homogeneous texture and no visible, identifiable unincorporated liquid. The gel is free-standing, elastic, and springy. It has a slightly pleasant odor and a mild, pleasant flavor. The taste is slightly salty.

[0270] Figure 14 A tissue analog based on pea globulin and prepared in Eppendorf tubes is described, which is extremely similar to a fat analog based on mung beans, except that it deforms less upon compression.

[0271] Figure 15 The strands of connective tissue analogues are shown, generated by using 70% ethanol at a 1:3 ratio in a syringe fitted with a 23-gauge needle (0.337 mm inner diameter). The solution is slowly squeezed from the bottom of a 5-inch-high container into an excess of 5 M NaCl solution. The ethanol-zein solution, being less dense than the NaCl solution, floats upward, pulling out the fibrous scaffold of the solidified zein. The NaCl is continuously stirred as the strands begin to develop, thus aiding in their elongation. The strands aggregate and form hard, dense masses.

[0272] Figure 16 The text describes the preparation of ground beef prototype patties using a combination of 62% (wt / wt) muscle analogues (62% (wt / wt) "dark muscle analogues" and 38% (wt / wt) "white muscle analogues"), 29% (wt / wt) adipose tissue analogues (derived from pea globulin and canola oil), and 5% (wt / wt) connective tissue analogues. Figure 16 Group A). ​​Ground beef patties were prepared by combining 62% muscle analogue (62% "dark muscle analogue" and 38% "white muscle analogue"), 29% adipose tissue analogue (derived from mung bean seed 8S protein and rice bran oil), and 5% connective tissue analogue. Figure 16 Group B). Ground beef patties were made by combining 71% (wt / wt) muscle tissue analogue (composed of 60% "white" muscle analogue and 40% "dark" muscle analogue) and 23% adipose tissue (derived from pea seed globulin protein and canola oil). Figure 16 Group C). Ground beef prototype patties were made by combining 67% "white" muscle analogue with 28% adipose tissue analogue (derived from pea globulin and rice bran oil). Figure 16 (Group D).

[0273] Figure 17The study depicted the preparation of ground beef patties using a combination of 62% (wt / wt) muscle tissue analogues (62% (wt / wt) "dark muscle analogues" and 38% (wt / wt) "white muscle analogues"), 29% (wt / wt) adipose tissue analogues (derived from pea globulin and canola oil), and 5% (wt / wt) connective tissue analogues. The left set shows the patties before cooking, and the right set shows the same patties after approximately 2 minutes of cooking. The observer described the odor of the cooked ground beef patties as distinctly "beefy." Detailed Implementation

[0274] This document describes methods and compositions for producing consumer products. These consumer products are intended for animal consumption. For example, they may be food suitable for human consumption. The consumer products may be approved by a suitable regulatory authority. They may be sold in grocery stores or prepared in restaurants, schools, hospitals, military facilities, prisons, shelters, long-term care facilities, similar to existing human food. The consumer products may also be food for domesticated animals. For example, dog food may be produced according to the present invention. The consumer products may also be food for wild animals. For example, they may be provided to non-domesticated carnivores.

[0275] The consumer products of this invention can counter, supplement, or replace animal-based foods. For example, the consumer products can be meat imitations made entirely from plant sources. The consumer products can be manufactured to mimic the cuts or appearance of meat currently sold. For example, the consumer products can be visually similar to or indistinguishable from ground beef or specific cuts of beef. Alternatively, the consumer products can be manufactured to have a unique appearance or shape. For example, the consumer products can contain patterns or lettering based on the structure of the consumer products. In some cases, the consumer products, after being prepared, resemble traditional meat products. For example, the consumer products can be produced to be larger than traditional beef cuts, but look identical to traditionally cooked meat after the consumer products are sliced ​​and cooked. In some embodiments, the consumer products may resemble the shape of traditional meat in two dimensions, but not in a third dimension. For example, the consumer products may resemble cuts of meat in two dimensions (e.g., when viewed from above), but may be much longer (or much thicker) than traditional cuts. Thus, in some embodiments, a composition is provided that can be repeatedly cut into products traditionally shaped with meat.

[0276] The consumer product may be made entirely from plant-based sources. In some cases, the consumer product may be made from organic sources. The consumer product may also be made from a combination of plant-based and animal-based sources. For example, the consumer product may be a ground beef product supplemented with the plant-based product of the present invention.

[0277] The consumer product may be made from locally sourced products. For example, the consumer product may be made from plants grown within a radius of the end consumer. For example, this radius may be 1, 10, 100, or 1000 miles. Therefore, in some embodiments, the present invention provides a method for producing a meat imitation that does not contain products that have been shipped 1, 10, 100, or 1000 miles prior to the production of the meat imitation.

[0278] This invention provides a method for producing a consistent quality from a consumer product when it is produced from various sources. Thus, for example, a plant-based meat imitation produced from local plants in Iowa, USA, will have a substantially similar taste, aroma, and texture to a plant-based meat imitation produced from local plants in Lorraine, France. This consistency allows for methods of advertising locally grown foods with consistent quality. This consistency can be caused by the concentration or purification of similar components from different locations. These components can be combined in predetermined ratios to ensure consistency. In some embodiments, using components from the same plant species (e.g., isolated or concentrated proteins and fats) enables a high degree of characteristic consistency. In some embodiments, using components from different plant species (e.g., isolated or concentrated proteins and fats) enables a high degree of characteristic consistency. In some embodiments, the same protein can be isolated from different plant species. In some embodiments, the invention provides a method comprising: isolating similar plant components from plant sources in different locations, assembling compositions provided therein in two locations, and smelling the compositions, wherein compositions assembled and sold in different geographical locations have consistent physical and chemical properties. In some embodiments, the separated components are derived from different plant populations in different locations. In some embodiments, one or more of the separated components are shipped to independent geographical locations.

[0279] The consumer product may require fewer resources to produce than consumer products produced from domesticated animals. Therefore, the present invention provides meat imitations that require less water or energy to produce than meat. For example, the consumer product may require less than about 10, 50, 100, 200, 300, 500, or 1000 gallons of water per pound of product. For the comparative example of beef, more than 2000 gallons of water per pound of meat may be required.

[0280] The consumer product may require less land to produce than meat products with similar protein contents. For example, the consumer product may require 30% or less of the land area required to produce meat products with similar protein contents.

[0281] The consumer product may offer health benefits compared to the animal products it replaces in the diet. For example, it may have less cholesterol or lower saturated fat content compared to similar meat products.

[0282] The consumer product may have animal welfare benefits compared to the animal products it replaces in the diet. For example, it may be produced without the need for restriction, force-feeding, premature weaning, disruption of mother-offspring interaction, or slaughter of animals for meat.

[0283] The consumer product may have a smaller "carbon footprint" than the meat product it replaces. For example, the consumer product may generate 1%, 5%, 10%, 25%, 50%, or 75% of the net greenhouse gas emissions caused by the animal product it replaces.

[0284] The consumer product may be a substitute for animal products or combinations of animal products that are forbidden by religious beliefs. For example, the consumer product may be pork chops permitted under kosher rules.

[0285] The consumer products may also be shipped in component form and manufactured or assembled in different locations. Local components, when available, can be used to produce the consumer products. Components not locally available can be used to supplement these components. This allows for methods of producing consumer products (e.g., meat imitations) using less shipping energy than meat. For example, local water can be used in combination with kits providing other components of the consumer products. Using local water will reduce shipping weight, thereby reducing costs and environmental impact.

[0286] The consumer product can be produced or assembled, wholly or partially, in areas where animal husbandry is impractical or prohibited. The consumer product can also be produced or assembled in urban environments. For example, kits can be provided to users to enable them to produce the consumer product. Users can use local water or plants from rooftop gardens (e.g., in Shanghai). In another instance, the consumer product can be produced on a spacecraft, space station, or lunar base. Therefore, the present invention provides methods and systems for producing meat replicas for space travel or for training in space travel. For example, the invention can be used for Earth-based training in space travel. The consumer product can also be produced on artificial platforms on islands or in the sea, where raising livestock is difficult or prohibited.

[0287] In some embodiments, the consumer product is designed to mimic the experience of consuming meat. The appearance, texture, and taste of the consumer product may make it similar to or indistinguishable from meat. Therefore, in some embodiments, the present invention provides a method for determining whether an animal or human can distinguish the consumer product from meat.

[0288] One method for determining whether a consumer product is comparable to meat is to a) define the properties of meat and b) determine whether the consumer product has similar properties. Testable meat properties include mechanical properties such as hardness, cohesiveness, brittleness, chewiness, adhesiveness, viscosity, elasticity, and tackiness. Testable meat properties also include geometric properties such as particle size and shape, and particle shape and orientation. Other properties may include moisture content and fat content. These properties can be described using terms such as: "soft," "firm," or "hard" to describe hardness; "crumbly," "crunchy," "brittle," "chewy," "tender," "tough," "short," "mealy," "pasty," or "gummy" to describe cohesion; "thin" or "viscous" to describe viscosity; and "plastic" or "elastic." The terms "elastic" (e.g., icy), "sticky," "tacky," or "gooey" (e.g., gooey), "gritty," "grainy," or "coarse" (e.g., coarse), "fibrous," "cellular," or "crystalline" (e.g., crystalline), "dry," "moist," "wet," or "watery" (e.g., watery), or "oily" or "greasy" (e.g., greasy) (are used to describe the properties of meat) are used to describe the elasticity; "sticky," "tacky," or "gooey" (e.g., fibrous), "cellular," or "crystalline" (e.g., crystalline), "dry," "moist," "wet," or "watery" (e.g., watery)," or "oily" or "greasy" (e.g., greasy) (are used to describe the fat content). Therefore, in one embodiment, a group of people can rate a type of meat (e.g., ground beef) based on the properties described for the meat. These ratings can be used as an indicator of the properties of the meat. The consumer product of the present invention can then be compared with meat to determine the degree of similarity between the consumer product and meat. In some cases, the properties of the consumer product are subsequently altered to make the consumer product more similar to meat. Therefore, in some embodiments, the consumer product receives a rating similar to that of meat based on human evaluation. In some embodiments, it is indistinguishable to humans from real meat.

[0289] In some embodiments, the individual being asked to identify the consumer product identifies it as a form of meat. In some embodiments, a property of the composition of the invention is that an animal (e.g., a human) would identify the composition as meat. In some embodiments, a human identifies the composition of the invention as having properties equivalent to meat. In some embodiments, one or more properties of meat are equivalent based on human perception. Such properties include testable properties. In some embodiments, a human identifies the consumer product of the invention as more like meat than meat substitutes found in the art.

[0290] In the embodiments, experiments can demonstrate that the consumer products are acceptable to consumers. A panel can be used to screen multiple consumer products described herein. Multiple human panel members can test multiple consumer product samples, i.e., natural meat compared to the consumer product compositions described herein. A mixture of lean and fatty meat can be used to standardize variables (e.g., fat content) to, for example, 20% fat. Fat content can be determined using the Babcock method for meat (SS Nielson, Introduction to the Chemical Analysis of Foods (Jones & Bartlett Publishers, Boston, 1994)). A mixture of ground beef prepared according to the procedures described herein with the consumer products of the present invention can be formulated.

[0291] Samples can be presented to panel members in an open consumer group in a small room under red or white light. Samples can be assigned as random three-digit numbers and rotated in anonymous voting positions to prevent bias. Panel members can rate the samples on softness, juiciness, texture, flavor, and overall acceptability using a pleasure scale of 1 = very averse, to 9 = very fond, with a median of 5 = neither dislike nor averse. Panel members can rinse their mouths between samples and be given the opportunity to comment on each sample.

