Minced meat imitation

Abstract

This document relates to ground meat simulations, and more particularly to plant-based products that mimic ground meat, including fiber degree, texture heterogeneity, beef taste, and a color transition from red to brown during cooking of ground meat. For example, provided herein are meat simulations that include proteins selected based on temperatures at which they gelatinize and / or denature to mimic the behavior and quality of meat during cooking.

Description

[0001] This application is a divisional application of the invention patent application with application number 201580026650.6, application date March 31, 2015, and invention name “Minced meat imitation”.

[0002] CROSS REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Application No. 61 / 973,181, filed on March 31, 2014, and U.S. Application No. 62 / 058,230, filed on October 1, 2014, the disclosures of which are incorporated by reference in their entirety. Technical Field

[0004] The present invention relates to meat replicas, such as ground meat replicas, and more particularly to plant-based products that mimic the texture, appearance, and sensory aspects of ground meat, including the texture, appearance, and sensory aspects of cooked and eaten ground meat, such as fiber content, texture heterogeneity, beefy flavor, and red to brown color transition during cooking of ground meat. The present invention also relates to compositions and methods for modifying the flavor of a food product or food replica, such as cheese or a meat replica. Background Art

[0005] Meat substitute compositions are typically extruded soy / cereal mixtures that cannot replicate the experience of cooking and eating meat. Common limitations of plant-based meat substitute products include a more homogeneous texture and mouthfeel compared to equivalent meat products. In addition, since most of these products must be pre-cooked and pre-incorporated with artificial flavorings and fragrances for sale, they cannot replicate the aroma, flavor, and other key features (such as texture and mouthfeel) associated with cooked or cooked meat, and they may also have added odors. Therefore, these products mainly attract a limited consumer base that has converted to vegetarianism, but cannot attract a larger consumer portion that is accustomed to eating meat. It would be useful to have an improved plant-based meat substitute that preferably replicates the fiber, texture, aroma, and flavor of meat during and / or after cooking. Summary of the invention

[0006] The present invention is based on methods and materials for making plant-based products that can mimic ground meat, including fiber content, texture heterogeneity, beefy or other meaty flavors, and a color transition from red to brown during cooking of ground meat, without off-flavors. For example, meat replicas are provided herein that include proteins selected based on temperature at which they gel and / or denature to replicate the behavior and quality of meat during cooking, i.e., firmness, syneresis (water release), chewy texture or mouthfeel. For example, the temperature of denaturation and gelation of the selected proteins in the meat replica can be similar to proteins commonly found in meat (e.g., actin and myosin). In addition, the plant-based products provided herein can include flavorings (e.g., flavorings, flavoring precursors, and / or flavoring compounds) that can provide a meaty flavor, so that the plant-based meat replica has a more natural flavor and does not have off-flavors. The present invention therefore also provides methods for making plant-based products containing such flavorings.

[0007] In one aspect, the present disclosure features a meat replica composition comprising about 5 wt% to about 88 wt% (e.g., about 40 wt% to about 88 wt%, about 45 wt% to about 60 wt%, or about 15 wt% to about 55 wt%) of meat paste; about 0 wt% to about 40 wt% (e.g., about 1 wt% to about 30 wt%, about 5 wt% to about 25 wt%, or about 15 wt% to about 25 wt%) of carbohydrate-based gel; about 5 wt% to about 35 wt% (e.g., about 10 wt% to about 15 wt%, about 12 wt% to about 18 wt%, about 13 wt% to about 19 wt%) of a meat replica composition; about 0 wt% to about 40 wt% (e.g., about 1 wt% to about 30 wt%, about 5 wt% to about 25 wt%, or about 15 wt% to about 25 wt%) of a carbohydrate-based gel; about 5 wt% to about 35 wt% (e.g., about 10 wt% to about 15 wt%, about 12 wt% to about 18 wt%, about or about 20% to about 25% by weight) of fat; about 0.00001% to about 10% by weight (e.g., about 3% to about 7% by weight, about 0.001% to about 2% by weight, or about 0.00001% to about 2% by weight) of flavoring agent; about 0% to about 15% by weight (e.g., about 2% to about 15% by weight or about 2% to about 10% by weight) of binder; and about 0.01% to about 4% by weight (e.g., about 0.05% to about 1% by weight, or about 0.5% to about 2% by weight) of heme protein and / or iron salt. The meat paste may include flavoring agent. The fat may include flavoring agent. The meat paste may be about 45% to about 60% by weight of the composition. The carbohydrate gel may be about 10% to about 25% by weight of the composition. The fat may be about 10% to about 15% by weight of the composition. The flavoring agent can be about 3% to about 7% by weight of the composition or about 0.001% to about 2% by weight. The flavoring agent can include one or more flavoring precursors, seasonings or flavoring compounds. The flavoring agent can be a combination of a seasoning and one or more flavoring precursors. The adhesive can be about 2% to about 10% by weight of the composition. The adhesive can include one or more proteins that have been chemically or enzymatically modified to improve its texture and / or flavor characteristics or to regulate its denaturation and gelling temperature. Heme-containing protein can be about 0.01% to about 2% by weight of the composition. The composition can include heme-containing protein and iron salts. The meat sticky mass can include separated vegetable protein, edible fiber components, optional flavoring agents and optional fat. The adhesive can be conglycinin.

[0008] In another aspect, the present disclosure features a meat replica composition comprising about 5% to about 80% by weight (e.g., about 20% to about 30% by weight) of a meat paste; about 5% to about 35% by weight (e.g., about 15% to about 25% by weight) of a fat; about 15% to about 40% by weight (e.g., about 15% to about 25% by weight) of an edible fiber component; about 0.1% to about 18% by weight (e.g., about 7% to about 18% by weight) of a carbohydrate-based gel; about 0% to about 10% by weight (e.g., about 0% to about 10% by weight) of a flavoring agent; about 0.5% to about 15% by weight (e.g., about 5% to about 15% by weight) of a binder; and about 0.1% to about 8% by weight (e.g., about 2% to about 8% by weight) of a heme-containing protein and / or an iron salt.

[0009] In another aspect, the present disclosure features a method of making a ground meat replica. The method may include (a) heating a viscous mass to a temperature ranging from 150°F to 250°F, the viscous mass comprising isolated plant protein, an optional edible fiber component, one or more optional flavoring agents, and an optional fat; (b) after heating, combining the viscous mass with a fat, the fat optionally containing a flavoring agent and / or isolated plant protein; and (c) combining the viscous mass of step (b) with a carbohydrate-based gel, an optional edible fiber component, an optional binder, a highly conjugated heterocycle complexed with iron ions and / or an iron salt, and one or more optional flavoring agents to make a ground meat replica. The method may further include breaking the viscous mass of step (b) into pieces prior to combining with the carbohydrate-based gel, the optional edible fiber component, the optional binder, the highly conjugated heterocycle complexed with iron ions and / or an iron salt, and one or more optional flavoring agents.

[0010] In another aspect, the present disclosure features a method for seasoning a meat paste. The method may include (a) combining a first highly conjugated heterocycle complexed with an iron ion and / or a first iron salt with one or more flavoring precursors and optionally a fat; (b) heating the mixture to form one or more flavoring compounds; and (c) making a paste comprising isolated plant protein, an optional edible fiber component, and the mixture of step (b). The method may further include (d) after heating, combining the paste with a fat, the fat optionally containing a flavoring agent and / or isolated plant protein; and (e) combining the paste of step (d) with a carbohydrate-based gel, an optional binder, a second highly conjugated heterocycle complexed with an iron ion and / or a second iron salt, and one or more optional flavoring agents to produce a ground meat replica. The method may further include breaking the paste of step (d) into pieces before combining with the carbohydrate-based gel, an optional binder, a second highly conjugated heterocycle complexed with an iron ion and / or a second iron salt, and one or more optional flavoring agents.

[0011] In another aspect, the present disclosure features a method of seasoning a meat dough, wherein the method includes (a) making a dough comprising an isolated vegetable protein, an optional edible fiber component, one or more optional flavoring agents, and an optional fat; (b) making a flavored fat by combining the fat with a highly conjugated heterocycle complexed with an iron ion and / or a first iron salt and one or more flavoring precursors and heating the mixture; and (c) after heating, combining the dough with the flavored fat. The method may further include combining the dough of step (c) with a carbohydrate-based gel, an optional binder, a second highly conjugated heterocycle complexed with an iron ion and / or a second iron salt, and one or more optional flavoring agents to make a ground meat replica. The method may further include fragmenting the dough of step (c) prior to combining with the carbohydrate-based gel, the optional binder, the second highly conjugated heterocycle complexed with an iron ion and / or a second iron salt, and one or more optional flavoring agents.

[0012] Also featured herein is a method of making a ground meat replica, wherein the method includes (a) combining an iron salt with one or more flavor precursors and optionally a fat; (b) heating the mixture to form one or more flavoring compounds; (c) making a viscous mass comprising isolated plant protein, an optional edible fiber component, and the mixture of step (b); (d) after heating, combining the viscous mass with a fat, the fat optionally containing a flavoring agent and / or isolated plant protein; and (e) combining the viscous mass of step (d) with a carbohydrate-based gel, an optional binder, an iron salt, an optional highly conjugated heterocycle complexed with an iron ion, and one or more optional flavoring agents to make the ground meat replica. The method may further include breaking the viscous mass of step (d) into pieces prior to combining with the carbohydrate-based gel, the optional binder, the iron salt, the optional highly conjugated heterocycle complexed with an iron ion, and the one or more optional flavoring agents. In some embodiments, the highly conjugated heterocycle complexed with an iron ion may be combined with the iron salt, one or more flavor precursors, and a fat, followed by heating the mixture.

[0013] In yet another aspect, the present disclosure features a method of making a ground meat replica. The method may include (a) making a viscous mass comprising an isolated plant protein, an optional edible fiber component, one or more optional flavoring agents, and an optional fat; (b) making a flavored fat by combining the fat with an iron salt and one or more flavor precursors and heating the mixture; (c) after heating, combining the viscous mass with the flavored fat; and (d) combining the viscous mass of step (c) with a carbohydrate-based gel, an optional binder, an iron salt, an optional highly conjugated heterocycle complexed with an iron ion, and one or more optional flavoring agents to make a ground meat replica. The method may further include fragmenting the viscous mass of step (c) prior to combining with the carbohydrate-based gel, the optional binder, the iron salt, the optional highly conjugated heterocycle complexed with an iron ion, and one or more optional flavoring agents. In some embodiments, the highly conjugated heterocycle complexed with an iron ion may be combined with the fat, the iron salt, and one or more flavor precursors, followed by heating the mixture.

[0014] In any of the methods or compositions described herein, the iron salt can be ferric gluconate, ferric chloride, ferric oxalate, ferric nitrate, ferric citrate, ferric ascorbate, ferrous sulfate, ferric pyrophosphate, or any other water soluble salt.

[0015] In any of the methods or compositions described herein, the heme-containing protein can be a non-animal heme-containing protein, such as a plant-derived heme-containing protein (eg, leghemoglobin). Additionally, in some embodiments, the heme-containing protein can be isolated or isolated and purified.

[0016] In any of the methods or compositions described herein, one or more of the flavoring precursors can be sugars, sugar alcohols, sugar acids, sugar derivatives, oils, free fatty acids, amino acids or derivatives thereof, nucleosides, nucleotides, vitamins, acids, peptides, phospholipids, protein hydrolysates, yeast extracts or mixtures thereof. For example, the flavoring precursor can be selected from the group consisting of glucose, fructose, ribose, arabinose, glucose-6-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, inositol, maltose, sucrose, maltodextrin, glycogen, nucleotide-binding sugars, molasses, phospholipids, lecithin, inosine, inosine monophosphate (IMP), guanosine monophosphate (GMP), pyrazine, adenosine monophosphate (AMP), lactic acid, succinic acid, glycolic acid, thiamine, creatine, pyrophosphate, vegetable oil, algae oil, sunflower oil, corn oil, soybean oil, palm fruit oil, palm kernel oil, safflower oil, linseed oil, rice bran oil, cottonseed oil, olive oil, Sunflower oil, canola oil, linseed oil, coconut oil, mango oil, free fatty acids, cysteine, methionine, isoleucine, leucine, lysine, phenylalanine, threonine, tryptophan, valine, arginine, histidine, alanine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, proline, serine, tyrosine, glutathione, amino acid derivatives, urea, pantothenic acid, ornithine, niacin, glycerol, citrulline, taurine, biotin, borage oil, fungal oil, black currant oil, betaine, beta carotene, B vitamins, N-acetyl L-cysteine, iron glutamate and peptone or mixtures thereof.

[0017] In any of the methods or compositions described herein, the isolated vegetable protein in the dough may include wheat gluten, dehydrin, albumin, globulin or zein, or mixtures thereof.

[0018] In any of the methods or compositions described herein, the optional edible fiber component may include plant fiber from carrots, bamboo, peas, broccoli, potatoes, sweet potatoes, corn, whole grains, alfalfa, kale, celery, celery root, parsley, cabbage, zucchini, green beans, kidney beans, black beans, red beans, white beans, beets, cauliflower, nuts, apple peels, oats, wheat, or psyllium, or mixtures thereof.

[0019] In any of the methods or compositions described herein, the edible fiber component may include an extruded mixture of isolated vegetable proteins. The extruded mixture may contain wheat gluten and isolated soy protein, and optionally may additionally contain flavorings (e.g., condiments such as yeast extract, protein hydrolysate or oil; flavoring compounds; or flavoring precursors). In some embodiments, the edible fiber component may be a solution-spun protein fiber (e.g., a solution-spun protein fiber containing prolamin such as zein, pea prolamin, kafirin, rye prolamin, hordein, oat protein, or a mixture thereof).

[0020] In any of the methods or compositions described herein, fat can be non-animal fat, animal fat, or a mixture of non-animal and animal fat. Fat can be algae oil, fungal oil, corn oil, olive oil, soybean oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed oil, rapeseed oil, canola oil, safflower oil, sunflower oil, linseed oil, palm oil, palm kernel oil, coconut oil, babassu oil, shea butter, mango fat, cocoa butter, wheat germ oil, borage oil, black currant oil, sea buckthorn oil, macadamia oil, saw palmetto oil, conjugated linoleic acid oil, arachidonic acid-rich oil, docosahexaenoic acid (DHA)-rich oil, eicosapentaenoic acid (EPA)-rich oil, palm stearic acid, sea buckthorn fruit oil, macadamia oil, saw palmetto oil or rice bran oil; or margarine or other hydrogenated fats. In certain embodiments, for example, fat is algae oil. Fat can contain flavorings and / or isolated vegetable protein (e.g., conglycinin).

[0021] In any of the methods or compositions described herein, the gluten may include a flavoring. In any of the methods or compositions, the non-animal fat in the gluten may include a flavoring. The flavoring may be selected from the group consisting of: vegetable extracts, fruit extracts, acids, antioxidants, carotenoids, lactones, and combinations thereof. The antioxidant may be epigallocatechin gallate. The carotenoid may be lutein, beta-carotene, zeaxanthin, trans-beta-apo-8'-carotenal, lycopene, or canthaxanthin. The vegetable extract may be from cucumber or tomato. The fruit extract may be from melon or pineapple.

[0022] In any of the methods or compositions described herein, the melting temperature of the carbohydrate-based gel may be between about 45° C. and about 85° C. The carbohydrate-based gel may include agar, pectin, carrageenan, konjac, alginate, chemically modified agarose, or mixtures thereof.

