Methods and compositions for consumables

Abstract

A method and composition relating to a plant-based meat substitute having characteristics similar to meat are provided herein.

Description

Technology Field

[0001] This application claims priority to U.S. application serial number 61 / 572,205 filed on July 12, 2011, the full text of which is incorporated herein by reference. Background Technology

[0002] Animal husbandry has serious negative environmental impacts. Currently, it is estimated that 30% of the Earth's surface is used exclusively for animal husbandry, and livestock accounts for 20% of total terrestrial animal biomass. Due to this massive scale, animal husbandry accounts for over 18% of net greenhouse gas emissions. Animal husbandry may be the largest source of human water pollution and is by far the world's greatest threat to biodiversity. It is estimated that if the global population could shift from a meat-based diet to a diet free of animal products, 26% of the Earth's surface would be utilized for other purposes. Furthermore, a shift to a plant-based diet would significantly reduce water and energy consumption.

[0003] Meat consumption has serious negative effects on human health. The health benefits of a vegetarian diet are widely established. If the population shifts to a vegetarian diet, there will be significant cost savings in healthcare.

[0004] Although the world's four major commodity crops (soybeans, corn, wheat, and rice) already supply more than 100% of the population's requirements for calories and protein (including all essential amino acids), hunger is a global problem.

[0005] Plant-based meat substitutes have failed to drive a shift toward vegetarian diets. Current technological advancements in meat substitute compositions involve the extrusion of soybean / grain mixtures, which produce products that fail to adequately replicate the cooking and eating experience of meat. A common limitation of these products is their lack of uniform texture and mouthfeel compared to equivalent meat products. Furthermore, because these products must be sold pre-cooked with minimal artificial flavorings, they fail to mimic the flavor, taste, and other key characteristics associated with cooked meat. Consequently, while these products have appealed primarily to a limited consumer base already practicing vegetarianism or veganism, they have not attracted the larger segment of consumers accustomed to meat consumption.

[0006] An improved method and composition are disclosed herein that more accurately mimic the characteristics evaluated for the production and consumption of meat and overcome the shortcomings and disadvantages of current meat substitutes.

[0007] In some aspects, the present invention provides a meat substitute composition that accurately mimics the taste, texture, or color of a meat product derived from an animal source, comprising a protein content, wherein one or more isolated and purified proteins account for at least 10% by weight of the protein content.

[0008] In one embodiment, the meat substitute composition accurately mimics the color of the meat product in its unprocessed state and in its cooked state after cooking.

[0009] In another embodiment, one or more isolated and purified proteins account for at least 25% of the protein contents by weight.

[0010] In another embodiment, one or more isolated and purified proteins account for 50% or more of the protein contents by weight.

[0011] In another embodiment, one or more isolated and purified proteins account for at least 75% of the protein contents by weight.

[0012] In another embodiment, one or more isolated and purified proteins account for 90% or more of the protein contents by weight.

[0013] In another embodiment, gluten does not account for more than 10% of the protein content by weight.

[0014] In another embodiment, each of the isolated, purified proteins is isolated and purified individually.

[0015] In another embodiment, the meat substitute composition comprises 1 to 7 types of isolated and purified proteins.

[0016] In another embodiment, the isolated and purified proteins of types 1-7 are each isolated from different plant species.

[0017] In some embodiments, the meat substitute comprises 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or fewer than 20 individual proteins isolated from one or more plant species.

[0018] In another embodiment, the protein contents include trace amounts of any other protein derived from one or more plant species.

[0019] In another embodiment, one or more isolated and purified proteins are hemoglobin, myoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, flavohemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, It is selected from the group consisting of pyruvate decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase (Rubisco activase), albumin, glycinin, cornglycinin, globulin, bisilin, cornalbumin, gliadin, glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein), proteome, secalin, extensin, gluten in tritheca, zein, any seed storage protein, oleosin, caloleosin, steroleosin or other oil body proteins, nutritional storage protein A, nutritional storage protein B, and mung bean seed storage 8S globulin.

[0020] In another embodiment, one or more isolated and purified proteins are not isolated from animals.

[0021] In another embodiment, one or more isolated and purified proteins are isolated from a single plant source.

[0022] In another embodiment, one or more isolated and purified proteins are isolated from multiple plant sources.

[0023] In another embodiment, one or more isolated, purified proteins are isolated from a genetically modified organism.

[0024] In some embodiments, the genetically modified organism is a genetically modified bacterium or yeast organism.

[0025] In some embodiments, the isolated, purified protein is formed into fibers.

[0026] In a specific embodiment, the fiber is similar to a skeletal muscle fiber.

[0027] In a more specific embodiment, the fiber is an asymmetric fiber.

[0028] In some embodiments, the meat substitute composition further comprises one or more isolated and purified iron-containing proteins.

[0029] In some embodiments, one or more isolated and purified iron-containing proteins are selected from the group consisting of hemoglobin, myoglobin, leghemoglobin, non-symbiotic hemoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, helgate globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin.

[0030] In a specific embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology to sequence 1. Sequence 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAKDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA.

[0031] In certain embodiments, the iron-containing protein comprises an amino acid sequence having at least 70% homology to sequence 2. Sequence 2: MIDQKEKELI KESWKRIEPN KNEIGLLFYA NLFKEEPTVS VLFQNPISSQ SRKLMQVLGI LVQGIDNLEG LIPTLQDLGR RHKQYGVVDS HYPLVGDCLL KSIQEYLGQG FTEEAKAAWT KVYGIAAQVM TAE. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 80% homology to sequence 2. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 90% homology to sequence 2. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 98% homology to sequence 2.

[0032] In certain embodiments, the iron-containing protein comprises an amino acid sequence having at least 70% homology to sequence 3. Sequence 3: MRKQPTVFEK LGGQAAMHAA VPLFYKKVLA DDRVKHYFKN TNMEHQAKQQ EDFLTMLLGG PNHYKGKNMA EAHKGMNLQN SHFDAIIENL AATLKELGVS DQIIGEAAKV IEHTRKDCLG K. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 80% homology to sequence 3. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 90% homology to sequence 3. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 98% homology to sequence 3.

[0033] In some embodiments, the isolated and purified proteins are assembled into one or more gels.

[0034] In some embodiments, the meat substitute composition further comprises one or more types of fat.

[0035] In a specific embodiment, one or more of the fats are derived from a plant source.

[0036] In another aspect, the present invention provides a meat substitute product derived from a non-animal source, comprising an indicator that indicates the progress of cooking from an unprocessed state to a cooked state.

[0037] In some embodiments, the indicator is a visual indicator that accurately mimics the color change of the meat product during the cooking process.

[0038] In one embodiment, the color change is a change from red to brown.

[0039] In one embodiment, the color change is a change from pink to white or yellowish-brown.

[0040] In one embodiment, the visual indicator changes from a translucent to an opaque color during the cooking process.

[0041] In some embodiments, the indicator is an olfactory indicator that indicates the progress of cooking.

[0042] In one embodiment, the olfactory indicator is one or more volatile odorants released during cooking.

[0043] In some embodiments, the indicator comprises one or more isolated, purified iron-containing proteins.

[0044] In a specific embodiment, one or more isolated, purified iron-containing proteins are in a reduced state before cooking.

[0045] In one embodiment, the one or more isolated and purified iron-containing proteins are selected from the group consisting of hemoglobin, myoglobin, leghemoglobin, non-symbiotic hemoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, cytochrome, helsecta globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin.

[0046] In a specific embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology to sequence 1. Sequence 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAKDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA.

[0047] In some embodiments, one or more isolated and purified iron-containing proteins are not isolated from animals. In some embodiments, the composition of the present invention does not contain any protein from animals.

[0048] In a specific embodiment, the one or more isolated and purified iron-containing proteins are isolated from one or more plant sources.

[0049] In a more specific embodiment, the one or more isolated, purified proteins are isolated from the root nodules, roots, seeds, leaves, or stems of the one or more plant sources.

[0050] In another specific embodiment, one or more of the plant sources are soybean or pea plants.

[0051] In one embodiment, the one or more plant sources include one or more legume plants.

[0052] In some embodiments, the one or more isolated and purified iron-retaining proteins in a reduced or oxidized state have a UV-VIS profile similar to that of a myoglobin protein derived from an animal source in an equivalent reduced or oxidized state.

[0053] In a specific embodiment, the difference between the peak absorbance wavelength of one or more isolated and purified iron-containing proteins and the peak absorbance wavelength of myoglobin derived from an animal source is less than 5%.

[0054] In some embodiments, one or more isolated, purified proteins are isolated from genetically modified organisms.

[0055] In one embodiment, the genetically modified organism is a genetically modified bacterium or yeast organism.

[0056] In some embodiments, the meat substitute product does not contain methylcellulose, carrageenan, caramel coloring, konjac powder, gum arabic, and acacia gum.

[0057] In a specific embodiment, the meat substitute product contains less than 1% wheat gluten.

[0058] In a more specific embodiment, the meat substitute product does not contain wheat gluten.

[0059] In another specific embodiment, the meat substitute product does not contain soy protein isolate.

[0060] In another specific embodiment, the meat substitute product does not contain soy protein concentrate.

[0061] In another specific embodiment, the meat substitute product does not contain soy protein.

[0062] In a more specific embodiment, the meat substitute product contains less than 5% carbohydrates.

[0063] In another specific embodiment, the meat substitute product does not contain tofu.

[0064] In some embodiments, the meat substitute product does not contain tofu and wheat gluten.

[0065] In some embodiments, the meat substitute product does not contain soy protein and wheat gluten.

[0066] In some embodiments, the meat substitute product does not contain animal products and contains less than 5% carbohydrates.

[0067] In some embodiments, the meat substitute product contains less than 1% cellulose.

[0068] In some embodiments, the meat substitute product contains less than 5% insoluble carbohydrates.

[0069] In some embodiments, the meat substitute product does not contain soy protein and contains less than 1% cellulose.

[0070] In some embodiments, the meat substitute product does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0071] In some embodiments, the meat substitute product does not contain wheat gluten and contains less than 1% cellulose.

[0072] In some embodiments, the meat substitute product does not contain wheat gluten and contains less than 5% insoluble carbohydrates.

[0073] In another aspect, the present invention provides a muscle tissue imitation that is similar in taste, texture, or color to equivalent muscle tissue derived from an animal source, comprising a protein content including one or more isolated and purified proteins.

[0074] In some embodiments, one or more isolated and purified proteins account for 50% or more of the protein content by weight. In some embodiments, one or more isolated and purified proteins account for 40% or more of the protein content by weight. In some embodiments, one or more isolated and purified proteins account for 30% or more of the protein content by weight. In some embodiments, one or more isolated and purified proteins account for 20% or more of the protein content by weight. In some embodiments, one or more isolated and purified proteins account for 10% or more of the protein content by weight.

[0075] In some embodiments, the one or more isolated and purified proteins account for at least 50% by weight of the contents of the composition. In some embodiments, the one or more isolated and purified proteins account for at least 40% by weight of the contents of the composition. In some embodiments, the one or more isolated and purified proteins account for at least 30% by weight of the contents of the composition. In some embodiments, the one or more isolated and purified proteins account for at least 20% by weight of the contents of the composition. In some embodiments, the one or more isolated and purified proteins account for at least 10% by weight of the contents of the composition. In some embodiments, the one or more isolated and purified proteins account for at least 5% by weight of the contents of the composition. In some embodiments, the one or more isolated and purified proteins account for at least 1% by weight of the contents of the composition.

[0076] In some embodiments, the protein contents are derived from one or more non-animal sources.

[0077] In a specific embodiment, one or more non-animal sources are plant sources.

[0078] In another specific embodiment, one or more non-animal sources are genetically modified yeast or bacteria.

[0079] In some embodiments, each of the above one or more isolated proteins is isolated and purified individually.

[0080] In some embodiments, one or more isolated proteins are hemoglobin, myoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, flavohemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate It is selected from the group consisting of decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase (Rubisco activase), albumin, glycinin, cornglycinin, globulin, bisilin, cornalbumin, gliadin, glutelin, glutenin, hordain, prolamin, phaseolin (protein), proteome, secalin, extensin, gluten in tritheca, zein, any seed storage protein, oleosin, caloleosin, steroleosin or other oil body proteins, nutritional storage protein A, nutritional storage protein B, and mung bean seed storage 8S globulin.

[0081] In one embodiment, the seed storage protein is mung bean 8S protein.

[0082] In some embodiments, the protein contents are suspended in a gel.

[0083] In some embodiments, the protein contents are in the form of a gel.

[0084] In one embodiment, the gel comprises isolated, purified crosslinking enzymes.

[0085] In some embodiments, the isolated, purified crosslinking enzyme is selected from the group consisting of transglutaminase, lysyl oxidase, and amine oxidase.

[0086] In a specific embodiment, the isolated, purified crosslinking enzyme is transglutaminase.

[0087] In some embodiments, the protein contents are assembled into fibers.

[0088] In a specific embodiment, the fibers are arranged isotropically.

[0089] In one embodiment, the fiber is an asymmetric fiber.

[0090] In some embodiments, the muscle tissue imitation further comprises one or more isolated and purified iron-containing proteins.

[0091] In some embodiments, one or more isolated and purified iron-containing proteins are selected from the group consisting of hemoglobin, myoglobin, leghemoglobin, non-symbiotic hemoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin.

[0092] In a specific embodiment, one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 70% homology to sequence 1. Sequence 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAKDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA. In a specific embodiment, one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 80% homology to sequence 1. In a specific embodiment, one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 90% homology to sequence 1. In a specific embodiment, one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 95% homology to sequence 1. In a specific embodiment, one or more isolated and purified iron-containing proteins comprise an amino acid sequence having at least 98% homology to sequence 1.

[0093] In a specific embodiment, the muscle tissue imitation comprises protein contents, wherein (i) one isolated and purified protein that is not an iron-containing protein accounts for 40-95% of the protein contents, (ii) one or more isolated and purified iron-containing proteins account for 1-20% of the protein contents, and (iii) one or more crosslinking reagents account for 0.1-35% of the protein contents.

[0094] In one embodiment, the protein content accounts for 5-50% of the imitation by weight or weight / volume.

[0095] In one embodiment, the isolated and purified protein is mung bean 8S protein.

[0096] In one embodiment, the one or more isolated and purified iron-containing proteins are leghemoglobin.

[0097] In one embodiment, the one or more crosslinking reagents are transglutaminases.

[0098] In some embodiments, the muscle tissue imitation does not contain methylcellulose, carrageenan, caramel color, konjac powder, gum arabic, and acacia gum.

[0099] In a specific embodiment, the muscle tissue imitation further contains less than 1% wheat gluten. In a specific embodiment, the muscle tissue imitation further contains less than 5% wheat gluten. In a specific embodiment, the muscle tissue imitation further contains less than 10% wheat gluten. In a specific embodiment, the muscle tissue imitation further contains less than 0.1% wheat gluten.

[0100] In a more specific embodiment, the muscle tissue imitation does not contain wheat gluten.

[0101] In another specific embodiment, the muscle tissue imitation does not contain soy protein isolate.

[0102] In another specific embodiment, the muscle tissue imitation does not contain soy protein concentrate.

[0103] In another specific embodiment, the muscle tissue imitation does not contain soy protein.

[0104] In a more specific embodiment, the muscle tissue imitation contains less than 5% carbohydrates.

[0105] In another specific embodiment, the muscle tissue imitation does not contain head.

[0106] In some embodiments, the muscle tissue imitation does not contain tofu and wheat gluten.

[0107] In some embodiments, the muscle tissue imitation does not contain soy protein and wheat gluten.

[0108] In some embodiments, the muscle tissue imitation does not contain animal products and contains less than 5% carbohydrates.

[0109] In some embodiments, the muscle tissue imitation contains less than 1% cellulose.

[0110] In some embodiments, the muscle tissue imitation contains less than 5% insoluble carbohydrates.

[0111] In some embodiments, the muscle tissue imitation does not contain soy protein and contains less than 1% cellulose.

[0112] In some embodiments, the muscle tissue imitation does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0113] In some embodiments, the muscle tissue imitation does not contain wheat gluten and contains less than 1% cellulose.