[0292] The results of this experiment can demonstrate significant differences (p<0.05) or similarities between conventional meat and the composition of the present invention.

[0293] These results demonstrate that the compositions of the present invention are acceptablely equivalent to real meat products. Furthermore, these results demonstrate that the compositions of the present invention are more preferred by the panel members compared to other commercially available meat substitutes. Therefore, in some embodiments, the present invention provides consumer products that are remarkably similar to conventional meat.

[0294] The consumer products of this invention may also have physical properties similar to those of conventional meat. In one embodiment, the force required to pierce a 1-inch-thick structure (e.g., a meat patty) made from the consumer product of this invention with a fixed-diameter steel bar is not significantly different from the force required to pierce a similar 1-inch-thick meat structure (e.g., ground beef patty) with a similar fixed-diameter steel bar. Therefore, this invention provides consumer products with physical strength properties similar to those of meat.

[0295] In some embodiments, the compositions of the present invention have cooking loss characteristics similar to those of meat. In one embodiment, a consumer product of the present invention having a fat and protein content similar to that of ground beef has the same size reduction upon cooking as real ground beef. The various compositions of consumer products described herein that can be matched with various meats can achieve similarity in terms of size loss characteristics.

[0296] In some embodiments, the consumer product is compared to real meat based on olfactory meter readings. In various embodiments, the olfactory meter can be used to assess odor concentration and odor threshold, odor threshold compared to a reference gas, hedonic rating scores to determine the degree of enjoyment, or the relative intensity of the odor. In some embodiments, the olfactory meter allows for the training and automated evaluation of expert panels. Therefore, in some embodiments, the consumer product is a product that yields similar or identical olfactory meter readings. In some embodiments, the similarity is sufficient to exceed a detection threshold perceived by humans.

[0297] Gas chromatography-mass spectrometry (GCMS) is a method that combines the characteristics of gas chromatography-liquid chromatography and mass spectrometry to separate and identify different substances within a test sample. In some embodiments, GCMS can be used to evaluate the properties of consumer products. For example, volatile chemicals can be separated from the headspace surrounding meat. These chemicals can be identified using GCMS. This yields a characteristic of the volatile chemicals in the headspace surrounding meat. In some cases, each peak of the GCMS can be further evaluated. For example, humans can rate the olfactory experience of the chemical that causes a particular peak. This information can be used to further optimize the characteristic. GCMS can then be used to evaluate the properties of the consumer product. GCMS characteristics can be used to optimize the consumer product.

[0298] Characteristic flavor and aromatic components are largely produced during the cooking process through chemical reactions of molecules, including amino acids, fats, and sugars found in plants and meats. Therefore, in some embodiments, the consumer product is tested for similarity to meat during or after cooking. In some embodiments, an olfactory map of cooked meat is created using human ratings, human assessments, olfactory meter readings, or GCMS measurements, or a combination thereof. Similarly, an olfactory map of the consumer product (e.g., a meat imitation) can be created. These maps can be compared to assess the similarity between the cooked consumer product and meat. In some embodiments, the olfactory map of the consumer product during or after cooking is similar to or indistinguishable from the olfactory map of cooked or being cooked meat. In some embodiments, the similarity is sufficient to exceed a detection threshold perceived by humans.

[0299] In one aspect, the present invention provides a meat substitute product (or, herein, a "consumer product") that is substantially or entirely composed of ingredients from non-animal sources, but mimics key characteristics associated with cooking and consuming an equivalent meat product from an animal. The equivalent meat product may be white or dark meat. The equivalent meat product may be derived from any animal. Non-limiting examples of animals used to obtain equivalent meat products include farmed animals such as cattle, sheep, pigs, chickens, turkeys, geese, ducks, horses, dogs, or hunted animals (whether wild or farmed) such as rabbits, deer, bison, buffalo, boars, snakes, pheasants, quails, bears, elk, antelopes, pigeons, wild pigeons, grouse, foxes, wild boars, goats, kangaroos, emus, alligators, crocodiles, turtles, groundhogs, marmots, opossums, partridges, squirrels, raccoons, whales, seals, ostriches, capybaras, coypus, guinea pigs, rats, mice, wild rats, any kind of insects or other arthropods, and marine prey such as fish, crabs, lobsters, oysters, muscles, scallops, abalone, squid, octopuses, sea urchins, tunicates, etc. Although many meat products are typically derived from the skeletal muscle of an animal, it should be understood that meat can also be derived from other muscles or organs of the same animal. In some embodiments, the equivalent meat product is a cut of meat derived from skeletal muscle. In other embodiments, the equivalent meat product is an organ, such as kidney, heart, liver, gallbladder, intestine, stomach, bone marrow, brain, thymus, lung, or tongue. Therefore, in some embodiments, the compositions of the present invention are consumer products similar to skeletal muscle or organs.

[0300] In some aspects, the present invention provides meat substitute products comprising one or more of the following compositions: a first composition comprising a muscle tissue imitation, a second composition comprising a fat tissue imitation, and / or a third composition comprising a connective tissue imitation, wherein the one or more compositions are combined in a manner that mimics the physical structure of meat. In other aspects, the present invention provides compositions of muscle tissue imitations (hereinafter referred to as "muscle imitations"), fat tissue imitations (hereinafter referred to as "fat imitations"), and connective tissue imitations (hereinafter referred to as "connective tissue imitations"). In some embodiments, the compositions and meat substitute products consist primarily or entirely of ingredients derived from non-animal sources. In alternative embodiments, the muscle, fat, and / or connective tissue imitations, or meat substitute products comprising one or more of the imitations, are partially derived from animal sources but supplemented with ingredients derived from non-animal sources. In other alternative embodiments, the present invention provides meat products substantially derived from animal sources but supplemented with one or more of muscle tissue imitations, fat imitations, and / or connective tissue imitations, wherein the imitations are substantially or entirely derived from non-animal sources. A non-limiting example of such meat products is an ultra-lean ground beef product supplemented with a non-animal fat imitation, which improves texture and mouthfeel while retaining the health benefits of lower-fat animal products. Such alternative embodiments produce products that more closely mimic the key characteristics associated with the preparation and consumption of meat, but at a lower cost and with less environmental impact, less animal welfare impact, or improved health benefits for consumers.

[0301] The physical texture of the meat substitute product can be manipulated by controlling the localization, organization, assembly, or orientation of the muscle, fat, and / or connective tissue imitations described herein. In some embodiments, the product is designed such that the imitations described herein are interconnected as in meat. In some embodiments, the consumer product is designed such that, after cooking, the imitations described herein are interconnected as in cooked meat. In some embodiments, one or more of the muscle, fat, and / or connective tissue imitations are combined in a manner that mimics the physical texture of different cuts or preparations of meat. In one example embodiment, the imitations are combined in a manner that approximates the physical texture of natural minced meat. In other embodiments, the imitations are combined in a manner that approximates different cuts of beef (e.g., sirloin, tenderloin, London steak, etc.).

[0302] Protein and protein sources

[0303] In some embodiments, any of the meat substitute product, muscle tissue imitation, fat imitation, or connective tissue imitation comprises one or more isolated and purified proteins. In some embodiments, the meat substitute product consists of one or more of a muscle imitation, fat imitation, and / or connective tissue imitation comprising one or more isolated and purified proteins. In other embodiments, the muscle imitation, fat imitation, and / or connective tissue imitation comprises one or more isolated and purified proteins. In some embodiments, about 0.1%, 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 of the protein component consists of one or more isolated and purified proteins. For the purposes of this document, “purified protein” will refer to a formulation in which, relative to the source material from which the specified protein was isolated, the cumulative abundance of the protein component by mass is reduced by 2 or more, 3 or more, 5 or more, 10 or more, 20 or more, 50 or more, 100 or more, or 1000 or more, or 1000 or more, except for the specified protein (which may be a single monomer or a multimeric protein).

[0304] In some embodiments, the one or more isolated and purified proteins are derived from non-animal sources. Non-limiting examples of non-animal sources include plants, fungi, bacteria, archaea, genetically modified organisms such as genetically modified bacteria or yeast, and chemical or in vitro synthesis. In specific embodiments, the one or more isolated and purified proteins are derived from plant sources. Non-limiting examples of plant sources include: cereals such as corn, maize, rice, wheat, barley, rye, triticale, and tamarisk; oilseeds including cottonseed, sunflower seeds, safflower seeds, and rapeseed; leafy greens such as lettuce, spinach, kale, green cabbage, radish, chard, mustard greens, dandelion leaves, cabbage, and head cabbage; and green substances generally not usable by humans, including biomass crops such as switchgrass, sorghum, other grasses, alfalfa, corn stalks, and other green materials generally harvested from plants. Discarded green matter, sugarcane leaves, and tree leaves; root crops such as cassava, sweet potato, potato, carrot, beet, and turnip; plants from the legume family such as clover, peas such as cowpea, English pea, yellow pea, green pea, beans such as soybean, broad bean, lima bean, kidney bean, chickpea, mung bean, black and white bean, lentil, lupin, bean, carob, soybean, and peanut, pea (Vigna genus), pendant bean (Penta genus), peanut, indigo, gum arabic, leucocephala, guar bean, and sesbania. Those skilled in the art will understand that proteins that can be isolated from any organism in the plant kingdom can be used in this invention.

[0305] Plant-rich proteins can be largely isolated from one or more plant sources, and are therefore an economical choice for any of the muscle, fat, connective tissue imitations, or meat substitute products. Thus, in some embodiments, the one or more isolated proteins comprise abundant proteins found in high concentrations in plants and capable of being largely isolated and extracted. In some embodiments, the abundant protein constitutes about 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, or 70% of the total protein content of the source plant. In some embodiments, the abundant protein constitutes about 0.5% to 10%, about 5% to 40%, about 10% to 50%, about 20% to 60%, or about 30% to 70% of the total protein content of the source plant. In some embodiments, the abundant protein constitutes about 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the total dry matter of the source plant. In some embodiments, the abundant protein constitutes about 0.5 to 5%, about 1 to 10%, about 5 to 20%, about 10 to 30%, about 15 to 40%, or about 20 to 50% of the total dry matter of the source plant.

[0306] In certain embodiments, the one or more isolated proteins comprise abundant proteins found in high concentrations in plant leaves. In some embodiments, the abundant protein constitutes about 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%, or 80% of the total protein content of the leaves of the source plant. In some embodiments, the abundant protein constitutes about 0.5% to 10%, about 5% to 40%, about 10% to 60%, about 20% to 60%, or about 30% to 70% of the total protein content of the leaves of the source plant. In certain embodiments, the one or more isolated proteins comprise ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO activator). RuBisCO is a protein particularly suitable for meat imitations due to its high solubility and is an amino acid composition with an optimal ratio of essential amino acids close to human nutrition. In a particular embodiment, the one or more isolated proteins comprise ribulose-1,5-bisphosphate carboxylase oxygenase activator (RuBisCO activator). In a particular embodiment, the one or more isolated proteins comprise nutrient storage proteins (VSPs).

[0307] In some embodiments, the one or more isolated proteins include abundant proteins found in high concentrations in plant seeds. In some embodiments, the abundant protein constitutes about 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%, or 90% or more of the total protein content of the seeds of the source plant. In some embodiments, the abundant protein constitutes about 0.5% to 10%, about 5% to 40%, about 10% to 60%, about 20% to 60%, or about 30% to 70% or >70% of the total protein content of the seeds of the source plant. Non-limiting examples of proteins found in high amounts in plant seeds include seed storage proteins, such as albumin, glycinin, conglycinin, globulin, pea globulin, conalbumin, gliadin, gluten, glutenin, barley gliadin, gliadin, bean globulin (protein), protein body, rye gliadin, wheat gluten, zein, any seed storage protein, oily protein, oily body calciprotein, oily body sterol protein or other oily body protein.