[0023] In any of the methods or compositions described herein, the ground meat replica may contain a binder. The binder may be an isolated plant protein (e.g., ribulose bisphosphate carboxylase, albumin, gluten, conglycinin, or a mixture thereof). The denaturation temperature of the binder may be between about 40°C and about 80°C. The binder may be a carbohydrate-based gel that becomes firm after cooking to 140°F to 190°F. The carbohydrate-based gel may contain methylcellulose, hydroxypropyl methylcellulose, guar gum, locust bean gum, xanthan gum, or a mixture thereof. The binder may be egg white protein or collagen.

[0024] In any of the methods or compositions described herein, the highly conjugated heterocycle complexed with the iron ion can be a heme moiety or a porphyrin, porphyrinogen, corrinoid, chlorin, bacteriochlorophyll, corphin, chlorophyllin, bacterioporphin, or isoporphin moiety complexed with the iron ion. The heme moiety can be a heme-containing protein (e.g., non-symbiotic hemoglobin, Hell's gate globin I, flavohemoprotein, leghemoglobin, heme-dependent peroxidase, cytochrome c peroxidase, or mammalian myoglobin). In some embodiments, the heme-containing protein can be leghemoglobin. Leghemoglobin can be from soybeans, peas, or cowpeas.

[0025] In another aspect, the present invention features a method of increasing meat flavor or masking off-flavors from plant materials in a food product. The method may include adding to the food product a concentration of 10% of the food product. -3 Up to 10 -11 One or more lactones, wherein the lactone is selected from the group consisting of tetrahydro-6-methyl-2H-pyran-2-one, δ-octanolactone, 5-ethyldihydro-2(3H)-furanone, butyrolactone, dihydro-5-pentyl-2(3H)-furanone, dihydro-3-methylene-2,5-furandione, l-valeryl lactone, tetrahydro-2H-pyran-2-one, 6-heptyltetrahydro-2H-pyran-2-one, γ-octanolactone, 5-hydroxymethyldihydrofuranone, In some embodiments, the lactone may be 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone, butyrolactone, gamma-octalactone, and delta-tetradecalactone. The food product may be a meat replica. The meat replica may be free of animal products.

[0026] The present invention also features a method of increasing meat flavor or masking off-flavors from plant materials in a food product, wherein the method comprises adding one or more carotenoids to the food product at a concentration between 0.00001% and 0.1% of the food product, wherein the carotenoid is selected from the group consisting of: beta-carotene, zeaxanthin, lutein, trans-beta-apo-8'-carotenal, lycopene, canthaxanthin, and combinations thereof. The food product can be a meat replica. The meat replica can be free of animal products.

[0027] In another embodiment, the disclosure features a method of increasing the meaty flavor of a meat replica. The method may include adding a vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract to a meat replica at a concentration of 0.0001% to 10% of the meat replica. The vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract may be a Cucumis juice, puree, or extract (e.g., a juice, puree, or extract from cucumber or melon). The method The vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract may be cooked or otherwise processed to denature the protein prior to being added to the meat replica. The meat replica may be free of animal products.

[0028] In another aspect, the disclosure features a food product or food replica containing a heme-containing protein and a concentration of 10% of the food product. -3 Up to 10 -11 One or more lactones, wherein the one or more lactones are selected from the group consisting of tetrahydro-6-methyl-2H-pyran-2-one, δ-octanolactone, 5-ethyldihydro-2(3H)-furanone, butyrolactone, dihydro-5-pentyl-2(3H)-furanone, dihydro-3-methylene-2,5-furandione, l-valeryl lactone, tetrahydro-2H-pyran-2-one, 6-heptyltetrahydro-2H-pyran-2-one, γ-octanolactone, 5-hydroxymethyldihydro-2(3H)-furanone, The one or more lactones may be 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone, butyrolactone, gamma-octalactone, and delta-tetradecalactone. The food product or food replica may be a meat replica. The meat replica may be free of animal products.

[0029] Also featured herein is a food product or food replica containing a heme-containing protein and one or more carotenoids at a concentration between 0.00001% and 0.1% of the food product, wherein the one or more carotenoids are selected from the group consisting of: beta-carotene, zeaxanthin, lutein, trans-beta-apo-8'-carotenal, lycopene, canthaxanthin, and combinations thereof. The food product or food replica can be a meat replica. The meat replica can be free of animal products.

[0030] In another aspect, the disclosure features a food product or food replica containing (a) a heme-containing protein, and (b) a vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract at a concentration of 0.0001% to 10% of the food product. The vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract can be a Cucumis juice, puree, or extract. The Cucumis juice, puree, or extract can be from cucumber or melon. The vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract can have been cooked or otherwise treated to denature the protein prior to being added to the food replica. For example, the vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract can have been heated to a temperature of about 60° C. to about 100° C. prior to being added to the food replica. The food product can be free of animal products.

[0031] In another aspect, the disclosure features a food replica containing a concentration of 10% of the food product. -3 Up to 10 -11 One or more lactones, wherein the one or more lactones are selected from the group consisting of tetrahydro-6-methyl-2H-pyran-2-one, δ-octanolactone, 5-ethyldihydro-2(3H)-furanone, butyrolactone, dihydro-5-pentyl-2(3H)-furanone, dihydro-3-methylene-2,5-furandione, l-valeryl lactone, tetrahydro-2H-pyran-2-one, 6-heptyltetrahydro-2H-pyran-2-one, γ-octanolactone, 5-hydroxymethyldihydro-2(3H)-furanone, The one or more lactones may be 5-ethyl-4-hydroxy-2-methyl-3 (2H)-furanone, butyrolactone, γ-octalactone and δ-tetradecalactone.

[0032] In yet another aspect, the disclosure features a food replica containing one or more carotenoids at a concentration between 0.00001% and 0.1% of the food product, wherein the one or more carotenoids are selected from the group consisting of β-carotene, zeaxanthin, lutein, trans-β-apo-8'-carotenal, lycopene, canthaxanthin, and combinations thereof.

[0033] Also featured herein is a food replica containing a vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract at a concentration of 0.0001% to 10% of the food product. The vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract can be a Cucumis juice, puree, or extract (e.g., a Cucumis juice, puree, or extract from cucumber or melon). The vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract can have been cooked or otherwise processed to denature proteins prior to being added to the food replica. For example, the vegetable juice, vegetable puree, vegetable extract, fruit juice, fruit puree, or fruit extract can have been heated to a temperature of about 60°C to about 100°C prior to being added to the food replica.

[0034] In some embodiments, the food replicas provided herein may be free of animal products, wheat gluten, soy protein, and / or tofu.

[0035] Any of the food replicas provided herein can contain one or more of a meat paste, a carbohydrate-based gel, a non-animal fat, and a binder.

[0036] Any of the food replicas provided herein can be a meat replica. Other materials and methods for making meat replicas can be found in, for example, US Publication No. 2014 / 0193547 and PCT Publications WO 2014 / 110532 and WO 2014 / 110539, each of which is incorporated herein by reference in its entirety.

[0037] Any of the food replicas provided herein can be a cheese replica. Cheese replicas can contain nut milks, cross-linking enzymes, or cheese starter cultures. Other materials and methods for making cheese replicas can be found in, for example, U.S. Publication No. 2014 / 0127358 and PCT Publication WO 2014 / 110540, both of which are incorporated herein by reference in their entirety.

[0038] In yet another aspect, the present disclosure features a ground meat replica containing (a) a paste containing isolated plant protein, an optional edible fiber component, one or more optional flavoring agents, and an optional fat; (b) a fat, the fat optionally containing a flavoring agent and / or isolated plant protein; and (c) a carbohydrate-based gel, a binder, a highly conjugated heterocycle complexed with iron ions and / or an iron salt, an optional edible fiber component, and one or more optional flavoring agents. The binder can be an isolated plant protein (e.g., ribulose bisphosphate carboxylase, albumin, gluten, conglycinin, or a mixture thereof). The denaturation temperature of the binder can be between about 40° C. and about 80° C.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the invention belongs. Although methods and materials similar to or equivalent to the methods and materials described herein can be used to practice the present invention, suitable methods and materials are described as follows. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, this specification (including definitions) shall prevail. In addition, materials, methods and examples are merely illustrative and are not intended to be limiting.

[0040] The details of one or more embodiments of the invention are described in the accompanying drawings and the following detailed description. Other features, objectives and advantages of the invention will be apparent from the detailed description and drawings, as well as from the claims. The word "comprising" in the claims may be replaced by "consisting essentially of" or "consisting of" according to standard practice in patent law. DETAILED DESCRIPTION

[0041] In general, provided herein are methods and materials for making meat replicas, including ground meat replicas (e.g., ground beef, ground chicken, ground turkey, ground lamb, or ground pork) and replicas of meat and fish cuts. In summary, provided herein are methods for making ground meat replicas, comprising preparing a meat replica dough (referred to herein as "meat dough") including an optional edible fiber component, combining the meat dough with a fat (typically non-animal fat, but it should be noted that animal fat can be used) that may optionally include flavorings and / or isolated vegetable protein, adding a carbohydrate gel, an optional edible fiber component, a binder, a highly conjugated heterocycle complexed with iron ions and / or an iron salt, and one or more flavorings to make the replica. After combining the meat dough with the fat, the mixture may be broken into smaller pieces before adding other ingredients.

[0042] The meat paste may incorporate an edible fiber component to help achieve a textural heterogeneity and fibrousness of the meat replica similar to that of ground meat (e.g., ground beef). Incorporating flavorings into multiple components of the meat replica (e.g., meat paste, edible fiber component, non-animal fat, or two or more of the assembled replicas) helps mimic the sensory properties of ground meat. In some embodiments, flavorings are incorporated into three components of the meat replica. In some embodiments, flavorings are incorporated into four components of the meat replica.

[0043] As described herein, the flavoring agent can be a flavoring precursor, a flavoring compound produced by the reaction of a flavoring precursor with iron, or a flavoring, such as an extract (e.g., a malt extract, a yeast extract, a vegetable or fruit extract, such as a cucumber extract or a melon extract, or a peptone) or a protein hydrolysate, such as a plant protein hydrolysate, a soy protein hydrolysate, a yeast protein hydrolysate, an algae protein hydrolysate, or a meat protein hydrolysate, or a natural or synthetic flavoring compound. The flavoring precursors can react, for example, with highly conjugated heterocycles complexed with iron ions or iron in iron salts, with each other, or with flavorings upon heating. Thus, in the meat replicas described herein, pre-cooked (i.e., reacted) flavoring components, uncooked flavoring precursors that can react (e.g., with iron salts and / or highly conjugated heterocycles complexed with iron ions or with each other) during the cooking of the replica, or flavorings or flavoring compounds that are introduced to the flavoring without reacting can be incorporated into the meat replica to reproduce the sensory experience of cooking and eating cooked ground meat. The flavor and / or aroma characteristics of the ground meat product can be adjusted by, among other things, the type and concentration of flavor precursors, reaction pH, cooking time, type and amount of iron complexes (e.g., heme cofactors such as heme-containing proteins or heme bound to non-peptide polymers or macromolecules), reaction temperature, and the amount of water activity in the product.

[0044] Highly conjugated heterocycles complexed with iron ions are referred to herein as iron complexes. Such iron complexes include a heme moiety or other highly conjugated heterocycles complexed with iron ions. "Heme" refers to iron (Fe) bound to the center of the porphyrin ring. 2+ or Fe 3+ ). Thus, the iron complex may be a heme moiety or a porphyrin, porphyrinogen, corrin, corrinoid, chlorin, bacteriochlorophyll, corrin, chlorophyllin, bacterioporphin or isoporphin moiety complexed with an iron ion. The heme moiety may be a heme cofactor, such as a heme-containing protein; a heme moiety bound to a non-peptide polymer or other macromolecule, such as a liposome, polyethylene glycol, carbohydrate, polysaccharide or cyclodextrin.

[0045] In some embodiments, the iron complex is an isolated and purified heme-containing protein. As used herein, the term "isolated and purified" with respect to a protein or protein fraction indicates that the protein or protein fraction has been separated from other components of a source material (e.g., other animal, plant, fungal, algal, or bacterial proteins) such that the protein or protein fraction is at least 50% (e.g., at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%) free of other components of the source material on a dry weight basis.

[0046] As used herein, an "enriched" protein or protein fraction composition is at least 2-fold (e.g., at least 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, or 100-fold) enriched in a protein or protein fraction relative to the source material.

[0047] The term "heme-containing protein" can be used interchangeably with "heme-containing polypeptide" or "heme protein" or "heme polypeptide" and includes any polypeptide that can covalently or non-covalently bind a heme moiety. In some embodiments, the heme-containing polypeptide is a globulin and can include a globulin fold, which includes a series of seven to nine alpha helices. Globulin-type proteins can be of any class (e.g., class I, class II, or class III), and in some embodiments, can transport or store oxygen. For example, the heme-containing protein can be a non-symbiotic hemoglobin or leghemoglobin. The heme-containing polypeptide can be a monomer, i.e., a single polypeptide chain, or can be a dimer, trimer, tetramer, and / or higher oligomer. Oxidized Fe 2+ The useful life of the unoxidized heme protein may be similar to that of myoglobin or may exceed that of myoglobin by 10%, 20%, 30%, 50%, 100% or more under the conditions of manufacturing, storing, handling or preparing for consumption of a consumer product containing the heme protein. 2+ The useful life of the heme-containing protein in a stable state may be similar to that of myoglobin or may exceed that of myoglobin by 10%, 20%, 30%, 50%, 100% or more under the conditions of manufacturing, storing, handling or preparing for consumption of a consumer product containing the heme protein.

[0048] Non-limiting examples of heme-containing polypeptides can include androstin, cytoglobulin, globulin E, globulin X, globulin Y, hemoglobin, myoglobin, invertebrate hemoglobin, beta hemoglobin, alpha hemoglobin, protoglobulin, cyanophycoglobulin, cytoglobulin, histaminin, neuroglobulin, hemoglobin, truncated hemoglobin (e.g., HbN or HbO), truncated 2 / 2 globulin, hemoglobin 3 (e.g., Glb3), cytochrome, or peroxidase.

[0049] Heme-containing proteins useful in the ground meat replicas described herein can be from mammals (e.g., farm animals such as cows, goats, sheep, pigs, bulls, or rabbits), birds, plants, algae, fungi (e.g., yeast or filamentous fungi), ciliates, or bacteria. For example, the heme-containing protein can be from a mammal, such as a farm animal (e.g., a cow, goat, sheep, pig, fish, bull, or rabbit) or a bird, such as a turkey or chicken. The heme-containing protein may be derived from a plant such as Nicotiana tabacum or Nicotiana sylvestris (tobacco); Zea mays (maize), Arabidopsis thaliana, a legume such as Glycine max (soybean), Cicer arietinum (chickpea or chickpea), a pea (Pisum sativum) (pea) variety such as garden pea or sweet pea, a common bean (Phaseolus vulgaris) variety such as green bean, black bean, kidney bean, white bean or pinto bean, a cowpea (Vigna unguiculata) variety (cowpea), mung bean (Vigna radiata) (mung bean), white lupin (Lupinus albus) (lupin) or alfalfa (Medicago sativa) (alfalfa); Brassica napus (canola); Triticum vulgaris (Triticum aestivum) (wheat); sps.) (wheat, including wheat kernels and spelt); Gossypium hirsutum (cotton); Oryza sativa (rice); Zizania sps. (wild rice); Helianthus annuus (sunflower); Beta vulgaris (sugar beets); Pennisetum glaucum (pearl millet); Chenopodium sp. (quinoa); Sesamum sp. (sesame); Linum usitatissimum (flax); or Hordeum vulgare (barley).The heme-containing protein can be isolated from a fungus, such as Saccharomyces cerevisiae, Pichia pastoris, Magnaporthe oryzae, Fusarium graminearum, Aspergillus oryzae, Trichoderma reesei, Myceliopthera thermophile, Kluyveramyces lactis, or Fusarium oxysporum. Heme-containing proteins can be isolated from bacteria such as Escherichia coli, Bacillus subtilis, Bacillus licheniformis, Bacillus megaterium, Synechocistis sp., Aquifex aeolicus, Methylacidiphiluminfernorum, or thermophilic bacteria such as Thermophilus spp. The sequences and structures of many heme-containing proteins are known. See, e.g., Reedy et al., Nucleic Acids Research, 2008, Vol. 36, Database issue D307-D313 and available on the World Wide Web at. http: / / hemeprotein.info / heme.php Obtained heme protein database.

