WO2023228574A1

WO2023228574A1 - Plant-based meat mimetics

Info

Publication number: WO2023228574A1
Application number: PCT/JP2023/013440
Authority: WO
Inventors: Sarah G. CORWIN; Joseph A. Formanek
Original assignee: Ajinomoto Co., Inc.
Priority date: 2022-05-27
Filing date: 2023-03-30
Publication date: 2023-11-30
Also published as: WO2023228575A1

Abstract

A plant protein-based food made of 15 to 85 wt% of at least one texturized vegetable protein, 0.35 to 45 wt% of a plant protein ingredient, and 0.0005 to 0.05 wt% transglutaminase, is described. The texturized vegetable protein and the plant protein ingredient may be derived from Bambara groundnut, black-eyed pea, dry bean, chickpea, fava bean, lentil, lupin, pea, pigeon pea, soybean, and/or wheat, and the plant protein-based food may be a lobster or other crustacean, fish, lamb, beef, chicken, or sausage mimetic.

Description

PLANT-BASED MEAT MIMETICS

The present invention relates to plant protein-based foods and methods of producing.

The “background” description provided herein is for the purpose of generally presenting the context of the disclosure. Work of the presently named inventors, to the extent it is described in this background section, as well as aspects of the description which may not otherwise qualify as prior art at the time of filing, are neither expressly nor impliedly admitted as prior art against the present invention.

There has been an increase in the demand for high protein foods made from plant proteins, especially those that function as meat substitutes. These foods are formulated to provide high protein contents and similar eating experiences as animal meat. However, such foods have had low consumer satisfaction rates, and developing a meat substitute that delivers acceptable texture and flavoring has been a challenge. Additionally, there has been a demand for meat substitutes beyond vegetarian mainstays such as veggie burgers and veggie dogs comprised primarily of fibrous vegetables.

A key ingredient for providing a meat-like texture in a meat substitute is a binding agent. Conventionally, egg white, methylcellulose, gluten, and other ingredients have been used as binders in plant protein-containing foods to give them meat-like texture (Patent Literatures 1 and 2).

However, these days, typical binding agents in meat substitutes have come under scrutiny due to the increasing sophistication of consumer tastes. A growing interest in vegan products and the avoidance of certain ingredients such as methylcellulose and gluten have increased the demand for plant protein-containing foods using different binders for the retention of texture and moisture. Further, these typical binders do not contribute significantly to a positive nutritional profile and/or may also be allergens.

Alternatively, small amounts of a protein sourced from plants, reacted with an active enzyme, may be used to hold ingredients together without having to rely on ingredients viewed as unfavorable binding agents. For example, transglutaminase is an enzyme that catalyzes the cross-linking of proteins and has been used as an enzyme for the adhesion of food materials. More recently, a method for producing plant protein-containing foods with excellent texture by adding transglutaminase and glucose oxidase to a soy-based hamburger has been disclosed (Patent Literature 3). Also, a method for producing a modified plant protein powder is known that includes adding a calcium compound and/or a magnesium compound, and transglutaminase to a plant protein-containing aqueous solution, and heating and drying the mixture (Patent Literature 4). However, higher qualities are required for high protein plant-based foods in recent years due to the increasing consumer preferences, and the development of a method for achieving both satisfactory cohesion and moisture retention while fulfilling the above-mentioned preferences is desired.

JPA2013-009617 US20090274814A1 US20190037883A1 JP2819797B2

One objective of the present invention is to provide a plant-based food that can be made from a variety of plant-based proteins. Another objective of the present invention is to provide plant protein-based foods with specific flavoring blends and textures and that can be used as a mimetic of lobster or other crustacean or shellfish, fish, lamb, beef, pork, or a variety of sausages (patties or links). The plant protein-based food is made from at least one plant protein ingredient, a texturized vegetable protein, and transglutaminase.