[0308] In some embodiments, the one or more isolated proteins include proteins that interact with lipids in a structure and help stabilize lipids. Without being bound by any particular theory, such proteins can improve the integration of lipids and / or fat imitations with other components of the meat substitute product, resulting in improved taste and texture of the final product. A non-limiting example of a plant protein that interacts with lipids is a protein of the olein family. Oleins are lipid-interacting proteins found in oil bodies of plants. Other non-limiting examples of plant proteins that can stabilize emulsions include seed storage proteins from the Great Northern Bean, albumin from peas, globulin from peas, 8S globulin from mung beans, and 8S globulin from common beans.

[0309] Muscle replicas

[0310] Many meat products contain a high proportion of skeletal muscle. Therefore, this invention provides a composition derived from a non-animal source that mimics or approximates key features of animal skeletal muscle. In another aspect, this invention provides a meat substitute product comprising a composition derived from a non-animal source that mimics or approximates animal skeletal muscle. Such compositions will be labeled herein as "muscle imitators." In some embodiments, the muscle imitator and / or the meat substitute product comprising the muscle imitator is partially derived from an animal source. In some embodiments, the muscle imitator and / or the meat substitute product comprising the muscle imitator is entirely derived from a non-animal source.

[0311] Many meat products contain a high proportion of striped skeletal muscle, where individual muscle fibers are organized primarily in an isotropic manner. Therefore, in some embodiments, the muscle replica comprises fibers that are organized in a manner that is somewhat isotropic. In some embodiments, the fibers comprise a protein component. In some embodiments, the fibers comprise about 1%, about 2%, about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 99%, or more than 99% protein component.

[0312] In some embodiments, the protein component comprises one or more isolated and purified proteins. For example, the one or more isolated and purified proteins may comprise 8S globulin from mung bean seeds or albumin or globulin fractions from pea seeds. These proteins provide examples of proteins with properties favorable for constructing meat imitations because they are capable of forming gels with a texture similar to animal muscle or adipose tissue. Examples and embodiments of the one or more isolated and purified proteins are described herein. The list of potential candidates herein is substantially open and may include RuBisCO, any major seed storage protein, proteins isolated from fungi, bacteria, archaea, viruses, or genetically engineered microorganisms, or synthesized in vitro. The proteins may be artificially engineered to mimic the physical properties of animal muscle tissue. In some embodiments, one or more isolated and purified proteins account for about 0.1%, 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 of the protein component by weight.

[0313] The skeletal muscle of animals (such as beef cattle) typically contains a significant amount of glycogen, which can constitute about 1% of the muscle tissue mass at slaughter. After slaughter, a portion of this glycogen continues to be metabolized, yielding byproducts including lactic acid, which contributes to a decrease in the pH of the muscle tissue—a desirable quality in meat. Glycogen is a branched polymer of glucose linked together by α (1->4) glycosidic bonds in a straight chain, with the branching points containing α (1->6) glycosidic bonds. Starch from plants, specifically amylopectin, is also a branched polymer of glucose linked together by α (1->4) glycosidic bonds in a straight chain, with the branching points containing α (1->6) glycosidic bonds, and can therefore be used as an analogue of glycogen in the construction of meat imitations. Thus, in some embodiments, the muscle or meat imitation comprises starch or pectin.

[0314] Other components of animal muscle tissue include sodium, potassium, calcium, magnesium, other metal ions, lactic acid, other organic acids, free amino acids, peptides, nucleotides, and sulfur-containing compounds. Therefore, in some embodiments, the muscle replica may include sodium, potassium, calcium, magnesium, other metal ions, lactic acid, other organic acids, free amino acids, peptides, nucleotides, and sulfur-containing compounds. In some embodiments, the concentrations of sodium, potassium, calcium, magnesium, other metal ions, lactic acid, other organic acids, free amino acids, peptides, nucleotides, and / or sulfur-containing compounds in the muscle replica or consumer product are within 10% of the concentrations found in the imitated muscle or meat.

[0315] In another aspect, the present invention provides a method for producing muscle replicas. In some embodiments, the composition is shaped into asymmetric fibers before being incorporated into the consumer product. In some embodiments, these fibers mimic muscle fibers. In some embodiments, the fibers are rotatable fibers. In other embodiments, the fibers are extruded fibers. Therefore, the present invention provides a method for producing asymmetric or rotatable protein fibers. In some embodiments, the fibers are formed by extruding protein components through an extruder. Extrusion methods are well known in the art and described in US6379738, US3693533, and US20120093994, which are incorporated herein by reference.

[0316] In some embodiments, extrusion can be performed using an MPF19 twin-screw extruder (APV Baker, Grand Rapids, Mich.) with a cooling die. The cooling die cools the extrudate before it returns to atmospheric pressure, thus substantially inhibiting the expansion or swelling of the final product. In the MPF19 unit, dry feed and liquid can be added separately and mixed in the barrel. Extrusion parameters can be, for example: screw speed 200 rpm, product temperature at the die 150°C, feed rate 23 g / min, and water flow rate 11 g / min. The product temperature can be measured during extrusion via a thermocouple at the end of the extrusion barrel. Observations can be made regarding the color, opacity, structure, and texture of each collected sample. The collected samples can be dried overnight at room temperature, and then ground into a fine powder (<60 mesh) using a Braun food grinder, if necessary. The pH value of the sample can be repeatedly measured using a 10% (w / v) slurry of the powdered sample in distilled water.

[0317] Fat imitation

[0318] Animal fat is essential to the experience of consuming cooked meat. Therefore, this invention provides a composition from a non-animal source that mimics key characteristics of animal fat. In another aspect, this invention provides a meat substitute product comprising a composition from a non-animal source that mimics animal fat. Such compositions will be labeled herein as "fat imitators." In some embodiments, the fat imitator and / or the meat substitute product comprising the fat imitator is partially derived from an animal source.

[0319] In some embodiments, the meat substitute product has a fat component. In some embodiments, the consumer product has a fat content of 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% fat. In some embodiments, the fat imitation comprises a gel in which fat droplets are suspended. In some embodiments, the gel is a soft, elastic gel comprising proteins and optionally carbohydrates. In a particular embodiment, the protein used for the gel is a plant or microbial protein. In some embodiments, the protein used for the fat imitation may include RuBisCO, any major seed storage protein, proteins isolated from fungi, bacteria, archaea, viruses, or genetically engineered microorganisms, or synthesized in vitro. The protein may be artificially designed to mimic the physical properties of animal fat.

[0320] The fat droplets used in some embodiments of the present invention may be derived from a variety of sources. In some embodiments, the source is non-animal. In specific embodiments, the source is plant-based. Non-limiting examples of oils include 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, algae oil, palm oil, palm kernel oil, coconut oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, rice bran oil, oils produced by bacteria, algae, archaea, or fungi, or genetically engineered bacteria, algae, archaea, or fungi, and triglycerides. Monoglycerides, diglycerides, sphingomyelin, glycolipids, lecithin, lysophosphatidylcholine, phosphatidic acid, lysophosphatidic acid, oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidin, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentanic acid, docosahexanoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid, or esters of the following substances: oleic acid, palmitoleic acid Palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic wax acid, caprylic acid, geranic acid, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentanic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid, or glycerides of the following substances: oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic wax acid, caprylic acid, geranic acid, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid, or triglyceride derivatives of the following substances: oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, geraniic acid, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid.

[0321] In some embodiments, the fat droplets are derived from pulp or seed oil. In other embodiments, the source may be yeast or mold. For example, in one embodiment, the fat droplets comprise those from *Morchella glabra* (…). Mortierella isabellina Triglycerides.

[0322] In some embodiments, vegetable oils are modified to resemble animal fats. Vegetable oils can be modified with flavorings or other agents to mimic the taste and aroma of meat during and after cooking. Therefore, some aspects of the invention relate to methods for testing the qualitative similarity between the cooking properties of animal fats and the cooking properties of vegetable oils in said consumer products.

[0323] In some embodiments, the fat imitation comprises a protein component comprising one or more isolated and purified proteins. The purified proteins contribute to the flavor and texture of the meat imitation. In some embodiments, the purified proteins stabilize emulsified fats. In some embodiments, the purified proteins can form a gel upon denaturation or enzymatic cross-linking, mimicking the morphology and texture of animal fats. Examples and embodiments of the one or more isolated and purified proteins are described herein. In certain embodiments, the one or more isolated proteins comprise proteins isolated from legumes. Non-limiting examples of legumes are described herein, but variations with other legumes are possible. In some embodiments, the legume is a pea plant. In some embodiments, the isolated and purified proteins stabilize the emulsion. In some embodiments, the isolated and purified proteins form a gel upon cross-linking or enzymatic cross-linking. In some embodiments, the isolated and purified proteins comprise seed storage proteins. In some embodiments, the isolated and purified proteins comprise albumin. In some embodiments, the isolated and purified proteins comprise globulin. In one specific embodiment, the isolated and purified protein is purified pea albumin protein. In another specific embodiment, the isolated and purified protein is purified pea globulin protein. In another specific embodiment, the isolated and purified protein is mung bean 8S globulin. In another specific embodiment, the isolated and purified protein is an oleogenous protein. In another specific embodiment, the isolated and purified protein is an oleobody calciprotein. In another specific embodiment, the isolated and purified protein is RuBisCO. In some embodiments, the protein component constitutes about 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more of the fat imitation by dry weight or total weight. In some embodiments, the protein component constitutes about 0.1 to 5%, about 0.5 to 10%, about 1 to 20%, about 5 to 30%, about 10 to 50%, about 20 to 70%, or about 30 to 90% or more of the fat imitation by dry weight or total weight. In some embodiments, the protein component constitutes a solution containing one or more isolated and purified proteins.

[0324] In some embodiments, the fat imitation comprises a cross-linking enzyme that catalyzes a reaction that produces covalent cross-links between proteins. The cross-linking enzyme can be used to generate or stabilize the desired structure and texture of the fat tissue imitation to mimic the desired texture of an equivalent animal fat. Non-limiting examples of cross-linking enzymes include, for example, transglutaminase, lysyl oxidase, or other amine oxidases (e.g., Pichia pastoris lysyl oxidase). In some embodiments, the cross-linking enzyme is isolated and purified from a non-animal source, examples and embodiments of which are described herein. In some embodiments, the fat imitation comprises at least 0.0001%, or at least 0.001%, or at least 0.01%, or at least 0.1%, or at least 1% (w / v) of the cross-linking enzyme. In a particular embodiment, the cross-linking enzyme is transglutaminase.

[0325] In another aspect, the present invention provides a method for producing fat imitations. In some embodiments, fat droplets are suspended in a gel. In some embodiments, the present invention provides a method for producing fat droplets suspended in a gel. The fat can be separated and homogenized. For example, an organic solvent mixture can be used to help mix the lipids. The solvent can then be removed. The lipids can then be frozen, lyophilized, or stored. Thus, in some aspects, the present invention provides a method for separating and storing selected lipids to have properties similar to animal fat. The lipid membrane or lipid cake can then be hydrated. Hydration can be achieved using stirring or temperature changes. Hydration can be carried out from a precursor solution to a gel. After hydration, the lipid suspension can be sonicated or extruded to further alter the properties of the lipids in solution.