[0050] In some embodiments, the non-symbiotic hemoglobin may be from any plant. In some embodiments, the non-symbiotic hemoglobin may be from a plant selected from the group consisting of soybean, sprouted soybean, alfalfa, golden flax, black bean, black-eyed pea, white bean, tobacco, pea, huihui bean, mung bean, cowpea, pinto bean, pea pod, quinoa, sesame, sunflower, wheat kernel, spelt, barley, wild rice and rice.

[0051] In some embodiments, the leghemoglobin can be soy, pea, or cowpea leghemoglobin.

[0052] In some embodiments, isolated plant proteins are used. As used herein, the term "isolated" with respect to a protein or protein fraction (e.g., a 7S fraction) indicates that the protein or protein fraction has been separated from other components of a source material (e.g., other animal, plant, fungal, algae, or bacterial protein) so that the protein or protein fraction is at least 2% (e.g., at least 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%) free of other components of the source material by dry weight. Thus, in some embodiments, the iron complex can be an isolated heme-containing protein (e.g., a plant heme-containing protein). Proteins can be separated based on their molecular weight, for example, by size exclusion chromatography, ultrafiltration through a membrane, or density centrifugation. In some embodiments, proteins can be separated based on their surface charge, for example, by isoelectric precipitation, anion exchange chromatography, or cation exchange chromatography. Proteins can also be separated based on their solubility, for example, by ammonium sulfate precipitation, isoelectric precipitation, surfactant, detergent or solvent extraction. Proteins can also be separated by their affinity for another molecule, using, for example, hydrophobic interaction chromatography, reactive dyes or hydroxyapatite. Affinity chromatography can also include the use of antibodies with specific binding affinity for heme-containing proteins, nickel nitriloacetic acid (NTA) of His-tagged recombinant proteins, lectins that bind to sugar moieties on glycoproteins, or other molecules that specifically bind to proteins.

[0053] Example 2 describes a method for isolating ribulose bisphosphate carboxylase from plants (e.g., spinach or alfalfa). The extraction process can be further improved by adding a reducing agent such as metabisulfite (about 2% w / v solution or more) to the initial extraction buffer and maintaining anaerobic conditions throughout the entire process and / or by adding 0.05-1% v / v cationic flocculants such as Superfloc 781G, Magnafloc LT 7989 (BASF) or Tramfloc 863A to the extraction buffer. The resuspended protein pellet from such a method will continue to perform after microfiltration at pH 7.0, providing the same color and having the same denaturation characteristics.

[0054] Example 4 describes a method for isolating conglycinin (also referred to as the 7S fraction) from plants such as soybeans. Other sources of 7S include seeds such as, but not limited to, peas, chickpeas, mung beans, kidney beans, broad beans, cowpeas, pine nuts, rice, corn, and sesame. Soluble protein can be extracted from defatted soy flour, and the mixture is then acidified (e.g., to pH 4.5) to precipitate the protein. The conglycinin can be redissolved and concentrated, for example, using ultrafiltration.

[0055] In certain embodiments, the separated protein is decolorized. For example, ribulose bisphosphate carboxylase concentrate can be decolorized (pH 7-9) by a column equipped with activated carbon. Pigment can be bound to the column, and ribulose bisphosphate carboxylase can be separated in the filtrate. Alternatively, ribulose bisphosphate carboxylase concentrate can be decolorized by cultivating the solution with FPX66 (Dow Chemicals) resin packaged in a column or batch mode. The slurry is cultivated for 30 minutes and then the liquid is separated from the resin. Pigment can be bound to the resin and ribulose bisphosphate carboxylase can be collected in the column flow-through.

[0056] In some embodiments, the isolated protein can be purified and decolorized as described in Example 3. See also "Methods for Extracting and Purifying Natural Proteins," US Serial No. 62 / 058,211, filed October 1, 2014.

[0057] In some embodiments, the isolated and decolorized plant protein can increase the shelf-life stability of the red color of a meat replica compared to a corresponding meat replica comprising an isolated, non-decolorized plant protein. In some embodiments, the decolorized protein provides an improvement in the flavor profile of a meat replica compared to that observed with a meat replica having a corresponding isolated, decolorized plant protein.

[0058] Heme-containing proteins or other proteins can also be produced recombinantly using polypeptide expression techniques (e.g., heterologous expression techniques using bacterial cells, insect cells, fungal cells (such as yeast), plant cells (such as tobacco, soybean or Arabidopsis), or mammalian cells). For example, leghemoglobin can be recombinantly produced in E. coli or Pichia pastoris as described in Example 1. In some cases, standard polypeptide synthesis techniques (e.g., liquid phase polypeptide synthesis techniques or solid phase polypeptide synthesis techniques) can be used to synthetically produce heme-containing proteins. In some cases, in vitro transcription-translation techniques can be used to produce heme-containing proteins.

[0059] In some embodiments, the meat replicas described herein consist essentially or entirely of ingredients derived from non-animal sources (e.g., plant, fungal, or microbial sources). In some embodiments, meat replicas may include one or more animal products. For example, meat replicas may be made from a combination of plant and animal sources.

[0060] Making meat imitations

[0061] Meat dough can be prepared by mixing separated vegetable protein and optional edible fiber component, optional seasoning and optional non-animal fat, adding aqueous components such as water or broth to the mixture and kneading or otherwise mixing to form dough. The aqueous component can be heated before being added to the mixture of vegetable protein and fiber component. Once the meat dough is formed, the meat dough is heated (e.g., steamed or boiled) to a temperature ranging from 150°F to 250°F (e.g., 160°F to 240°F, 170°F to 230°F, 180°F to 220°F or 190°F to 212°F). For example, the meat dough can be steamed by placing it in a rice cooker, a steam cabinet or a channel steamer. The meat dough can be heated by applying dry heat, such as by placing it in a bread machine or oven, or by immersing it in hot water or broth. Boiling in broth can improve the flavor of the meat dough because the beneficial flavor and odor masking agent can be absorbed into the dough. Textural properties can also be modulated by the choice of cooking method.

[0062] As used herein, the term "isolated plant protein" indicates that the plant protein (e.g., heme-containing protein, wheat gluten, dehydrin, albumin, globulin, conglycinin, glycinin, or zein, or a mixture thereof) or a plant protein fraction (e.g., 7S fraction) has been separated from other components of a source material (e.g., other animal, plant, fungal, algal, or bacterial proteins) such that the protein or protein fraction is at least 2% (e.g., at least 5%, 10%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%) free of other components of the source material on a dry weight basis. For example, wheat gluten may be used alone or in combination with one or more other proteins (e.g., dehydrin). Dehydrin may be particularly useful for enhancing the juiciness and texture of ground meat replicas. In some embodiments, meat replicas can be formulated to be gluten-free, and for example, a blend of corn starch, tapioca flour, rice flour, and guar gum can replace wheat gluten in the meat dough.

[0063] The edible fiber component can be an extruded mixture of plant fibers, isolated plant proteins (e.g., wheat gluten or other isolated plant proteins such as gluten, albumin, legumin, faba bean globulin, confibulin, glycinin, and protein isolates such as from any seed or legume (including soybeans, peas, lentils, etc.) or solution-spun protein fibers. In some embodiments, the solution-spun protein fibers are prolamine solution-spun protein fibers. Prolamine proteins can be from any plant source (e.g., corn or peas) and can include zein, prolamine, kafirin, secalin, hordein, or oat protein. The texture of the ground meat product (e.g., meat patties) depends on the properties of the edible fiber component, such as fiber size and tensile strength. As described herein, an extruded mixture of isolated plant proteins or solution-spun protein fibers can be referred to as a connective tissue replica, and the fiber size and tensile strength of the connective tissue replica can be controlled by co-variation of extrusion parameters such as temperature, throughput, and die size. For example, a combination of lower extrusion temperature, medium / low throughput, and smaller dies favors making highly fibrous tissues with low tensile strength, whereas higher extrusion temperature, greater throughput, and larger dies favors making low fibrous tissue replicas with very high tensile strengths.

[0064] The fiber size and tensile strength of the connective tissue replicas can also be adjusted by changing the composition of the extrusion mixture. For example, by increasing the ratio of isolated plant protein (e.g., soy protein such as conglycinin) to wheat gluten to 3:1 w / w, and simultaneously reducing the moisture content of the extrusion mixture to 50%, connective tissue replicas with thinner fibers and greater tensile strength can be produced.

[0065] The texture of the meat paste can also be adjusted by adding cream of tartar to the preparation. For example, a meat paste preparation containing cream of tartar can be more viscous, with a shape factor similar to ground beef after grinding to make it easier to shape. 0.05% to 2.5% (e.g., 0.5%) of cream of tartar can be added.

[0066] The appearance of the ground meat replica can be adjusted by chopping the edible fiber component into pieces of desired size and shape. In some embodiments, the edible fiber component can be chopped using a commercially available chopper, such as a Cuisineart chopper / grinder, UM 12 with a blunt knife attachment, Comitrol chopper (Urschel Laboratories, Indiana), or similar chopper. The fiber size can be adjusted by the type of chopper, the choice of blades and the type of screen, and by adjusting the chopping time, thereby mimicking the fibrous appearance of meat.

[0067] In other embodiments, the edible fiber component can be separated into fibers by combing using a handheld combing machine, such as a Pat Green combing machine. By changing the size and spacing of the pins on the combing drum, the size of the fibers can be adjusted to mimic the fibrous appearance of meat.

[0068] In other embodiments, the edible fiber component may be prepared by pushing it through rollers (e.g., The fibers are separated by gentle chopping using, for example, a blunt knife on a UM 12 machine. By varying the number of rollers and the spacing between the rollers, the size of the fibers can be adjusted to mimic the fibrous appearance of meat.

[0069] The fiber size, tensile strength, and appearance of the connective tissue replica can be adjusted to mimic a specific ground meat product (eg, ground beef or different beef cuts that can be ground).

[0070] In some embodiments, the edible fiber component includes soluble or insoluble plant fiber. For example, plant fibers from carrots, bamboo, peas, broccoli, potatoes, sweet potatoes, corn, whole grains, alfalfa, kale, celery, celery root, parsley, cabbage, zucchini, green beans, French beans, black beans, red beans, white beans, beets, cauliflower, nuts, apple peels, oats, wheat or psyllium or mixtures thereof can be used as edible fiber components.

[0071] In some embodiments, the edible fiber component may include compounds that prevent the occurrence of off-flavors during the extrusion process. The high temperature and low humidity conditions to which the extruded mixture is exposed during the extrusion process result in the formation of compounds associated with cereal flavors, wood flavors, nutty flavors, rubber flavors, and other off-flavors. Including certain classes of compounds, such as antioxidants or carotenoids, can help reduce the formation of off-flavor compounds. For example, the extruded mixture may include canthaxanthin to prevent the occurrence of granular off-flavors. Carotenoids can be about 0% by weight to about 1% by weight of the edible fiber component.

[0072] In some embodiments, the meat dough is formed using approximately equal proportions of the isolated vegetable protein and edible fiber components. It should be understood that the ratios may be varied as desired to adjust the properties of the final product.

[0073] In some embodiments, broth (such as flavored broth) can be used for the meat dough.For example, the meat dough can be formed using approximately equal proportions of isolated vegetable protein and broth.

[0074] In some embodiments, the flavored broth comprises a flavored mixture formed by pre-reacting (cooking) the flavoring precursors before adding them to the meat paste. The flavoring precursor molecules or compositions can be added to the pre-reaction mixture in a purified form and / or can be derived from ingredients in the uncooked meat paste that contain and / or are enriched in one or more of the specific flavoring precursors or compositions, including, for example, coconut oil, cysteine, glucose, ribose, thiamine, algae oil, lactic acid and or yeast extract. The resulting flavor and / or aroma characteristics can be adjusted, among other things, by the type and concentration of the flavoring precursors, the reaction pH, the cooking time, the cooking temperature, the type and amount of iron complexes (e.g., iron-containing proteins, heme cofactors such as heme-containing proteins, or ferrous chlorophyllin) or iron salts (ferric gluconate), the reaction temperature, and the amount of water activity in the product. The flavored broth may contain non-animal products (e.g., plants) or it may be a combination of animal and non-animal precursors (e.g., lard). Seasoning bouillons can impart flavor to consumable food products, producing the taste and aroma of beef, bacon, pork, lamb, goat, turkey, duck, deer, yak, bison, chicken or desired meaty notes.

[0075] In some embodiments, the flavored broth can be made by combining an iron complex (e.g., an isolated heme-containing protein) and / or an iron salt (e.g., ferric gluconate, ferric chloride, ferric oxalate, ferric nitrate, ferric citrate, ferric ascorbate, ferrous sulfate, ferric pyrophosphate, or any other water-soluble salt) with one or more flavoring precursors and a fat (e.g., non-animal fat), and heating the mixture to obtain a flavored broth containing one or more flavoring compounds. Suitable flavoring precursors include sugars, sugar alcohols, sugar derivatives, free fatty acids, triglycerides, alpha-hydroxy acids, dicarboxylic acids, amino acids and their derivatives, nucleosides, nucleotides, vitamins, peptides, phospholipids, lecithin, pyrazine, creatine, pyrophosphate, and organic molecules. For example, sugars, sugar alcohols, sugar acids and sugar derivatives may include glucose, fructose, ribose, sucrose, arabinose, glucose-6-phosphate, fructose-6-phosphate, fructose 1,6-bisphosphate, inositol, maltose, mannose, glycerol, molasses, maltodextrin, glycogen, galactose, lactose, ribitol, gluconic acid, glucuronic acid, amylose, amylopectin or xylose. Free fatty acids may include caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alpha linolenic acid, gamma linolenic acid, arachidic acid, arachidonic acid, behenic acid, eicosapentaenoic acid, petroselinic acid or erucic acid. Triglycerides can include fatty acid esters of caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alpha linolenic acid, gamma linolenic acid, arachidic acid, arachidonic acid, behenic acid, eicosapentaenoic acid, petroselinic acid or erucic acid. Amino acids and derivatives thereof can include cysteine, cystine, cysteine ​​sulfoxide, allicin, selenocysteine, methionine, isoleucine, leucine, lysine, phenylalanine, threonine, tryptophan, 5-hydroxytryptophan, valine, arginine, histidine, alanine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, proline, serine, tyrosine, ornithine, carnosine, citrulline, carnitine, ornithine, theanine and taurine. Phospholipids can include multiple amphiphilic molecules comprising fatty acids, glycerol and polar groups.The fatty acid is selected from the group consisting of oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caprylic acid, caprylic acid, caprylic acid, pelargonic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, 18:2 conjugated linoleic acid, conjugated oleic acid; or oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caprylic acid, caprylic acid, caprylic acid, pelargonic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, 18:2 conjugated linoleic acid, or conjugated oleic acid. The invention relates to a glyceride of oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caprylic acid, caprylic acid, caprylic acid, geranic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid; or a triglyceride derivative of oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caprylic acid, caprylic acid, caprylic acid, geranic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, 18:2 conjugated linoleic acid or conjugated oleic acid. In certain embodiments, the polar group is selected from the group consisting of choline, ethanolamine, serine, phosphoric acid, glycerol-3-phosphate, inositol and inositol phosphate.