The present invention provides:
(1) A plant protein-based food, comprising: 15 to 85 wt% texturized vegetable protein; 0.35 to 45 wt% plant protein ingredient; and 0.0005 to 0.05 wt% transglutaminase, each weight percent relative to a total weight of the texturized vegetable protein, the plant protein ingredient, and the transglutaminase,
wherein the plant protein ingredient is at least one selected from the group consisting of a flour, a protein concentrate, or a protein isolate.
(2) The plant protein-based food of (1), which is substantially free of one or more of methylcellulose, carrageenan, casein, egg white, glucose oxidase, phospholipase, cereal grain flour, gluten, cellulose, gelatin, and collagen.
(3) The plant protein-based food of (1) or (2), which is substantially free of animal-derived ingredients.
(4) The plant protein-based food of any one of (1) to (3), wherein the plant protein ingredient is derived from at least one selected from the group consisting of Bambara groundnut, black-eyed pea, dry bean, chickpea, fava bean, lentil, lupin, pea, pigeon pea, soybean, and wheat.
(5) The plant protein-based food of any one of (1) to (4), wherein the dry bean is at least one selected from the group consisting of mung bean, navy bean, and pinto bean.
(6) The plant protein-based food of any one of (1) to (5), wherein the plant protein ingredient is derived from at least one selected from the group consisting of Bambara groundnut, black-eyed pea, chickpea, fava bean, mung bean, pea, pinto bean, lupin, pigeon pea, and red lentil.
(7) The plant protein-based food of any one of (1) to (6), wherein the plant protein ingredient comprises 19 to 100 wt% protein relative to a total weight of the protein ingredient.
(8) The plant protein-based food of any one of (1) to (7), wherein the plant protein ingredient comprises 80 to 100 wt% protein relative to a total weight of the protein ingredient.
(9) The plant protein-based food of any one of (1) to (8), wherein the texturized vegetable protein is made from at least one selected from the group consisting of wheat, soy, pea, dry bean, broad bean, chickpea, cowpea, pigeon pea, lentil, Bambara groundnut, and lupin.
(10) The plant protein-based food of any one of (1) to (9), wherein the texturized vegetable protein is made from pea.
(11) The plant protein-based food of any one of (1) to (10), further comprising 30 to 75 wt% water relative to a total weight of the plant protein-based food.
(12) The plant protein-based food of any one of (1) to (11), further comprising 2 to 35 wt% of one or more oils and/or fats relative to a total weight of the plant protein-based food.
(13) The plant protein-based food of (12), wherein the one or more oils and/or fats is selected from the group consisting of sunflower seed oil, canola oil, corn oil, vegetable oil, coconut oil, palm oil, and algal oil.
(14) The plant protein-based food of any one of (1) to (13), which is substantially free of soy.
(15) The plant protein-based food of any one of (1) to (14), which is a chorizo, pork, beef, chicken, sausage patty, sausage link, lamb, lobster, shrimp, crab, crustacean, calamari, or fish mimetic.
(16) The plant protein-based food of any one of (1) to (15), wherein the plant protein-based food comprises:
15 to 85 wt% of at least one texturized vegetable protein;
0.35 to 45 wt% of one or more plant protein ingredients;
0.0005 to 0.01 wt% transglutaminase;
0.3 to 1.5 wt% table salt;
0.5 to 12 wt% of one or more seasonings and/or flavorings other than table salt;
30 to 75 wt% water; and
2 to 35 wt% of one or more oils and/or fats, each weight percent relative to a total weight of the plant protein-based food.
(17) A method of making the plant protein-based food of (16), the method comprising: mixing at least one texturized vegetable protein, one or more plant protein ingredients, transglutaminase, table salt, and one or more seasonings and/or flavorings other than table salt to form a first mixture; mixing water with the first mixture to form a second mixture; mixing oil and/or fat with the second mixture to form a third mixture; and maintaining the third mixture at a temperature in a range of 1 to 65 °C for a time in a range of 30 s to 72 h to produce the plant protein-based food.
(18) The method of (17), further comprising, before the maintaining, subjecting the third mixture to a vacuum to remove air bubbles.
(19) The method of (17) or (18), further comprising cooling the plant protein-based food to a temperature in a range of -80 to 6 °C.
(20) A method of making a plant protein-based food of any one of (1) to (16), the method comprising: mixing the texturized vegetable protein, plant protein ingredient, transglutaminase, and water to produce a first mixture; and maintaining the first mixture at a temperature in a range of 1 to 15 °C for a time in a range of 15 min to 72 h.
(21) The method of (20), further comprising shaping the first mixture between the mixing and the maintaining.
(22) The method of (20) or (21), further comprising, after the maintaining, cooking the first mixture at a temperature in a range of 100 to 250 °C for a time in a range of 1 to 20 minutes.