[0114] In some embodiments, the muscle tissue imitation does not contain wheat gluten and contains less than 5% insoluble carbohydrates.

[0115] In some embodiments, the muscle tissue imitation does not contain methylcellulose, carrageenan, caramel color, konjac powder, gum arabic, and acacia gum.

[0116] 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.

[0117] In a more specific embodiment, the meat imitation does not contain wheat gluten.

[0118] In another specific embodiment, the meat imitation does not contain soy protein isolate.

[0119] In another specific embodiment, the meat imitation does not contain soy protein concentrate.

[0120] In another specific embodiment, the meat imitation does not contain soy protein.

[0121] In a more specific embodiment, the meat imitation contains less than 5% carbohydrates.

[0122] In another specific embodiment, the meat imitation does not contain tofu.

[0123] In some embodiments, the meat imitation does not contain tofu and wheat gluten.

[0124] In some embodiments, the meat imitation does not contain soy protein and wheat gluten.

[0125] In some embodiments, the meat imitation does not contain animal products and contains less than 5% carbohydrates.

[0126] 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.

[0127] In some embodiments, the meat imitation contains less than 5% insoluble carbohydrates.

[0128] In some embodiments, the meat imitation does not contain soy protein and contains less than 1% cellulose.

[0129] In some embodiments, the meat imitation does not contain soy protein and contains less than 5% insoluble carbohydrates.

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

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

[0132] In another aspect, the present invention provides an adipose tissue imitation comprising a gelled emulsion comprising a protein solution in which a fat droplet is suspended.

[0133] In some embodiments, the fat droplets are derived from a non-animal source.

[0134] In some embodiments, the fat droplet is composed of one or more vegetable oils.

[0135] In some embodiments, one or more vegetable oils are 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, bacteria, algae, archaea or fungi or genetically modified bacteria, oil produced by algae, archaea or fungi, triglycerides, monoglycerides, diglycerides, sphingosides, glycolipids, lecithin, lysolecithin, phosphatidic acid, lysophosphatidic acid, oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, mystoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 Eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, ester of 18:2 conjugated linoleic acid or conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, It is selected from the group consisting of glycerol esters of 18:2 conjugated linoleic acid or conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, and triglyceride derivatives of 18:2 conjugated linoleic acid or conjugated oleic acid.

[0136] In one embodiment, the one or more vegetable oils are rice bran oil or canola oil.

[0137] In some embodiments, the protein solution comprises one or more isolated, purified proteins.

[0138] In some embodiments, one or more isolated, purified proteins account for 75% or more of the protein in the protein solution.

[0139] In some embodiments, one or more isolated, purified proteins are derived from non-animal sources.

[0140] In some embodiments, the non-animal source is a plant source.

[0141] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0142] In some embodiments, each of the above one or more isolated proteins is isolated and purified individually.

[0143] In some embodiments, one or more isolated proteins are hemoglobin, myoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, flavohemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate It is selected from the group consisting of decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase (Rubisco activase), albumin, glycinin, cornglycinin, globulin, bisilin, cornalbumin, gliadin, glutelin, glutenin, hordain, prolamin, phaseolin (protein), proteome, secalin, extensin, gluten in tritheca, zein, any seed storage protein, oleosin, caloleosin, steroleosin or other oil body proteins, nutritional storage protein A, nutritional storage protein B, and mung bean seed storage 8S globulin.

[0144] In some embodiments, one or more isolated, purified proteins are albumin protein, seed storage protein, or pea globulin protein.

[0145] In a specific embodiment, the albumin protein is isolated pea albumin protein.

[0146] In some embodiments, the seed storage protein is mung bean 8S protein.

[0147] In some embodiments, the gelling emulsion comprises a protein solution comprising 1-3 types of isolated and purified proteins and occupying 30-70% of the volume of the emulsion; a vegetable oil occupying 30-70% of the volume of the emulsion; and an isolated, purified crosslinking enzyme occupying 0.5-5% of the emulsion by weight / volume, wherein the vegetable oil is emulsified in the protein solution and the emulsion is formed into a gel by the crosslinking enzyme.

[0148] In another embodiment, the gelling emulsion comprises a protein solution comprising 1-3 types of isolated and purified proteins and occupying 1-30% of the volume of the emulsion; a vegetable oil occupying 70-99% of the volume of the emulsion; and an isolated, purified crosslinking enzyme occupying 0.5-5% of the emulsion by weight / volume, wherein the vegetable oil is emulsified in the protein solution and the emulsion is formed into a gel by the crosslinking enzyme.

[0149] In some embodiments, the fat imitation further includes a cross-linking enzyme.

[0150] In some embodiments, the crosslinking enzyme is transglutaminase.

[0151] In some embodiments, one of the 1-3 isolated and purified proteins is mung bean 8S protein, pea albumin protein, or pea globulin protein.

[0152] In a specific embodiment, the vegetable oil is rice bran oil or canola oil.

[0153] In some embodiments, the adipose tissue imitation does not contain methylcellulose, carrageenan, caramel coloring, konjac powder, gum arabic, and acacia gum.

[0154] In a specific embodiment, the fat tissue imitation contains less than 1% wheat gluten.

[0155] In a more specific embodiment, the fat tissue imitation does not contain wheat gluten.

[0156] In another specific embodiment, the adipose tissue imitation does not contain soy protein isolate.

[0157] In another specific embodiment, the adipose tissue imitation does not contain soybean protein concentrate.

[0158] In another specific embodiment, the adipose tissue imitation does not contain soy protein.

[0159] In a more specific embodiment, the adipose tissue imitation contains less than 5% carbohydrates.

[0160] In another specific embodiment, the fat tissue imitation does not contain tofu.

[0161] In some embodiments, the fat tissue imitation does not contain tofu and wheat gluten.

[0162] In some embodiments, the adipose tissue imitation does not contain soy protein and wheat gluten.

[0163] In some embodiments, the adipose tissue imitation does not contain animal products and contains less than 5% carbohydrates.

[0164] In some embodiments, the adipose tissue imitation contains less than 1% cellulose.

[0165] In some embodiments, the adipose tissue imitation contains less than 5% insoluble carbohydrates.

[0166] In some embodiments, the adipose tissue imitation does not contain soy protein and contains less than 1% cellulose.

[0167] In some embodiments, the adipose tissue imitation does not contain soy protein and contains less than 5% insoluble carbohydrates.

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

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

[0170] In another aspect, the present invention provides a connective tissue imitation comprising a protein content comprising one or more isolated, purified proteins, wherein the protein content is assembled into a structure similar to the texture and visual appearance of connective tissue or skin.

[0171] In some embodiments, the protein contents are derived from a non-animal source.

[0172] In some embodiments, the non-animal source is a plant source.

[0173] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0174] In some embodiments, one or more isolated proteins account for 50% or more of the protein content by weight.

[0175] In some embodiments, the isolated and purified protein of the one type accounts for 90% or more of the protein content by weight.

[0176] In some embodiments, each of the above one or more isolated proteins is isolated and purified individually.

[0177] In some embodiments, one or more isolated proteins are hemoglobin, myoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, flavohemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate It is selected from the group consisting of decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase (Rubisco activase), albumin, glycinin, cornglycinin, globulin, bisilin, cornalbumin, gliadin, glutelin, glutenin, hordain, prolamin, phaseolin (protein), proteome, secalin, extensin, gluten in tritheca, zein, any seed storage protein, oleosin, caloleosin, steroleosin or other oil body proteins, nutritional storage protein A, nutritional storage protein B, and mung bean seed storage 8S globulin.

[0178] In some embodiments, one or more isolated and purified proteins are prolamin family proteins.

[0179] In some embodiments, one or more isolated and purified proteins are zein.

[0180] In some embodiments, the protein contents are suspended in a gel.

[0181] In some embodiments, the gel comprises isolated, purified crosslinking enzymes.

[0182] In some embodiments, the isolated, purified crosslinking enzyme is selected from the group consisting of transglutaminase, lysyl oxidase, and amine oxidase.

[0183] In some embodiments, the isolated, purified crosslinking enzyme is transglutaminase.

[0184] In some embodiments, the protein contents are formed into fibers.

[0185] In some embodiments, the fiber is produced by an extrusion process.

[0186] In some embodiments, the fiber is stabilized by protein crosslinking.

[0187] In some embodiments, the fiber contains isolated, purified crosslinking enzymes.

[0188] In some embodiments, the isolated, purified crosslinking enzyme is selected from the group consisting of transglutaminase, lysyl oxidase, and amine oxidase.

[0189] In some embodiments, the isolated, purified crosslinking enzyme is transglutaminase.

[0190] 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, the adipose tissue imitation, and / or the connective tissue imitation are assembled in a manner close to the physical organization of meat.

[0191] In some embodiments, the meat substitute product comprises two or more of the muscle imitation, fat tissue imitation, and connective tissue imitation.

[0192] In some embodiments of the meat substitute product, the muscle imitation accounts for 40-90% of the product by weight, the fat tissue imitation accounts for 1-60% of the product by weight, and the connective tissue imitation accounts for 1-30% of the product by weight.

[0193] In some embodiments, the meat substitute comprises 60-90% water; 5-30% protein content; and 1-20% fat or fat imitation, wherein the protein content comprises one or more isolated, purified plant proteins.

[0194] In some embodiments, the protein contents are derived from a non-animal source.

[0195] In some embodiments, the non-animal source is a plant source.

[0196] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0197] In some embodiments, at least 50% of the protein content by weight is one or more isolated purified proteins.

[0198] In some embodiments, each of the above one or more isolated proteins is individually isolated and purified from different plant species.

[0199] In some embodiments, one or more of the isolated proteins are hemoglobin, myoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, flavohemoglobin, ribosomal protein, actin, hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, It is selected from the group consisting of pyruvate decarboxylase, actin, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase (Rubisco activase), albumin, glycinin, cornglycinin, globulin, bisilin, cornalbumin, gliadin, glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein), proteome, secalin, extensin, gluten in tritheca, zein, any seed storage protein, oleosin, caloleosin, steroleosin or other oil body proteins, nutritional storage protein A, nutritional storage protein B, and mung bean seed storage 8S globulin.

[0200] In some embodiments, the meat substitute product further comprises one or more isolated and purified iron-containing proteins.

[0201] In some embodiments, the one or more isolated and purified iron-containing proteins are selected from the group consisting of hemoglobin, myoglobin, leghemoglobin, non-symbiotic hemoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, helgate globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin. In some embodiments, the iron-containing protein comprises an amino acid sequence having at least 70% homology to sequence 1. [SEQ ID NO: 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAKDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA].

[0202] In some embodiments, the meat substitute product does not contain methylcellulose, carrageenan, caramel coloring, konjac powder, gum arabic, and acacia gum.

[0203] In a specific embodiment, the meat substitute product contains less than 1% wheat gluten.

[0204] In a more specific embodiment, the meat substitute product does not contain wheat gluten.

[0205] In another specific embodiment, the meat substitute product does not contain soy protein isolate.

[0206] In another specific embodiment, the meat substitute product does not contain soy protein concentrate.

[0207] In another specific embodiment, the meat substitute product does not contain soy protein.

[0208] In a more specific embodiment, the meat substitute product contains less than 5% carbohydrates.

[0209] In another specific embodiment, the meat substitute product does not contain tofu.

[0210] In some embodiments, the meat substitute product does not contain tofu and wheat gluten.

[0211] In some embodiments, the meat substitute product does not contain soy protein and wheat gluten.

[0212] In some embodiments, the meat substitute product does not contain animal products and contains less than 5% carbohydrates.

[0213] In some embodiments, the meat substitute product contains less than 1% cellulose.

[0214] In some embodiments, the meat substitute product contains less than 5% insoluble carbohydrates.

[0215] In some embodiments, the meat substitute product does not contain soy protein and contains less than 1% cellulose.

[0216] In some embodiments, the meat substitute product does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0217] In some embodiments, the meat substitute product does not contain wheat gluten and contains less than 1% cellulose.

[0218] In some embodiments, the meat substitute product does not contain wheat gluten and contains less than 5% insoluble carbohydrates.

[0219] In another aspect, the present invention provides a food product formed for consumption by animals, comprising one or more isolated, purified iron-containing proteins.

[0220] In some embodiments, one or more isolated, purified iron-containing proteins are derived from non-animal sources.

[0221] In some embodiments, the non-animal source is a plant source.

[0222] In some embodiments, the plant source comprises one or more legume plants.

[0223] In some embodiments, one or more of the legumes are soybean or pea plants.

[0224] In some embodiments, the non-animal source is genetically modified yeast or bacteria.

[0225] In some embodiments, the iron-containing protein is selected from the group consisting of hemoglobin, myoglobin, leghemoglobin, non-symbiotic hemoglobin, chlorocruorin, erythrocruorin, neuroglobin, cytoglobin, protoglobin, truncated 2 / 2 globin, HbN, cyanoglobin, HbO, Glb3, and cytochrome, helgate globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin.

[0226] In one embodiment, the iron-containing protein comprises an amino acid sequence having at least 70% homology to sequence 1. [Sequence 1: MVAFTEKQDALVSSSFEAFKANIPQYSVVFYTSILEKAPAAKDLFSFLANGVDPTNPKLTGHAEKLFALVRDSAGQLKASGTVVADAALGSVHAQKAVTDPQFVVVKEALLKTIKAAVGDKWSDELSRAWEVAYDELAAAIKKA].

[0227] In some embodiments, the food product does not contain methylcellulose, carrageenan, caramel color, konjac powder, gum arabic, and acacia gum.

[0228] In a specific embodiment, the food product contains less than 1% wheat gluten.

[0229] In a more specific embodiment, the food product does not contain wheat gluten.

[0230] In another specific embodiment, the food product does not contain soy protein isolate.

[0231] In another specific embodiment, the food product does not contain soybean protein concentrate.

[0232] In another specific embodiment, the food product does not contain soy protein.

[0233] In a more specific embodiment, the food product contains less than 5% carbohydrates.

[0234] In another specific embodiment, the food product does not contain tofu.

[0235] In some embodiments, the food product does not contain tofu and wheat gluten.

[0236] In some embodiments, the food product does not contain soy protein and wheat gluten.

[0237] In some embodiments, the food product does not contain animal products and contains less than 5% carbohydrates.

[0238] In some embodiments, the food product contains less than 1% cellulose.

[0239] In some embodiments, the food product contains less than 5% insoluble carbohydrates.

[0240] In some embodiments, the food product does not contain soy protein and contains less than 1% cellulose.

[0241] In some embodiments, the food product does not contain soy protein and contains less than 5% insoluble carbohydrates.

[0242] In some embodiments, the food product does not contain wheat gluten and contains less than 1% cellulose.

[0243] In some embodiments, the food product does not contain wheat gluten and contains less than 5% insoluble carbohydrates.

[0244] 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 the one or more proteins into a physical organization close to the physical organization of meat.

[0245] In another aspect, the present invention provides a method for manufacturing a muscle tissue imitation, comprising isolating and purifying one or more proteins; and assembling the one or more proteins into a physical organization close to the physical organization of skeletal muscle.

[0246] In another aspect, the present invention provides a method for producing an adipose tissue imitation, 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 reagents.

[0247] In another aspect, the present invention provides a method for producing a connective tissue imitation, comprising isolating and purifying one or more proteins; and precipitating the one or more proteins, wherein the precipitation produces one or more proteins that form a physical structure similar to the physical organization of connective tissue.

[0248] In some embodiments, the precipitation comprises solubilizing one or more proteins in a first solution; and extruding the first solution into a second solution, wherein one or more proteins are insoluble in the second solution, and the extrusion induces precipitation of one or more proteins.

[0249] In another aspect, the present invention provides a food product formed for consumption by animals, comprising one or more isolated, purified iron-containing proteins.

[0250] In another aspect, the present invention provides a food product formed for human consumption comprising one or more isolated, purified iron-containing proteins.

[0251] In another aspect, the present invention provides a food product formed for consumption by animals, comprising one or more isolated, purified iron-containing proteins. In another aspect, the present invention provides a food product formed for consumption by humans, comprising one or more isolated, purified iron-containing proteins.