[0076] Nucleosides and nucleotides may include inosine, inosine monophosphate (IMP), guanosine, guanosine monophosphate (GMP), adenosine or adenosine monophosphate (AMP). Vitamins may include thiamine, vitamin B2, vitamin B9, vitamin C, 4-aminobenzoic acid, choline, niacin, vitamin B8, vitamin B12, biotin, betaine, vitamin A, beta carotene, vitamin D, vitamin B6 or vitamin E. Acids such as acetic acid, caffeic acid, glycolic acid, aspartic acid, pantothenic acid; alpha hydroxy acids such as lactic acid or glycolic acid; tricarboxylic acids such as citric acid; or dicarboxylic acids such as succinic acid or tartaric acid. Peptides and protein hydrolysates may include glutathione, plant protein hydrolysates, soy protein hydrolysates, wheat protein hydrolysates, corn protein hydrolysates, yeast protein hydrolysates, algae protein hydrolysates and meat protein hydrolysates. Extracts may include malt extract, yeast extract or peptone.

[0077] For example, in some embodiments, broth can be made by combining an iron complex (e.g., an isolated and purified heme-containing protein such as leghemoglobin) and / or an iron salt (e.g., ferric gluconate, ferric chloride, ferric oxalate, ferric nitrate, ferric citrate, ferric ascorbate, ferrous sulfate, ferric pyrophosphate, or any other water-soluble salt) with one or more flavoring precursors (e.g., a precursor mixture shown in Table 2 or Table 13) and a fat (e.g., non-animal fat), and heating the mixture to obtain a flavored broth containing one or more flavoring compounds. Non-animal fats can include oils of plant origin, algae oils, or oils from bacteria or fungi. Suitable plant-derived oils include coconut oil, mango oil, sunflower oil, cottonseed oil, safflower oil, rice bran oil, cocoa butter, palm kernel oil, palm fruit oil, palm oil, soybean oil, rapeseed oil, canola oil, corn oil, sesame oil, walnut oil, almond oil, linseed, jojoba oil, castor, grape seed oil, peanut oil, olive oil, borage oil, algae oil, fungal oil, black currant oil, babassu oil, shea butter, mango fat, wheat germ oil, black currant oil, sea buckthorn oil, macadamia oil, saw palmetto oil, conjugated linoleic acid oil, arachidonic acid-rich oil, docosahexaenoic acid (DHA)-rich oil, eicosapentaenoic acid (EPA)-rich oil or margarine. Oil can be hydrogenated (e.g., hydrogenated vegetable oil) or non-hydrogenated. Oil fractions can also be used, such as stearin (e.g., palm stearin) or olein. For example, non-animal fat can be coconut oil or a combination of coconut oil and stearin. In some embodiments, the fat may contain non-animal (eg, plant) products, or it may be a combination of animal and non-animal precursors (eg, lard) or just animal fat.

[0078] In some embodiments, a flavored broth can be made by combining water, non-animal fats such as coconut oil, and flavoring agents such as acids (e.g., lactic acid), carotenoids (e.g., lutein), or antioxidants, and heating the mixture to make the broth.

[0079] After heating the meat sticks as described above, the non-animal fat, optionally containing a flavoring agent, may be combined with the meat sticks. Typically, the meat sticks are cooled (e.g., to room temperature) before combining the meat sticks with the non-animal fat. The non-animal fat may be flavored by combining the non-animal fat with an iron complex or iron salt and one or more flavoring precursors (described above) and heating the mixture to produce a flavoring compound. The heated mixture may be cooled so that the non-animal fat may solidify. Before the mixture solidifies, one or more additional non-animal fats (e.g., algae oil), one or more masking agents (e.g., lactones such as butyrolactone, delta-tridecalactone, gamma-decanolide, delta-dodecalactone, gamma-octalactone, dihydro-5-methyl-2(3H)-furanone, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone, delta-tetradecanolide, or a combination thereof) or one or more flavoring compounds (e.g., acetoin, carotenoids, antioxidants, vegetable or fruit juices, purees, or extracts) may be added to improve the flavor of the non-animal fat. In some embodiments, a combination of 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone, butyrolactone, gamma-octalactone, and / or delta-tetradecanolide may be used as masking agents. Adding one or more lactones (e.g., at a concentration of 10 -3 Up to 10 -10 ) can reduce off-flavors perceived as grainy, eggy, bitter, cardboard, liver, or mushroom and increase desirable flavors such as creamy, buttery, caramel, fatty, umami, and fruity. For example, combinations of two, three, or four lactones can be used to mask characteristics such as bitterness. In addition, concentrations of 10 -3 Up to 10 -11Lactones between 5-hydroxy-1-butyl-1-olactone and 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone can be used to provide desired flavors to meat replicas, such as creamy, buttery, caramel, fatty, umami, fruity, tallow, and meaty aromas. Thus, lactones can be used as masking agents or as flavoring agents. Lactones can act as masking agents in other products, including, but not limited to, dairy replicas, such as milk, cheese, and yogurt; or protein supplements, such as protein bars and protein powders. The combination of lactones can provide unique flavor characteristics that are important in forming meaty flavors (e.g., tallow and sweet aromas) in food products such as meat replicas or in providing beefy flavors to non-beef food products. When lactones are added to products, meat replicas are improved in terms of overall liking and meaty ratings. In some embodiments, for example, a combination of butyrolactone, delta-tetradecalactone, and 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone can be used to provide meaty flavors. Lactones can be added to vegetable oils to make fat taste more like animal fat and increase the stickiness in the mouth. Lactones can also be added to increase the sweetness of the product without changing the sugar content. It should be noted that agents such as lactones and carotenoids can be used for flavoring food replicas (e.g., plant-based food replicas) including meat or cheese replicas, and can also be used to modify the flavor of food products such as meat and cheese (e.g., to increase meat or cheese flavor).

[0080] In some embodiments, carotenoids such as β-carotene, zeaxanthin, lutein, trans-β-apo-8'-carotenal, lycopene, and canthaxanthin can be used to control the formation of desired flavors and prevent the formation of undesirable flavors in food products such as plant-based food products (e.g., the meat replicas described herein). Carotenoids can be used to reduce plant off-flavors in other food products, including dairy replicas. It has been found that each type of carotenoid has different properties in forming desired flavors and controlling off-flavors. See Examples 18 and 26. Carotenoids can increase sweetness and fat notes to improve meat replicas when added between 0.00001% and 0.1%.

[0081] Carotenoids can be added to meat replicas by adding them to a flavoring emulsion or a flavoring broth. Carotenoids can be added before or after cooking. Between 0.00001% and 0.1% carotenoids can be added. When carotenoids are added before cooking, they can serve as a base for a reaction flavoring mixture that forms a flavor before it is added to a meat replica. Carotenoids also alter the pathways by which other flavors are produced by acting as antioxidants. By adding carotenoids, a flavoring emulsion can have improved flavoring qualities; reduced oxidative off-flavors (waxy, fishy, ​​pigmented), reduced other off-flavors (earthy, mushroomy, cereal, beany), and increased sweetness, fat, meatiness, and umami. Each carotenoid has a different resulting flavor profile. For example, adding lycopene to a flavoring emulsion before cooking produces a bland flavor, while beta-carotene is very tasty with the added fat and meatiness compared to the control. The flavor profile of adding carotenoids before cooking has a greater effect on the flavor profile. When carotenoids are added after cooking, they can still have a beneficial effect, especially in reducing the off-flavors produced during storage. Other flavor precursor molecules in the seasoning emulsion or seasoning broth have an impact on the effect of carotenoids. The resulting flavor and / or aroma characteristics can be adjusted, in particular, by the type and concentration of the seasoning precursor, the reaction pH, the cooking time, the cooking temperature, the type and amount of iron complexes (e.g., heme cofactors such as heme proteins, or ferrous chlorophyllin) or iron salts (ferric gluconate), the reaction temperature, and the amount of water activity in the product, all of which change how carotenoids change the flavor characteristics. Specific examples include how carotenoids can reduce or prevent the formation of flavor compounds produced in vegetable oils, especially when metals are present in the oil source. Carotenoids, when added to seasoning emulsions with fats and oils containing polyunsaturated fatty acids (such as linoleic acid, gamma linoleic acid, DHA and EPA), can prevent the off-flavors of fishy, ​​pigmented and vegetable flavors and help produce meaty and sweet flavors.

[0082] Specifically, carotenoids can reduce grain taste, wood taste, earthy smell, mushroom taste, plant taste and oxidation taste. Carotenoids can be added to different parts of plant products to have different effects. Carotenoids can reduce or prevent the formation of off-flavor compounds produced in wheat flour including wheat gluten. For example, lutein can be added to raw meat sticky dough and reduce the overall flavor intensity, reducing the grain taste, wood taste and oxidation taste in cooked meat sticky dough and final products. In some cases, these changes in flavor characteristics are supported by the reduction of specific seasoning compounds as seen by SPME gas chromatography-mass spectrometry (GC-MS), and in other cases, there is no change in seasoning compounds, but it is observed that the grain characteristics are reduced, indicating that carotenoids work by changing the chemical reactions that occur and by masking specific flavors. In addition, carotenoids are added to meat sticky dough so that the sample is described as more fat and sweeter than the control without carotenoids. The main compounds reduced with lutein included oxidized flavor compounds such as alcohols and aldehydes, including (Z)-2-nonenal, (E,E)-2,4-nonadienal, and 1-penten-3-ol; in addition, sulfur-containing compounds including methyl mercaptan, 2-acetylthiazole, and dimethyl sulfide were reduced with lutein; many of these compounds were also described as grainy and oxidized by trained flavor scientists using gas chromatography-olfactometry (GCO).

[0083] Antioxidants such as epigallocatechin gallate (EGCG) can also be used to reduce off-flavors in food products such as plant-based products (e.g., meat replicas). Antioxidants such as EGCG found in green tea extract (and which can be purified therefrom) can be added at 0.0001% to 0.1%. Antioxidants including EGCG can also be added to meat pie and change the flavor profile of the cooked meat pie and consumables produced from the pie. As described by trained flavor scientists and confirmed using GCMS, EGCG reduces the overall flavor of the pie and particularly reduces off-flavors such as cereal and oxidized flavors.

[0084] Vegetables or fruits (juices, purees or extracts) can be added to meat replicas to increase the perceived meaty flavor (e.g., meaty taste) and likeability of the product, as well as to increase the perceived fat and fat mouth adhesion. Additionally, it can cause the taster to increase salivation when consuming the product, resulting in an increase in the perceived juiciness of the meat replica. The type of meaty flavor enhanced by the vegetable or fruit depends on the type and processing. Examples include beef tallow flavors enhanced under cooking added by cucumbers and melons; sweet tang, grilled meat flavors, and savory flavors added by honeydew melon; sweet tang and umami flavors added by pineapple and savory red meat flavors added by tomatoes.

[0085] Vegetables or fruits can be added to meat replicas in the form of juices, purees, extracts produced by pressing, juicing, steam distillation, pressure distillation, solvent-assisted flavor extraction, or other methods. The vegetables or fruits can be uncooked or unprocessed, or can be cooked or otherwise processed (e.g., by pasteurization or by enzyme inactivation) to denature proteins (e.g., lipoxygenase). The flavor profile (the amount of meaty and off-flavors, including the fresh or vegetable flavors of fruits or vegetables) can change according to cooking or other processing, and depends on the amount and process of cooking or other processing. Many flavors in fruit and vegetable extracts, purees, and juices are produced by enzymes. These enzymes can produce desirable or undesirable flavors, and the desired flavor depends on the application of the extract and juice. Selecting the appropriate type of fruit or vegetable and processing allows the production of flavors suitable for meat replicas. In addition, during processing, it may be necessary to inactivate enzymes that can cause off-flavors. A specific enzyme in an extract that can produce off-flavors when added to a meat replica is lipoxygenase, which is particularly active in fruit and vegetable peels. Destroying the epidermis can increase lipoxygenase activity. Therefore, inactivation of the enzyme before cutting the fruit or vegetable peel can help reduce odor. When producing fruit and vegetable extracts, mud or juice, the enzyme can be inactivated by heating above 60°C, high pressure pasteurization or enzyme inhibition. In some embodiments, for example, lipoxygenase can be inhibited by adding inhibitors such as epigallocatechin gallate (EGCG) or by adding other redox active enzymes. In some embodiments, the whole fruit or vegetable can be cooked or processed before penetrating the epidermis, or cooking can occur after cutting the product. Cooking or other processing can be fast (several minutes) or long (several hours). When cooking is used, the temperature can be slightly increased from room temperature to more than 120°C at normal pressure. For example, fruits or vegetables can be cooked at a temperature of 60-100°C (e.g., 70-80°C, 80-90°C or 90-100°C). The process may include blending, leaching and or pressing. In some cases, seeds can be removed, or seeds can be retained.

[0086] For example, cucumber puree added to a meat replica can provide additional beef tallow flavor, but can also bring out green vegetable flavors. When the fruit is first cooked, several compounds that cause the fragrance and strong cucumber flavors are reduced, including (but not limited to) 2-nonenal and 2,6-nonadienal. In addition, buttery, fatty, and beef tallow flavors increase, which may come from increased lactone concentrations as seen by SPME GC-MS. Cooking of tomatoes also enhances meaty flavors while reducing fragrance and tomato flavors.

[0087] Fruit or vegetable flavoring liquids can be added to different components of the product, such as to meat paste before cooking, to fat emulsions after or before cooking, to gelled matrices, to fully assembled products, or to unreacted flavored broths. Extracts can be added from 0.0001% up to 10% purees and juices.

[0088] Acids, such as lactic acid, can be added to meat pastes to lower the pH and change the flavor reactions that occur under cooking and processing. The pH of beef is about 5.5; to achieve a meat paste at pH 5.5, additional acidity is required. Lactic acid brings the desirable umami, sour flavor found in beef.

[0089] In other embodiments, non-animal fats may include isolated plant proteins. For example, the emulsion can be made by combining an aqueous solution of an oil of plant origin, an algae oil, or an oil from bacteria or fungi and an optional flavoring agent with an isolated plant protein (e.g., conglycinin from soybeans), then homogenizing the mixture using, for example, a high-speed homogenizer and heating it at 90° C. for a short time (e.g., 5 minutes). The physical properties of the emulsion, such as melting temperature, firmness, brittleness, color, can be adjusted by using different types of isolated proteins, changing protein concentration, oil-water ratio, homogenization speed, heating temperature, and heating time. For example, an emulsion with a high oil-water ratio and a low protein concentration is more brittle and melts more easily, while an emulsion with a lower oil-water ratio and a higher protein concentration is softer, less brittle, more viscous, and melts at a higher temperature.

[0090] In some embodiments, emulsions can be made by combining an oil of plant origin, algae oil, or oil from bacteria or fungi and optional flavorings with an aqueous solution of an isolated protein (e.g., soybean conglycinin) brought to pH>10 (e.g., pH 12) using, for example, sodium hydroxide. Agitation, stirring, or homogenization of this mixture results in the formation of an emulsion. After the emulsion is formed, the pH can be adjusted to a neutral or acidic pH by adding, for example, hydrochloric acid or lactic acid. The physical properties of these emulsions can be controlled by varying the protein type, protein concentration, pH level during homogenization, homogenization speed, and oil-to-water ratio.

[0091] In other embodiments, the emulsion can be made by mixing an aqueous solution of an oil of plant origin, an algae oil, or an oil from bacteria or fungi, a salt, and a flavoring agent (e.g., a flavoring precursor) and an emulsifier. For example, monoglycerides / diglycerides, lecithin, phospholipids, Tween surfactants, sodium stearoyl lactylate, or DATEM (diacetyl tartaric acid ester of monoglycerides) can be used as emulsifiers. The physical properties of these emulsions can be controlled by changing the emulsifier type and concentration, the homogenization speed, and the oil-water ratio.