[0326] In some embodiments, the fat imitation is assembled to approximate the fatty tissue found in meat. In some embodiments, some or all components of the fat imitation are suspended in a gel. In various embodiments, the gel may be a protein gel, hydrogel, organic gel, or dry gel. In some embodiments, the gel may be thickened to a desired consistency using polysaccharide- or protein-based reagents. For example, fecula, arrowroot powder, corn starch, katakuri starch, potato starch, sago, tapioca starch, alginate, guar gum, locust bean gum, xanthan gum, collagen, protein, red algae gum, gelatin, agar, carrageenan, cellulose, methylcellulose, hydroxymethylcellulose, acacia gum, konjac, starch, pectin, amylopectin, or proteins derived from legumes, grains, nuts, other seeds, leaves, algae, bacteria, or fungi may be used alone or in combination to form the architecture or structure of the consumer product.

[0327] In certain embodiments, the fat imitation is an emulsion comprising a solution of one or more proteins and one or more fats, wherein the proteins and fats are suspended therein in droplet form. In some embodiments, the emulsion is stabilized into a gel by one or more cross-linking enzymes. In other specific embodiments, the one or more proteins in the solution are isolated and purified proteins. In other specific embodiments, the isolated and purified proteins comprise a purified fraction rich in pea albumin. In other specific embodiments, the isolated and purified proteins comprise a purified fraction rich in pea globulin. In other specific embodiments, the isolated and purified proteins comprise a purified fraction rich in mung bean 8S globulin. In other specific embodiments, the isolated and purified proteins comprise a fraction rich in RuBisCO. In other specific embodiments, the one or more fats are derived from plant-based oils. In other specific embodiments, the one or more fats are derived from one or more of the following substances: 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, algae oil, palm oil, palm kernel oil, coconut oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, rice bran oil, or derived from bacteria, algae, archaea, fungi, or genetically engineered bacteria. Oils produced by algae, archaea, or fungi, including triglycerides, monoglycerides, diglycerides, sphingomyelin, glycolipids, lecithin, lysophosphatidylcholine, phosphatidic acid, lysophosphatidic acid, oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidin, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentanic acid, docosahexaenoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid, or in the form of... Esters of the following substances: oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic wax acid, caprylic acid, pelargonidin, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentanic acid, docosahexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid, or glycerides of the following substances: oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic wax acid, caprylic acid, pelargonidin, undecanoic acid. The fat is linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, dodecanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid, or triglyceride derivatives of the following substances: oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristone acid, caprylic acid, caprylic acid, caprylic acid, pelargonidin, undecanoic acid, linoleic acid, 20:1 arachidic acid, arachidonic acid, eicosopentonic acid, dodecanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid. In other specific embodiments, one or more of the fats are rice bran oil. In another specific embodiment, one or more of the fats are rapeseed oil.In other specific embodiments, the cross-linking enzyme is transglutaminase, lysyl oxidase, or other amine oxidase. In other specific embodiments, the cross-linking enzyme is transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion containing a protein solution of purified pea albumin, the protein solution being emulsified with 40 to 80% rice bran oil and stabilized into a gel with 0.5 to 5% (weight / volume) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion containing a protein solution of partially purified mung bean 8S globulin, the protein solution being emulsified with 40 to 80% rice bran oil and stabilized into a gel with 0.5 to 5% (weight / volume) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion containing a protein solution of partially purified mung bean 8S globulin, the protein solution being emulsified with 40 to 80% canola oil and stabilized into a gel with 0.5 to 5% (weight / volume) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion containing a protein solution of purified pea albumin, the protein solution being emulsified with 40 to 80% rice bran oil and stabilized into a gel with 0.0001 to 1% (weight / volume) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion containing a protein solution of partially purified mung bean 8S globulin, the protein solution being emulsified with 40 to 80% rice bran oil and stabilized into a gel with 0.0001 to 1% (weight / volume) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion containing a protein solution of partially purified mung bean 8S globulin, the protein solution being emulsified with 40 to 80% canola oil and stabilized into a gel with 0.0001 to 1% (weight / volume) transglutaminase.

[0328] connective tissue imitations

[0329] Animal connective tissue provides key textural features that are an important component of the experience of eating meat. Therefore, this invention provides a composition derived from a non-animal source that mimics the key features of animal connective tissue. In another aspect, this invention provides a meat substitute product comprising a composition derived from a non-animal source that mimics the important textural and visual features of animal connective tissue. Such compositions will be labeled herein as "connective tissue imitators." In some embodiments, the connective tissue imitators and / or meat substitute products comprising the connective tissue imitators are partially derived from an animal source.

[0330] Animal connective tissue is generally classified into fascial and cartilaginous types. Fascial tissue is highly fibrous, resistant to elongation (with a high modulus of elasticity), and has a high protein content, moderate water content (approximately 50%), and low to zero fat and polysaccharide content. Therefore, the present invention provides a connective tissue imitation that mimics the key characteristics of fascial tissue. In some embodiments, the connective tissue imitation comprises approximately 50% protein by total weight and approximately 50% protein by liquid weight, and has a low fat and polysaccharide component.

[0331] The protein contents of most fascial connective tissues are primarily composed of collagen. Collagen is characterized by high proportions of proline and alanine and is assembled into characteristic elongated fibrils or rod-shaped flexible structures. Gliadin is a family of proteins found in non-animal (e.g., plant) sources. Gliadin is highly abundant in plants and is similar to collagen in its amino acid composition. Among the proteins we tested for this purpose, gliadin is particularly advantageous because of its lower cost and its ability to readily form fibers or flakes when rotated or extruded. Non-limiting examples of gliadin family proteins include, for example, zein (found in maize), and include barley gliadin from barley, wheat gliadin from wheat, rye gliadin from rye, extensin, sorghum gliadin from sorghum, and oat gliadin from oats. In fascial connective tissue, the gliadin family of proteins, alone or in combination, confirms the suitability of the protein components due to their high abundance, similarity to collagen (high proportions of proline and alanine) in the overall amino acid composition, and their ability to be processed into membranes and fibers. In addition to zein (found in maize), these proteins include barley gliadin from barley, wheat gliadin from wheat, rye gliadin from rye, extensin, sorghum gliadin from sorghum, and oat gliadin from oats. Other proteins may be necessary to supplement gliadin in order to achieve the target specifications for physicochemical and nutritional properties. The list of potential candidates here is substantially open and may include RuBisCO, any major seed storage protein, proteins isolated from fungi, bacteria, archaea, viruses, or genetically engineered microorganisms, or synthesized in vitro. The protein can be artificially designed to mimic the physical properties of animal connective tissue, animal-derived collagen or recombinant collagen, and extensin (a hydroxyproline-rich glycoprotein abundant in cell walls, such as in Arabidopsis thaliana, whose monomers are "collagen-like" rod-shaped, flexible molecules). The protein can be artificially designed to mimic the physical properties of animal connective tissue.

[0332] Methods for forming fascial connective tissue will be as those practiced in the art, and will favor methods that, prior to final formation, produce fibrous or fibrous structures, individually or in combination, continuously or in parallel, by biological, chemical, or physical methods. These methods may include extrusion or rotation.

[0333] Cartilaginous tissue is macroscopically homogeneous and compressibility resistant, with a high water content (up to 80%), a low protein (collagen) content, and a high polysaccharide (proteoglycan) content (approximately 10% each).

[0334] In terms of composition, cartilaginous connective tissue will be very similar to fascia-type tissue, with the relative ratio of each adjusted to more closely mimic 'meat' connective tissue.

[0335] The method for forming cartilaginous connective tissue will be similar to that for fascia-type connective tissue, but will be biased towards producing isotropic homogeneous structures.

[0336] Fat can be suspended in a gel. In some embodiments, the present invention provides a method for producing fat droplets suspended in a protein gel. The fat can be isolated from plant tissue and emulsified. Emulsification can be achieved using high-speed blending, homogenization, stirring, or temperature changes. The lipid suspension can be sonicated or extruded to further alter the properties of the lipids in solution. In some embodiments, other components of the consumer product are added to the solution, followed by the addition of a gelling agent. In some embodiments, a cross-linking agent (e.g., transglutaminase or lysyl oxidase) is added to bond the components of the consumer product. In other embodiments, the gelling agent is added and the lipid / gel suspension is subsequently combined with other components of the consumer product. In fascial connective tissue, the prolysin family of proteins, alone or in combination, confirms the suitability of the protein components due to their high abundance, similarity to collagen (high proportions of proline and alanine) in a global amino acid composition, and their ability to undergo processing for membrane and fiber formation. Besides zein (found in maize), these proteins include barley gliadin from barley, wheat gliadin from wheat, rye gliadin from rye, extensin, sorghum gliadin from sorghum, and oat gliadin from oats. Other proteins may be necessary to supplement gliadin in order to achieve the target specifications for physicochemical and nutritional properties. The list of potential candidates here is largely open and may include any major seed storage protein, collagen or recombinant collagen from animals, and extensin (hydroxyproline-rich glycoproteins abundant in the cell wall, such as in Arabidopsis, whose monomers are "collagen-like" rod-shaped flexible molecules).

[0337] In some embodiments, some or all of the components of the consumer product are suspended in a gel. In various embodiments, the gel may be a hydrogel, an organic gel, or a dry gel. The gel may be thickened using polysaccharide- or protein-based reagents. For example, residues, arrowroot powder, corn starch, katakuri starch, potato starch, sago, tapioca starch, alginate, guar gum, locust bean gum, xanthan gum, collagen, protein, red algae gum, gelatin, agar, carrageenan, cellulose, methylcellulose, hydroxymethylcellulose, acacia gum, konjac, starch, pectin, amylopectin, or proteins derived from legumes, grains, nuts, other seeds, leaves, algae, bacteria, or fungi may be used, alone or in combination, to form the architecture or structure of the consumer product. Enzymes that catalyze reactions that produce covalent cross-links between proteins may also be used, alone or in combination, to form the architecture or structure of the consumer product. For example, transglutaminase, lysyl oxidase, or other amine oxidases (e.g., Pichia pastoris lysyl oxidase (PPLO)) may be used alone or in combination to form the architecture or structure of the consumer product. In some embodiments, multiple gels with different components are combined to form the consumer product. For example, a gel containing plant-based proteins may be combined with a gel containing plant-based fats. In some embodiments, the fibers or strands of proteins are oriented parallel to each other and then immobilized in situ by coating with a gel containing plant-based fats.

[0338] The compositions of the present invention can be heated to become fluffy or expanded according to methods well known in the art, such as frying, baking, microwave heating, heating in a forced ventilation system, heating in a wind tunnel, etc.

[0339] In some embodiments, multiple gels with different components are combined to form the consumer product. For example, a gel containing plant-based proteins may be combined with a gel containing plant-based fats. In some embodiments, the fibers or filaments of the proteins are oriented parallel to each other and then immobilized in situ by coating them with a gel containing plant-based fats.

[0340] In some embodiments, the meat imitation contains no animal products, less than 1% wheat gluten, no methylcellulose, no carrageenan, no caramel color, no konjac flour, no gum arabic, and no acacia gum.

[0341] In some embodiments, the meat imitation does not contain the following substances: animal products, wheat gluten, methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0342] In some embodiments, the meat imitation does not contain the following substances: animal products, soy protein isolate, wheat gluten, methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0343] In some embodiments, the meat imitation does not contain the following substances: animal products, soy protein concentrate, wheat gluten, methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0344] In some embodiments, the meat imitation does not contain animal products, soy protein, wheat gluten, methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, or acacia gum.

[0345] In some embodiments, the meat imitation does not contain the following substances: animal products, tofu, wheat gluten, methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, and acacia gum.