[0252] All disclosures and patent applications mentioned in this specification are incorporated herein by reference to the same extent as where each individual disclosure or patent application is specifically and individually indicated to be included by reference. Brief explanation of the drawing

[0253] The patent or application file contains one or more drawings produced in color. A copy of this patent or patent application publication containing the color drawings will be provided by the relevant office upon the billing and payment of necessary fees. Novel features of the present invention are described particularly in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by referring to the following detailed description, which describes exemplary embodiments utilizing the principles of the present invention, and the following attached drawings: Figure 1 illustrates a comparison of aligned GCMS profiles (mirror orientations) of identical muscle-mimicking samples cooked in the presence of leghemoglobin (LHb, right profile) or ferric ions (left profile). In this example, the vertical axis represents the retention time in the gas chromatography separation step, and the horizontal axis represents the m / z ratio of ions generated by the fragmentation of the corresponding separated volatile compounds. Selected compounds that appeared differentially in the two samples are labeled on the right side of the figure. Figure 2 shows a portion of the root of a pea plant (Pisum sativum) with root nodules, incised to demonstrate the red color imparted by the leghemoglobin contained within. The incised root nodules appear red. Figure 3 shows leghemoglobin isolated from 1 oz of pea root. The usual red color from meat is evident in the color photograph. Figure 4 shows that leghemoglobin from various species is homologous and has similar color characteristics. In Figure 4, Panel A shows SDS_PAGE gels of dissolved root nodules from three leguminous species (1) broad beans (2) peas (3) and soybeans. Arrows indicate leghemoglobin, respectively. Panel B shows the similarity of UV-VIS spectral profiles of leghemoglobin from two different plant species (broad beans and soybeans). Figure 5 shows a comparison of reduced (heme iron 2+) and oxidized (heme iron 3+) soybean leghemoglobin (Figure 5 Panel A) and horse myoglobin (Figure 5 Panel B), demonstrating the similarity of the UV-VIS absorption profiles of the two proteins. The inventors purified soybean leghemoglobin from soybean root nodules using the protocol described herein. The purified horse myoglobin was purchased from SigmaAldrich. Soybean leghemoglobin (Figure 5 Panel A) and horse myoglobin (Figure 5 Panel B) were reduced using 1 mm sodium bisulfite. The UV-VIS absorption spectra of heme Fe3+ (blue line – higher peak in Figures 5 and 6) and heme Fe2+ (red line) of soybean leghemoglobin (Figure 5 Panel A) and horse myoglobin (Figure 5 Panel B) are shown. The inset shows an enlarged view of the UV-VIS spectrum in the 450 nm to 700 nm range. (Figure 5, Panel C) The image of a 10 µl liquid droplet of a 40 mg / ml solution of heme-Fe3+ soybean leghemoglobin (left droplet) shows a characteristic greenish-red color, and the image of a 40 mg / ml solution of heme-Fe2+ soybean leghemoglobin (right droplet) shows a characteristic red color, and (right image) corresponds to a sample of equine myoglobin. Figure 6 illustrates an example of the successful reduction of heme iron in leghemoglobin using sodium bisulfite and titanium citrate. In Figure 6, the UV-VIS spectrogram of purified soybean leghemoglobin in the oxidized (+3) state is represented by the blue curve in each panel (the blue curve has a higher peak in the main graph). The red curve in each panel represents the UV-VIS spectrum of the same leghemoglobin species after reduction to the (+2) state (red line) by adding 1 mM sodium bisulfite (Panel A) or 0.24% (wt / v) titanium citrate (Panel B) in 20 mM potassium phosphate, pH 7.3, and 100 mM sodium chloride buffer. The inset shows an enlarged view of the UV-VIS spectrum in the 450–700 nm region. In this example, leghemoglobin was purified from soybean root nodules using 60 / 90% ammonium sulfate fractionation and exchanged with 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride buffer. The sodium bisulfite stock solution was prepared by dissolving 100 mM sodium bisulfite in 1 mM sodium hydroxide in water. The titanium citrate stock solution was prepared by mixing 20% ​​(wt / v) Ti-chloride in hydrochloric acid with 0.2 M sodium citrate (1:10 v / v). The pH was adjusted to pH 7.0 using sodium carbonate. Figure 7 illustrates an example of the leghemoglobin purification flow from soybean root nodules. The figure shows different soybean leghemoglobin purification steps: (Lane 1) soybean root nodule lysate; (Lane 2) SDS-PAGE fractionation of soybean root nodule lysate purified by 60 / 90% (wt / v) ammonium sulfate fractionation. It shows the protein contents of a 90% ammonium sulfate fractionated protein pellet resuspended in 20 mM potassium phosphate pH 7.4, 100 mM sodium chloride, and 1 mM EDTA buffer; the protein from the 90% ammonium sulfate pellet was further purified by anion-exchange chromatography (FFQ GE Healthcare) in 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride. Leghemoglobin collected in the floss is shown in lane 3. The anion-exchange floss was fractionated using size-exclusion chromatography (Sephacryl S-100 GE Healthcare), and the resulting leghemoglobin fraction is shown in lane 4. The leghemoglobin content at various purification steps was determined by determining the fraction of leghemoglobin bands on the SDS-PAGE gel in each sample using ImageDoc analysis software (BioRad). The purity (partial abundance) of leghemoglobin at each stage of the purification step was: lysate: 32.7% (lane 1), 60 / 90% (wt / v) ammonium sulfate fractionation: 78% (lane 2), anion-exchange chromatography: ~83% (lane 3), and size-exclusion chromatography: up to ~95% (lane 4). Figure 8 shows stained SDS-PAGE gel analysis of (A) soybean leghemoglobin expressed and purified using recombinant protein technology and (B) soybean leghemoglobin purified from soybean root nodules. (A) Recombinant soybean leghemoglobin A possessing a His-tag and a TEV protease His-tag removal site was expressed in E. coli BL21 strain and purified using His-tag affinity chromatography (Talon resin, CloneTech). The left lane contains the molecular weight standard, and the right lane contains the purified recombinant soybean leghemoglobin A (arrow). The estimated 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 the molecular weight standard, and the right lane contains purified soybean leghemoglobin A (arrow). Mass spectroscopic analysis of the purified material determined that all four types of soybean leghemoglobin isoforms were present and were full length (data not presented). The estimated molecular weight (MW) of the soybean leghemoglobin isoforms is in the range of MW 15.4 to 15.8 kDa. Figure 9 shows an example of six cubes of a commercial meat analogue (Quorn chicken analogue) with sides of approximately 1 cm, of which four (left and bottom right) were immersed in a solution of approximately 10 mg / ml soybean leghemoglobin in 20 mM potassium phosphate pH 7.4 and 100 mM NaCl; and the remaining two (top right) were immersed in the same buffer without leghemoglobin. It is evident from the color photographs that the deep pink color of the leghemoglobin-injected cubes contrasts with the yellowish-brown color of the non-injected cubes. FIG. 10 shows four leghemoglobin-infused cubes of Quon chicken analogue being cooked in a pan at 350°C. Two lower cubes were flipped over to expose the baked surfaces, which turned brown. In the upper two cubes, the heated parts turned grayish-brown, while the cooler top surfaces retained their pink color. In some embodiments, a heme-containing solution, such as a leghemoglobin solution, was injected into the product until the product reached the color of raw meat. Figure 11 illustrates the mixture of 43 ml of mung bean protein solution (150 mg / ml in dialysis buffer) with 37 ml of leghemoglobin solution (46.5 mg / ml leghemoglobin and 20 mg / ml other soybean root nodule protein in 20 mM potassium phosphate, 100 mM NaCl, pH 7.3). 20 ml of transglutaminase solution (20% w / w) was added, the solution was thoroughly mixed, separated into two 50 ml Falcon tubes, and incubated overnight at room temperature. The final protein concentrations were 65 mg / ml for mung bean protein, 18 mg / ml for leghemoglobin, and 91 mg / ml for total protein. Figure 12 shows a "white" muscle analog prepared by mixing 43 ml of mung bean protein solution (150 mg / ml) with 45 ml of a 11.7 mg / ml solution of leghemoglobin and a 0.8% (wt / v) transglutaminase solution. 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. Figure 13 illustrates a mung bean-based lipid tissue analog, prepared in an Eppendorf tube, and formed an opaque gel of grayish-white, smooth, and uniform texture with no visually identifiable liquid incorporated into the gel. The gel stood upright and was elastic and resilient. The gel has a slightly pleasant aroma and a soft, pleasant flavor. The taste is mildly salty. Figure 14 illustrates a pea globulin-based tissue analog, prepared in an Eppendorf tube that is very similar to a mung bean-based fat analog, except that it has less oil upon compression. Figure 15 shows a connective-tissue analog strand produced using a 1:3 ratio in 70% ethanol, loaded into a syringe equipped with a 23-gauge needle (ID 0.337 mm). The solution was slowly extruded from the bottom of a 5-inch-high vessel into an excess of 5 M NaCl solution. The ethanol-zein solution, which is less dense than the NaCl solution, floated to the top and pulled out the fibrous strands of solidified zein. The NaCl was continuously stirred, and the strands began to unfold, assisting in strand elongation. The strands bundled together and became a hard, high-density mass. FIG. 16 shows a ground beef round patty (Fig. 16 Panel A) prepared by combining a 62% (wt / wt) muscle analog (62% (wt / wt) "dark muscle analog" and 38% (wt / wt) "white muscle analog"), a 29% (wt / wt) adipose tissue analog (from pea globulin and canola oil), and a 5% (wt / wt) connective tissue analog. Ground beef round patties were prepared by combining 62% muscle analogs (62% "dark muscle analogs" and 38% "white muscle analogs"), 29% adipose tissue analogs (from mung bean seed 8S protein and rice bran oil), and 5% connective tissue analogs (Figure 16, Panel B). Ground beef round patties were prepared by combining 71% (wt / wt) muscle tissue analogs (composed of 60% "white" muscle analogs and 40% "dark" muscle analogs) and 23% adipose tissue (from pea seed globulin protein and canola oil) (Figure 16, Panel C). Ground beef round patties were prepared by combining 67% "white" muscle analogs with 28% adipose tissue analogs (from pea globulin and rice bran oil) (Figure 16, Panel D). Figure 17 illustrates a ground beef patty analog prepared by combining a 62% (wt / wt) muscle tissue analog (62% (wt / wt) "dark muscle analog" and a 38% (wt / wt) "muscle analog"), a 29% (wt / wt) adipose tissue analog (from pea globulin and canola oil), and a 5% (wt / wt) connective tissue analog. The left panel shows the patty before cooking, and the right panel shows the same patty after cooking for about 2 minutes. The observer described the flavor of the ground beef imitation during cooking as distinctly "beef-like." Specific details for implementing the invention

[0254] Methods and compositions for the production of consumer goods are described herein. The consumer goods may be intended for animal consumption. For example, the consumer goods may be food suitable for human consumption. The consumer goods may be approved by a suitable regulatory authority. The consumer goods may be sold in grocery stores similar to existing human food, or manufactured in restaurants, schools, hospitals, military facilities, prisons, shelters, and long-term care facilities. The consumer goods may also be feed for livestock. For example, dog food may be produced according to the present invention. The consumer goods may also be feed for wild animals. For example, the consumer goods may be provided to non-domesticated predatory animals.

[0255] The consumer goods of the present invention may compete with, supplement, or replace animal-based foods. For example, the consumer goods may be meat imitations manufactured entirely from plant sources. The consumer goods may be manufactured to mimic the cuts or appearances of meat currently on the market. For example, the consumer goods may be visually similar to or indistinguishable from ground beef or a specific cut of beef. Alternatively, the consumer goods may be manufactured with a unique appearance or look. For example, the consumer goods may contain patterns or characters based on the structure of the consumer goods. In some cases, the consumer goods look like conventional meat products after their manufacture. For example, a consumer goods may be produced that is larger than a conventional cut of beef but looks identical to conventional cooked meat after being cut and heated. In some embodiments, the consumer goods may resemble the shape of conventional meat in two dimensions but not in three dimensions. For example, the consumer goods may resemble a cut of meat in two dimensions (e.g., when viewed from above), but may be much longer (or thicker) than a conventional cut. Accordingly, in some embodiments, a composition is provided that can be repeatedly cut into conventional meat-shaped products.

[0256] Consumer goods may be manufactured entirely from plant-based sources. In some cases, consumer goods may be manufactured from organic sources. Consumer goods may also be manufactured from a combination of plant-based and animal-based sources. For example, consumer goods may be a liver beef product supplemented with the plant-based product of the present invention.

[0257] Consumer goods may be manufactured from local products. For example, consumer goods may be manufactured from plants grown within a specific radius of the ultimate consumer. The radius could be, for example, 1, 10, 100, or 1,000 miles. Accordingly, in some embodiments, the present invention provides a method for producing a meat imitation that does not contain products transported more than 1, 10, 100, or 1,000 miles prior to the production of the meat imitation.

[0258] The present invention provides a method for producing consistent characteristics from consumer goods when produced from various sources. Thus, for example, a plant-based meat substitute produced from local plants in Iowa, USA, will have substantially similar taste, smell, and texture to a plant-based meat substitute produced from local plants in Lorraine, France. This consistency takes into account the advertising method of locally grown food having consistent characteristics. Consistency may result from the concentration or purification of similar ingredients from different locations. These ingredients may be combined in specific ratios to ensure consistency. In some embodiments, a high degree of characteristic consistency is made possible by using ingredients derived from the same plant species (e.g., isolated or concentrated proteins and fats). In some embodiments, a high degree of characteristic consistency is made possible by using ingredients derived from different plant species (e.g., isolated or concentrated proteins and fats). In some embodiments, the same protein may be isolated from different plant species. In some embodiments, the present invention provides a method comprising isolating similar plant components from plant sources at different locations, assembling the composition provided herein at both locations, and selling the composition, wherein the composition assembled and sold at different geographical locations has consistent physical and chemical properties. In some embodiments, the isolated components are from different plant populations at different locations. In some embodiments, one or more of the isolated components are transported to individual geographical locations.

[0259] Consumer goods may require fewer resources for production than consumer goods produced from domesticated animals. Accordingly, the present invention provides a meat substitute that requires less water or energy than meat for production. For example, consumer goods may require less than about 10, 50, 100, 200, 300, 500, or 1,000 gallons of water per pound of consumer goods. For comparison, beef may require more than 2,000 gallons of water per pound of meat.

[0260] Consumer goods may require less land for production than meat products containing similar protein content. For example, consumer goods may require less than 30% of the land area required to produce meat products containing similar protein content.

[0261] Consumer goods can have health benefits when replaced in the diet compared to animal products. For example, they may have less cholesterol or lower levels of saturated fat than similar meat products.

[0262] Consumer goods can provide animal welfare benefits when they replace animal products in the diet. For example, they can be produced without requiring confinement, force-feeding, early weaning, disruption of mother-offspring interaction, or the slaughter of animals for their meat.

[0263] Consumer goods can have a smaller "carbon footprint" than the meat products they replace. For example, consumer goods can cause 1%, 5%, 10%, 25%, 50%, or 75% of the greenhouse gas emissions attributable to the animal products they replace.

[0264] Consumer goods can provide alternatives to animal products or combinations of animal products whose consumption is prohibited by religious beliefs. For example, the consumer goods could be kosher pork chops.

[0265] Consumer goods can also be transported as ingredients and produced or assembled at different locations. Where available, local ingredients can be used in the production of consumer goods. These can be supplemented with ingredients that are not locally available. This considers a method of producing consumer goods, such as meat substitutes, using less energy for transport than is required for meat. For example, local water can be used in combination with kits that provide other ingredients for the consumer goods. Using local water will reduce transport weight, thereby reducing costs and environmental impact.

[0266] Consumer goods may be produced or assembled wholly or partially in areas where animal husbandry is impossible or not permitted. Consumer goods may be produced or assembled in an urban environment. For example, a kit that enables a user to produce consumer goods may be provided to the user. The user may, for example, use local water in Shanghai or plants from a rooftop garden. In another example, consumer goods may be produced on a spacecraft, a space station, or a lunar base. Accordingly, the present invention provides a method and system for producing meat substitutes for use in space travel or training therefor. For example, the present invention may be used for Earth-based training for space travel. Consumer goods may also be produced on islands where raising livestock is difficult or prohibited, or on artificial platforms at sea.