[0092] The solidified, optionally seasoned, and / or protein-containing fat may be combined with the meat dough, and the mixture of the meat dough and the non-animal fat may be broken into smaller pieces, for example, by chopping, grinding, cutting, mincing, shearing, or tearing. In some embodiments, shear may be applied to the dough while heating so that the dough becomes firm during the cooking process and eventually breaks into pieces. Thus, another step for breaking into pieces will not be required.

[0093] A carbohydrate gel and an optional binder may be added to the dough-fat mixture. Carbohydrate gels are also useful for creating the texture of meat replicas and providing juiciness to the final product without making it soggy. Typically, carbohydrate gels with a melting temperature between about 45° C. and about 85° C. are used. Non-limiting examples of suitable carbohydrate gels include agar, pectin, carrageenan, konjac (also known as glucomannan), alginates, chemically modified agarose, or mixtures thereof.

[0094] The adhesive can be a separated vegetable protein or carbohydrate gel. Non-limiting examples of suitable vegetable proteins include ribulose bisphosphate carboxylase, albumin, gluten, glycinin, conglycinin, legumin, globulin, pea globulin, conalbumin, gliadin, gluten, glutenin, hordein, prolamin, phaseol, protein body, secalin, wheat gluten, zein, oleosin, caleosin, steroleosin or a mixture thereof (e.g., albumin fraction). Vegetable protein can be obtained from any source, including soybean, pea or lentil. In certain embodiments, suitable adhesives can be obtained from non-plant sources. For example, in some embodiments, ovalbumin or collagen can be used as adhesives.

[0095] When the binder is a protein, it is beneficial that the denaturation temperature of the protein is less than the melting temperature of the carbohydrate-based gel. For example, a suitable protein binder (e.g., ribulose bisphosphate carboxylase, albumin, soy conglycinin, or gluten, or mixtures thereof) may have a denaturation temperature between about 40° C. and about 80° C. This allows the carbohydrate-based gel to melt after the protein binder denatures and binds the meat replica together, providing a better texture and forming a meat replica.

[0096] In some embodiments, the protein used as a binder may be chemically or enzymatically modified to improve its texture and / or flavoring properties. For example, a food-grade enzyme such as papain may be used to partially proteolyze the protein to produce a better water release profile during gelation and cooking. In some embodiments, the protein used as a binder may be chemically or enzymatically modified to adjust the denaturation and gelation temperature of the protein, for example, to achieve a specific gelation temperature (e.g., 52° C. to mimic myosin or 68° C. to mimic actin). In some cases, proteins such as proteases may be used to reduce bitterness that may be present in purified protein fractions.

[0097] In some embodiments, the binder is a carbohydrate gel. For example, a carbohydrate gel that becomes firm after cooking to 140°F to 190°F (e.g., 150°F to 180°F). Non-limiting examples of carbohydrate gels include methylcellulose, modified starches (such as hydroxypropyl methylcellulose), guar gum, locust bean gum, xanthan gum, or mixtures thereof.

[0098] Additionally, iron complexes and / or iron salts and flavorings can be added to the meat replica. The iron complexes and / or iron salts can be the same or different than the iron complexes and / or iron salts used for meat paste, connective tissue replicas, or non-animal fat flavoring. The flavoring can be a flavoring precursor or a mixture of flavoring precursors (described above) so that upon cooking of the meat replica, the iron complexes and / or iron salts and the flavoring precursors can react and produce a flavoring compound. The flavoring can also be a flavoring, such as yeast extract, hydrolyzed protein; or a flavoring compound. Flavoring compounds may include, for example, phenylacetic acid, (E,E)-2,4-nonadienal, water-soluble aquaresin onion, oil-soluble onion, p-cresol, acetone acetate, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, (E,E)-2,4-octadienal, 2-methyl-1-butanethiol, 2-methyl-3-furanyl tetrasulfide, ethyl 2-mercaptopropionate, 2-mercapto-3-butanol (mixture of isomers), n-decane -d22, oil-soluble garlic, thiothiazole, thioacetic acid ester, mercapto-3-butanol, spiromeat, 1-penten-3-one, 2-methyl-3-furanthiol, 2-methyl-3-tetrahydrofuranthiol, oleic acid, dipropyl trisulfide, difurfuryl disulfide, methylcyclopentenol ketone, 3-methylthiohexanal, butyric acid, butyrolactone, 5-methyl-2(3H)-furanone, furanone, 1-(1H-pyrrol-2-yl)-ethanone, hexanoic acid and combinations thereof. Additional flavor compounds are commercially available from companies such as Sigma Aldrich (St. Louis, MO), Penta Manufacturing Co. (Fairfield, NJ), Advanced Biotech (Totowa, NJ), Firmenich (Meyrin, Switzerland), Givaudan (Vernier, Switzerland), International Flavors and Fragrances (New York, NY), and Wild Flavors (Erlanger, KY).

[0099] In some embodiments, seasonings such as table salt (e.g., sodium chloride or potassium chloride), garlic or herbs (e.g., rosemary, thyme, basil, sage, or mint), emulsifiers (e.g., lecithin), additional fiber (e.g., zein or inulin), minerals (e.g., iodine, zinc, and / or calcium), meat shelf-life extenders (e.g., carbon monoxide, nitrites, sodium metabisulfite, Bombal, vitamin E, rosemary extract, green tea extract, catechins, and other antioxidants) can be incorporated into meat replicas.

[0100] The meat replicas described herein may also include natural colorants, such as turmeric or beet juice; or artificial colorants, such as azo dyes, triphenylmethane, xanthenes, quinine, indigo, titanium dioxide, Red No. 3, Red No. 40, Blue No. 1, or Yellow No. 5; or any combination of natural and / or artificial colorants.

[0101] Any replica described herein can be shaped into a desired use, such as into meat patties, meatloaf, whitefish, meatballs, or nuggets, and used in any type of food product that would use ground meat, such as as a burrito filling, or in a casserole, sauce, topping, soup, stew, meatballs, or meatloaf. In some embodiments, a meat replica can be formed into, for example, meatballs or nuggets, and then coated with breadcrumbs, rice, or flour (e.g., oatmeal or coconut flour) for convenience.

[0102] Meat imitations

[0103] The meat replicas described herein can include about 5 wt% to about 88 wt% (e.g., about 10 wt% to about 40 wt%, about 25 wt% to about 35 wt%, about 40 wt% to about 88 wt%, or 45 wt% to about 60 wt%) of meat replica paste; about 0 wt% to about 40 wt% (e.g., about 15 wt% to about 25 wt%) of carbohydrate-based gel; about 3 wt% to about 35 wt% of non-animal fat (e.g., about 10 wt% to about 15 wt%); about 0.00001 wt% to about 10 wt% of flavoring; about 0 wt% to about 15 wt% (e.g., about 2 wt% to about 15 wt% or about 2 wt% to about 10 wt%) of binder; and about 0.01 wt% to about 4 wt% (e.g., about 0.05 wt% to about 1 wt% or about 0.2 wt% to about 2 wt%) of iron complexes (e.g., heme-containing proteins) and / or iron salts. The amount of flavoring can vary depending on the type of flavoring. In some embodiments, the flavoring agent can be about 0.5% to about 7% of the meat replica. For example, the flavoring agent (such as a mixture of flavoring precursors) can be about 0.5% to about 7% of the meat replica (e.g., about 1% to about 3%; about 3% to about 6%; about 4% to about 7%). In some embodiments, the flavoring agent (such as a flavoring compound) can be about 0.00001% to about 2% of the meat replica.

[0104] As described herein, one or more, two or more, three or more, or four or more components may include a flavoring. For example, a meat paste may include a flavoring in the edible fiber component (e.g., a flavoring compound produced by combining an iron complex or iron salt with one or more flavoring precursors and heating) or may include a flavoring (such as yeast extract). Non-animal fats may also include a flavoring (e.g., a flavoring compound produced by combining an iron complex or iron salt with one or more flavoring precursors and heating). Replicas may also include an iron complex or iron salt and one or more flavoring precursors, which may react when the replica is cooked, thereby enhancing the sensory experience of cooking the replica. Additionally, the replica may include a flavoring or flavoring compound.

[0105] In some embodiments, the components are produced at a desired particle size and then compressed together for 5 minutes to 24 hours (e.g., 10 minutes to 2 hours, 1 to 4 hours, 4 to 8 hours, 6 to 12 hours, or 12 to 24 hours) to allow the components to adhere to the meat replica. The meat replica can then be ground to replicate the properties of ground meat. The meat replica can be compressed into any desired form to replicate the shape and density of, for example, steaks, tenderloins, chops, or fillets. The meat replica can also be further processed into processed meats, such as sausages.

[0106] The present invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0107] Examples

[0108] Example 1

[0109] Isolation and purification of leghemoglobin

[0110] The nucleic acid of encoding soybean leghemoglobin C2 (Uniprot KBP02236) with N-terminal His6 epitope tag and TEV cleavage site is cloned into pJexpress401 carrier (DNA2.0) and transformed into Escherichia coli BL21. Transformed cells are grown by the batch feed fermentation supplemented with kanamycin (kanamycin), 0.1mM ferric chloride and 10 μg / ml 5-aminolevulinic acid. Expression is induced by 0.3mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and cells are grown for 24 hours at 30 ℃. Cells are concentrated by centrifugation and resuspended in 20mM potassium phosphate pH7.8, 100mM NaCl. Cells are dissolved by high pressure homogenization and purified by centrifugation and microfiltration. Use IMAC agarose fast flow resin (GE Healthcare) filled with zinc to purify leghemoglobin from soluble lysate. The bound leghemoglobin was eluted from the resin with 500mM potassium dihydrogen phosphate, 100mM NaCl. The purified leghemoglobin was neutralized and concentrated using ultrafiltration. The concentrated leghemoglobin was reduced with 20mM sodium dithionite. The sodium dithionite was removed by diafiltration. The leghemoglobin concentration was determined by soret peak absorbance and adjusted to 60-70mg / ml. The final leghemoglobin product was frozen in liquid nitrogen, lyophilized and stored at -20°C. The purity (partial abundance) of the leghemoglobin was analyzed by SDS-PAGE and was determined to be ~80%. Analysis of the UV-VIS spectrum (250-700nm) showed that the spectral characteristics were consistent with leghemoglobin loaded with heme.

[0111] Soybean leghemoglobin C2 and eight Pichia pastoris heme biosynthesis genes (listed in Table 1) were cloned into the Pichia pastoris expression vector pJA (BioGrammatics Inc.; Carlsbad, CA) under the control of the pAOX1 methanol-inducible promoter. Pichia pastoris strain Bg11 (BioGrammatics Inc.; Carlsbad, CA) was transformed with the linearized plasmids, and stable integrants were selected by antibiotic resistance.

[0112] Table 1

[0113]

[0114] Transformed Pichia cells were grown by batch feed fermentation and induced to express leghemoglobin with methanol at 30°C for 120 hours. Cells were concentrated by centrifugation, resuspended in water and dissolved by high pressure homogenization. Solids were removed by treatment with Tramfloc 863A, centrifugation and 0.2 μm microfiltration (Koch Membrane Systems). Soluble lysates were concentrated and diafiltered with water using 3kDa ultrafiltration (Spectrum Laboratories). The prepared lysates were partially purified to a final purity of ~40% using HPA25L anion exchange resin (Mitsubishi). The partially purified leghemoglobin solution was reconstituted by concentration using 3kDa ultrafiltration (Spectrum Laboratories) and water diafiltration and further purified using Q fast flow anion exchange resin (GE Lifesciences). The final leghemoglobin product was concentrated and frozen at -20°C using 3kD ultrafiltration. The final product was -80% pure and contained 80 g / L leghemoglobin.

[0115] Example 2

[0116] Isolation of ribulose bisphosphate carboxylase

[0117] One kilogram of fresh spinach leaves was macerated in a Vita-prep 3 blender (Vitamix Corp, Cleveland, OH) at a ratio of 1:1 with potassium phosphate buffer (pH 7.4) containing 0.1M NaCl. The extraction was performed for 10 minutes at the highest setting (3HP motor). The temperature was maintained below 30°C. The pH was adjusted to 7.4 using a 10M NaOH solution after grinding. The homogenate was centrifuged at 3500g for 5 minutes to simulate large-scale (GEA Westfalia decanter GCE-345 at a feed rate of about 1 gpm) conditions. The centrifuge pellet was discarded. Next, the centrate (about 1.6 L) was microfiltered using a 0.2 μm modified polyethersulfone (mPES) membrane (KrosFlo K02E20U-05N, Spectrum Laboratories Inc. Rancho Dominguez, CA) in hollow fiber form. The retentate (about 0.25 L) was diafiltered using about 1.5 L of extraction buffer. The permeate (~3 L) of this filtration step was concentrated to about 0.1 L using a 10 kDa mPES membrane (MiniKros N02E010-05N, Spectrum Laboratories Inc. Rancho Dominguez, CA). The pH of the protein concentrate was about 7.4. A concentrated acid solution (such as 6 M hydrochloric acid) was slowly added to the concentrate to reduce the pH to 5. The mixture was vigorously stirred for 30 minutes using a magnetic stirring plate or homogenizer, and then centrifuged at 3500 g for 5 minutes to obtain an off-white centrifuge cake and a brown centrifuge filtrate. The centrifugal filtrate is discarded and the protein centrifugal block is washed with deionized water. The centrifugal block is resuspended in 0.05-0.1L deionized water. The solution is vigorously mixed into a uniform slurry and the pH is slowly raised to 11 using a concentrated alkali solution (such as 10M sodium hydroxide). The resulting solution is a transparent yellow color. Then, the pH is reduced to 9 to maintain a transparent mixture. The product is dried using a spray dryer, or frozen and dried using a freeze dryer. This material is analyzed using a Leco FP-528 nitrogen combustion analyzer (Leco, St. Joseph, MI, Michigan) by the AAOC method (AOAC, 2000). Protein is calculated according to % nitrogen × 6.25 and is calculated to be 86% protein. The product obtained is slightly decolorized and retains low temperature denaturation properties.

[0118] Example 3

[0119] Isolation and Decolorization of Ribulose Bisphosphate Carboxylase

[0120] One kilogram of fresh spinach leaves were softened in a Vita-prep 3 blender (Vitamax, Cleveland, Ohio) at a ratio of 1:1 (w / w) with potassium phosphate buffer (pH 7.4) containing 8% (w / v) PEG (Carbowax Sentry PEG 8000; Dow Chemicals, Midland, MI) and 0.1% (w / v) cationic flocculant (863A; Tramfloc, Inc., Houston, TX). The extraction was performed at the highest setting (3HP motor) for 3 minutes, always maintaining the temperature below 30°C. The pH was adjusted to 7.4 using 10M NaOH solution after grinding. Homogenate uses table centrifuge (Allegra X15R, SX4750 rotor; Beckman Coulter, Inc., Pasadena, CA) at 3500g for centrifugation for 5 minutes.Discard centrifugal block and collect supernatant (about 1.6L) separately.Seven hydrate magnesium sulfate salt (K+S KALI Co., Ltd., Kassel, Germany) is added in supernatant to reach 1M concentration.Solution is thoroughly mixed and uses table centrifuge (Allegra X15R, SX4750 rotor; Beckman Coulter) at 5451g for centrifugation for 3 minutes.Three layers are formed in centrifuge bottle, and remaining green solid is separated (about 0.1L) with centrifugal block form.Separate PEG layer (about 0.3L) and form top layer, selective fractionation colored compound and odorous compound. The remaining transparent product in the middle layer is then microfiltered using a 0.2 μm modified polyethersulfone (mPES) membrane (Spectrum Laboratories) in the form of a hollow fiber. The retentate (about 0.25 L) is filtered using about 0.75 L of a 1 M magnesium sulfate solution. The permeate (about 3 L) of this filtration step is concentrated to about 0.1 L using a 70 kDa mPES membrane (Spectrum Laboratories). It is further filtered in 5 steps with about 0.5 L of deionized water. The pH of the protein concentrate is about 7 and the conductivity is less than 5 mS / cm. The resulting protein concentrate is a transparent light yellow. The product is dried using a spray dryer or frozen and dried using a freeze dryer. This material is analyzed using a standard 660 nm Pierce protein analysis and SDS gel density determination method. The dried solid is analyzed using an IR moisture analyzer. The flocculant and PEG concentrations in the final product are analyzed using a titration method. The protein concentration is about 91% (w / w), and the total solids are about 95% (w / w). The PEG and flocculant concentrations were analyzed to be less than 0.2% (w / w). The product purity was over 90%, and the recovery rate in the whole process was over 90%.The obtained product is decolorized and retains low temperature denaturation properties.