[0346] In some embodiments, the meat imitation does not contain animal products, tofu, or wheat gluten.

[0347] In some embodiments, the meat imitation does not contain animal products, soy protein, or wheat gluten.

[0348] In some embodiments, the meat imitation does not contain methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, or acacia gum.

[0349] In some embodiments, the meat imitation contains no animal products and contains less than 5% carbohydrates.

[0350] In some embodiments, the meat imitation does not contain animal products, soy protein, wheat gluten, methylcellulose, carrageenan, caramel color, konjac flour, gum arabic, or acacia gum, and contains less than 5% carbohydrates.

[0351] In some embodiments, the meat imitation contains no animal products and contains less than 1% cellulose.

[0352] In some embodiments, the meat imitation does not contain animal products and contains less than 5% insoluble carbohydrates.

[0353] In some embodiments, the meat imitation contains no animal products, no soy protein, and less than 1% cellulose.

[0354] In some embodiments, the meat imitation contains no animal products, no soy protein, and less than 5% insoluble carbohydrates.

[0355] In some embodiments, the meat imitation contains no animal products, no wheat gluten, and less than 1% cellulose.

[0356] In some embodiments, the meat imitation contains no animal products, no wheat gluten, and less than 5% insoluble carbohydrates.

[0357] The percentages of different components can also be controlled. For example, non-animal-based substitutes of muscle, adipose tissue, connective tissue, and blood components can be combined in different ratios and physical structures to most closely resemble the appearance and feel of meat. The various components can also be arranged to ensure consistency across bites of the consumer product. The components can be arranged to ensure the consumer product does not generate waste. For example, traditional cuts of meat may have parts that are not normally eaten, while meat imitations can modify meat by excluding these inedible parts. Such modifications allow for the consumption of the entire product produced or shipped, reducing waste and shipping costs. Alternatively, meat imitations may include inedible parts to simulate the experience of eating meat. Such parts may include bone, cartilage, connective tissue, or other materials commonly referred to as gristle, or materials that mimic these components. In some embodiments, the consumer product may contain simulated inedible parts of meat products, which are designed to provide secondary functions. For example, simulated bone may be designed to distribute heat during cooking, allowing the consumer product to cook faster or more evenly than meat. In other embodiments, the simulated skeleton may also help maintain the consumer product at a constant temperature during shipment. In other embodiments, the simulated inedible portion may be biodegradable.

[0358] In some embodiments, the meat substitute composition is free of animal protein and comprises between 10% and 30% protein, between 5% and 80% water, and between 5% and 70% fat, containing one or more isolated and purified proteins. In a particular embodiment, the meat substitute composition contains transglutaminase.

[0359] In some embodiments, the consumer product contains components that mimic the composition of meat. The main component of meat is typically skeletal muscle. Skeletal muscle typically consists of approximately 75% water, 19% protein, 2.5% intramuscular fat, 1.2% carbohydrates, and 2.3% other soluble non-protein substances. These soluble non-protein substances include organic acids, sulfur-containing compounds, nitrogen-containing compounds (e.g., amino acids and nucleotides), and inorganic substances (e.g., minerals). Therefore, some embodiments of the invention provide the consumer product with an approximate composition that mimics this composition. For example, in some embodiments, the consumer product is a plant-based meat imitation that may contain approximately 75% water, 19% protein, 2.5% fat, 1.2% carbohydrates, and 2.3% other soluble non-protein substances. In some embodiments, the consumer product is a plant-based meat imitation that contains between 60 and 90% water, 10 to 30% protein, 1 to 20% fat, 0.1 to 5% carbohydrates, and 1 to 10% other soluble non-protein substances. In some embodiments, the consumer product is a plant-based meat imitation comprising 60 to 90% water, 5 to 10% protein, 1 to 20% fat, 0.1 to 5% carbohydrates, and 1 to 10% other soluble non-protein substances. In some embodiments, the consumer product is a plant-based meat imitation comprising 0 to 50% water, 5 to 30% protein, 20 to 80% fat, 0.1 to 5% carbohydrates, and 1 to 10% other soluble non-protein substances. In some embodiments, the imitation contains between 0.01% and 5% heme protein by weight. In some embodiments, the imitation contains between 0.01% and 5% leghemoglobin by weight. Some meats also contain myoglobin (a heme protein), which constitutes a large portion of the red color and iron content of some meats. In some embodiments, the imitation contains between 0.01% and 5% heme protein by weight. In some embodiments, the imitation contains between 0.01% and 5% leghemoglobin by weight. It should be understood that these percentages are variable in meat, and the meat imitations can be produced to approximate the natural variations in meat. Additionally, in some cases, the present invention provides modified meat imitations that typically contain these components in non-natural percentages. For example, a meat imitation can be produced with a higher-than-typical average fat content. The percentages of these components can also be altered to enhance other desired properties.

[0360] In some cases, a meat imitation is designed to have component percentages similar to cooked meat when cooked. Therefore, in some embodiments, the uncooked consumer product has different component percentages than uncooked meat, but when cooked, the consumer product is similar to cooked meat. For example, a meat imitation may be made with a higher water content than typical raw meat, but when cooked in a microwave, the resulting product has component percentages similar to those of cooked meat.

[0361] In some embodiments, the consumer product is a meat imitation with a lower water content than typical meat. In some embodiments, the invention provides a method for hydrating the meat imitation to give it contents similar to meat. For example, a meat imitation with a lower water content than meat, such as 1%, 10%, 20%, 30%, 40%, or 50% water, is hydrated to approximately 75% water. In some embodiments, once hydrated, the meat imitation is subsequently cooked for human consumption.

[0362] The consumer product may contain a protein component. In some embodiments, the protein content of the consumer product is 10%, 20%, 30%, or 40%. In some embodiments, the protein content of the consumer product is similar to that of meat. In some embodiments, the protein content of the consumer product is greater than that of meat. In some embodiments, the consumer product has less protein than meat.

[0363] The proteins in the consumer product may be derived from multiple sources or a combination of sources. Non-animal sources may provide some or all of the proteins in the consumer product. Non-animal sources may include vegetables, fruits, nuts, grains, algae, bacteria, or fungi. The proteins may be isolated or concentrated from one or more of these sources. In some embodiments, the consumer product is a meat imitation containing proteins obtained solely from non-animal sources.

[0364] In some embodiments, the protein is shaped into asymmetric fibers for incorporation into the consumer product. In some embodiments, these fibers mimic muscle fibers. In some embodiments, the protein is a rotatable fiber. Therefore, the present invention provides a method for producing asymmetric or rotatable protein fibers. In some embodiments, the consumer product contains one or more proteins having all the amino acids found in proteins essential for human nutrition. In some embodiments, the protein added to the consumer product is supplemented with amino acids.

[0365] Indicators for cooking meat

[0366] The release of odorants during cooking is an important aspect of meat consumption. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked beef. In some embodiments, the consumer product, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked pork. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked bacon. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked chicken. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked lamb. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked fish. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that, when cooked, produces an odor that is recognizable to humans as the typical odor of cooked turkey. In some embodiments, the consumer product is a meat imitation composed primarily or entirely of ingredients from non-animal sources, having odorants released during cooking. In some embodiments, the consumer product is a meat imitation composed primarily or entirely of ingredients from non-animal sources, having odorants produced by chemical reactions occurring during cooking. In some embodiments, the consumer product is a meat imitation composed primarily or entirely of ingredients from non-animal sources, containing a mixture of proteins, peptides, amino acids, nucleotides, sugars, and polysaccharides and fats in a combination and spatial configuration that allows these compounds to undergo chemical reactions during cooking to produce odorants and flavor compounds. In some embodiments, the consumer product is a meat imitation composed primarily or entirely of ingredients from non-animal sources, having volatile or unstable odorants released during cooking. In some embodiments, the consumer product is a method of preparing a meat imitation, wherein the meat imitation composed primarily or entirely of ingredients from non-animal sources is heated to release volatile or unstable odorants.

[0367] Flavoring agents released during meat cooking are generated through reactions involving the following substances as reactants: fats, proteins, amino acids, peptides, nucleotides, organic acids, sulfur-containing compounds, sugars, and other carbohydrates. In some embodiments, flavoring agents bound during meat cooking are identified and located close to each other in the consumer product, such that the flavoring agents bind when the consumer product is cooked. Therefore, in some embodiments, the characteristic flavor and aroma components are generated during the cooking process through chemical reactions involving amino acids, fats, and sugars found in plants and meats. Thus, in some embodiments, the characteristic flavor and aroma components are largely generated during the cooking process through chemical reactions involving one or more amino acids, fats, peptides, nucleotides, organic acids, sulfur-containing compounds, sugars, and other carbohydrates found in plants and meats.

[0368] Some reactions that generate flavorings released during meat cooking can be catalyzed by iron, specifically heme iron from myoglobin. Therefore, in some embodiments, some of the characteristic flavor and aroma components are generated during the cooking process via iron-catalyzed chemical reactions. In some embodiments, some of the characteristic flavor and aroma components are generated during the cooking process via heme-catalyzed chemical reactions. In some embodiments, some of the characteristic flavor and aroma components are generated during the cooking process via heme iron-catalyzed chemical reactions from legume hemoglobin. In some embodiments, some of the characteristic flavor and aroma components are generated during the cooking process via heme iron-catalyzed chemical reactions from heme proteins.

[0369] Evidence that the presence of leghemoglobin favorably enhances the odor of meat simulants: Muscle simulants containing pea flour, sunflower oil, and glucose were heated at 140°C for 10 minutes in a sealed container equipped with solid-phase microextraction (SPME) fibers in the presence of reduced leghemoglobin (LHb) or a mixture of iron (Fe3+), sodium, and EDTA (EFS). These fibers contained polydimethylsiloxane (PDMS) to adsorb volatile compounds for analysis by GC-MS. Analysis of GC-MS data from multiple simulants revealed consistent differences between LHb and EFS samples. Non-limiting examples of unique or abundant compounds found in LHb samples include 2-octanone, 2-methylfuran (which is generally associated with the odor of cooked meat), and many other unidentified compounds. For example, Figure 1This figure shows a comparison of aligned GCMS distributions (mirror-oriented) of otherwise identical muscle replica samples cooked in the presence of bean-leuthemin (LHb, right-hand distribution) or ferric ions (left-hand distribution). In this figure, the vertical axis represents the residence time during the gas chromatography separation step, and the horizontal axis represents the m / z ratio of the ions produced by fragmenting the corresponding separated volatile compounds. Selected compounds that show differences in the two samples are marked on the right side of the figure.

[0370] Color indicator

[0371] The color of meat is an important part of the experience of cooking and eating it. For example, cuts of beef are characteristically red when raw and gradually turn brown during cooking. As another example, white meat (such as chicken or pork) is characteristically pink when raw and gradually turns white or light brown during cooking. The amount of color change is used to indicate the cooking progress of beef and to quantify cooking time and temperature to produce the desired level of doneness. In some aspects, the present invention provides a non-meat meat substitute product that provides a visual indicator of cooking progress. In some embodiments, the visual indicator is a color indicator that undergoes a color change during cooking. In a particular embodiment, the color indicator mimics the color change of a cut of meat from raw to cooked. In other particular embodiments, the color indicator turns the meat substitute product red before cooking to indicate a raw state and turns the meat substitute product brown during cooking. In other particular embodiments, the color indicator turns the meat substitute product pink before cooking to indicate a raw state and turns the meat substitute product white or brown during cooking.