[0267] In some embodiments, the consumer product is designed to mimic the experience of eating meat. The appearance, texture, and taste of the consumer product may be similar to or indistinguishable from meat. Accordingly, in certain embodiments, the present invention provides a method for determining whether an animal or a human can distinguish the consumer product from meat.

[0268] One method of determining whether a consumer product is comparable to meat is to a) define the characteristics of the meat and b) determine whether the consumer product possesses similar characteristics. The characteristics of the meat that may be tested include mechanical properties, such as hardness, cohesiveness, brittleness, chewiness, gumminess, viscosity, elasticity, and adhesion. Additionally, the characteristics of the meat that may be tested include geometric properties, such as particle size and shape, and particle shape and orientation. Additional characteristics may include moisture content and fat content. These characteristics are described as "soft," "hard," or "hard" (description of hardness); "brittle," "crisp," "fragile," "poorly chewable," "soft," "tough," "crude," "splitting," "soft," or "sticky" (description of cohesiveness); "loose" or "viscous" (description of viscosity); "plastic" or "elastic" (description of elasticity); It may be described using terms such as "sticky," "adhesive," or "chewy" (description of adhesion); "rough," "coarse," or "rough" (description of particle shape and size); "fibrous," "cellular," or "crystalline" (description of particle shape and orientation); "dry," "moist," "wet," or "watery" (description of moisture content); or "oily" or "greasy" (description of fat content). Accordingly, in one embodiment, a group of people may be asked to grade a specific meat, e.g., ground beef, according to the characteristics describing the meat. These gradings may be used as indicators of the meat's characteristics. Subsequently, the consumer product of the present invention may be compared to meat to determine how similar the consumer product is to meat. In some cases, the characteristics of the consumer product are subsequently modified so that the consumer product becomes more similar to meat. Thus, in some embodiments, the consumer product is graded as similar to meat according to human evaluation. In some embodiments, the consumer product is indistinguishable from actual meat by humans.

[0269] In some embodiments, subjects are asked to identify the identity of the consumer product as meat. In some embodiments, one characteristic of the composition of the invention is that an animal, e.g., a human, will identify the composition as meat. In some embodiments, a human identifies the composition of the invention as having characteristics equivalent to meat. In some embodiments, one or more characteristics of meat are equivalent according to human perception. These characteristics include characteristics that can be tested. In some embodiments, a human identifies the consumer product of the invention as being more similar to meat than meat substitutes found in the art.

[0270] In one embodiment, an experiment may be conducted to determine whether the consumer product is acceptable to consumers. Various consumer products described herein may be screened using a panel. Numerous human panelists may test various consumer product samples, namely natural meat versus the consumer product composition described herein. Variables, such as fat content, may be standardized to, for example, 20% using a lean and fatty meat mix. Fat content may be determined using a Babcock for meat methods (see SS Nielson, Introduction to the Chemical Analysis of Foods (Jones & Bartlett Publishers, Boston, 1994)). A mixture of the consumer product of the present invention and ground beef prepared according to the procedure described herein may be devised.

[0271] Panelists may be served samples in a booth under red light or white light during a discussion-based consumer panel. Random three-digit numbers may be assigned to the samples, and voting positions may be rotated to prevent bias. Panelists may be asked to evaluate the samples for tenderness, juiciness, texture, flavor, and overall acceptability using a preference scale ranging from 1=extremely bad to 9=extremely good (median 5=neither good nor bad). Panelists may be encouraged to rinse their mouths with water between samples and provided with an opportunity to express their opinions on each sample.

[0272] The results of this experiment may indicate a significant difference (p<0.05) or similarity between conventional meat and the composition of the present invention.

[0273] These results will demonstrate that the composition of the present invention is deemed acceptablely equivalent to actual meat products. Additionally, these results may demonstrate that the composition of the present invention is preferred by panelists over other commercially available meat substitutes. Accordingly, in some embodiments, the present invention provides a consumer product that is significantly similar to conventional meat.

[0274] The consumer goods of the present invention may also have physical characteristics similar to conventional meat. In one embodiment, the force required to penetrate a 1-inch thick structure (e.g., a patty) made of the consumer goods of the present invention with a fixed-diameter steel rod does not differ significantly from the force required to penetrate a similar 1-inch thick meat structure (e.g., a ground beef patty) with a similar fixed-diameter steel rod. Thus, the present invention provides a consumer goods having physical strength characteristics similar to meat.

[0275] In some embodiments, the compositions of the present invention have cooking loss characteristics similar to meat. In one embodiment, the consumer product of the present invention, having fat and protein contents similar to ground beef, has the same size reduction as when cooked with actual ground beef. Similar similarity in size loss profiles can be obtained for various compositions of the consumer product described herein that are comparable to various meats.

[0276] In some embodiments, the consumer product is compared to actual meat based on olfactory readings. In various embodiments, the olfactory may be used to evaluate odor concentration and odor threshold, the odor threshold compared to a reference gas, a preference scale score to determine the degree of appreciation, or the relative intensity of the odor. In some embodiments, the olfactory allows for automated evaluation and training of an expert panel. Thus, in some embodiments, the consumer product is a product that elicits similar or identical olfactory readings. In some embodiments, the similarity is sufficient to exceed the detection threshold of human perception.

[0277] Gas Chromatography-Mass Spectrometry (GCMS) is a method that combines the characteristics of gas-liquid chromatography and mass spectrometry to separate and identify various substances within a test sample. In some embodiments, GCMS can be used to evaluate the characteristics of consumer goods. For example, volatile chemicals can be isolated from the headspace surrounding meat. These chemicals can be identified using GCMS. This generates a profile of the volatile chemicals in the headspace surrounding the meat. In some cases, each peak of the GCMS can be further evaluated. For example, humans can grade their experience of smelling the chemicals causing a specific peak. This information can be used to further refine the profile. Subsequently, GCMS can be used to evaluate the characteristics of consumer goods. The GCMS profile can be used to refine the consumer goods.

[0278] Characteristic flavor and aromatic components are generated during the cooking process by chemical reaction molecules, including amino acids, fats, and sugars, which are found in most meats as well as plants. Accordingly, in some embodiments, the consumer product is tested for similarity to meat during or after cooking. In some embodiments, an olfactory map of the cooked meat is generated using human grading, human evaluation, olfactory readings, or GCMS measurements, or a combination thereof. Similarly, an olfactory map of the consumer product, e.g., a meat imitation, may be generated. By comparing these maps, the similarity of the cooked consumer product to cooked meat can be evaluated. In some embodiments, the olfactory map of the consumer product during or after cooking is similar to or indistinguishable from the olfactory map of the cooked or cooking meat. In some embodiments, the similarity is sufficient to exceed the detection threshold of human perception.

[0279] In one aspect, the present invention provides a meat substitute (alternatively referred to herein as a “consumer product”) composed of ingredients derived substantially or entirely from non-animal sources, but reproducing the key characteristics associated with the cooking and consumption of an equivalent meat product derived from animals. The equivalent meat product may be white meat or dark meat. The equivalent meat product may be derived from any animal. Non-limiting examples of animals used to derive equivalent meat products include farm animals, e.g., cattle, sheep, pigs, chickens, turkeys, geese, ducks, horses, dogs, or game animals (whether wild or farm animals), e.g., rabbits, deer, bison, buffalo, boars, snakes, pheasants, quails, bears, elk, antelopes, pigeons, rock pigeons, grouse, foxes, wild boars, goats, kangaroos, emus, alligators, crocodiles, turtles, woodchucks, marmots, opossums, partridges, squirrels, raccoons, whales, seals, ostriches, capybaras, nutrias, guinea pigs, rats, mice, field mice, any various insects or other arthropods, seafood, e.g., fish, crabs, lobsters, oysters, mussels, scallops, abalone, squid, octopuses, sea urchins, sea cucumbers, etc. It is understood that while many meat products are typically derived from the skeletal muscle of an animal, meat can also be derived from other muscles or organs of an animal. In some embodiments, an equivalent meat product is a cut of meat derived from skeletal muscle. In other embodiments, an equivalent meat product is an organ, e.g., kidney, heart, liver, gallbladder, intestine, stomach, bone marrow, brain, thymus, lung, tongue. Thus, in some embodiments, the composition of the present invention is a consumer product similar to skeletal muscle or organ.

[0280] In some aspects, the present invention provides a meat substitute product comprising one or more of a first composition comprising a muscle tissue imitation, a second composition comprising an adipose tissue imitation, and / or a third composition comprising a connective tissue imitation, wherein one or more compositions are combined in a manner that reproduces the physical organization of meat. In other aspects, the present invention provides compositions for a muscle tissue imitation (referred herein as “muscle imitation”), an adipose tissue imitation (referred herein as “fat imitation”), and a connective tissue imitation (referred herein as “connective tissue imitation”). In some embodiments, the composition and the meat substitute product consist of ingredients derived primarily or entirely from non-animal sources. In an alternative embodiment, a meat substitute product comprising muscle, fat and / or connective tissue imitation, or one or more of said imitations, is partially derived from an animal source but supplemented with ingredients derived from a non-animal source. In another alternative embodiment, the present invention provides a meat product derived substantially from an animal source but supplemented with one or more of muscle tissue imitations, fat imitations, and / or connective tissue imitations, said imitations derived substantially or entirely from a non-animal source. A non-limiting example of such a meat product is an ultra-lean liver beef product supplemented with a non-animal-derived fat imitation that improves texture and mouthfeel while preserving the health benefits of low-animal-fat consumer goods. This alternative embodiment produces a product that more closely replicates the key characteristics associated with the manufacture and consumption of meat but is less costly, less associated with environmental impact, has fewer conflicts with animal welfare, or has improved health benefits for consumers.

[0281] The physical organization 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 so that the imitations described herein assemble with one another as in meat. In some embodiments, the consumer product is designed so that the imitations described herein assemble with one another as in cooked meat after cooking. In some embodiments, one or more of the muscle, fat, and / or connective tissue imitations are combined in a manner that replicates the physical organization of various cuts or the preparation of meat. In a best embodiment, the imitations are combined in a manner close to the physical organization of natural liver meat. In other embodiments, the imitations are combined in a manner close to various cuts of beef, e.g., particularly ribeye, filet mignon, and London broil.

[0282] Protein and protein sources

[0283] In some embodiments, any meat substitute, muscle tissue imitation, fat imitation, or connective tissue imitation comprises one or more isolated, purified proteins. In some embodiments, the meat substitute consists of one or more muscle imitations, fat imitations, and / or connective tissue imitations comprising one or more isolated, purified proteins. In other embodiments, the muscle imitation, fat imitation, and / or connective tissue imitation comprises one or more isolated, 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, purified proteins. For the purposes of this document, “purified protein” will refer to a preparation in which the cumulative abundance by mass of a protein component other than the specified protein, which may be a single monomeric or polymeric protein species, is reduced by more than 2 times, more than 3 times, more than 5 times, more than 10 times, more than 20 times, more than 50 times, more than 100 times, or more than 1000 times compared to the source material from which the specified protein was isolated.

[0284] In some embodiments, one or more isolated, 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, chemicals, or in vitro synthesis. In certain embodiments, one or more isolated, purified proteins are derived from plant sources. Non-limiting examples of plant sources include grains, e.g., corn, maize, rice, wheat, barley, rye, triticale, teff; oilseeds, e.g., cottonseed, sunflower seed, safflower seed, rapeseed; leafy vegetables, e.g., lettuce, spinach, kale, collard greens, turnip leaves, Swiss chard, mustard greens, dandelion leaves, broccoli, cabbage; thallus not typically consumed by humans, e.g., biomass crops, e.g., switchgrass, miscanthus, sorbum, other vegetables, alfalfa, corn stalks; thallus typically discarded from harvested plants, sugarcane leaves, tree leaves; root crops, e.g., cassava, sweet potato, potato, carrot, beet, turnip; legumes, e.g., clover; peas, e.g., cowpea, pea, yellow pea, green pea; beans, e.g., soybean, broad bean, lima bean, kidney bean. Includes garbanzo beans, mung beans, pinto beans, lentils, lupins, mesquite, carob, soybeans and peanuts, vetch (Vetchia), stylosanthes (Stylosanthes), Arachis, Indigofera, Acacia, Leucaena, Siamophysis, and Sesbania. Those skilled in the art will understand that proteins that can be isolated from any organism of the plant kingdom may be used in the present invention.

[0285] Proteins abundant in plants can be isolated in large quantities from one or more source plants and thus become an economical choice for use in any muscle, fat, connective tissue imitation or meat substitute product. Accordingly, in some embodiments, one or more isolated proteins comprise proteins abundant that are found in high levels in plants and can be isolated and purified in large quantities. In some embodiments, the proteins abundant comprise 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% of the total protein content of the source plants. In some embodiments, the protein-rich material comprises about 0.5-10%, about 5-40%, about 10-50%, about 20-60%, or about 30-70% of the total protein content of the source plant. In some embodiments, the protein-rich material comprises 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 weight of the dry matter of the source plant. In some embodiments, the protein-rich material comprises about 0.5-5%, about 1-10%, about 5-20%, about 10-30%, about 15-40%, or about 20-50% of the total weight of the dry matter of the source plant.

[0286] In certain embodiments, one or more isolated proteins comprise an abundant protein found in high levels in the leaves of a plant. In some embodiments, the abundant protein comprises 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%, and 80% of the total protein content of the leaves of the source plant. In some embodiments, the abundant protein comprises about 0.5-10%, about 5%-40%, about 10%-60%, about 20%-60%, or about 30-70% of the total protein content of the leaves of the source plant. In certain embodiments, one or more isolated proteins comprise ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco activase). Rubisco is a protein particularly useful for meat imitations due to its high solubility and amino acid composition that approaches the optimal ratio of essential amino acids for human nutrition. In certain embodiments, one or more isolated proteins comprise ribulose-1,5-bisphosphate carboxylase oxygenase activase (Rubisco activase). In certain embodiments, one or more isolated proteins comprise a nutrient storage protein (VSP).

[0287] In some embodiments, one or more isolated proteins comprise an abundant protein found in high levels in the seeds of a plant. In some embodiments, the abundant protein comprises 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 comprises about 0.5-10%, about 5%-40%, about 10%-60%, about 20%-60%, or about 30-70% or >70% of the total protein content of the seeds of the source plant. Non-limiting examples of proteins found at high levels in plant seeds are seed storage proteins, e.g., albumin, glycinin, cornglycinin, globulin, bicylin, cornalbumin, gliadin, glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein), proteome, secalin, tritheca, gluten, zein, any seed storage protein, oleosin, caloleosin, steroleosin, or other oil body proteins.

[0288] In some embodiments, one or more isolated proteins include proteins that interact with lipids within their structure and help stabilize lipids. Without being bound by any particular theory, these proteins can improve the fusion of lipids and / or fat imitations with other ingredients of meat substitute products, thereby inducing improved texture and mouthfeel of the final product. Non-limiting examples of lipid-interacting plant proteins are the oleosin family of proteins. Oleosins are lipid-interacting proteins found in the oil bodies of plants. Other non-limiting examples of plant proteins capable of stabilizing emulsions include seed storage proteins from Great Northern beans, albumin from peas, globulin from peas, 8S globulin from mung beans, and 8S globulin from kidney beans.

[0289] Muscle imitation

[0290] Numerous meat products contain a high proportion of skeletal muscle. Accordingly, the present invention provides a composition derived from a non-animal source that mimics the key characteristics of animal skeletal muscle or is similar thereto. In another aspect, the present invention provides a meat substitute product comprising a composition derived from a non-animal source that mimics animal skeletal muscle or is similar thereto. Such compositions will be labeled herein as "muscle imitations." In some embodiments, the muscle imitations and / or meat substitute products containing muscle imitations are partially derived from an animal source. In some embodiments, the muscle imitations and / or meat substitute products containing muscle imitations are entirely derived from a non-animal source.