[0121] Example 4

[0122] Separation of Soluble Conglycinin from Soybean

[0123] The soluble conglycinin fraction (7S fraction) of soybean protein was obtained by the following method: 1 kg defatted soybean flour (CHS PDI 90) was mixed with 10 L of deionized water in a container equipped with an overhead mixer. After the powder mass was dispersed, the pH of the slurry was adjusted to 8 with 2N NaOH. The mixture was stirred at 4°C for 1 hour to extract all soluble proteins. Next, 2N H 2 SO 4 The pH of the mixture was adjusted to 5.8 and mixed for another hour at 4°C. Next, the mixture was centrifuged at 10,000 g for 10 minutes in a JLA 8.1 rotor (JHC centrifuge, Beckman Coulter) to remove insoluble carbohydrates and protein (soybean globulin). 2N H 2 SO 4 The soluble supernatant was further acidified to pH 4.5 and mixed at 4°C for 1 hour. Next, the acidified mixture was centrifuged at 10,000 g for 10 minutes to collect the precipitated protein, and the supernatant containing lipoxygenase, soy lecithin and trypsin inhibitor was discarded. The conglycinin in the pH 4.5 precipitated protein fraction was re-dissolved by resuspending the pellet in 4 volumes of water (approximately 2 L) and adjusting to pH 8 using 2N NaOH. The mixture was stirred at 4°C for 1 hour. 2N H 2 SO 4 The pH of the mixture was lowered to 5.8 again to minimize co-purification of contaminating proteins. The mixture was centrifuged at 15000 g for 20 minutes to collect soluble conglycinin in the supernatant. The conglycinin fraction was concentrated using ultrafiltration (70 kDa mPES ultrafiltration membrane, 2600 sq. cm, Spectrum Laboratories). The resulting protein solution (approximately 0.5 L at 10% protein concentration) contained 55-65% pure conglycinin and gelled at 65° C. The protein was then lyophilized and stored at room temperature until used to make meat replicas.

[0124] Example 5

[0125] Preparation of meat bouillon broth for pre-seasoning meat bouillon

[0126] The dough broth was formed by mixing 1× Precursor Mix 1 (see Table 2), 0.5% leghemoglobin (LegH, isolated and purified as described in Example 1), and 18% refined, bleached and deodorized (RBD) coconut oil (from Shay and company, Milwaukie, OR), and stirring while the solution was heated until boiling, followed by simmering for 10 minutes. This solution was called "dough broth" and was used to form the meat dough of Example 10. Incubation of the coconut oil with the LegH and the precursor mix imparted a savory or meaty flavor to the broth, including caramel, fatty, beefy, nutty, sulfurous, metallic, buttery, sweet, savory, and umami.

[0127] Table 2 Composition of magic mixture

[0128]

[0129] Example 6

[0130] Preparation of flavored infused fat replicas

[0131] The flavored fat replica was formed by mixing a solution of 0.5% LegH (from Example 1), 1× Precursor Mix 1 (Table 1), and 30% RBD coconut oil (Shea and Company, Milwaukee, OR), and stirring the mixture while heating until boiling, then simmering for 10 minutes. The solution was cooled to allow the oil to solidify. Once the oil solidified, it was separated from the aqueous layer and used to prepare the burgers described in Example 11. Incubation of the coconut oil with the LegH and precursor mix allowed the flavors to infuse the oil, including savory, meaty, beef fat, slightly sweet, and sulfurous.

[0132] Example 7

[0133] Preparation of "soft connective" tissue replicas

[0134] Soft connective tissue replica is made from soy protein isolate ( EX38 (Solae), active wheat gluten (131100, Guisto's Specialty Foods, San Francisco, CA) and water were used to prepare. A Nano 16 extruder (Leistritz Advanced Technologies Corp., Somerville, NJ) with a custom cooling die (round, ID 6.5 mm, length 300 mm), a cooling water circulator and a high-pressure water pump (Optos, Eldex Laboratories Inc.) was used.

[0135] Fifty (50) g ​​of isolated soy protein and 50 g of wheat gluten flour were thoroughly mixed by hand mixing and tumbling for 5 minutes and then loaded into the loading tube of the batch feeder of the extruder. The dry mixture was supplied to the extruder at a rate of 2.4 g / min. Water was supplied to the second zone of the extruder barrel by a pump at a rate of 3.6 ml / min. The screw speed of the extruder was maintained at 120 RPM. The temperature gradient was set along the extruder barrel as follows: feed zone -25°C, zone 1 -30°C, zone 2 -60°C, zone 3 -110°C, zone 4 -110°C. The mold plate was neither actively heated nor cooled. The cooling mold was cooled by a cooling water circulator so that the mold was maintained at 24°C.

[0136] The soft connective tissue replica produced by this method is off-white and highly fibrous / stringy, with a neutral taste and flavor. The tensile strength of this material is low and comparable to that of roast beef tenderloin.

[0137] Example 8

[0138] Preparation of "tough fibrous connective" tissue replicas

[0139] To prepare a tough fibrous connective tissue replica, 50 g of isolated soy protein and 50 g of wheat gluten flour were thoroughly mixed by hand mixing and tumbling for 5 minutes and loaded into the loading tube of the batch feeder of the extruder. The dry mixture was supplied to the extruder at a rate of 3.6 g / min. Water was supplied to the second zone of the extruder barrel by a pump at a rate of 5.4 ml / min. The screw speed of the extruder was maintained at 120 RPM. The temperature gradient was set along the extruder barrel as follows: feed zone -25°C, zone 1 -37°C, zone 2 -61°C, zone 3 -135°C, zone 4 -135°C. The die plate was neither actively heated nor cooled. The cooling die was cooled by a cooling water circulator so that the die was maintained at 26°C.

[0140] The tough fibrous connective tissue replica produced by this method is light brown in color and is a fibrous / layered material with a neutral taste and flavor. The tensile strength of this material is high and comparable to that of cooked beef tendon.

[0141] Example 9

[0142] Preparation of pre-seasoned "soft connective" tissue

[0143] To prepare the flavored soft connective tissue replica, 50 g soy protein isolate, 50 g wheat gluten flour, 1 g yeast extract No. 9 (Flavor house Inc., X11020) and yeast extract No. 21 (Biospringer 1405 / 40MG1) were thoroughly mixed by hand mixing and tumbling for 5 minutes, loaded into the loading tube of the batch feeder of the extruder and extruded as described in Example 7. The pre-seasoned soft connective tissue had a savory taste with increased flavor complexity and reduced off-flavors compared to the soft connective tissue produced in Example 7.

[0144] Example 10

[0145] Preparation of "Meat Paste"

[0146] The ground beef replica "meat paste" was prepared using the following ingredients:

[0147] a. Vital wheat gluten (No. 131100, Guitu Specialty Foods, San Francisco, California)

[0148] b. Soft connective tissue replica (see Example 7, the pre-seasoned soft connective tissue of Example 9 can also be used)

[0149] c. Sticky broth (see Example 5)

[0150] A 100 g portion of meat dough is prepared as follows. First, 25 g of soft connective tissue replica is manually chopped lengthwise into approximately 1 inch long pieces. The chopped soft connective tissue replica is combined with 25 g of dry wheat gluten in a mixing bowl and gently shaken by hand to mix well. In another container, 50 mL of dough broth is boiled and simmered for 10 minutes. The hot dough broth is added to the dry gluten-connective tissue replica mixture and blended in a stand mixer (e.g., a stand mixer with a dough mixing attachment). Professional 600 Series 6-Quart Bowl Stand Mixer Model KP26M1XER was set on speed 2) and kneaded for 30 seconds to form a meat dough.

[0151] After kneading, the meat dough was formed into a thick mass and transferred to another vessel for steaming. The meat dough was steamed (in an Aroma rice cooker model ARC-1030SB) until the internal temperature reached approximately 200°F and maintained at that temperature for an additional 20 minutes. After steaming, the dough was transferred to a container on ice to cool it to room temperature. The steamed meat dough can also be stored at 4°C for up to one week at this point. Before forming a beef patty replica, the steamed meat dough was manually torn into smaller pieces of approximately 1 cubic inch. The mixture is now ready for forming a beef patty replica (described in Examples 11 and 12).

[0152] Example 11

[0153] Assembling and cooking the burgers

[0154] Replica burgers containing the ingredients in Table 3 were prepared. A 1% agar preparation was made by adding 1 g of agar powder (Trade 6410, Now Foods Bloomingdale, IL) to 99 ml of water in a glass beaker. The agar was completely dissolved by heating the mixture to 100° C. while stirring, and then cooled in an ice bath for 20-30 minutes until a firm gel was formed. The gel was then transferred to a coffee grinder ( Model CUIDCG-20N) and grind for 20 seconds to break it into small pieces for mixing.

[0155] Table 3 Composition of hamburgers

[0156]

[0157] The meat paste (Example 10) and flavored coconut oil (Example 6) were mixed manually in a bowl. Typical batch sizes were 100 g to 2000 g. The mixture was then blended using a stand mixer ( Professional 600 Series 6-Quart Bowl Lift Stand Mixer Model KP26M1XER and Food grinder model FGA, St. Joseph, MI) was ground at speed setting 1. The mixture was fed by a screw conveyor past rotating knives mounted in front of a fixed orifice plate. The ground tissue was collected in a bowl.

[0158] Next, the following ingredients were added in the ratios shown in Table 3: 1% agar preparation, ribulose bisphosphate carboxylase (approximately 50% ribulose bisphosphate carboxylase by weight), 16× Precursor Mix 2, and LegH (350-650 mg / g). The ingredients were added in the order listed here and after each addition, the materials were gently mixed. Next, thirty (30) g or 90 g portions of the ground tissue were manually formed into round patty shapes. Typical dimensions for 30 g patties are 50 mm×12 mm. Typical dimensions for 90 g patties are 70 mm×18 mm. During assembly, grinding, and forming, keep all materials cold (4-15° C.). Refrigerate the patties until cooking. The patties are cooked on a preheated (325-345° F.) non-stick long-handled skillet and heated to an internal temperature of 160° F., while turning over every 2 minutes. Typical cooking times range from 12 to 15 minutes. The appearance, texture and flavor of the cooked patties were similar to ground beef as judged by a trained sensory panel. In addition to cooking in patty form, the unformed material can be used in a variety of dishes such as burrito fillings, casseroles, sauces, toppings, soups, stews or meatloaf.

[0159] Example 12

[0160] Adding flavoring molecules to burgers

[0161] Replica burgers containing the ingredients in Table 4 were prepared.

[0162] Table 4 Composition of hamburgers

[0163]

[0164] Unseasoned meat paste (wheat gluten, unseasoned soft connective tissue and water) and coconut oil were manually mixed and ground in a bowl as described in Example 11. Next, the following ingredients were added in the ratios shown in Table 4: 1% agar preparation, ribulose bisphosphate carboxylase (approximately 50% by weight), 16× Precursor Mix 2, and LegH (350-650 mg / g). The flavoring compound and garlic oil were diluted to 1×10 -2 , followed by addition at the concentrations listed in Table 4. The ingredients were added in the order listed here and after each addition, the materials were gently mixed. Next, 100 g portions of the ground tissue were manually formed into round patty shapes. During assembly, grinding and forming, all materials were kept cold (4-15° C.). The patties were cooked on a preheated (325-345° F.) non-stick long-handled frying pan and heated to an internal temperature of 160° F., while turning over every 2 minutes. The patties were typically cooked for 12 to 15 minutes. The appearance, texture and flavor of the cooked patties were similar to ground beef. The flavor of these patties was not as deep as that of burgers formed with pre-seasoned sticky dough and fat, however, as judged by a trained sensory panel, these burgers had an additional flavor associated with beef.

[0165] Example 13

[0166] Preparation of solution-spun corn protein fibers for connective tissue replicas

[0167] Solution spun corn protein fiber was produced using corn protein meal (Prairie Gold Inc., Bloomington, IL), ethanol (190 proof Everclear by Luxco), sodium hydroxide (Fisher Scientific), glycerol (Fisher Scientific), and water. Fifty (50) g ​​corn protein meal, ten (10) g glycerol, thirty-six (36) g ethanol, and four (4) g water were mixed in a glass jar using a homogenizer for 5 minutes. The pH of the solution was adjusted to 7.0 with a 1 M solution of sodium hydroxide in ethanol. The solution was loaded into a 1 ml syringe with a 30 gauge needle. The syringe was mounted on a syringe pump (New Era Syringe Pumps, Inc.) with the needle pointing downward and mounted vertically above a custom fiber winder with a Delrin spool. The winding rod was set to rotate at 3 RPM.

[0168] The syringe pump was set to 0.12 ml / h and started. When a drop of solution formed at the end of the needle, it was picked up with a spatula and stretched into a fiber. The fiber end was touched to the winding rod until it adhered. Next, a heating fan directed to the winding rod at the fiber attachment position was turned on to help the fiber dry. The fiber was wound until the syringe was empty, after which the syringe was reloaded and the above procedure was repeated. After winding, the fiber was pre-cured in a 110°C oven for 1 hour and then finished by baking at 175°C for 5 minutes.

[0169] The zein fiber obtained by this process is a translucent yellowish fiber, 60-80 microns thick as measured by optical microscopy. It is very flexible in air and water, maintaining a high tensile strength (10-15 MPa) similar to animal connective tissue even after being soaked in water for several hours.

[0170] Example 14

[0171] Preparation of meat bouillon broth with iron gluconate for pre-seasoning meat bouillon

[0172] The dough broth was formed by mixing 1× Precursor Mix 1 (see Table 2), 1 mM ferric gluconate, and 18% refined, bleached, and deodorized (RBD) coconut oil (Shea and Company), and stirring while the solution was heated until boiling, followed by simmering for 30 minutes. This solution is called "ferric gluconate dough broth" and can be used in place of the "dough broth" used in the meat dough of Example 10. Incubation of the coconut oil with the ferric gluconate and the precursor mix imparts a savory and or meaty flavor to the broth, including porky, beefy, sulfurous, metallic, sweet, savory, and umami flavors to the broth.

[0173] Example 15

[0174] Preparation of flavored infused fat replicas with iron gluconate

[0175] A flavored fat replica containing ferric gluconate was formed by mixing 0.25% LegH solution, 1 mM ferric gluconate, 1× Precursor Mix 1 (Table 1), and 30% RBD coconut oil (Shea and Company) and stirring the mixture while heating until boiling, then simmering for 10 minutes. The solution was cooled to 4° C. to allow the oil to solidify. Once the oil solidified, it was separated from the aqueous layer and used in place of the flavored fat replica to prepare the burgers described in Example 13. Incubation of the coconut oil with LegH, ferric gluconate, and precursor mix allowed flavors to infuse the oil, including savory, meaty, beef fat, sweet, metallic, and sulfur.