[0372] The primary determinant of the nutritional definition of meat color is the concentration of ferritin in the meat. In the skeletal muscle component of meat products, one of the main ferritins is myoglobin. It is estimated that white chicken contains less than 0.05%; pork and veal contain 0.1% to 0.3%; veal contains 0.4% to 1.0%; and older beef contains 1.5% to 2.0%. Therefore, in some embodiments, the consumer product is a meat imitation containing ferritin. In some embodiments, the meat imitation contains about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2%, or more than about 2% ferritin by dry weight or gross weight. In some cases, the ferritin has been isolated and purified from its source. In other cases, the ferritin has not been isolated and purified. In some cases, the ferritin is from an animal source or a non-animal source, such as plants, fungi, or genetically modified organisms, such as bacteria or yeast. In some cases, the ferritin is myoglobin. In some embodiments, the consumer product is a plant-based meat imitation fortified with animal myoglobin. Thus, for example, a calf meat imitation may have about 0.4 to 1% myoglobin. In some cases, the ferritin is leghemoglobin. In some embodiments, the consumer product is a plant-based meat imitation fortified with leghemoglobin. Thus, for example, a calf meat imitation may have about 0.4 to 1% leghemoglobin. In some cases, the ferritin is cytochrome. In some embodiments, the consumer product is a plant-based meat imitation fortified with cytochrome. Thus, for example, a calf meat imitation may have about 0.4 to 1% cytochrome.

[0373] Another example of ferritin is hemoglobin, an iron-containing oxygen-binding protein found in the red blood cells of vertebrates. Hemoglobin is similar in color to myoglobin. In some embodiments, the present invention provides a method for preserving and recycling blood from animal husbandry to enhance the color of a consumer product. For example, blood is preserved from a slaughterhouse, and the hemoglobin from said blood is used to enhance the color of the consumer product. In some aspects, said consumer product is a plant-based meat imitation containing hemoglobin.

[0374] Other iron-containing proteins are naturally occurring. In some embodiments, the consumer product contains an iron-containing protein that is not myoglobin. In some embodiments, the consumer product does not contain myoglobin. In some embodiments, the consumer product does not contain hemoglobin. In some embodiments, the consumer product is a meat imitation containing an iron-containing protein other than myoglobin or hemoglobin.

[0375] Examples of ferritin include hemoglobin, myoglobin, neuroglobin, cytoglobin, legume hemoglobin, non-symbiotic hemoglobin, hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavin hemoglobin. In various embodiments, these ferritin are added to the consumer product to alter its visual properties or iron content. In some embodiments, the consumer product comprises heme proteins (e.g., hemoglobin, myoglobin, neuroglobin, cytoglobin, legume hemoglobin, non-symbiotic hemoglobin, hellgate globin I, bacterial hemoglobin, ciliate myoglobin, and flavin hemoglobin).

[0376] Leghemoglobin, structurally and physically similar to myoglobin, is readily available as an unused byproduct of commercial leguminous crops such as soybeans and peas. In the United States, the leghemoglobin content in the roots of these crops exceeds the myoglobin content of all red meat consumed in the US. In some embodiments, the consumer product is a meat imitation primarily or entirely composed of non-animal-derived ingredients, including muscle tissue imitations, adipose tissue imitations, connective tissue imitations, and leghemoglobin. In some embodiments, the consumer product is a meat imitation primarily or entirely composed of non-animal-derived ingredients containing heme proteins. In some embodiments, the consumer product is a meat imitation primarily or entirely composed of non-animal-derived ingredients and contains leghemoglobin. In some embodiments, the consumer product is a meat imitation primarily or entirely composed of non-animal-derived ingredients containing members of the hemoglobin protein family. In some embodiments, the consumer product is a meat imitation primarily or entirely composed of non-animal-derived ingredients with a high iron content derived from heme proteins. In some embodiments, the iron content is similar to that of meat. In some embodiments, the consumer product has a distinctive meat-red color provided by leghemoglobin.

[0377] Leghemoglobin is used in some embodiments as an indicator of the end of cooking of the consumer product. Therefore, one embodiment of the invention is a method of cooking a consumer product comprising detecting leghemoglobin that migrates from the interior of the consumer product to its surface during cooking. Another embodiment of the invention is a method of cooking a consumer product comprising detecting a color change from red to brown during cooking.

[0378] Heme proteins are used in some embodiments as indicators of the end of cooking of the consumer product. Therefore, one embodiment of the invention is a method of cooking a consumer product comprising detecting legheme proteins that migrate from the interior of the consumer product to its surface during cooking. Another embodiment of the invention is a method of cooking a consumer product comprising detecting a color change from red to brown during cooking.

[0379] Heme proteins from the following groups have been used in some embodiments as indicators of the end of cooking for consumer products: hemoglobin, myoglobin, neuroglobin, cytoglobin, legume hemoglobin, non-symbiotic hemoglobin, hellgospherin I, bacterial hemoglobin, ciliate myoglobin, and flavohemoglobin. Therefore, one embodiment of the invention is a method of cooking a consumer product comprising detecting legume hemoglobin that migrates from the interior of the consumer product to its surface during cooking. Another embodiment of the invention is a method of cooking a consumer product comprising detecting a color change from red to brown during cooking.

[0380] Food products containing isolated and purified leghemoglobin

[0381] In some embodiments, leghemoglobin may be added to meat to enhance its properties. For example, a solution containing leghemoglobin may be injected into raw or cooked meat. In another instance, a leghemoglobin solution may be dripped over meat or the consumer product of the present invention to enhance its appearance. In one embodiment, leghemoglobin is used to enhance the advertising, photography, or visual text of food products (e.g., meat or meat substitutes).

[0382] Sources of leghemoglobin in soybeans

[0383] In some embodiments, the present invention provides a method for obtaining leghemoglobin from plants. Leghemoglobin can be obtained from a variety of plants. Various leguminous species and varieties, such as soybean, broad bean, lima bean, cowpea, English pea, yellow pea, lupin, common bean, chickpea, peanut, alfalfa, wild pea hay, clover, lespedeza, and black-and-white pea, contain nitrogen-fixing root nodules (e.g., root nodules from pea plants) in which leghemoglobin plays a key role in controlling oxygen concentration. Figure 2 ). Figure 3 This shows a 100 ml solution of leghemoglobin isolated from 30 g of pea root nodules. Leghemoglobin from different species are homologues and have similar color properties. Figure 4 ).exist Figure 4In Group A, SDS-PAGE gels of dissolved root nodules from the following three leguminous species are shown: (1) broad bean, (2) English pea, and (3) soybean. Arrows indicate the corresponding leghemoglobin. It should be noted that leghemoglobin is the most abundant soluble protein in each soluble sample. Group B shows the similarity of the UV-Vis spectral distributions of leghemoglobin from two different plant species (broad bean and soybean). We purified leghemoglobin from root nodules of broad bean (green curve) and soybean (red curve) using the protocol described elsewhere in this specification. The UV-Vis spectra of the two proteins show heme iron in a reduced (+2) state. It should be noted that they almost perfectly overlap, consistent with their visually identical red color. The heme iron in the corresponding leghemoglobin was reduced to the +2 oxidation state by incubating broad bean and soybean leghemoglobin in 10 mM sodium bisulfite in a 20 mM potassium phosphate buffer at pH 7.4 and 100 mM sodium chloride buffer. Sodium bisulfite was then removed from the legume hemoglobin solution using gel size exclusion chromatography. The inset shows a magnified view of the UV-Vis spectrum in the 450 nm to 700 nm region. Some plant species express several legume hemoglobin isoforms (for example, soybean has four). Small changes in the precise amino acid sequence can alter the total charge of the protein at a specific pH and can change the precise structural conformation of the heme group in legume hemoglobin. Differences in the structural conformation of the heme group in different legume hemoglobins can affect the oxidation and reduction rates of heme iron. These differences can contribute to the color and flavor-producing properties of different legume hemoglobins.

[0384] Leghemoglobin has an absorption spectrum and visual appearance that are almost identical to myoglobin derived from animal muscle. Figure 5 Showing reduced (heme iron 2+) and oxidized (heme iron 3+) soybean leghemoglobin ( Figure 5 Group A) and equine cardiac myoglobin ( Figure 5 The comparison of Group B shows the similarity of the UV-Vis absorption distribution of the two proteins. We purified soybean leghemoglobin from soybean root nodules using the protocol described here. Purified horse myoglobin was purchased from Sigma-Aldrich. Soybean leghemoglobin was reduced with 1 mm sodium bisulfite ( Figure 5 Group A) and horse myoglobin ( Figure 5 Group B). Showing soybean leghemoglobin ( Figure 5 Group A) and horse myoglobin ( Figure 5 The UV-Vis absorption spectra of heme Fe3+ (blue line) and heme Fe2+ (red line) in group B). The inset shows a magnified view of the UV-Vis spectrum in the 450 nm to 700 nm region. Figure 5Group C), the image of a 10 μl droplet (left droplet) of a 40 mg / ml solution of soybean leghemoglobin in the Fe3+ state shows a characteristic rust-red color, and the image of a 10 μl droplet (right droplet) of a 40 mg / ml solution of soybean leghemoglobin in the Fe2+ state shows a characteristic red color. The characteristic rust-red color and the characteristic red color are present in and correspond to the sample of horse myoglobin (right image).

[0385] In other embodiments, leghemoglobin may be derived from non-plant sources, such as organisms, such as bacteria or yeast that have been genetically modified to express high levels of leghemoglobin.

[0386] The oxidation state of iron ions in leghemoglobin is crucial to its color. Leghemoglobin with heme iron in the +2 oxidation state is bright red, while that with heme iron in the +3 oxidation state is brownish-red. Therefore, in using leghemoglobin as a source of red color in meat imitations, it is desirable to reduce the heme iron from the +3 state to the +2 state. A reducing agent can be used to convert the heme iron in leghemoglobin from the oxidized (+3) state to the reduced (+2) state. An example of successfully reducing the heme iron in leghemoglobin using sodium bisulfite and titanium citrate is shown in [reference needed]. Figure 6 In. Figure 6 In the images, the UV-Vis spectra of purified soybean leghemoglobin in the oxidized (+3) state of heme iron are represented by the blue curves in each group. The red curves in each group represent the UV-Vis spectra of the same leghemoglobin species reduced to the (+2) state (red line) after addition of (Group A) 1 mM sodium bisulfite or (Group B) a 20 mM potassium phosphate buffer (pH 7.3) containing 0.24% (wt / v) titanium citrate. The inset shows a magnified view of the UV-Vis spectrum in the 450 to 700 nm region. For this example, leghemoglobin was purified from soybean root nodules using 60 / 90% ammonium sulfate fractionation, and the leghemoglobin was exchanged into a 20 mM potassium phosphate buffer (pH 7.4) containing 100 mM sodium chloride. A sodium bisulfite stock solution was prepared by dissolving 100 mM sodium bisulfite in 1 mM sodium hydroxide in water. A titanium citrate stock solution was prepared from 20% (wt / v) titanium chloride in hydrochloric acid (by mixing it with 0.2 M sodium citrate (1:10 v / v)). The pH was adjusted to 7.0 using sodium carbonate.