[0291] Many meat products contain a high proportion of striated skeletal muscle in which individual muscle fibers are organized primarily in an isotropic manner. Accordingly, in some embodiments, muscle imitations contain fibers organized isotropically to a certain extent. In some embodiments, the fibers contain a protein component. In some embodiments, the fibers contain 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 of a protein component.

[0292] In some embodiments, the protein component comprises one or more isolated, purified proteins. For example, one or more isolated, purified proteins may comprise 8S globulin from mung bean seeds or albumin or globulin fractions from pea seeds. These proteins provide examples of proteins having properties advantageous for constructing meat imitations due to their ability to form a gel with a texture similar to animal muscle or adipose tissue. Examples and embodiments of one or more isolated, purified proteins are described herein. A list of potential candidates is essentially disclosed herein and may include proteins isolated from Rubisco, any major seed storage protein, fungi, bacteria, archaea, viruses, or genetically modified microorganisms, or synthesized in vitro. Proteins may be artificially designed to mimic the physical properties of animal muscle tissue. Proteins may be artificially designed to mimic the physical properties of animal muscle tissue. In some embodiments, one or more isolated, 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 by weight of the protein components.

[0293] Skeletal muscle of animals, such as beef cattle, typically contains a significant amount of glycogen that can constitute approximately 1% of the muscle tissue mass at the time of slaughter. After slaughter, some of this glycogen continues to be metabolized to produce products containing lactic acid, which lowers the pH of the muscle tissue and contributes to making the meat of desirable quality. Glycogen is a branched polymer of glucose in which the linear chains are linked together by alpha (1->4) glycosidic bonds and the branching points contain alpha (1->6) glycosidic bonds. Starch from plants, particularly amylopectin, is also a branched polymer of glucose in which the linear chains are linked together by alpha (1->4) glycosidic bonds and the branching points contain alpha (1->6) glycosidic bonds, and thus can be used as an analogue of glycogen in the construction of meat imitations. Accordingly, in some embodiments, the muscle or meat imitation contains starch or pectin.

[0294] Additional 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 compounds. Thus, in some embodiments, the muscle imitation may include sodium, potassium, calcium, magnesium, other metal ions, lactic acid, other organic acids, free amino acids, peptides, nucleotides, and sulfur compounds. In some embodiments, the concentration of sodium, potassium, calcium, magnesium, other metal ions, lactic acid, other organic acids, free amino acids, peptides, nucleotides, and / or sulfur compounds in the muscle imitation or consumer product is within 10% of the concentration found in the muscle or meat being imitated.

[0295] In another aspect, the present invention provides a method for manufacturing muscle imitations. In some embodiments, the composition is formed into asymmetric fibers before being incorporated into a consumable. In some embodiments, these fibers mimic muscle fibers. In some embodiments, the fibers are spun fibers. In other embodiments, the fibers are extruded fibers. Thus, the present invention provides a method for producing asymmetric or spun protein fibers. In some embodiments, the fibers are formed by extruding a protein component through an extruder. Extrusion methods are widely known in the art and are described in US6379738, US3693533, and US20120093994 (incorporated herein by reference).

[0296] In some embodiments, extrusion may be performed using an MPF19 twin-screw extruder with a cooling die (APV Baker, Grand Rapids, Michigan). The cooling die returns the extruder to atmospheric pressure after cooling, thereby substantially suppressing enlargement or puffing of the final product. In the MPF19 apparatus, the dry feed and liquid may be added separately and mixed in the barrel. Extrusion parameters may be, for example: a screw speed of 200 rpm, a product temperature at the die of 150°C, a feed rate of 23 g / min, and a water flow rate of 11 g / min. The product temperature may be measured during extrusion by a thermocouple at the end of the extrusion barrel. Observations may be made for color, opacity, structure, and texture for each collected sample. After the collected samples are optionally dried overnight at room temperature, they may be ground into a fine powder (<60 mesh) using a Braun food grinder. The pH of the sample can be measured in two sets using a 10% (w / v) slurry of the powder sample in distilled water.

[0297] fat imitation

[0298] Animal fat is important in the eating experience of cooked meat. Accordingly, the present invention provides a composition derived from a non-animal source that replicates the key characteristics of animal fat. In another aspect, the present invention provides a meat substitute product comprising a composition derived from a non-animal source that replicates animal fat. Such compositions will be labeled herein as "fat imitations." In some embodiments, fat imitations and / or meat substitute products comprising fat imitations are partially derived from an animal source.

[0299] In some embodiments, the meat substitute product has a fat component. In some embodiments, the fat content of the consumer product is 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 droplets of fat are suspended. In some embodiments, the gel is a soft, elastic gel containing protein and optionally carbohydrates. In certain embodiments, the protein used in the gel is a plant or microbial protein. In some embodiments, the protein used in the fat imitation will include Rubisco, any major seed storage protein, fungi, bacteria, archaea, viruses, or genetically modified microorganisms, or proteins synthesized in vitro. The protein may be artificially designed to mimic the physical properties of animal fat.

[0300] The fat droplets used in some embodiments of the present invention may be from various sources. In some embodiments, the source is a non-animal source. In certain embodiments, the source is a plant source.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, bacteria, algae, archaea or fungi or genetically modified bacteria, oils produced by algae, archaea or fungi, triglycerides, monoglycerides, diglycerides, sphingosides, glycolipids, lecithin, lysolecithin, phosphatidic acid, lysophosphatidic acid, oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, mystoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 Eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, ester of 18:2 conjugated linoleic acid or conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, It includes glycerol esters of 18:2 conjugated linoleic acid or conjugated oleic acid, or triglyceride derivatives of oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid.

[0301] In some embodiments, the fat droplets are derived from pulp or seed oil. In other embodiments, the source may be yeast or fungus. For example, in one embodiment, the fat droplets are Mortierella isabella ( Mortierella isabellina It contains triglycerides derived from ).

[0302] In some embodiments, vegetable oil is modified to resemble animal fat. Vegetable oil may be modified by flavoring agents or other agents to reproduce the taste and smell of meat during and after cooking. Accordingly, some aspects of the present invention include a method for testing the qualitative similarity between the cooking properties of animal fat and the cooking properties of vegetable oil in a consumer product.

[0303] In some embodiments, the fat imitation comprises a protein component comprising one or more isolated, purified proteins. The purified proteins contribute to the flavor and texture of the meat imitation. In some embodiments, the purified proteins can stabilize the emulsified fat. In some embodiments, the purified proteins can form a gel upon denaturation or enzymatic crosslinking, which mimics the appearance and texture of animal fat. Examples and embodiments of one or more isolated, purified proteins are described herein. In specific embodiments, one or more isolated proteins comprise proteins isolated from legumes. Non-limiting examples of legumes are described herein, but variations using other legumes are possible. In some embodiments, the legume is a pea plant. In some embodiments, the isolated, purified proteins stabilize the emulsion. In some embodiments, the isolated, purified proteins form a gel upon crosslinking or enzymatic crosslinking. In some embodiments, the isolated, purified proteins comprise seed storage proteins. In some embodiments, the isolated, purified protein comprises albumin. In some embodiments, the isolated, purified protein comprises globulin. In certain embodiments, the isolated, purified protein is purified pea albumin protein. In another specific embodiment, the isolated, purified protein is purified pea globulin protein. In another specific embodiment, the isolated, purified protein is mung bean 8S globulin. In another specific embodiment, the isolated, purified protein is oleosin. In another specific embodiment, the isolated, purified protein is caloleosin. In another specific embodiment, the isolated, purified protein is Rubisco. In some embodiments, the protein component is about 0.1% by dry weight or total weight, 0.It comprises 5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more of a fat imitation. In some embodiments, the protein component comprises about 0.1-5%, about 0.5-10%, about 1-20%, about 5-30%, about 10-50%, about 20-70%, or about 30-90% or more of a fat imitation based on dry weight or total weight. In some embodiments, the protein component comprises a solution containing one or more isolated, purified proteins.

[0304] In some embodiments, the fat imitation comprises a crosslinking enzyme that catalyzes a reaction causing covalent crosslinking between proteins. The crosslinking enzyme may be used to create or stabilize the desired structure and texture of the fat tissue imitation to mimic the desired texture of an equivalent desirable animal fat. Non-limiting examples of the crosslinking enzyme include, for example, transglutaminase, lysyl oxidase, or other amine oxidases (e.g., Pichia pastoris lysyl oxidase). In some embodiments, the crosslinking enzyme is isolated and purified from a non-animal source, and examples and embodiments thereof 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% (wt / vol) of the crosslinking enzyme. In certain embodiments, the crosslinking enzyme is transglutaminase.

[0305] In another aspect, the present invention provides a method for manufacturing a fat imitation. 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 isolated and homogenized. For example, an organic solvent mixture may be used to aid in mixing the lipids. Subsequently, the solvent may be removed. At that point, the lipids may be frozen, freeze-dried, or stored. Thus, in some aspects, the present invention provides a method for isolating and storing lipids selected to have characteristics similar to animal fats. Subsequently, the lipid film or cake may be hydrated. Hydration may be achieved using stirring or temperature changes. Hydration may occur from a precursor solution to a gel. After hydration, the lipid suspension may be sonicated or extruded to further modify the properties of the lipids in the solution.

[0306] In some embodiments, the fat imitation is assembled closely to the organized fat tissue of the meat. In some embodiments, some or all of the components of the fat imitation are suspended in the gel. In various embodiments, the gel may be a proteinaceous gel, a hydrogel, an organogel, or a xerogel. In some embodiments, the gel may be thickened to a desired viscosity using a polysaccharide or protein-based agent. For example, a gel can be thickened to form an architecture or structure for a consumer product by using starch, arrowroot, corn starch, Katakuri starch, potato starch, sago, tapioca, arginine, guar gum, locust bean gum, xanthan gum, collagen, egg white, furcellalan, 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, either alone or in combination.

[0307] In certain embodiments, the fat imitation is an emulsion comprising a solution of one or more proteins and one or more fats suspended as droplets. In some embodiments, the emulsion is stabilized into a gel by one or more crosslinking enzymes. In more specific embodiments, one or more proteins in the solution are isolated, purified proteins. In more specific embodiments, the isolated, purified proteins comprise a purified pea albumin-enriched fraction. In another more specific embodiment, the isolated, purified proteins comprise a purified pea globulin-enriched fraction. In another more specific embodiment, the isolated, purified proteins comprise a purified mung bean 8S globulin-enriched fraction. In a more specific embodiment, the isolated, purified proteins comprise a Rubisco-enriched fraction. In another specific embodiment, one or more fats are derived from plant-based oils.In a more specific embodiment, one or more fats are 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, bacteria, algae, archaea or fungi or genetically modified bacteria, oils produced by algae, archaea or fungi, triglycerides, monoglycerides, diglycerides, sphingosides, glycolipids, lecithin, lysolecithin, phosphatidic acid, lysophosphatidic acid, oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, mystoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 Eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, ester of 18:2 conjugated linoleic acid or conjugated oleic acid, or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, It is derived from one or more of the following: glycerol esters of 18:2 conjugated linoleic acid or conjugated oleic acid, or triglyceride derivatives of oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, caprylic acid, pelargonic acid, undecanic acid, linoleic acid, 20:1 eicosanic acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid, or 18:2 conjugated linoleic acid or conjugated oleic acid. In a more specific embodiment, one or more fats are rice bran oil.In another specific embodiment, one or more fats are canola oil. In another specific embodiment, the crosslinking enzyme is transglutaminase, lysyl oxidase, or other amine oxidase. In a more specific embodiment, the crosslinking enzyme is transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion comprising a protein solution of purified pea albumin emulsified with 40-80% rice bran oil and gel-stabilized with 0.5-5% (wt / vol) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion comprising a protein solution of partially purified mung bean 8S globulin emulsified with 40-80% rice bran oil and gel-stabilized with 0.5-5% (wt / vol) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion comprising a protein solution of partially purified mung bean 8S globulin emulsified with 40-80% canola oil and gel-stabilized with 0.5-5% (wt / vol) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion comprising a protein solution of purified pea albumin emulsified with 40-80% rice bran oil and gel-stabilized with 0.0001-1% (wt / vol) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion comprising a protein solution of partially purified mung bean 8S globulin emulsified with 40-80% rice bran oil and gel-stabilized with 0.0001-1% (wt / vol) transglutaminase. In a specific embodiment, the fat imitation is a high-fat emulsion comprising a protein solution of partially purified mung bean 8S globulin emulsified with 40-80% canola oil and stabilized into a gel with 0.0001-1% (wt / vol) transglutaminase.

[0308] connective tissue imitation

[0309] Animal connective tissue provides key textural characteristics that are an important component of the meat-eating experience. Accordingly, the present invention provides a composition derived from a non-animal source that reproduces the key characteristics of animal connective tissue. In another aspect, the present invention provides a meat substitute product comprising a composition derived from a non-animal source that reproduces the important textural and visual characteristics of animal connective tissue. Such compositions will be labeled herein as "connective tissue imitations." In some embodiments, the connective tissue imitations and / or meat substitute products comprising the connective tissue imitations are partially derived from an animal source.

[0310] Animal connective tissue can generally be divided into fascia-type and cartilage-type tissues. Fascia-type tissue is highly fibrous and resistant to elongation (having a high elastic modulus), has a high protein content, a moderate moisture content (approximately 50%), and a low-to-low fat and polysaccharide content. Accordingly, the present invention provides a connective tissue imitation that reproduces the key characteristics of fascia-type tissue. In some embodiments, the connective tissue imitation comprises approximately 50% protein and approximately 50% liquid by weight based on total weight, and has low fat and polysaccharide components.

[0311] The protein contents of most fascia-type connective tissues consist primarily of collagen. Collagen is characterized by a high fraction of proline and alanine and is also assembled into characteristic elongated fibrils or rod-shaped, flexible structures. Prolamin is a family of proteins found in non-animal sources, such as plant sources. Prolamin is highly abundant in plants and has an amino acid composition similar to collagen. Among the proteins tested by the inventors for this purpose, prolamin was particularly advantageous due to its low cost and the ability to easily form fibers or sheets during spinning or extrusion. Non-limiting examples of prolamin family proteins include, for example, zein (found in corn), and these include hordain from barley, gliadin from wheat, secalin and extensin from rye, capyrin from sorbum, and avenin from oats. In fascia-type connective tissue, the prolamin family of proteins is highly abundant, individually or in combination, and demonstrates suitability as a protein component because its overall amino acid composition is similar to collagen (high proportions of proline and alanine) and it is easy to process into films and fibers. In addition to zein (found in corn), these include hordain from barley, gliadin from wheat, secalin and extensin from rye, capyrin from sorbum, and avenin from oats. Other proteins may require supplementation with prolamins to achieve target specifications for physicochemical and nutritional properties. A list of potential candidates is essentially disclosed herein and may include proteins isolated from Rubisco, any major seed storage protein, fungi, bacteria, archaea, viruses, or genetically modified microorganisms, or synthesized in vitro. Proteins may be artificially designed to mimic the physical properties of animal connective tissue. Animal-derived or recombinant collagen, extensin (a hydroxyproline-rich glycoprotein abundant in the cell wall, e.g., Arabidopsis thaliana, whose monomer is a "collagen-like" rod-shaped flexible molecule).Proteins can be artificially designed to mimic the physical properties of animal connective tissue.

[0312] Methods for forming fascia-type connective tissue will be the same as those practiced in the art for producing fibrous or fiber-like structures by biological, chemical, or physical means, individually or in combination, sequentially or simultaneously, prior to final formation. These methods may include extrusion or spinning.

[0313] Cartilage-type tissue is macroscopically homogeneous, resistant to compression, and has a high water content (80% or less), a low protein (collagen) content, and a higher polysaccharide (proteoglycan) content (approximately 10% each).

[0314] In terms of composition, cartilage-type connective tissue will be very similar to fascia-type tissue, each having relative ratios adjusted to more closely mimic 'meat' connective tissue.

[0315] The method of forming cartilage-type connective tissue will be similar to that for fascia-type connective tissue, but is more convenient for producing an isotropic and homogeneous structure.