The foregoing paragraphs have been provided by way of general introduction and are not intended to limit the scope of the following claims. The described embodiments, together with further advantages, will be best understood by reference to the following detailed description.

Embodiments of the present disclosure will now be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments are shown.

The present disclosure will be better understood with reference to the following definitions. As used herein, the words “a” and “an” and the like carry the meaning of “one or more,” or “at least one.” Within the description of this disclosure, where a numerical limit or range is stated, the endpoints are included unless stated otherwise. It will be further understood that the terms “comprises” and/or “comprising,” when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.

As used herein, the words “about,” “approximately,” or “substantially similar” may be used when describing magnitude and/or position to indicate that the value and/or position described is within a reasonable expected range of values and/or positions. For example, a numeric value may have a value that is ± 0.1% of the stated value (or range of values), ± 1% of the stated value (or range of values), ± 2% of the stated value (or range of values), ± 5% of the stated value (or range of values), ± 10% of the stated value (or range of values), ± 15% of the stated value (or range of values), or ± 20% of the stated value (or range of values). Within the description of this disclosure, where a numerical limit or range is stated, the endpoints are included unless stated otherwise. Also, all values and subranges within a numerical limit or range are specifically included as if explicitly written out.

As used here, a “percent difference” between two values refers to the absolute difference between the two values, divided by the average of the two values, all multiplied by 100.

As used herein, the phrase “substantially free,” unless otherwise specified, refers to a composition/material which contains less than 1 wt%, preferably less than 0.5 wt%, preferably less than 0.1 wt%, preferably less than 0.05 wt%, preferably less than 0.001 wt%, preferably less than 0.0005 wt%, preferably 0 wt% of a particular component, relative to a total weight of the composition/material.

According to a first aspect, the present invention relates to a plant protein-based food, comprising, a plant protein ingredient, a texturized vegetable protein, and transglutaminase. The plant protein-based food may comprise 0.35 to 75 wt%, 1 to 70 w%, 5 to 65 wt%, 10 to 60 wt%, 15 to 50 wt%, 17 to 45 wt%, 20 to 40 wt%, 22 to 38 wt%, 25 to 35 wt%, or about 21 wt%, or about 25 wt%, or about 32 wt% plant protein ingredient; 15 to 99 wt%, 20 to 95 wt%, 25 to 92 wt%, 30 to 90 wt%, 35 to 85 wt%, 40 to 84 wt%, 45 to 83 wt%, 50 to 82 wt%, 55 to 81 wt%, 60 to 80 wt%, or 65 to 80 wt%, or about 74 wt%, or about 79 wt%, or about 68 wt% texturized vegetable protein; and 0.000015 to 0.30 wt%, 0.0001 to 0.01 wt%, 0.0005 to 0.05 wt%, 0.0008 to 0.02 wt%, 0.001 to 0.015 wt%, 0.001 to 0.01 wt%, 0.002 to 0.008 wt%, or about 0.002 wt%, or about 0.006 wt%, or about 0.007 wt%, or about 0.01 wt% transglutaminase, where each weight percentage is relative to a combined weight of the plant protein ingredient, the texturized vegetable protein, and the transglutaminase.