[0387] Leghemoglobin can be purified from root nodules of leguminous plants, such as pea or soybean root nodules. Figure 1 Showing pea root nodules, Figure 2(Showing leghemoglobin isolated from pea root nodules). Root nodules from soybeans were thoroughly washed to remove soil and foreign root tissue, and then dissolved in 20 mM potassium phosphate (pH 7.4), 100 mM sodium chloride, 1 mM EDTA, and 1 mM ascorbic acid. Nodules were dissolved by grinding using a Vitamix blender. For some samples, polyvinylpyrrolidone was added at 30 wt / v to aid in the removal of small phytophenolic molecules that mediate the oxidation of heme iron in leghemoglobin. The nodule dissolution was fractionated using ammonium sulfate in two steps, first by adding ammonium sulfate to 60 wt / v. The nodules were discarded, and the supernatant was brought to 90 wt / v ammonium sulfate. Leghemoglobin was collected as precipitated nodules in the 90% ammonium sulfate fraction. The ammonium sulfate-precipitated leghemoglobin is resuspended in 20 mM potassium phosphate, 1 mM EDTA, and 50 mM sodium chloride, and the ammonium sulfate is removed by dialysis or size exclusion chromatography in the same buffer. In some cases, this is the final purification step, but in others, anion exchange chromatography (FFQ GE Healthcare) (which is sometimes followed by size exclusion chromatography (Sephacryl S-100, GE Healthcare)) is used to further purify the leghemoglobin. Soybean leghemoglobin from 90% ammonium sulfate pellets was loaded onto different buffer-type anion exchange columns (FFQ or DEAE, GE Healthcare) containing 0 to 100 mM sodium chloride at pH 7.4, 20 mM Tris at pH 8, 20 mM sodium tetraborate at pH 9.8, 20 mM sodium chloride, 20 mM sodium carbonate at pH 9, and 20 mM sodium chloride) and purified either in a flow-through manner or using sodium chloride (0 to 1 M salt gradient). An example of a purified flow of leghemoglobin from soybean root nodules is presented in… Figure 7The figure shows SDS-PAGE fractions of different soybean leghemoglobin purification steps: (lane 1) soybean root nodule lysate; (lane 2) soybean root nodule lysate purified by fractionation with 60 / 90% (wt / v) ammonium sulfate. The figure also shows the protein contents of protein globules fractionated with 90% ammonium sulfate, which were resuspended in 20 mM potassium phosphate pH 7.4, 100 mM sodium chloride, 1 mM EDTA buffer; the protein from the 90% ammonium sulfate globules was further purified by anion exchange chromatography (FFQ GE Healthcare) with 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride. Leghemoglobin collected in the effluent is shown in lane 3. Size exclusion chromatography (Sephacryl S-100 GE Healthcare) was used to fractionate the anion exchange effluent, and the resulting leghemoglobin fraction is shown in lane 4. ImageDoc analysis software (Bio-Ray) was used to determine the fraction of the leghemoglobin band on the SDS-PAGE gel in the corresponding samples to identify the contents of leghemoglobin at different purification steps. The purity (partial abundance) of leghemoglobin at the corresponding steps in the purification process was as follows: dissolved 32.7% (lane 1), 60 / 90% (wt / v) ammonium sulfate fraction 78% (lane 2), anion exchange chromatography approximately 83% (lane 3), and size exclusion chromatography approximately 95% (lane 4).

[0388] Leghemoglobin can also be produced by genetically engineering the bacteria or fungi that produce it. An illustrative example is shown below. Figure 8 middle. Figure 8 The following are SDS-PAGE gel analyses of stained materials: (A) soybean leghemoglobin expressed and purified using recombinant protein technology, and (B) soybean leghemoglobin purified from soybean root nodules. (A) Recombinant soybean leghemoglobin A, carrying a His tag and a TEV protease His tag removal site, was expressed in *E. coli* strain BL21 and purified using His tag affinity chromatography (Taren resin, Clone Technologies). The left lane contains molecular weight standards, and the right lane contains purified recombinant soybean leghemoglobin A (arrow). The expected molecular weight of recombinant soybean leghemoglobin A is 17.1 kDa. (B) SDS-PAGE gel of soybean leghemoglobin purified from root nodules. The left lane contains molecular weight standards, and the right lane contains purified soybean leghemoglobin A (arrow). Mass spectrometry analysis of the purified material confirmed the presence of all four soybean leghemoglobin isoforms in full length (data not shown). The expected molecular weight (MW) of soybean leghemoglobin isoforms is in the range of 15.4 to 15.8 kDa.

[0389] Soybean hemoglobin, purified from soybean and broad bean root nodules, was tasted by a group of volunteers and described as tasting like blood in each case.

[0390] Leghemoglobin can be isolated from root nodules of leguminous plants (such as soybean, broad bean, cowpea, lima bean, chickpea, pea, lupin, birdsfoot root (lotus japonicus), or other leguminous plants). Root nodules (e.g., those from pea plants) Figure 1 The soluble proteins, including leghemoglobin, are obtained and homogenized in aqueous solution and recovered after insoluble substances are removed by precipitation or filtration. Leghemoglobin can be purified by selective precipitation and / or chromatography and / or using molecules with a specific affinity for leghemoglobin. Figure 2 (This shows a 100 ml solution of bean hemoglobin isolated from 30 g of pea root nodules).

[0391] Heme proteins, such as soy leghemoglobin, can be combined with other plant-based meat imitation components. In some embodiments, the heme protein is captured in a gel containing other components (e.g., lipids and / or proteins). In some aspects, multiple gels are combined with a non-gel-based heme protein. In some embodiments, the heme protein is combined with other compounds of the consumer product to ensure that the heme protein can diffuse throughout the consumer product. In some embodiments, the consumer product is produced in a solution containing heme proteins (e.g., a soy leghemoglobin solution). In some embodiments, the consumer product is immersed in a solution containing heme proteins (e.g., a soy leghemoglobin solution) for 1, 5, 10, 15, 20, or 30 hours. In some embodiments, the consumer product is immersed in a solution containing heme (e.g., a soy leghemoglobin solution) for 1, 5, 10, 15, 30, or 45 minutes.

[0392] Figure 9 This image shows six examples of commercially available meat analogues (plant-based broiler analogues), each approximately 1 cm in diameter at the side. Four of these (left and lower right) have been soaked in a solution of 20 mM potassium phosphate (pH 7.4) and 100 mM NaCl containing approximately 10 mg / ml soybean leghemoglobin; the remaining two (upper right) have been soaked in the same buffer solution without soybean leghemoglobin. Note the striking contrast between the deep pink of the leghemoglobin-infused blocks and the pale brown of the uninfused blocks in the color photograph.

[0393] Figure 10Four blocks of plant-based chicken analogue, infused with bean leghemoglobin, are shown cooking in a pan at 350°C. The two lower blocks have been flipped to expose the roasted, browned surface. Note that in the two upper blocks, the heated portion has turned grayish-brown, while the cooler top surface remains pink. In some embodiments, a heme-containing solution (e.g., bean leghemoglobin solution) is injected into the consumer product until it resembles the color of uncooked meat.

[0394] Given the use of heme proteins in coloring consumer products, this would be suitable for detecting whether a product contains a specific heme protein. Therefore, in some embodiments of the invention, a method for determining whether a product contains heme proteins is included. Methods for detecting proteins are well known in the art. For example, ELISA, or adjacent junction tests, or luninex tests, or western blot analysis can be performed to determine the presence of leghemoglobin in food products (e.g., meat or meat imitations). In one embodiment, the detection method is performed to determine whether leghemoglobin has been used to modify meat.

[0395] Example

[0396] This article describes an exemplary muscle replica composition comprising one or more isolated and purified plant proteins.

[0397] Protein purification of the components of the counterfeit product

[0398] Mung bean seeds and dried green pea seeds were purchased in powder form and used to purify the corresponding seed storage proteins. RuBisCo was purified from fresh alfalfa plants. Protein composition at individual fractionation steps was monitored by SDS-PAGE, and protein concentration was measured by standard UV-Vis and Pierce tests.

[0399] Mung bean 8S globulin: Mung bean powder was resuspended in 50 mM potassium phosphate buffer (pH 7) and 0.5 M NaCl at a ratio of 1:4 (wt / v) and the mixture was incubated for 1 hour. Insoluble material was separated by centrifugation, and the protein in the supernatant was fractionated by adding 50% (wt / v) followed by 90% (wt / v) ammonium sulfate in two steps. The protein precipitated in the 90% fraction contained mung bean 8S globulin, and this protein was stored at -20°C until further use.

[0400] Pea albumin: Dried green pea seed powder was resuspended in 50 mM sodium acetate buffer (pH 5) at a ratio of 1:10 (wt / v) and incubated for 1 hour. Insoluble material was separated by centrifugation, and proteins in the supernatant were fractionated by two steps of 50% (wt / v) followed by 90% (wt / v) ammonium sulfate precipitation. The ammonium sulfate solution was stirred for 1 hour, and the ammonium sulfate-precipitated protein was removed by centrifugation. The protein of interest was precipitated in 90% (wt / v) ammonium sulfate. The pellets were stored at -20°C until further use.

[0401] Pea globulin: Dried green pea seed powder was resuspended in 20 mM potassium phosphate buffer (pH 8), 0.4 M sodium chloride at a ratio of 1:10 (wt / v) and stirred for 1 hour. After centrifugation, the supernatant was subjected to ammonium sulfate fractionation. First, the supernatant was brought to 50% (wt / v) ammonium sulfate, and the precipitated protein was removed. Second, the 50% (wt / v) ammonium sulfate supernatant was brought to 80% (wt / v) ammonium sulfate saturation. The protein globules formed by 80% (wt / v) ammonium sulfate fractionation contained the globulin of interest. The globules were stored at -20°C until further use.

[0402] RuBisCO: RuBisCO was fractionated from alfalfa leafy plants (or other green plants, such as soybean plants, spinach, etc.) by first grinding the leaves in a blender with 4 volumes of cold 50 mM potassium phosphate buffer pH 7.4 buffer (with (in the lab) or without (in the field) 0.5 M NaCl + 2 mM DTT + 1 mM EDTA). The resulting slurry was centrifuged to remove debris, and the supernatant (crude solution) was used for further purification. Proteins in the crude solution were fractionated by adding ammonium sulfate to 30% (wt / v) saturation. The solution was stirred for 1 hour and then centrifuged. The pellets from this step were discarded, and ammonium sulfate was added back to the supernatant to 50% (wt / v) ammonium sulfate saturation. The solution was centrifuged again after stirring for 1 hour. The pellets from this step contained RuBisCO, and the pellets were kept at -20°C until use.

[0403] Obtain plant protein.

[0404] 8S protein from mung bean seeds was purified by ammonium sulfate fractionation as described. The protein globules were resuspended in 20 mM potassium phosphate (pH 7.4) and 0.5 M sodium chloride, and the ammonium sulfate was removed by dialysis relative to the same buffer. Any precipitate was removed by centrifugation at 16,000 g for 10 minutes, and the protein was concentrated to the desired concentration. Pea globulin was purified by ammonium sulfate fractionation as described. The protein globules were resuspended in 20 mM potassium phosphate (pH 7.4) and 0.4 M sodium chloride, and the ammonium sulfate was removed by dialysis relative to the same buffer. Any precipitate was removed by centrifugation at 16,000 g for 10 minutes, and the protein was concentrated to the desired concentration. Pea albumin was purified by ammonium sulfate fractionation as described. The protein globules were resuspended in 20 mM potassium phosphate (pH 7.4) and 0.1 M sodium chloride, and the ammonium sulfate was removed by dialysis relative to the same buffer. Any precipitate was removed by centrifugation at 16,000 g for 10 minutes, and the protein was concentrated to the desired concentration.