[0316] Fats can be suspended in a gel. In some embodiments, the present invention provides a method for producing droplets of fat suspended in a proteinaceous gel. Fats can be isolated from plant tissue and emulsified. Emulsification may utilize high-speed blending, homogenization, stirring, or temperature changes. The properties of the lipids in the solution may be further modified by sonicating or extruding the lipid suspension. At this point, in some embodiments, a gelling agent is added after other components of the consumable are added to the solution. In some embodiments, a crosslinking agent (e.g., transglutaminase or lysyl oxidase) is added to bind the components of the consumable. In other embodiments, after adding the gelling agent, the lipid / gel suspension is combined with additional components of the consumable. In fascia-type connective tissue, the prolamin family of proteins is highly abundant individually or in combination thereof, and since the overall amino acid composition is similar to collagen (high proportions of proline and alanine) and is easy to process into films and fibers, it demonstrates suitability for protein components. In addition to zein (found in corn), these include hordain from barley, gliadin from wheat, secalin from rye, extensin, capyrin from sorbum, and avenin from oats. Other proteins may require supplementation with prolamin to achieve target specifications for physicochemical and nutritional properties. A list of potential candidates is essentially disclosed herein and may include any major seed storage protein, animal-derived or recombinant collagen, and extensin (a hydroxyproline-rich glycoprotein abundant in the cell wall, e.g., Arabidopsis thaliana, whose monomer is a "collagen-like" rod-shaped flexible molecule).

[0317] In some embodiments, some or all of the components of the consumer product are suspended in the gel. In various embodiments, the gel may be a hydrogel, an organogel, or a xerogel. The gel may be made viscous using a polysaccharide or protein-based agent. For example, the gel may be thickened to form an architecture or structure for the consumer product by using, alone or in combination, starch, arrowroot, corn starch, katakuri starch, potato starch, sago, tapioca, arginine, guar gum, locust bean gum, xanthan gum, collagen, egg white, furcellaran, 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. Enzymes that catalyze reactions causing covalent crosslinking between proteins may also be used alone or in combination to form an architecture or structure for 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 an architecture or structure for the consumer product. In some embodiments, various gels having various components are combined to form the consumer product. For example, a gel containing a plant-based protein may be associated with a gel containing a plant-based fat. In some embodiments, the stings or fibers of the proteins are oriented parallel to each other and subsequently fixed in place by the application of a gel containing a plant-based fat.

[0318] The composition of the present invention may be expanded or enlarged by heating, such as frying, baking, microwave heating, heating in a forced ventilation system, heating in a ventilation tunnel, etc., according to methods widely known in the art.

[0319] In some embodiments, various gels having various components are combined to form a consumer product. For example, a gel containing a plant-based protein may be associated with a gel containing a plant-based fat. In some embodiments, strings or fibers of the protein are oriented parallel to each other and are then fixed in place by the application of a gel containing a plant-based fat.

[0320] In some embodiments, the meat imitation does not contain animal products, contains less than 1% wheat gluten, and does not contain methylcellulose, carrageenan, caramel color, and konjac powder, gum arabic, and acacia gum.

[0321] In some embodiments, the meat imitation does not contain animal products, wheat gluten, methylcellulose, carrageenan, caramel coloring, and konjac flour, gum arabic, and acacia gum.

[0322] In some embodiments, the meat imitation does not contain animal products, soy protein isolates, wheat gluten, methylcellulose, carrageenan, caramel coloring, and konjac powder, gum arabic, and acacia gum.

[0323] In some embodiments, the meat imitation does not contain animal products, soy protein concentrate, wheat gluten, methylcellulose, carrageenan, caramel color, and konjac powder, gum arabic, and acacia gum.

[0324] In some embodiments, the meat imitation does not contain animal products, soy protein, wheat gluten, methylcellulose, carrageenan, caramel color, and konjac powder, gum arabic, and acacia gum.

[0325] In some embodiments, the meat imitation does not contain animal products, tofu, wheat gluten, methylcellulose, carrageenan, caramel coloring, and konjac powder, gum arabic, and acacia gum.

[0326] In some embodiments, the meat imitation does not contain animal products, tofu, or wheat gluten.

[0327] In some embodiments, the meat imitation does not contain animal products, soy protein, or wheat gluten.

[0328] In some embodiments, the meat imitation does not contain methylcellulose, carrageenan, caramel coloring, konjac powder, gum arabic, and acacia gum.

[0329] In some embodiments, the meat imitation does not contain animal products and contains less than 5% carbohydrates.

[0330] In some embodiments, the meat imitation does not contain animal products, soy protein, wheat gluten, methylcellulose, carrageenan, caramel coloring and konjac powder, gum arabic and acacia gum, and contains less than 5% carbohydrates.

[0331] In some embodiments, the meat imitation does not contain animal products and contains less than 1% cellulose.

[0332] In some embodiments, the meat imitation does not contain animal products and contains less than 5% insoluble carbohydrates.

[0333] In some embodiments, the meat imitation does not contain animal products or soy protein and contains less than 1% cellulose.

[0334] In some embodiments, the meat imitation does not contain animal products or soy protein and contains less than 5% insoluble carbohydrates.

[0335] In some embodiments, the meat imitation does not contain animal products or wheat gluten and contains less than 1% cellulose.

[0336] In some embodiments, the meat imitation does not contain animal products or wheat gluten and contains less than 5% insoluble carbohydrates.

[0337] The percentages of various components can also be controlled. For example, non-animal-based substitutes for muscle, adipose tissue, connective tissue, and blood components can be combined in various non- and physical organization types to most closely approximate the appearance and feel of meat. Various components can also be arranged to ensure consistency between bites of the consumer product. Components can be arranged to ensure that no waste is generated from the consumer product. For example, while a typical cut of meat may typically contain inedible parts, meat imitations can improve upon meat by excluding these inedible parts. Such improvements result in reduced waste and transportation costs for all products manufactured and transported for consumption. Alternatively, meat imitations may include inedible parts to mimic the experience of consuming meat. These parts may include bone, cartilage, connective tissue, or other materials commonly referred to as cartilage, or materials that simulate these components. In some embodiments, the consumer product may contain simulated inedible parts of a meat product designed to serve a secondary function. For example, the mock bone may be designed to disperse heat during cooking, allowing the consumer product to be cooked faster or more uniformly than meat. In another embodiment, the mock bone may also function to maintain the consumer product at a constant temperature during transport. In another embodiment, the mock inedible part may be biodegradable.

[0338] In some embodiments, the meat substitute composition does not contain animal protein, comprises 10-30% protein, 5-80% water, and 5-70% fat, and comprises one or more isolated, purified proteins. In certain embodiments, the meat substitute composition comprises transglutaminase.

[0339] In some embodiments, the consumer product contains ingredients that mimic the composition of meat. The major component of meat is typically skeletal muscle. Skeletal muscle typically consists of approximately 75 percent water, 19 percent protein, 2.5 percent intramuscular fat, 1.2 percent carbohydrates, and 2.3 percent other soluble non-protein materials. These include organic acids, sulfur compounds, nitrogen-containing compounds, such as amino acids and nucleotides, and inorganic materials, such as minerals. Accordingly, some embodiments of the present invention provide a consumer product that mimics an approximation of this composition. For example, in some embodiments, the consumer product is a plant-based meat imitation, which may comprise approximately 75 percent water, 19 percent protein, 2.5 percent fat, 1.2 percent carbohydrates; and 2.3 percent other soluble non-protein materials. In some embodiments, the consumable is a plant-based meat imitation comprising 60-90% water, 10-30% protein, 1-20% fat, 0.1-5% carbohydrates; and 1-10% other soluble non-protein material. In some embodiments, the consumable is a plant-based meat imitation comprising 60-90% water, 5-10% protein, 1-20% fat, 0.1-5% carbohydrates; and 1-10% other soluble non-protein material. In some embodiments, the consumable is a plant-based meat imitation comprising 0-50% water, 5-30% protein, 20-80% fat, 0.1-5% carbohydrates; and 1-10% other soluble non-protein material. In some embodiments, the imitation contains 0.01% to 5% by weight of heme protein. In some embodiments, the imitation contains 0.01% to 5% by weight of leghemoglobin. Some meats also contain myoglobin, a heme protein, which accounts for most of the red and iron content of some meats. In some embodiments, the imitation contains 0.01% to 5% by weight of heme protein. In some embodiments, the imitation contains 0.It contains 0.1% to 5% by weight of leghemoglobin. It is understood that these percentages may vary from meat and that meat imitations can be produced to be close to the natural variations of meat. Additionally, in some cases, the present invention provides improved meat imitations containing these components in typically non-natural percentages. For example, meat imitations may be produced to exceed the typical average fat content. The percentages of these components may also be changed to increase other desirable properties.

[0340] In some cases, meat imitations are designed so that the percentage of ingredients becomes similar to cooked meat upon cooking. Thus, in some embodiments, the uncooked product has a different percentage of ingredients than uncooked meat, but upon cooking, the product becomes similar to cooked meat. For example, the meat imitation may be manufactured to have a higher moisture content than the typical moisture content of unprocessed meat, but the product produced when cooked in a microwave has a percentage of ingredients similar to meat cooked over a fire.

[0341] In some embodiments, the consumer product is a meat imitation having a moisture content lower than the typical moisture content of meat. In some embodiments, the present invention provides a method for hydrating a meat imitation to induce the meat imitation to have a content similar to that of meat. For example, a meat imitation having a moisture content lower than that of meat, e.g. 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.

[0342] 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 higher than that of meat. In some embodiments, the consumer product has a lower protein content than meat.

[0343] Proteins in consumer goods may be derived from various sources or combinations thereof. Non-animal sources may provide some or all of the proteins in consumer goods. Non-animal sources may include vegetables, fruits, nuts, grains, algae, bacteria, or fungi. Proteins may be isolated or concentrated from one or more of these sources. In some embodiments, consumer goods are meat imitations containing proteins obtained solely from non-animal sources.

[0344] In some embodiments, the protein is formed into asymmetric fibers for incorporation into a consumer product. In some embodiments, these fibers mimic muscle fibers. In some embodiments, the protein is a spun fiber. Accordingly, the present invention provides a method for producing asymmetric or spun protein fibers. In some embodiments, the consumer product contains a protein or proteins having all amino acids found in proteins essential for human nutrition. In some embodiments, the protein added to the consumer product is supplemented with amino acids.

[0345] Indicator for cooked meat

[0346] The release of flavoring agents 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 produce a flavor that can be recognized by humans as typical cooked beef when cooked. In some embodiments, the consumer product produces a flavor that can be recognized by humans as typical cooked pork when cooked. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that produce a flavor that can be recognized by humans as typical cooked bacon when cooked. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that produce a flavor that can be recognized by humans as typical cooked chicken when cooked. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that produce a flavor that can be recognized by humans as typical cooked lamb when cooked. In some embodiments, the consumer product is a meat imitation composed entirely of non-animal products that produce a flavor that can be recognized by humans as typical cooked fish when cooked. In some embodiments, the consumer product is a meat imitation composed entirely of a non-animal product that produces a flavor that can be recognized by humans as a typical cooked turkey when cooked. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources that contain an odorant released upon cooking. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources that contain an odorant produced by a chemical reaction occurring upon cooking.In some embodiments, the consumable is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, containing a mixture of proteins, peptides, amino acids, nucleotides, sugars and polysaccharides and fats in a combination and spatial arrangement that allows these compounds to undergo chemical reactions during cooking to produce odorants and flavor-generating compounds. In some embodiments, the consumable is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, containing volatile or unstable odorants that are released upon cooking. In some embodiments, the consumable is a method for producing a meat imitation composed of ingredients derived primarily or entirely from non-animal sources that release volatile or unstable odorants when heated.

[0347] The odorants released during the cooking of meat are generated by reactions in which reactant fats, proteins, amino acids, peptides, nucleotides, organic acids, sulfur compounds, sugars, and other carbohydrates may be involved. In some embodiments, the odorants combined during the cooking of meat are isolated and placed near each other within the consumer product so that the odorants combine during the cooking of the consumer product. Thus, in some embodiments, characteristic flavor and aromatic components are generated during the cooking process by chemical reactions involving amino acids, fats, and sugars found in meat as well as plants. Thus, in some embodiments, characteristic flavor and aromatic components are generated during the cooking process by chemical reactions involving one or more amino acids, fats, peptides, nucleotides, organic acids, sulfur compounds, sugars, and other carbohydrates found mostly in meat as well as plants.

[0348] Some reactions that generate odorants released during the cooking of meat can be catalyzed by iron, particularly the heme iron of myoglobin. Thus, in some embodiments, some of the characteristic flavor and aromatic components are produced during the cooking process by chemical reactions catalyzed by iron. In some embodiments, some of the characteristic flavor and aromatic components are produced during the cooking process by chemical reactions catalyzed by heme. In some embodiments, some of the characteristic flavor and aromatic components are produced during the cooking process by chemical reactions catalyzed by the heme iron in leghemoglobin. In some embodiments, some of the characteristic flavor and aromatic components are produced during the cooking process by chemical reactions catalyzed by the heme iron in heme proteins.

[0349] Evidence that the presence of leghemoglobin contributes favorably to the flavor of meat imitations is as follows: Muscle imitations containing pea flour, sunflower oil, and glucose were heated at 140°C for 10 minutes in a sealed container holding solid-phase microextracted (SPME) fibers in the presence of reduced leghemoglobin (LHb) or a mixture of iron (Fe3+), sodium, and EDTA (EFS). These fibers contain polydimethylsiloxane (PDMS) that adsorbs volatile compounds upon analysis by GC-MS. Analysis of GC-MS data from various imitations reveals consistent differences between LHb and EFS samples. Non-limiting examples of compounds found exclusively or more abundantly in LHb samples include: 2-octanone, 2-methylfuran (often associated with the flavor of cooked meat), and several unresolved compounds. For example, Fig. 1 illustrates a comparison of aligned GCMS profiles (mirror orientations) of otherwise identical muscle-mimicking samples cooked in the presence of leghemoglobin (LHb, right profile) or ferric ions (left profile). In this example, the vertical axis represents the retention time in the gas chromatography separation step, and the horizontal axis represents the m / z ratio of ions generated by the fragmentation of the corresponding separated volatile compounds. Selected compounds that appeared differentially in the two samples are labeled on the right side of the figure.

[0350] Color indicator

[0351] The color of meat is an important part of the cooking and eating experience. For example, a cut of beef has a characteristic red color in its raw state and gradually changes to brown during cooking. As another example, white meats, such as chicken or pork, have a characteristic pink color in their raw state and gradually change to a white or brownish color during cooking. The amount of color change indicates the cooking progress of the beef and is used to determine the cooking time and temperature to produce a state of desired doneness. In some aspects, the present invention provides a non-meat-based 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 certain embodiments, the color indicator reproduces the color change of the meat cut as the meat progresses from a raw state to a cooked state. In more specific embodiments, the color indicator imparts a pre-cooked red color to the meat substitute product indicating a raw state and induces the meat substitute product to change to brown during cooking. In other specific embodiments, the color indicator imparts a pre-cooked pink color to the meat substitute product indicating a raw state and induces the meat substitute product to change to white or brown during cooking.

[0352] The primary determinant of the nutritional regulation of meat color is the concentration of iron-retaining proteins within the meat. In the skeletal muscle components of meat products, one of the major iron-retaining proteins is myoglobin. White chicken meat contains less than 0.05%; pork and veal contain 0.1-0.3%; young beef contains 0.4-1.0%; and old beef contains 1.5-2.0%. Thus, in some embodiments, the consumer product is a meat imitation containing iron-retaining proteins. In some embodiments, the meat imitation contains iron-retaining protein of 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% based on dry weight or gross weight. In some cases, the iron-retaining protein is isolated and purified from a source. In other cases, the iron-retaining protein is not isolated and purified. In some cases, the source of the iron-retaining protein is 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 iron-retaining protein is myoglobin. In some embodiments, the consumer product is a plant-based meat imitation to which animal myoglobin has been added. Thus, for example, the imitation of young beef may have about 0.4-1% myoglobin. In some cases, the iron-retaining protein is leghemoglobin. In some embodiments, the consumer product is a plant-based meat imitation to which leghemoglobin has been added. Thus, for example, the imitation of young beef may have about 0.4-1% leghemoglobin. In some cases, the iron-retaining protein is cytochrome. In some embodiments, the consumer product is a plant-based meat imitation to which cytochrome has been added. Thus, for example, the imitation of young beef may have about 0.It can have 4-1% cytochrome.