Plant protein ingredient

The plant protein ingredient is in the form of a powder and may be a flour (such as a bean flour or a grain flour, or a starch), a protein concentrate, or a protein isolate. Any of the plant protein ingredients and texturized vegetable proteins used in the present invention may be independently derived from at least one protein source selected from the group consisting of corn, rice, almond, peanut, pumpkin seed, canola, barley, pistachio, walnut, pine nut, chestnut, Brazil nut, sesame, amaranth, pecan nut, rapeseed, buckwheat, potato, sweet potato, asparagus, broccoli, avocado, flaxseed, chia seed, oat, wheat, teff, buckwheat, millet, hemp seed, carob, sunflower seed, quinoa, soy, pea, fava bean, dry bean, fava bean, cowpea, pigeon pea, lentil, Bambara groundnut, lupin, or any pulse or legume designated by the Food and Agriculture Organization of the United Nations (FAO). Such FAO pulses and legumes include but are not limited to dry beans (FAOSTAT code 0176, Phaseolus spp. including several species now in Vigna); fava bean (FAOSTAT code 0181, Vicia faba, also known as faba bean, fava, or broad bean), pea (FAOSTAT code 0187, Pisum spp.); chickpea (FAOSTAT code 0191, Cicer arietinum, also known as garbanzo or Bengal gram), cowpea (FAOSTAT code 0195, Vigna unguiculata, also known as black-eyed pea or blackeye bean); pigeon pea (FAOSTAT code 0197, Cajanus cajan, also known as Arhar dal, Toor dal, cajan pea, Congo bean, or gandules), lentil (FAOSTAT code 0201, Lens culinaris), Bambara groundnut (FAOSTAT code 0203, Vigna subterranean, also known as earth pea and African yellow bean, Bambara bean, or Bambara pea), and lupin (FAOSTAT code 0210, Lupinus spp.). Lentils include but are not limited to brown lentils, green lentils, black beluga lentils, puy lentils, water lentils, red lentils, and yellow lentils. Dry beans include but are not limited to adzuki beans, Anasazi beans, appaloosa beans, black turtle beans, black gram, calypso beans, cranberry beans, dragon tongue beans, green bean, flageolet beans, kidney beans, lima beans, moth beans, mung beans, navy beans, pea beans, peruano beans, pink beans, pinto beans, rattlesnake beans, sulphur beans, tongue of fire beans, wax beans, white beans, yellow beans, yellow eye beans, and varieties thereof. Any of the above may be sprouted or germinated before processing into a flour, starch, protein concentrate, protein isolate, etc. Though not plants, microbial protein sources such as yeasts, fungi, and bacteria may be used as a protein ingredient. The plant protein ingredients listed above may be derived from the same protein source as the texturized vegetable protein or from a different protein source.

In one embodiment, the plant protein ingredients used in the plant protein-based food of the present invention are each independently derived from at least one protein source selected from the group consisting of soy, pea, chickpea, pinto, fava bean, pigeon pea, black-eyed pea, mung bean, lentil, Bambara groundnut, and lupin.

In one embodiment, the plant protein ingredient comprises at least 19 wt%, at least 20 wt%, at least 25 wt%, at least 35 wt%, at least 40 wt%, at least 45 wt%, at least 50 wt%, at least 55 wt%, at least 60 wt%, at least 65 wt%, at least 70 wt%, at least 75 wt% protein relative to the weight of the plant protein ingredient.

In one embodiment, the plant protein ingredient is considered a plant protein isolate and may have a protein concentration of at least 80 wt%, at least 85 wt%, at least 90 wt%, at least 95 wt%, or 100 wt% relative to the weight of the plant protein concentrate. In one embodiment the plant protein ingredient may comprise a blend of two or more vegetable protein sources or may be derived from two or more vegetable protein sources.

In one embodiment, soy protein may be present in the plant protein-based food as the plant protein ingredient and/or the texturized vegetable protein. The soy protein may be present as a soy-protein containing composition. This soy protein-containing composition is not particularly limited as long as it contains soybean-derived protein. It may be purified soybean protein itself, or may contain other soybean-derived components. For example, proteins obtained by processing soybeans, such as soy flour, soymilk powder, concentrated soybean protein, isolated soybean protein, fibrous soybean protein, and extracted soybean protein, can be used. Specific examples include, but are not limited to, New FUJIPRO 4500, FUJIPRO FR (both from Fuji Oil Co., Ltd.), in addition to those listed in Table 1.