[0405] Constructing muscle tissue analogs

[0406] 8S protein from mung bean seeds was purified by ammonium sulfate fractionation as described above. To prepare the gel, 200 g of pellets were dissolved in 400 ml of dialysis buffer (20 mM potassium phosphate, 400 mM NaCl, pH 7.3), and the resulting solution was dialyzed against 5 l of dialysis buffer for 6 hours, with the dialysis buffer being replaced twice with fresh buffer. The protein solution was centrifuged at 12,000 g for 15 minutes to remove debris. The protein was concentrated by dialyzing against 5 l of a 30% w / w solution of PEG 8000 (polyethylene glycol, molecular weight 8000) in dialysis buffer for 36 hours. The final protein concentration was 150 mg / ml.

[0407] Legumin was purified from soybean root nodules. Leguminous root nodules were washed to remove soil and foreign root tissue, and then dissolved in 20 mM potassium phosphate (pH 7.4), 100 mM sodium chloride, 1 mM EDTA, and 1 mM ascorbic acid. The nodules were dissolved by grinding them using a juicer blender. Insoluble material was separated by centrifugation. The nodule solution was fractionated using ammonium sulfate in two steps: first, ammonium sulfate was added to 60% wt / v and the solution was incubated at 4°C for 1 hour. The nodules were discarded, and the supernatant was incubated at 90% wt / v ammonium sulfate for 12 hours at 4°C. Legumin was collected as precipitated nodules in the 90% ammonium sulfate fraction and resuspended in 20 mM potassium phosphate, 1 mM EDTA, and 100 mM sodium chloride. SDS-PAGE gel analysis confirmed that the protein solution contained 70% legumin and 30% other nodule proteins. Ammonium sulfate was removed using size exclusion chromatography with the same buffer solution. Soybean leghemoglobin was concentrated by dialysis for 48 hours with 30% PEG 8000 (polyethylene glycol, molecular weight 8000) in 100 mM sodium chloride and 20 mM potassium phosphate at pH 7.3. The total protein concentration was 57 mg / ml. UV-Vis spectroscopy indicated that soybean leghemoglobin was in the oxidized heme iron state. Therefore, soybean leghemoglobin was incubated with 5 mM sodium bisulfite for 5 minutes, and the sodium bisulfite was removed using size exclusion chromatography with a 20 mM potassium phosphate and 100 mM sodium chloride buffer solution. Soybean leghemoglobin was further concentrated to 35.4 mg / ml. UV-Vis spectroscopy confirmed that soybean leghemoglobin was in the reduced heme iron state.

[0408] The transglutaminase was purchased from Activa TI (Ajimoto). A stock solution (20% wt / v) was prepared in 20 mM potassium phosphate buffer (pH 7.3) and 100 mM sodium chloride buffer.

[0409] In order to prepare "dark" muscle tissue analogs ( Figure 11 43 ml of mung bean protein solution (150 mg / ml in dialysis buffer) was mixed with 37 ml of soybean leghemoglobin solution (46.5 mg / ml soybean leghemoglobin and 20 mg / ml other soybean rhizonoproteins in 20 mM potassium phosphate, 100 mM NaCl, pH 7.3). 20 ml of transglutaminase solution (20% w / w) was added, and the solution was thoroughly mixed. The solution was then aliquoted into two 50 ml Falkenberg tubes and incubated overnight at room temperature. The final protein concentrations were 65 mg / ml mung bean protein, 18 mg / ml soybean leghemoglobin, and 91 mg / ml total protein.

[0410] A “white” muscle analogue was prepared by mixing 43 ml of mung bean protein solution (150 mg / ml) with 45 ml of 11.7 mg / ml bean hemoglobin solution and 0.8% (wt / v) transglutaminase solution. Figure 12 The final protein concentrations were 63 mg / ml for mung bean protein, 5.2 mg / ml for leghemoglobin, and 68 mg / ml for total protein.

[0411] The “dark” muscle tissue analogue forms a dark chocolate-colored, smooth, homogeneous, opaque gel with a reflective surface and a small amount (<1 ml) of dark red venous blood-colored liquid on top. The gel is free-floating, elastic but fragile, and visually resembles a thin Jell-O jelly. The gel has a moderate odor and noticeably contains clearly discernible beans and blood. The flavor is controlled by the noticeable beans and iron / blood, with a weak herbal and medicinal / chemical flavor. The taste is salty with a long, lingering bloody finish.

[0412] The “white” muscle tissue analogue is very similar, but has a much lighter, cappuccino-like color. Furthermore, it is more fragile, with a compressive strength two to three times lower.

[0413] adipose tissue analogues

[0414] The following preparation of an adipose tissue analog using the 8S globulin fraction of mung bean was performed: 15 ml of mung bean protein solution (150 mg / ml in dialysis buffer) was mixed with 15 ml of rice bran oil. 6 ml of transglutaminase solution (20% w / w) was added, and the solution was thoroughly emulsified using a vrinator (VWR) at speed 2. The emulsion was aliquoted into 1.6 cm Eppendorf tubes and incubated overnight at room temperature. Subsequently, the tubes were heated in a heating block at 95°C for 5 minutes and then allowed to cool to room temperature on the lab bench. The final concentration was 75 mg / ml mung bean protein and 50% w / w oil.

[0415] Adipose tissue analogs using pea globulin (100 mg / ml) were prepared using the same method. Additionally, adipose tissue analogs were prepared in bulk from pea globulin and rice bran or canola oil using the same method, but without aliquoting the emulsion into Eppendorf tubes. Instead, the emulsion in 50 ml Falkenberg tubes was spun overnight on a nutator and then incubated at 90°C for 30 minutes.

[0416] Adipose tissue analog based on mung beans and prepared in Eppendorf tubes ( Figure 13It forms a grayish-white, smooth, homogeneous, opaque gel with no visible or identifiable unincorporated liquid. The gel is free-floating, elastic, and soft. It has a slightly pleasant odor and a mild, pleasant flavor. The taste is slightly salty.

[0417] Adipose tissue analog based on pea globulin and prepared in Eppendorf tubes ( Figure 14 It is very similar to mung bean-based fat analogs, except that it deforms less when compressed.

[0418] The adipose tissue analog prepared in 50 ml Falk tubes was similar in appearance, texture and odor, but was actually softer (twice as soft as canola oil and three times as soft as rice bran oil, according to compressibility measurements).

[0419] connective tissue analogues

[0420] Connective tissue analog prototypes were developed using zein protein from 100% yellow corn gluten flour or from commercially available sources such as Amazein (PrairieGold, Bloomington, IL). The zein protein was dissolved in 70 to 90% ethanol at a desired ratio of 1:3 to 1:5 (solid:solution). The zein protein was precipitated in a controlled manner, for example by altering the pH, to impart the desired, manipulated physicochemical properties to the zein structure. Figure 15 The strands of connective tissue analogues are shown, generated by using 70% ethanol at a 1:3 ratio in a syringe fitted with a 23-gauge needle (0.337 mm inner diameter). The solution is slowly squeezed from the bottom of a 5-inch-high container into an excess of 5 M NaCl solution. The ethanol-zein solution, being less dense than the NaCl solution, floats upward, pulling out the fibrous scaffold of the solidified zein. The NaCl is continuously stirred as the strands begin to develop, thus aiding in their elongation. The strands aggregate and form hard, dense masses.

[0421] The prototype of the minced beef is made from a gel of plant protein and vegetable oil.

[0422] Ground beef patties were prepared by combining 62% (wt / wt) muscle analogue (62% (wt / wt) "dark muscle analogue" and 38% (wt / wt) "white muscle analogue"), 29% (wt / wt) adipose tissue analogue (derived from pea globulin and canola oil), and 5% (wt / wt) connective tissue analogue. Figure 16Group A). ​​Ground beef patties were prepared by combining 62% muscle analogue (62% "dark muscle analogue" and 38% "white muscle analogue"), 29% adipose tissue analogue (derived from mung bean seed 8S protein and rice bran oil), and 5% connective tissue analogue. Figure 16 Group B). Ground beef patties were made by combining 71% (wt / wt) muscle tissue analogue (composed of 60% "white" muscle analogue and 40% "dark" muscle analogue) and 23% adipose tissue (derived from pea seed globulin protein and canola oil). Figure 16 Group C). Ground beef prototype patties were made by combining 67% "white" muscle analogue with 28% adipose tissue analogue (derived from pea globulin and rice bran oil). Figure 16 (Group D).

[0423] In another test, the effect of cooking the ground beef patties was evaluated by grilling them in a 350℉ skillet. The mixture consisted of 62% (wt / wt) muscle tissue analogue (62% (wt / wt) "dark muscle analogue" and 38% (wt / wt) "white muscle analogue"), 29% (wt / wt) adipose tissue analogue (derived from pea globulin and canola oil), and 5% (wt / wt) connective tissue analogue (…). Figure 17 To create patty analogues of ground beef, the left set shows the patties before cooking, and the right set shows the same patties after cooking for approximately 2 minutes. The observer described the smell of the cooked ground beef analogues as distinctly "beef-like."

Claims

1. A meat imitation product comprising: From 0.01% to 5% by weight of at least one heme-containing protein from a non-animal source, wherein the heme-containing protein is leghemoglobin; sugar; Sulfur-containing compounds; and 10% or more plant protein by weight; The meat replica comprises muscle replicas and adipose tissue replicas assembled in a manner similar to the physical structure of meat, wherein the muscle replicas comprise 40 to 90% of the meat replicas by weight, and the adipose tissue replicas comprise 1 to 60% of the meat replicas by weight. The meat imitations described therein do not contain animal products, and, The meat imitation precisely simulates the color of raw meat before cooking, precisely simulates the color of cooked meat after cooking, and produces a meat-like aroma during cooking.

2. The meat imitation according to claim 1, wherein the meat imitation contains between 0.05% and 2% by weight of heme protein.

3. The meat imitation according to any one of claims 1-2, wherein the heme protein comprises an amino acid sequence having at least 70% homology with SEQ ID NO 1.

4. The meat imitation according to any one of claims 1-2, wherein the heme protein comprises an amino acid sequence having at least 80% homology with SEQ ID NO 1.

5. The meat imitation according to any one of claims 1-2, wherein the heme protein comprises an amino acid sequence having at least 90% homology with SEQ ID NO 1.

6. The meat imitation according to any one of claims 1-2, wherein the plant protein is selected from the group consisting of: ribosomal proteins, hexokinase, lactate dehydrogenase, fructose-1,5-bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO), ribulose- 1,5-bisphosphate carboxylase oxygenase activator (RuBisCO activator), albumin, soybean globulin, con-soybean globulin, globulin, pea globulin, con-albumin, gliadin, gluten, glutenin, barley gliadin, gliadin, bean globulin, protein body, rye gliadin, extensin, wheat gluten, zein, oil protein, oil body calciprotein, oil body sterol protein, nutrient storage protein A, nutrient storage protein B, and mung bean seed storage 8S globulin.

7. The meat imitation according to any one of claims 1-2, wherein the sugar is glucose.

8. The meat imitation according to any one of claims 1-2, wherein the meat imitation is characterized by one or more of the following: free of methylcellulose, free of carrageenan, free of caramel color, free of konjac flour, free of gum arabic, free of acacia gum, containing less than 1% wheat gluten, free of soy protein isolate, free of soy protein concentrate, free of soy protein, containing less than 5% carbohydrates, free of tofu, containing less than 1% cellulose, or containing less than 5% insoluble carbohydrates.

9. The meat imitation according to any one of claims 1-2, wherein the meat imitation further comprises fat from a non-animal source.

10. The meat imitation according to any one of claims 9, wherein the fat is one or more fats 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, algae oil, palm oil, palm kernel oil, coconut oil, babassu oil, shea butter, mango butter, cocoa butter, wheat germ oil, and rice bran oil.

11. The meat imitation according to any one of claims 1-2, wherein the meat imitation further comprises nucleotides.

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