[0353] Another example of an iron-retaining protein 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 storing and recycling blood from animal farms to supplement the color of consumer goods. For example, blood is stored from a slaughterhouse, and hemoglobin from the blood is used to enhance the color of consumer goods. In some aspects, the consumer goods are plant-based meat substitutes containing hemoglobin.

[0354] Additional iron-containing proteins exist in nature. In some embodiments, the consumer product contains an iron-containing protein other than 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 substitute containing an iron-containing protein other than myoglobin or hemoglobin.

[0355] Examples of iron-containing proteins include hemoglobin, myoglobin, neuroglobin, cytoglobin, leghemoglobin, non-symbiotic hemoglobin, helgate globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin. In various embodiments, these iron-containing proteins are added to consumer products to alter the visual characteristics or iron content of the consumer products. In some embodiments, the consumer products contain heme proteins (e.g., hemoglobin, myoglobin, neuroglobin, cytoglobin, leghemoglobin, non-symbiotic hemoglobin, helgate globin I, bacterial hemoglobin, ciliated myoglobin, flavohedron hemoglobin).

[0356] Leghemoglobin, which is similar in structure and physical properties to myoglobin, is readily available as an unused byproduct of commercial legume crops (e.g., soybeans, peas). The leghemoglobin in the roots of these crops within the United States exceeds the myoglobin content of all red meat consumed in the United States. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, including muscle tissue imitation, adipose tissue imitation, connective tissue imitation, and leghemoglobin. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, containing heme protein. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, containing leghemoglobin. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, containing a member of the globin protein family. In some embodiments, the consumer product is a meat imitation composed of ingredients derived primarily or entirely from non-animal sources, having a high iron content from heme protein. In some embodiments, the iron content is similar to that of meat. In some embodiments, the consumer product has the distinctive red color of meat, such as the color provided by leghemoglobin.

[0357] In some embodiments, leghemoglobin is used as an indicator that the cooking of the consumer product is complete. Accordingly, one embodiment of the present invention is a method for cooking a consumer product comprising detecting leghemoglobin that has migrated from the interior to the surface of the consumer product when the product is cooked. Another embodiment of the present invention is a method for cooking a consumer product comprising detecting a color change from red to brown when the product is cooked.

[0358] In some embodiments, heme protein is used as an indicator that the cooking of the consumer product is complete. Accordingly, one embodiment of the present invention is a method for cooking a consumer product comprising detecting leghemoglobin that has migrated from the interior to the surface of the consumer product when the product is cooked. Another embodiment of the present invention is a method for cooking a consumer product comprising detecting a color change from red to brown when the product is cooked.

[0359] In some embodiments, heme proteins from the group consisting of hemoglobin, myoglobin, neuroglobin, cytoglobin, leghemoglobin, non-symbiotic hemoglobin, helgate globin I, bacterial hemoglobin, ciliated myoglobin, and flavohedron hemoglobin are used as indicators that the cooking of a consumer product is complete. Accordingly, one embodiment of the present invention is a method for cooking a consumer product comprising detecting leghemoglobin that has migrated from the interior of the consumer product to the surface when the product is cooked. Another embodiment of the present invention is a method for cooking a consumer product comprising detecting a change in color from red to brown when the product is cooked.

[0360] Food products containing isolated, purified leghemoglobin

[0361] In some embodiments, leghemoglobin is added to meat to enhance its characteristics. For example, a leghemoglobin-containing solution may be injected into raw or cooked meat. In another example, a leghemoglobin solution is dropped onto meat or the consumer product of the present invention to enhance its appearance. In one embodiment, advertising, photography, or videography of food products, such as meat or meat substitutes, is enhanced by leghemoglobin.

[0362] Sources of leghemoglobin

[0363] In some embodiments, the present invention provides a method for obtaining leghemoglobin from plants. Leghemoglobin can be obtained from various plants. Various leguminous species and varieties thereof, e.g., soybeans, broad beans, lima beans, cowpeas, peas, yellow peas, lupins, kidney beans, garbanzo beans, peanuts, alfalfa, vetch hay, clover, lespedeza, and pinto kidney beans, contain nitrogen-fixing root nodules in which leghemoglobin plays a major role in regulating oxygen concentration (e.g., root nodules from pea plants, Fig. 2). Fig. 3 shows 100 ml of a leghemoglobin solution isolated from 30 grams of pea root nodules. Leghemoglobin from various species is homologous and has similar color characteristics (Fig. 4). In FIG. 4, Panel A shows the SDS-PAGE gel of lysed root nodules of three leguminous plant species: (1) broad beans, (2) peas, and (3) soybeans. Arrows indicate leghemoglobin, respectively. Note that leghemoglobin is the most abundant soluble protein in each lysate. Panel B shows the similarity of the UV-VIS spectral profiles of leghemoglobin from two different plant species (broad beans and soybeans). The inventors purified leghemoglobin from broad bean (green curve) and soybean (red curve) root nodules using protocols described elsewhere in the specification. The UV-VIS spectra of both proteins show that the heme iron is in a reduced (+2) state. Note that they overlap almost perfectly and visually match to the same red color. The heme iron in each leghemoglobin was reduced to a +2 oxidation state by incubating broad bean and soybean leghemoglobins with 10 mM sodium bisulfite in 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride buffer. Subsequently, sodium bisulfite was removed from the leghemoglobin solution using gel-exclusion chromatography. The inset shows an enlarged view of the UV-VIS spectrum in the 450 nm to 700 nm region.Some plant species express multiple leghemoglobin isoforms (for example, soybeans have four types of leghemoglobin isoforms). Small variations in the precise amino acid sequence can alter the overall charge of a protein at a specific pH and modify the precise structural conformation of the iron-containing heme group within leghemoglobin. Differences in the structural conformation of the heme group in various leghemoglobins can affect the oxidation and reduction rates of heme iron. These differences can contribute to the color and flavor characteristics of various leghemoglobins.

[0364] Leghemoglobin has substantially the same absorbance spectrum and visual appearance as myoglobin from animal muscle. Figure 5 shows a comparison of reduced (heme iron 2+) and oxidized (heme iron 3+) soybean leghemoglobin (Figure 5 Panel A) and equine cardiac myoglobin (Figure 5 Panel B), demonstrating the similarity of the UV-VIS absorption profiles of the two proteins. The inventors purified soybean leghemoglobin from soybean root nodules using the protocol described herein. Purified equine myoglobin was purchased from Sigma-Aldrich. Soybean leghemoglobin (Figure 5 Panel A) and equine myoglobin (Figure 5 Panel B) were reduced using 1 mm sodium bisulfite. UV-VIS absorption spectra of heme Fe3+ (blue line) and heme Fe2+ (red line) of soybean leghemoglobin (Figure 5 Panel A) and equine myoglobin (Figure 5 Panel B) are shown. The inset shows an enlarged view of the UV-VIS spectrum in the 450 nm to 700 nm region. (Figure 5 Panel C) The image of a 10 µl liquid droplet of a 40 mg / ml solution of soybean leghemoglobin in the heme-Fe3+ state (left droplet) shows a characteristic greenish-red color, the image of a 40 mg / ml solution of soybean leghemoglobin in the heme-Fe2+ state (right droplet) shows a characteristic red color, and (right image) corresponds to a sample of equine myoglobin.

[0365] In another embodiment, leghemoglobin may be supplied from a non-plant source, for example, from an organism such as bacteria or yeast that has been genetically modified to express high levels of leghemoglobin.

[0366] The oxidation state of iron ions within leghemoglobin is important for its color. Leghemoglobin with heme iron in a +2 oxidation state appears as a vivid red color, whereas leghemoglobin with heme iron in a +3 oxidation state appears as a brownish-red color. Therefore, when 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. The heme iron within leghemoglobin can be converted from the oxidized (+3) state to the reduced (+2) state by a reducing agent. An example of the successful reduction of heme iron in leghemoglobin using sodium bisulfite and titanium citrate is shown in Fig. 6. In Fig. 6, the UV-VIS spectrogram of purified soybean leghemoglobin with heme iron in an oxidized (+3) state is shown as a blue curve in each panel. In each panel, the red curve represents the UV-VIS spectrum of the same leghemoglobin species after reduction to the (+2) state (red line) by adding 1 mM sodium bisulfite (Panel A) or 0.24% (wt / v) titanium citrate (Panel B) in 20 mM potassium phosphate, pH 7.3, 100 mM sodium chloride buffer. The inset shows an enlarged view of the UV-VIS spectrum in the 450–700 nm region. In this example, leghemoglobin was purified from soybean root nodules using 60 / 90% ammonium sulfate fractionation and exchanged with 20 mM potassium phosphate, pH 7.4, 100 mM sodium chloride buffer. The sodium bisulfite stock solution was prepared by dissolving 100 mM sodium bisulfite in 1 mM sodium hydroxide in water. The titanium citrate stock solution was prepared by mixing 20% ​​(wt / v) Ti-chloride in hydrochloric acid with 0.2M sodium citrate (1:10 v / v). The pH was adjusted to pH 7.0 using sodium carbonate.

[0367] Leghemoglobin can be purified from leguminous root nodules, such as those of peas or soybeans (Fig. 1 shows pea root nodules, and Fig. 2 shows leghemoglobin isolated therefrom). After thoroughly washing soybean root nodules to remove soil and irrelevant root tissue, the root nodules were dissolved in 20 mM potassium phosphate at pH 7.4, 100 mM sodium chloride, 1 mM EDTA, and 1 mM ascorbic acid. The root nodules were dissolved by grinding them using a Vitamix blender. For some samples, a polyvinylpyrrolidone polymer was added at 30% wt / v to assist in the removal of plant phenolic small molecules that mediate the oxidation of the heme-iron in leghemoglobin. The root nodule lysate was fractionated in two steps using ammonium sulfate, with the addition of 60% wt / v ammonium sulfate. The pellet was discarded, and the supernatant was prepared with 90% wt / v ammonium sulfate. Leghemoglobin was collected as a pellet precipitated in the 90% ammonium sulfate fraction. The ammonium sulfate-precipitated leghemoglobin was resuspended in 20 mM potassium phosphate, 1 mM EDTA, 50 mM sodium chloride, and ammonium sulfate, and removed by dialysis or size-exclusion chromatography in the same buffer. In some cases, this was the final purification step, whereas in others, the leghemoglobin was further purified using anion-exchange chromatography (FFQ GE Healthcare), often followed by size-exclusion chromatography (Sephakril S-100, GE Healthcare). Soybean leghemoglobin from 90% ammonium sulfate pellets in various buffers (20 mM potassium phosphate pH 7.4, containing 0 to 100 mM sodium chloride, 20 mM Tris pH 8, containing 0 to 100 mM sodium chloride, 20 mM sodium borax pH 9.It was loaded onto an anion exchange column (FFQ or DEAE, GE Healthcare) in pH 8, 20 mM sodium chloride, 20 mM sodium carbonate, pH 9, 20 mM sodium chloride, and purified using flow-through or sodium chloride (0-1 M salt gradient). An example of the leghemoglobin purification stream from soybean root nodules is shown in Fig. 7. The figure shows different soybean leghemoglobin purification steps: (Lane 1) soybean root nodule lysate; (Lane 2) SDS-PAGE fractionation of soybean root nodule lysate purified by 60 / 90% (wt / v) ammonium sulfate fractionation. It shows the protein contents of the 90% ammonium sulfate fractionated protein pellet resuspended in 20 mM potassium phosphate, pH 7.4, 100 mM sodium chloride, and 1 mM EDTA buffer; Protein from the 90% ammonium sulfate pellet was further purified by anion-exchange chromatography (FFQ GE Healthcare) in 20 mM potassium phosphate pH 7.4 and 100 mM sodium chloride. The leghemoglobin collected in the floss is shown in lane 3. The anion-exchange floss was fractionated using size-exclusion chromatography (Sephakril S-100 GE Healthcare), and the resulting leghemoglobin fraction is shown in lane 4. The leghemoglobin content at various purification steps was determined by determining the fraction of leghemoglobin bands on the SDS-PAGE gel in each sample using ImageDoc analysis software (Biorad). The purity (partial abundance) of leghemoglobin at each stage of the purification step was: lysate: 32.7% (lane 1), 60 / 90% (wt / v) ammonium sulfate fractionation: 78% (lane 2), anion-exchange chromatography: ~83% (lane 3), and size-exclusion chromatography: up to ~95% (lane 4).

[0368] Leghemoglobin can also be produced by genetically modifying bacteria or fungi to produce it. An exemplary example is shown in Fig. 8. Fig. 8 shows stained SDS-PAGE gel analysis of (A) soybean leghemoglobin expressed and purified using recombinant protein technology and (B) soybean leghemoglobin purified from soybean root nodules. (A) Recombinant soybean leghemoglobin A, possessing a His-tag and a TEV protease His-tag removal site, was expressed in E. coli BL21 strain and purified using His-tag affinity chromatography (Talon resin, Clontech). The left lane contains the molecular weight standard, and the right lane contains the purified recombinant soybean leghemoglobin A (arrow). The estimated 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 the molecular weight standard, and the right lane contains purified soybean leghemoglobin A (arrow). Mass spectroscopic analysis of the purified material determined that all four types of soybean leghemoglobin isoforms were present and were full length (data not presented). The estimated molecular weight (MW) of the soybean leghemoglobin isoforms is in the range of 15.4 to 15.8 kDa.

[0369] Leghemoglobin purified from soybeans and broad bean root nodules, respectively, was tasted by a panel of volunteers, and in each case, it was described as tasting like blood.

[0370] Leghemoglobin can be isolated from root nodules of leguminous plants, such as soybeans, broad beans, cowpeas, lima beans, kabanjo beans, peas, lupins, Lotus japonica, or other leguminous plants. Root nodules (e.g., root nodules from pea plants, Fig. 1) are obtained, homogenized in an aqueous solution, and after removing insoluble substances by precipitation or filtration, soluble proteins including leghemoglobin are recovered. Leghemoglobin can be purified by selective precipitation and / or chromatography and / or the use of molecules having a specific affinity for leghemoglobin. (Fig. 2 shows 100 ml of a solution of leghemoglobin isolated from 30 grams of pea root nodules).

[0371] Heme proteins, such as leghemoglobin, may be combined with other plant-based meat imitation ingredients. In some embodiments, the heme protein is captured in a gel containing other ingredients, such as lipids and / or proteins. In some aspects, various gels are combined with non-gel-based heme proteins. In some embodiments, the combination of the heme protein and other compounds of the consumer product is performed to ensure that the heme protein can diffuse through the consumer product. In some embodiments, the consumer product is immersed in a heme-protein-containing solution, such as a leghemoglobin solution. In some embodiments, the consumer product is immersed in a heme protein-containing solution, such as a leghemoglobin solution, for 1, 5, 10, 15, 20, or 30 hours. In some embodiments, the consumer product is immersed in a heme-containing solution, such as a leghemoglobin solution, for 1, 5, 10, 15, 30, or 45 minutes.

[0372] Figure 9 shows examples of six cubes of a commercial meat analogue (Queen chicken analogue) with sides of approximately 1 cm, of which four (left and bottom right) were immersed in a solution of approximately 10 mg / ml soybean leghemoglobin in 20 mM potassium phosphate pH 7.4 and 100 mM NaCl; and the remaining two (top right) were immersed in the same buffer without leghemoglobin. Note that the deep pink color of the leghemoglobin-infused cubes contrasts with the yellowish-brown color of the non-infused cubes.

[0373] FIG. 10 shows four leghemoglobin-infused cubes of Quon chicken analogue being cooked in a pan at 350°C. Two lower cubes were flipped over to expose the baked surfaces, which turned brown. Note that in the upper two cubes, the heated parts turned grayish-brown, while the cooler top surface retained its pink color. In some embodiments, a heme-containing solution, such as a leghemoglobin solution, was injected into the product until the product reached the color of raw meat.