[0176] Example 16

[0177] Preparation of meat paste containing cream of tartar

[0178] Meat dough was prepared using the ingredients shown in Table 5 as follows.

[0179] Table 5 Composition of meat paste

[0180]

[0181] First, water, coconut oil, 1M lactic acid solution, and hydrolyzed vegetable protein are mixed and heated to 60°C to make a broth. Heating is performed to melt and aid in the distribution of the coconut oil. Gluten powder (vital wheat gluten No. 131100, Guitu Specialty Foods, San Francisco, California) and cream of tartar are mixed in another container. Next, the warm broth is added to the dry mixture and stirred with a stand mixer (e.g., a 10000 Blend Mixer with a sticky mixing attachment). Professional 600 series 6 quart bowl lift stand mixer model KP26M1XER, set at speed 2) knead for 30 seconds to form a meat dough. After kneading, the meat dough is formed into a thick mass and transferred to another vessel for steaming. The meat dough is steamed in an aroma rice cooker model ARC-1030SB until the internal temperature reaches approximately 88°C. After steaming, the dough is transferred to a container on ice to cool it to 4°C. Tartar changes the texture of the dough in a favorable manner. When compared to the meat dough of Example 10, the meat dough of this example is more sticky, the appearance factor after grinding is more similar to ground beef, and has improved handling characteristics to make it easier to shape and form meat patties.

[0182] Example 17

[0183] Preparation of meat paste containing lutein

[0184] Meat dough was prepared using the ingredients shown in Table 6 as follows.

[0185] Table 6 Composition of meat paste

[0186]

[0187] First, water, coconut oil (Shea and Company, Milwaukee, OR), and lutein (FloraGLO Lutein 20% SAF, DSM Nutritional Products, Overland Park, KS) were mixed and heated to greater than 25°C to make a broth. Heating was performed to melt and aid in the distribution of the coconut oil and lutein. Next, the warm broth was added to the gluten flour (vital wheat gluten). LF, ADM, Chicago, IL) and using a vertical mixer (e.g., a 400-degree mixer with a dough mixing attachment) set at speed "2". The mixture was kneaded for 30 seconds using a professional 600 series 6 quart bowl stand mixer model KP26M1XER) to form a meat dough. As in Example 16, after kneading, the meat dough was formed into a thick mass and transferred to another vessel for steaming, and then transferred to a container on ice to cool it to 4°C. A control batch of meat dough was made as described above, except that lutein was not added. The meat dough containing lutein was described as having less grain taste and closer to beef than the control meat dough.

[0188] Example 18

[0189] Reducing cereal and off-flavors by adding carotenoids

[0190] Meat dough was prepared using the ingredients shown in Table 7 as follows.

[0191] Table 7 Composition of meat paste

[0192]

[0193] First, melted coconut oil (50°C) was mixed with carotenoids, which were then mixed into the water. The broth was stirred vigorously, and then the wheat gluten powder was quickly added to the broth and mixed thoroughly with a spoon. The resulting raw meat dough was transferred to a metal mold or glass beaker for steaming as described in Example 16, and then transferred to a container on ice to cool it to 4°C.

[0194] The addition of carotenoids changes the flavor of the paste in a favorable manner; when compared to meat paste without carotenoids (Example 10), five trained flavor scientists described the meat paste with carotenoids as having less cereal taste, less oxidized taste, and less off-flavor overall in four tastings. Table 8 provides the summary sensory results of a group of samples with different carotenoids evaluated by five trained flavor scientists for cereal taste. The trained flavor scientists rated the samples from 1 to 5 for cereal taste, with 1 being the lowest cereal taste and 5 being the highest. The reduction of off-flavors in meat paste with lutein is supported by SPME gas chromatography-mass spectrometry (GC-MS) data. In addition, carotenoids (lycopene, β-carotene, zeaxanthin, canthaxanthin, and lutein) were added to the meat paste, resulting in the samples being described as more fat and sweeter.

[0195] By adding lutein to the meat paste, most flavor compounds were reduced depending on the concentration of lutein. See Table 9; the lutein concentrations were none, 0.0005%, and 0.005%. The main compounds reduced with carotenoids included oxidized flavor compounds such as alcohols and aldehydes, including (Z)-2-nonenal, (E,E)-2,4-nonadienal, and 1-penten-3-ol; in addition, sulfur-containing compounds were reduced, including methyl mercaptan, 2-acetylthiazole, and dimethyl sulfide; many of these compounds were also described as grainy and oxidized by trained flavor scientists using gas chromatography-olfactometry (GCO).

[0196] Table 8 Sensory simulation of meat agglomeration with reduced grain flavor by adding carotenoids

[0197]

[0198] Table 9

[0199] Flavor compounds affected by the addition of carotenoids to wheat gluten flour during cooking.

[0200] Data collected.

[0201]

[0202]

[0203] Example 19

[0204] Reducing grainy and off-flavor notes by adding antioxidants Meat dough was prepared using the ingredients shown in Table 10 as follows.

[0205] Table 10 Composition of meat paste

[0206]

[0207] EGCG (epigallocatechin gallate) was dissolved in water, and then melted coconut oil (50° C.) was mixed into the water. The broth was stirred vigorously, and then the wheat gluten powder was quickly added to the broth and mixed thoroughly with a spoon. The resulting raw meat dough was transferred to a metal mold or glass beaker for steaming as described in Example 16, and then transferred to a container on ice to cool it to 4° C.

[0208] The reduction of off-flavors in meat paste as described by trained flavor scientists by the addition of EGCG is supported by SPMEGC-MS data. The GCMS data showed that multiple flavoring compounds were no longer detectable or reduced by at least 2-fold, including compounds 2-pentyl-furan, 6-methyl-5-heptan-2-one, 1-penten-3-ol, 2-penten-1-ol, methyl-pyrazine, butyraldehyde, 5-ethyl-2(5H)-furanone, 5-ethyldihydro-2(3H)-furanone, 2-nonenal, phenylacetaldehyde, and 3,5-octadien-2-one, described as grainy and oxidized by GCO.

[0209] Example 20

[0210] Preparation of meat paste with reduced grain taste by washing wheat gluten

[0211] Meat dough was prepared using the ingredients shown in Table 11 as follows.

[0212] Table 11 Composition of meat paste

[0213]

[0214] Wheat gluten flour (ADM Low flavoring active wheat gluten) is slowly stirred into a solution containing 10× washing solution (50 mM NaCl) and then mixed thoroughly to prevent lumps from forming. The solution is placed on ice for five minutes, during which time the wheat gluten settles to the bottom. A second washing step is then performed by removing the first washing solution and stirring into 10× fresh washing solution. The second solution is discarded and a final wash with tap water is then performed. The water wash solution is discarded and the washed wheat gluten is then measured to determine the exact amount of water incorporated. Water is added or pressed out so that the weight of the wheat gluten dough is equal to the measured amount of initial wheat gluten flour and the theoretical amount of water. Molten coconut oil is added and the dough is kneaded manually for 30 seconds to incorporate the oil. The formed raw meat dough is transferred to a metal mold or glass beaker for steaming as in Example 16 and then transferred to a container on ice to cool it to 4°C.

[0215] The washing step changes the flavor of the paste in a favorable manner; when compared to unwashed meat paste, five trained flavor scientists described the washed meat paste as having less cereal, less oxidized, and less off-flavor overall in four tastings. The reduction in off-flavors in the washed meat paste is supported by SPME GC-MS coupled to GCO comparing unwashed and washed meat pastes. In the washed meat paste, flavor compounds are reduced, including oxidized flavor compounds such as alcohols and aldehydes; and specific compounds, including 1-(2-furanyl)-ethanone, methyl-pyrazine, valeric acid, 3-methyl-1-butanol, 2,3-butanedione, benzyl alcohol, (E,E)3,5-octadien-2-one, (E)-2-nonenal, (E,E)-2,4-decadienal, and 1-octen-3-one, which are determined by GCO as odor active compounds and these compounds are reduced or not detected in the washed meat paste.

[0216] Example 21

[0217] Preparation of blood agar

[0218] Prepare blood agar using the ingredients shown in Table 12.

[0219] Table 12 Composition of blood agar

[0220]

[0221] Agar powder (Now Foods, Bloomingdale, IL) was dissolved in a mixture of lactic acid and seasoning broth (made as in Example 5, except that 10% coconut oil and Magic Mix 1 of Table 13 were used) by heating to 100°C in a stirred beaker. The solution was cooled to 65°C by immersion in an ice bath. 17× liquid Magic Mix (Magic Mix 2 of Table 13) and leghemoglobin, both at 4°C, were then added, causing the temperature of the mixture to drop to 50°C. It is important that the mixture be cooled before the leghemoglobin is added to prevent denaturation of the leghemoglobin. Dry ribulose bisphosphate carboxylase was then added and the mixture was stirred vigorously by hand. It is important that the temperature be between 40°C and 60°C when the ribulose bisphosphate carboxylase is added. If the temperature is too high, the ribulose bisphosphate carboxylase may denature and will not be able to act as a coagulant during cooking of the final product. If the temperature is too low, the agar will solidify and prevent the creation of a homogenous mixture.

[0222] Table 13 Composition of magic mixture

[0223]

[0224] Example 22

[0225] Preparation of blood agarPrepare blood agar using the ingredients shown in Table 14.

[0226] Table 14 Composition of blood agar

[0227]

[0228]

[0229] Agar powder (Agar 100, TIC Gums, White Marsh, MD) was dissolved in a mixture of water and lactic acid by heating to at least 91°C in a stirred beaker. Heating was performed to completely dissolve the agar. The solution was then cooled to 50-70°C, and a premix of leghemoglobin (Pichia expressed, Example 1) and 17× Liquid Magic Mix (Table 12) was added. If the temperature is too high, the leghemoglobin may denature and will not function as intended for flavoring reaction chemistry. If the temperature is too low, the agar will solidify and prevent the creation of a homogenous mixture. The mixture was then stirred and further cooled to 4-25°C. The finished product had a ketchup-like appearance and texture.

[0230] Example 23

[0231] Preparation of fat replica emulsions with improved melting, adhesion and mouthfeel properties

[0232] To prepare one hundred (100) g of fat replica, 1 g of dry precursor mixture 1 (Table 12) was dissolved in 18.8 ml of water and the pH was adjusted to 6 with concentrated NaOH solution. A frozen solution of leghemoglobin (5.5%) was added to the precursor solution and placed on a stirring hot plate maintained at 160° C. with a rotation speed of 250 RPM.

[0233] In another container, thirty-five (35) g of coconut oil (Shea and Company, Milwaukee, Oregon) and thirty-five (35) g of palm stearin were melted together in a water bath at 60° C. The molten oil mixture was slowly (about 12 ml / min) added to the solution of precursor and leghemoglobin while increasing the stirring rate to 450 RPM.

[0234] After adding oil, the resulting thick emulsion was maintained at the same temperature and stirring rate for 23 minutes. Then, the emulsion was transferred to a 600 ml beaker and placed on ice and placed in a refrigerator for rapid cooling. When the emulsion reached 25 ° C, 0.35 g of algae plant oil and 0.35 g of acetoin were added to the emulsion and quickly mixed with a spatula. The lactones used to improve seasoning and mask odors (as described in Example 31) were added in the following amounts: 5-ethyl-4-hydroxy-2-methyl-3 (2H)-furanone was added to 2.5*10 -5% final concentration, add butyrolactone to reach 2.5*10 -8 % final concentration, and δ-tetradecalactone was added to reach 5*10 -9 The emulsion was homogenized for 2.5 minutes using a handheld homogenizer at setting 6. The emulsion was incubated at 4°C until it was completely solidified.

[0235] After solidification, the fat replica emulsion is off-white to slightly brown, waxy solid at room temperature, with a flavor characterized as savory, meaty, bloody, and chicken fat-like. When incorporated into ground beef replica, the viscosity of the replica increases, as does the ability of the replica to be manipulated and shaped.

[0236] Example 24

[0237] Preparation of fat replica emulsion stabilized by soybean conglycinin

[0238] To prepare 100 g of fat replica, 1.5 g of isolated soy conglycinin powder from Example 4 was dissolved in 28.5 ml of water and placed on a hot stirring plate. In another container, 70 g of coconut oil (Shea and Company) was melted in a 60° C. water bath. Under constant stirring, the molten oil mixture was slowly (about 12 ml / min) added to the solution of purified protein. The resulting emulsion was heated to a temperature of 90° C. and maintained at this temperature for 5 minutes. Next, the emulsion was transferred to a 600 ml beaker and placed on ice and placed in a refrigerator for rapid cooling. When the emulsion reached 25° C., 0.35 g of algae plant oil was added to the emulsion and mixed quickly with a spatula. Lactones for improving flavoring and masking off-flavors (as described in Example 31) were added in the following amounts: 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone was added to 2.5*10 -5 % final concentration, add butyrolactone to reach 2.5*10 -8 % final concentration, and δ-tetradecalactone was added to reach 5*10 -9 The emulsion was homogenized at setting 6 for 2.5 minutes using a handheld homogenizer and kept in a 4°C refrigerator until it was completely solidified. After solidification, the fat replica emulsion was white to slightly off-white, solid at room temperature, and had a light, very neutral flavor and texture characterized as similar to extracted beef tallow.

[0239] Example 25

[0240] Preparation of fat replica emulsions stabilized by soybean conglycinin via pH shift method

[0241] To prepare 100 g of fat replica, 0.5 g of isolated soybean globulin powder from Example 4 was dissolved in 29.5 ml of water in a beaker. The pH of the protein solution was adjusted to 12 using 2M sodium hydroxide solution. In another container, 70 g of coconut oil (Shea and Company) was melted in a 60°C water bath. Under constant stirring, the molten oil mixture was slowly (about 12 ml / min) added to the solution of purified protein. 0.35 g of algae plant oil was added to the emulsion and mixed quickly with a spatula. The pH of the protein solution was adjusted to 12 using 2M sodium hydroxide solution, and the emulsion was homogenized for 30 seconds (thirty seconds) using a handheld homogenizer at setting 6 and kept at 4°C until it was completely solidified. After solidification, the fat replica emulsion was white to slightly off-white, solid at room temperature, with a light, very neutral flavor and a texture similar to cottage cheese.

[0242] Example 26

[0243] Adding carotenoids to flavor emulsions to enhance meaty flavor

[0244] Each carotenoid (canthaxanthin, β-carotene, lutein, or lycopene) (Shea and Company, Milwaukee, OR) was dissolved in coconut oil or water at 10%, depending on solubility. The carotenoids were added to the flavored emulsion prior to cooking by mixing a solution of 0.5% LegH, 1× Precursor Mix 1 (Table 1), 30% RBD coconut oil, and the respective 10% carotenoid solution (final carotenoid concentration in the emulsion was 0.025%) and stirring the mixture while heating until boiling, then simmering for 10 minutes. 0.7% additional algae oil was added to form additional precursors for flavoring as described in Example 32. Lactones for improving flavor and masking off-flavors (as described in Example 31) were added in the following amounts: 5-ethyl-4-hydroxy-2-methyl-3(2H)-furanone was added to 2.5*10 -5 % final concentration, add butyrolactone to reach 2.5*10 -8 % final concentration, and δ-tetradecalactone was added to reach 5*10 -9 The emulsion was homogenized at setting 6 for 2.5 minutes using a handheld homogenizer and then incubated at 4°C until it was completely solidified. The emulsion was then added to taco meat prepared as described in Example 30. The samples with different carotenoids were then compared to a control with no carotenoid added. The samples were evaluated by at least five trained flavor scientists. The results are summarized in Table 15.