[0374] Given the usefulness of heme proteins in the color of consumer goods, it would be useful to detect whether a product contains a specific heme protein. Accordingly, in some embodiments, the present invention includes a method for determining whether a product contains a heme protein. Methods for detecting proteins are widely known in the art. For example, ELISA, proximity-ligation assays, luminex assays, or Western blot analysis may be performed to determine whether leghemoglobin is present in food products, such as meat or meat imitations. In one embodiment, the detection method is performed to determine whether meat has been altered into leghemoglobin.

[0375] Examples

[0376] Exemplary muscle imitation compositions comprising one or more isolated, purified plant proteins are described herein.

[0377] Protein purification of the components of the imitation

[0378] Mung bean seeds and dried green pea seeds were purchased as ground powder and used for the purification of their respective seed storage proteins. Rubisco was purified from fresh alfalfa plants. Protein compositions at the individual fractionation stage were monitored by SDS-PAGE, and protein concentrations were measured by standard UV-VIS and Pierce assay methods.

[0379] Mung bean 8S globulin: Mung bean flour 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 in two stages by the addition of ammonium sulfate: 50% (wt / v) followed by 90% (wt / v). The protein precipitated from the 90% fraction containing mung bean 8S globulin was stored at -20°C until further use.

[0380] Pea Albumin: Dried green pea seed powder was resuspended in 50 mM sodium acetate buffer at a 1:10 (wt / v) ratio in pH 5 and incubated for 1 hour. Insoluble material was separated by centrifugation, and the protein in the supernatant was fractionated in two stages by ammonium sulfate precipitation: 50% (wt / v) followed by 90% (wt / v). The ammonium sulfate solution was stirred for 1 hour, and the ammonium sulfate-precipitated protein was removed by centrifugation. The protein of interest precipitated in 90% (wt / v) ammonium sulfate. The pellet was stored at -20°C until further use.

[0381] Pea Globulin: Dried green pea seed powder was resuspended in 20 mM potassium phosphate buffer, pH 8, and 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 converted to 50% (wt / v) ammonium sulfate, and the precipitated protein was removed. Second, the 50% (wt / v) ammonium sulfate supernatant was saturated with 80% (wt / v) ammonium sulfate. The protein pelleted with 80% (wt / v) ammonium sulfate contained the globulin of interest. The pellet was stored at -20°C until further use.

[0382] Rubisco: First, Rubisco was fractionated from alfalfa greens (or other green plants, e.g., soybeans, spinach, etc.) by grinding the leaves in a blender with 4 volumes of cold 50 mM K-phosphate buffer pH 7.4 buffer (containing 0.5 M NaCl + 2 mM DTT + 1 mM EDTA (in the laboratory) or without (in the field)). The resulting slurry was centrifuged to remove debris, and the supernatant (crude lysate) was used for a further purification step. Proteins within the crude lysate were fractionated by adding ammonium sulfate to a saturation of 30% (wt / v). The solution was stirred for 1 hour and then centrifuged. The pellet from this step was discarded, and additional ammonium sulfate was added to the supernatant to a saturation of 50% (wt / v). The solution was stirred for 1 hour and then centrifuged again. The pellets from this stage contained Rubisco and were maintained at -20°C until use.

[0383] Obtaining plant protein.

[0384] Mung bean seed 8S protein was purified by ammonium sulfate fractionation as described. The pellet was resuspended in 20 mM potassium phosphate (pH 7.4) and 0.5 M sodium chloride, and ammonium sulfate was removed by dialysis against the same buffer. Any precipitate was removed by centrifugation at 16,000 g for 10 minutes, and the protein was concentrated to the target concentration. Pea globulin was purified by ammonium sulfate fractionation as described. The protein pellet was resuspended in 20 mM potassium phosphate (pH 7.4) and 0.4 M sodium chloride, and ammonium sulfate was removed by dialysis against the same buffer. Any precipitate was removed by centrifugation at 16,000 g for 10 minutes, and the protein was concentrated to the target concentration. Pea albumin was purified by ammonium sulfate fractionation as described. The protein pellet was resuspended in 20 mM potassium phosphate at pH 7.4 and 0.1 M sodium chloride, and ammonium sulfate was removed by dialysis against the same buffer. Any precipitate was removed by centrifugation at 16,000 g for 10 minutes, and the protein was concentrated to the target concentration.

[0385] Construction of muscle tissue analogs

[0386] Mung bean seed 8S protein was purified by ammonium sulfate fractionation as described above. For gel preparation, 200 g of the pellet was 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 (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.

[0387] Leghemoglobin was purified from soybean root nodules. After washing the leguminous root nodules to remove soil and irrelevant root tissue, the root nodules were dissolved in 20 mM potassium phosphate (pH 7.4), 100 mM sodium chloride, 1 mM EDTA, and 1 mM ascorbic acid. The root nodules were dissolved by grinding them using a juicer blender. Insoluble substances were separated by centrifugation. The root nodule lysate was fractionated in two stages using ammonium sulfate; 60% wt / v ammonium sulfate was added, and the solution was incubated at 4°C for 1 hour. The pellet was discarded, and the supernatant was prepared with 90% wt / v ammonium sulfate and incubated at 4°C for 12 hours. Leghemoglobin was collected as a pellet precipitated from the 90% ammonium sulfate fraction and resuspended in 20 mM potassium phosphate, 1 mM EDTA, and 100 mM sodium chloride. SDS-PAGE gel analysis determined that the protein solution contained 70% leghemoglobin and 30% other root nodule proteins. Ammonium sulfate was removed using size-exclusion chromatography in the same buffer. Leghemoglobin was concentrated by dialyzing against 30% PEG 8000 (polyethylene glycol, molecular weight 8000) in 20 mM potassium phosphate, pH 7.3, and 100 mM sodium chloride for 48 hours. The total protein concentration was 57 mg / ml. The UV-VIS spectrum suggested that the leghemoglobin was in a heme-iron oxidized state. Therefore, leghemoglobin was incubated with 5 mM sodium bisulfite for 5 minutes, and the sodium bisulfite was removed using size-exclusion chromatography in 20 mM potassium phosphate and 100 mM sodium chloride buffer. The leghemoglobin was further concentrated to 35.4 mg / ml. UV-VIS spectral analysis confirmed that the leghemoglobin was in a heme-iron reduced state.

[0388] Transglutaminase was obtained commercially from Activa TI (Activa TI, Azimoto). The stock solution (20% wt / v) was prepared in 20 mM potassium phosphate pH 7.3, 100 mM sodium chloride buffer.

[0389] To prepare "dark" muscle tissue analogs (Fig. 11), 43 ml of mung bean protein solution (150 mg / ml in dialysis buffer) was mixed with 37 ml of leghemoglobin solution (46.5 mg / ml leghemoglobin and 20 mg / ml other soybean root nodule protein in 20 mM potassium phosphate, 100 mM NaCl, pH 7.3). 20 ml of transglutaminase solution (20% w / w) was added, the solution was thoroughly mixed, separated into two 50 ml Falcon tubes, and incubated overnight at room temperature. The final protein concentrations were 65 mg / ml for mung bean protein, 18 mg / ml for leghemoglobin, and 91 mg / ml for total protein.

[0390] A "white" muscle analog (Fig. 12) was prepared by mixing 43 ml of mung bean protein solution (150 mg / ml) with 45 ml of a 11.7 mg / ml leghemoglobin solution and a 0.8% (wt / v) transglutaminase solution. 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.

[0391] The "dark" muscle tissue analog formed an opaque gel of dark chocolate color with a smooth, uniform texture, featuring a glossy surface and a small amount (< 1 ml) of dark red, venous blood-colored liquid on top. The gel stood upright, was elastic but crumbly, and resembled a thin Jell-O in appearance. The gel had a distinctly recognizable intermediate flavor profile of blood and soy. The flavor profile was dominated by soy and iron / blood characteristics, with a weak vegetable and medicinal / chemical flavor. The taste was salty with a long blood aftertaste.

[0392] The "white" muscle tissue analog was very similar but much brighter cappuccino-like in color. It was also 2-3 times less resistant to compression and more brittle.

[0393] Adipose tissue analogue

[0394] Adipose tissue analogs were prepared using the mung bean 8S globulin fraction as follows: 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 homogenizer (VWR) at speed #2. The emulsion was aliquoted into 1.6 Eppendorf tubes and incubated overnight at room temperature. Subsequently, the tubes were heated on a heating block at 95°C for 5 minutes and allowed to cool to room temperature on a bench. The final concentration was 75 mg / ml of mung bean protein and 50% w / w oil.

[0395] Adipose tissue analogs were prepared by the same method using pea globulin (100 mg / ml). Additionally, adipose tissue analogs were prepared in bulk from pea globulin and rice bran or canola oil by the same method, except that the emulsion was aliquoted into an Eppendorf tube. Instead, the emulsion in a 50 ml Falcon tube was spun overnight on a Nutator and subsequently incubated at 90°C for 30 minutes.

[0396] The adipose tissue analog prepared in an Eppendorf tube based on mung beans (Fig. 13) formed an opaque, grayish-white gel with a smooth, uniform texture and no visually identifiable liquid incorporated into the gel. The gel stood upright and was elastic and resilient. The gel had a slightly pleasant flavor and a soft, pleasant taste. The taste was mildly salty.

[0397] A fat tissue analog prepared in an Eppendorf tube based on pea globulin (Fig. 14) was very similar to the mung bean-based fat analog, except that it had less oil when compressed.

[0398] The adipose tissue analogs prepared in 50 ml Falcon tubes were similar in appearance, texture, and flavor, but were substantially softer (based on compressibility measurements, 2 times softer for canola oil and 3 times softer for rice bran oil).

[0399] connective tissue analogues

[0400] A connective tissue analog prototype was developed using zein protein supplied from 100% yellow corn gluten wheat or from commercial sources, such as flaxseed (Prairie Gold, Bloomington, Illinois). The zein protein was solubilized in 70–90% ethanol at a preferred ratio of 1:3 to 1:5 (solid:solution). The zein protein was precipitated in a controlled manner, for example by changes in pH, to produce large zein structures with physicochemical properties that could be manipulated if desired. For example, Fig. 15 shows a connective-tissue analog strand produced using a 1:3 ratio in 70% ethanol, loaded into a syringe with a 23-gauge needle (ID 0.337 mm). The solution was slowly extruded from the bottom of a 5-inch-high vessel into an excess of 5 M NaCl solution. The ethanol-zein solution, which has a lower density than the NaCl solution, floated to the top and pulled out the fibrous strands of the solidified zein. The NaCl was continuously stirred, and as the strands began to unfold, it aided in strand elongation. The strands bundled together and became a hard, high-density mass.

[0401] A prototype of a ground beef imitation was prepared from a gel of vegetable protein and vegetable oil.

[0402] Ground beef round patties were prepared by combining 62% (wt / wt) muscle analogs (62% (wt / wt) "dark muscle analogs" and 38% (wt / wt) "white muscle analogs"), 29% (wt / wt) adipose tissue analogs (from pea globulin and canola oil), and 5% (wt / wt) connective tissue analogs (Figure 16, Panel A). Ground beef round patties were prepared by combining 62% muscle analogs (62% "dark muscle analogs" and 38% "white muscle analogs"), 29% adipose tissue analogs (from mung bean seed 8S protein and rice bran oil), and 5% connective tissue analogs (Figure 16, Panel B). Ground beef round patties were prepared by combining 71% (wt / wt) muscle tissue analogs (composed of 60% "white" muscle analogs and 40% "dark" muscle analogs) and 23% adipose tissue (from pea seed globulin protein and canola oil) (Figure 16, Panel C). Ground beef round patties were prepared by combining 67% "white" muscle analogs with 28% adipose tissue analogs (from pea globulin and rice bran oil) (Figure 16, Panel D).

[0403] In additional tests, the cooking effects of the ground beef imitation patties were evaluated by grilling them on a 350°F pan. Ground beef patty analogs were prepared by combining 62% (wt / wt) muscle tissue analogs (62% (wt / wt) "dark muscle analogs" and 38% (wt / wt) "muscle analogs"), 29% (wt / wt) adipose tissue analogs (from pea globulin and canola oil), and 5% (wt / wt) connective tissue analogs (Fig. 17). The left panel shows the patty before cooking, and the right panel shows the same patty after cooking for about 2 minutes. The observer described the flavor of the ground beef imitation during cooking as distinctly "beef-like."

Claims

Claim 1 A muscle imitation comprising fibers comprising 0.01% to 5% by weight of heme-containing protein, one or more sugar compounds, one or more sulfur compounds, and one or more non-animal proteins; and a meat imitation product comprising an adipose tissue imitation comprising modified plant protein and one or more non-animal fats. Claim 2 A meat imitation product according to paragraph 1, wherein the fibers are similar to skeletal muscle fibers. Claim 3 A meat imitation product according to claim 1, wherein the fibers include asymmetric fibers. Claim 4 A meat imitation product according to claim 1, wherein the fiber comprises isotropic fibers. Claim 5 A meat imitation product according to claim 1, wherein the fiber comprises extruded fiber. Claim 6 A meat imitation product according to paragraph 1, wherein the fiber comprises spun fiber. Claim 7 A meat imitation product according to paragraph 1, wherein the fibers are suspended in a gel. Claim 8 In paragraph 7, a meat imitation product in which the gel contains plant-based fat. Claim 9 A meat imitation product according to claim 7, wherein the gel comprises one or more of starch, arrowroot, corn starch, Katakuri starch, potato starch, sago, tapioca, arginine, guar gum, locust bean gum, xanthan gum, furcellalan, agar, carrageenan, cellulose, methylcellulose, hydroxymethylcellulose, acacia gum, konjac, starch, pectin, amylopectin, and protein, and said protein is derived from legumes, grains, nuts, seeds, leaves, algae, bacteria, or fungi. Claim 10 A meat imitation product according to claim 1, wherein the fiber contains a cross-linking enzyme or is stabilized by protein cross-linking. Claim 11 A meat imitation product according to claim 1, wherein the fibers are oriented parallel to each other or formed into sheets. Claim 12 A meat imitation product according to claim 1, wherein the fiber contains 5 weight percent or more of one or more non-animal proteins. Claim 13 A meat imitation product according to claim 1, wherein the muscle imitation accounts for 40% to 90% by weight of the meat imitation product and the fat tissue imitation accounts for 1% to 60% by weight of the meat imitation product. Claim 14 In claim 1, one or more non-animal proteins are ribosomal proteins, actin, hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase, phosphofructokinase, triose phosphate isomerase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase, pyruvate decarboxylase, translation elongation factor, ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), Rubisco activase, albumin, glycinin, cornglycinin, globulin, bisilin, cornalbumin, gliadin, glutelin, glutenin, hordain, prolamin, phaseolin, proteome, Meat imitation product selected from the group consisting of secalin, extensin, tritisea gluten, zein, seed storage protein, oleosin, caloleosin, steroleosin or other oil body proteins, nutritional storage protein A, nutritional storage protein B, and mung bean seed storage 8S globulin. Claim 15 A meat imitation product according to claim 1, further comprising a connective tissue imitation, wherein the muscle imitation, the fat tissue imitation, and the connective tissue imitation are assembled in a manner close to the physical organization of meat. Claim 16 A meat imitation product according to claim 1, wherein the fat tissue imitation further comprises non-animal protein. Claim 17 A meat imitation product according to claim 1, wherein one or more non-animal fats are selected from 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. Claim 18 In paragraph 1, a meat imitation product that does not contain animal products. Claim 19 A meat imitation product according to claim 1, wherein the muscle imitation and fat tissue imitation are assembled in a manner close to the physical organization of ground meat or a cut of meat. Claim 20 In paragraph 1, a meat imitation product that is a white meat imitation product, a dark meat imitation product, a red meat imitation product, a seafood meat imitation product, or an organ meat imitation product. Claim 21 A meat imitation product according to claim 1, wherein the color of the meat imitation product in an unprocessed state is pink, red, or pink to red, and the color of the meat imitation product in a cooked state is light yellowish-brown, brown, or light yellowish-brown to brown. Claim 22 In paragraph 1, a meat imitation product in which the heme-containing protein is leghemoglobin or myoglobin.

Images