[0245] Table 15

[0246] Flavor description of burritos tasted with flavored emulsions prepared with different carotenoids

[0247]

[0248] *Slightly

[0249] § Numbers in parentheses indicate the number of tasters with the stated response.

[0250] Example 27

[0251] Assembling and cooking the burgers

[0252] Replica burgers containing the ingredients in Table 16 were prepared.

[0253] Table 16

[0254] Composition of hamburger

[0255]

[0256] The frozen meat dough was blended using a stand mixer fitted with a food grinder attachment ( Professional 600 Series 6-Quart Bowl Lift Stand Mixer Model KP26M1XER and Food grater model FGA, St. Joseph, Michigan) at speed setting 1. In this device, the material is fed by a screw conveyor through a rotating knife mounted in front of a fixed orifice plate. Soft connective tissue is minced using a universal machine (UM-12, Stephen Machinery GmbH, Schwarzenbeck, Germany) equipped with a blunt knife and running at a slow speed for 20-30 seconds. In a single step process, the seasoned emulsion is frozen to -20°C and then shredded with a mini chopper ( Chop using a mini chopper (Cuisinart, Stamford, CT). Approximately 400 g of the emulsion was placed in a mini chopper and processed for 60 seconds on the chop setting to produce pieces 1-3 mm long.

[0257] The ground meat paste, chopped soft connective tissue and seasoned emulsion pieces are then mixed. During mixing, the mixture is kept at -5 to 4°C to prevent fat melting. Blood agar is then added and mixed until it is thoroughly incorporated. The total batch size is 1kg. Next, 50g or 150g portions of ground tissue are manually formed into round patty shapes. The typical dimensions of a 50g patty are 55mm×15mm. The typical dimensions of a 150g patty are 100mm×22mm. The patties are refrigerated until cooking. The patties are cooked on a preheated (325-345°F) non-stick long-handled frying pan and heated to an internal temperature of 160°F, while turning over every 2 minutes. The appearance, texture and flavor of the cooked patties are similar to ground beef. In addition to cooking in patty form, the unformed material can also be used in a variety of dishes, such as burrito fillings, casseroles, sauces, toppings, soups, stews or meat loaves.

[0258] Example 28

[0259] Assembling and cooking the burgers

[0260] Replica burgers containing the ingredients in Table 17 were prepared.

[0261] Table 17 Composition of hamburger

[0262]

[0263] The frozen meat dough was blended using a stand mixer fitted with a food grinder attachment ( Professional 600 Series 6-Quart Bowl Lift Stand Mixer Model KP26M1XER and Food grinder model FGA, St. Joseph, Michigan) at speed setting 1. Soft connective tissue was minced using a universal grinder (UM-12, Stephan Maschinenbau, Schwarzenbeck, Germany) equipped with a blunt knife and running at slow speed for 20-30 seconds. The seasoned emulsion was frozen to -20°C and then The ground meat paste, chopped soft connective tissue, seasoned emulsion pieces and dried soy conglycinin were then mixed. During mixing, the mixture was kept at -5 to 4°C to prevent fat melting. The hydrating liquid (a 1:1 mixture of leghemoglobin and 17× liquid magic mix as described in Example 22) was then added and the mixture was kept at 4°C for a minimum of 15 minutes to hydrate the dried soy conglycinin. Finally, blood agar was added and mixed until it was thoroughly incorporated. The total batch size was 200g. Next, 50g portions of the ground tissue were manually formed into round patty shapes. The typical dimensions of a 50g patty are 55mm×15mm. The patties were refrigerated until cooking. The patties are cooked on a preheated (325-345°F) non-stick skillet and heated to an internal temperature of 170°F, flipping every minute. The cooked patties resemble ground beef in appearance, texture and flavor. In addition to cooking in patty form, the unformed material can be used in a variety of dishes such as burrito fillings, casseroles, sauces, toppings, soups, stews or meatloaf.

[0264] Example 29

[0265] Assembly and cooking of burgers with 10% meat paste

[0266] Replica burgers containing the ingredients in Table 18 were prepared.

[0267] Table 18 Composition of hamburger

[0268]

[0269] The frozen meat dough was blended using a stand mixer fitted with a food grinder attachment ( Professional 600 Series 6-Quart Bowl Lift Stand Mixer Model KP26M1XER and Food grinder model FGA, St. Joseph, Michigan) at speed setting 1. Soft connective tissue was minced using a universal grinder (UM-12, Stephan Maschinenbau, Schwarzenbeck, Germany) equipped with a blunt knife and running at slow speed for 20-30 seconds. The seasoned emulsion was frozen to -20°C and then The ground meat paste, chopped soft connective tissue, seasoned emulsion pieces and dried soy conglycinin were then mixed. During mixing, the mixture was kept at -5 to 4°C to prevent fat melting. The hydrating liquid (a 1:1 mixture of leghemoglobin and 17× liquid magic mix as described in Example 22) was then added and the mixture was kept at 4°C for a minimum of 15 minutes to hydrate the dried soy conglycinin. Blood agar was finally added and mixed until it was thoroughly incorporated. The total batch size was 200 g. Fifty (50) g ​​portions of the ground tissue were then manually formed into round patty shapes. The typical dimensions of a 50 g patty were 55 mm×15 mm. The patties were refrigerated until cooked. The patties were cooked on a preheated (325-345°F) non-stick long-handled skillet and heated to an internal temperature of 170°F, while turning over every minute. The appearance, texture and flavor of the cooked patties are similar to ground beef. In addition to cooking in patty form, the unformed material can be used in a variety of dishes such as burrito fillings, casseroles, sauces, toppings, soups, stews or meatloaf.

[0270] Example 30

[0271] Assembling and cooking the "taco meat"

[0272] Preparation of replica "burrito meat" containing the ingredients in Table 19

[0273] Table 19 Composition of hamburger

[0274]

[0275] As described in Example 28, the frozen meat paste was ground, the soft connective tissue was minced, and the seasoned emulsion was minced. The ground meat paste, minced soft connective tissue, and seasoned emulsion pieces were then mixed. During mixing, the mixture was kept at -5 to 4°C to prevent fat melting. The hydrating liquid (a 1:1 mixture of leghemoglobin and 17× liquid magic mix as described in Example 22) and blood agar were then added and mixed. The total batch size was 20 g. The mixed tissue was then cooked to 160°F on a preheated (325-345°F) non-stick long-handled skillet. The appearance, texture, and flavor of the cooked tissue were similar to ground beef. The appearance of the material was similar to taco meat; in the absence of 7S protein, the meat was not firm and stuck together a lot.

[0276] Example 31

[0277] Using lactones as masking agents

[0278] The lactones were diluted in water or oil, depending on solubility. The diluted lactones were then added to the flavored emulsion (Example 23) and homogenized as indicated in Table 20. The final concentrations of the lactones are given in Table 20. The flavored emulsions were added to the final 20% of ground meat (e.g., taco meat) (all components of a meat replica without ribulose bisphosphate carboxylase). The flavored emulsions were mixed with meat paste, connective tissue, magic mix, and heme as indicated in Example 17, but without ribulose bisphosphate carboxylase. The ground meat was then tested for overall taste, reduction of any off-flavors, and overall improvement by five trained flavor scientists. The summarized results are indicated in Table 20. Addition of specific lactones and lactone combinations resulted in a reduction in off-flavors, including cereal, egg, bitter, liver, and mushroom. Specific masking properties, such as bitterness, require unique combinations. Lactones also add desirable creamy, buttery, caramel, fatty, umami, and fruity flavors.

[0279] Table 20 Sensory data on the addition of lactones as masking agents in meat replicas

[0280]

[0281]

[0282] Example 32

[0283] Add polyunsaturated fats for meaty flavor

[0284] Algal oil (DSM life's omega 45 02412-0100) was added to the flavored emulsion (Example 23) and then homogenized to a final concentration of 0.07% in the meat replica. The flavored emulsion was added to the replica as described in Example 27. As described by a trained flavor scientist, the addition of algal oil increased the tallow, fatty, and overall meaty flavor of the replica.

[0285] The addition of algae oil increased the precursors required for the production of fat flavoring molecules, including eicosapentaenoic acid and docosahexaenoic acid. As detected by SPME gas chromatography-mass spectrometry (GC-MS), the addition of algae oil to the precursor mixture and hemoglobin produced flavors compared to the control without algae oil, including nonane, (E,E)-3,5-octadien-2-one, 1-heptene-3-ol, 1-pentene-3-one, 2-propylfuran, n-caprylic acid vinyl ester, 3-ethyl-2-methyl-1,3-hexadiene, 1-ethyl-5-methylcyclopentene, trans-2-(2-pentenyl)furan, 1-pentene-3-one. -3-ol, 4,7-dimethyl-undecane, 1-octanol, 3-ethyl-pyridine, 3-ethylcyclopentanone, (Z)-2-octen-1-ol, 2-n-heptylfuran, (Z)-2-decenal, hexanoic acid, (E,E)-2,4-nonadienal, 6-methyl-2-heptanone, (Z)-2-heptenal, (E,E)-2,4-heptadienal, 1-hexanol, (E,E)-2,4, decadienal, (E,Z)-2,6-nonadienal and 1-octen-3-ol.

[0286] Example 33

[0287] Generating Cucumis sativus slurry for meat replicas

[0288] In order to produce the boiled cucumber slurry, use the intact, non-permeable, crisp variety cucumbers (the skin is not peeled or otherwise damaged). The water bath is heated to 80-90°C, the intact cucumbers are placed in the water bath and cooked until the internal temperature of the cucumbers is balanced with the temperature of the water bath, about 30 minutes for this example. Then, the cucumbers are taken out of the bath and the skin of the cucumbers is completely removed from the pulp. The pulp is blended with seeds and then sieved to separate any larger particles. The blended pulp is then used as a slurry and added to the meat replica. This same method is used for other varieties of the genus Cucumis, including honeydew melon and cantaloupe.

[0289] Example 34

[0290] Production of Cucumis sativus extracts using solvent-assisted flavor extraction

[0291] The extract is produced using solvent-assisted flavor extraction (SAFE) and water as the solvent. SAFE works by pulling the flavoring compounds out of the material under pressure and slightly elevated temperatures.

[0292] By removing all the epidermis of crisp variety cucumbers and cutting the cucumbers into pieces, then blending with magic bullet (magicbullet) to produce extract. Then, the cucumber slurry is poured into the sample inlet of the SAFE glassware under the pressure of Edwards 12 floor vacuum pump (Edwards12floor vacuum), and the temperature is set at 40 ℃ with a water pump and a warm water bath for the sample round-bottom flask. A small amount of (2-4mL sample) cucumber slurry is placed in the sample round-bottom flask. The slurry immediately presents boiling when it moves to the sample round-bottom flask. When visual boiling stops, more sample volume is added to the sample round-bottom flask. This continues until all samples are entered into the SAFE glassware device. Once all samples are exhausted, the sample keeps extracting when the water bath reaches 40 ℃, and then extracts for 20 minutes. Along with the extraction, the round-bottom flask is collected and immersed in liquid nitrogen and the finger-shaped cold inlet is filled with liquid nitrogen. Then the extract is collected from the collection round-bottom flask.

[0293] Example 35

[0294] Cucumis liquids are added to meat replicas to increase meaty and fatty notes by adding to a gelling matrix

[0295] Blood agar as outlined in Example 22 was made as described, except that Cucumis liquid was added in place of deionized water. Cucumis liquid was one of the following: (i) a commercial water extract of cantaloupe with 2% deionized water added, (ii)-(v) a slurry of honeydew (6.25%) or cucumber (3.2%), cooked or uncooked, as described in Example 33, or (vi) a solvent-assisted flavor extract (pressurized distillate) of cucumber as described in Example 34. The addition of these cucumber and melon extracts brought some elements of the meaty flavor profile to enhance the overall liking and meaty flavor of the replica. Many of the compounds were aldehydes, lactones, many of which are found in beef, as shown by SPME GC-MS and confirmed by a trained flavor scientist using GCO as odor active compounds. Compounds that are similar between beef and Cucumis include, but are not limited to, nonanal, 2-decenal, 2-nonenal, 2-heptenal, 2,6-nonadienal, 2,4-decadienal, 2-undecenal, 2-octenal, 2-nonenal, dodecanal, 2,4-heptadienal, 2,6-nonadienal, 2,4-nonadienal, 2,4-octadienal, decanal, 5-(methylenecyclopropyl)-pentanal, 6-nonadienal, 3,7-Dimethyl-1,6-octadien-3-ol, 2-nonen-1-ol, 3-nonen-1-ol, 3,5-octadien-2-one, 2,3-butanedione, 2-methyl-cyclopentanone, 2-butanone, á-ionone, 6-octen-2-one, dihydro-5-pentyl-2(3H)-furanone, L-menthone, vinyl caprylate, 4-methyloctanoic acid and vinyl acetate.

[0296] When blood agar with extracts was added to meat replicas as described in Example 30, sweet aroma, fatty, and in some cases, tallow and beef flavors increased, see Table 21 for a full description of the additional flavors not found in controls and blind controls by five trained flavor scientists. Additionally, the addition of these melon and cucumber extracts was observed to also reduce the perception of off-flavors, including grainy, earthy, woody, and astringent.

[0297] Three of these extracts were tested in a pro forma panel with 8 trained panelists and compared to a control replica indicated as is, a blinded control 80:20 beef not indicated as a control, and three additional samples with Cucumis sativus liquids (cooked cucumber slurry tested at 0.37% final taco meat, cooked honeydew melon slurry tested at 0.73% final taco meat, and cantaloupe extract (from TREATT) at 0.24% final taco meat). The results showed that all three samples were rated as higher in fat and sweet flavors, while earthy, cereal, astringent, and clear off-flavors were reduced. The cantaloupe extract had an additional off-flavor that tasted different from beef than the sweet flavored control. The other two samples, cooked honeydew melon slurry and cucumber slurry made as described in Example 33, were both rated as higher in meat and fat than the two controls, and did not have the additional off-flavors.

[0298] Table 21 Flavor Descriptions of Addition of Cucumis Sativus Liquids to Meat Replicas

[0299]

[0300]

[0301] Other embodiments

[0302] It should be understood that although the invention has been described in conjunction with its detailed description, the foregoing description is intended to illustrate rather than limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the appended claims.

Claims

1. A meat replica composition comprising: (a) from about 5% to about 88% by weight of a meat paste; (b) from about 0% to about 40% by weight of a carbohydrate gel; (c) from about 5% to about 35% by weight of non-animal fat; (d) from about 0.00001% to about 10% by weight of a flavoring agent; (e) from about 0% to about 15% by weight of a binder; and (f) from about 0.01% to about 4% by weight of a heme-containing protein and / or an iron salt.

2. The composition according to claim 1, wherein the meat dough comprises a flavoring agent.

3. A composition according to claim 1 or 2, wherein the non-animal fat comprises a flavouring agent.

4. The composition of any one of claims 1 to 3, wherein the meat dough is about 45% to about 60% by weight of the composition.

5. The composition of any one of claims 1 to 4, wherein the carbohydrate gel is about 10% to about 25% by weight of the composition.

6. The composition of any one of claims 1 to 5, wherein the non-animal fat is from about 10% to about 15% by weight of the composition.

7. The composition of any one of claims 1 to 6, wherein the flavoring agent is from about 3% to about 7% by weight of the composition.

8. A composition according to any one of claims 1 to 7, wherein the flavouring agent comprises one or more flavour precursors.

9. The composition of any one of claims 1 to 7, wherein the flavoring agent is from about 0.001% to about 2% by weight of the composition.

10. The composition of claim 9, wherein the flavoring agent comprises a flavoring or a flavoring compound.

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