In one embodiment, wheat protein may be present in the plant protein-based food as the plant protein ingredient and/or the texturized vegetable protein. The wheat protein may be present as a wheat protein-containing composition. The wheat protein-containing composition is not particularly limited as long as it contains a wheat-derived protein. It may be purified wheat protein itself, or may contain other wheat-derived components. For example, proteins obtained by processing wheat, such as whole wheat flour, semolina flour, cake flour, high-gluten bread flour, white flour, wheat starch, wheat bran, vital wheat gluten, concentrated wheat protein, isolated wheat protein, fibrous wheat protein, and extracted wheat protein, can be used. Specific examples include, but are not limited to, various wheat flours (of various companies), wheat gluten SWP-5A (Amylum), wheat gluten A-Glu CV (Glico Nutrition Co., Ltd.), and those listed in Table 1.

In one embodiment, pea protein may be present in the plant protein-based food as the plant protein ingredient and/or the texturized vegetable protein. The pea protein may be contained in the food as a pea protein-containing composition. The pea protein-containing composition is not particularly limited as long as it contains a pea-derived protein. It may be purified pea protein itself, or may contain other pea-derived components. For example, proteins obtained by processing pea, such as pea flour, pea protein concentrates, pea protein isolates, and pea protein hydrolysates, may be used. Specific examples include, but are not limited to, MGP Proterra 2100 granule (MGP Ingredients), NUTRALYS S85F (Roquette), and those listed in Table 1.

In one embodiment, chickpea protein may be present in the plant protein-based food as the plant protein ingredient and/or the texturized vegetable protein. The chickpea protein may be contained in the food as a chickpea protein-containing composition. The chickpea protein-containing composition is not particularly limited as long as it contains a chick pea derived protein. It may be purified chickpea protein itself, or may contain other chickpea-derived components. For example, proteins obtained by processing chickpea, such as chickpea flour, chickpea protein concentrates and chickpea protein isolates, can be used. Specific examples include, but are not limited to, ORPROTEIN CP-AC (Organo Food Tech Corporation) and those listed in Table 1.

In other related embodiments other plant proteins may be present in the plant protein-based food as the plant protein ingredient and/or the texturized vegetable protein. The plant protein may be contained in the food as a plant protein-containing composition. The plant protein-containing composition is not particularly limited as long as it contains the plant protein. It may be a starch or flour, a purified protein, a protein concentrate, a protein isolate, or some other plant-protein derived component, including but not limited to those listed in Table 1.

Texturized vegetable protein

Texturized vegetable protein products such as those described above are known in the art and are typically prepared by heating a mixture of protein material along with water under mechanical pressure in a cooker extruder and extruding the mixture through a die. Upon extrusion, the extrudate generally expands to form a fibrous cellular structure as it enters a medium of reduced pressure (usually atmospheric). The TVP may be in the form of granules, strips, slices, flakes, chunks, crumbles, and short noodles. Preferably the TVP is in the form of granules or crumbles. In one embodiment, the TVP granules or crumbles have an average longest dimension (e.g., length or diameter) in a range of 0.2 to 2 cm, 0.4 to 1.5 cm, 0.5 to 1.2 cm, or 0.5 to 1.0 cm.

Transglutaminase

Transglutaminase (TG) is an enzyme having an activity for catalyzing an acyl group transfer reaction using a glutamine residue in a protein as a donor, and a lysine residue in the same or different protein as a receptor. Transglutaminase may be derived from various organisms such as mammals, fishes, and microorganisms, and any of these may be used. Preferably the transglutaminase is derived from a microorganism. Specific examples of the transglutaminase include the transglutaminase preparation derived from a microorganism and marketed with the brand name of “Activa(registered)” from Ajinomoto Co., Inc. In one embodiment, the transglutaminase may be obtained diluted in a mixture with a starch or starch derivative to protect the activity of the transglutaminase and provide ease of handling. The starch or starch derivative includes but is not limited to maltodextrin, dextrin, glucose, sucrose, fructose, beta-glucan, cellulose, and/or corn starch. Preferably the starch or starch derivative does not influence the taste, texture, or other properties of the final product. The transglutaminase may be present in the mixture with the starch or starch derivative at a weight percentage in a range of 0.1 to 5 wt%, 0.5 to 2 wt%, or about 1 wt% relative to a total weight. The transglutaminase may be obtained as a preparation such as ACTIVA TI, ACTIVA YG, ACTIVA GS, or ACTIVA RM, each obtained from Ajinomoto. Preferably the transglutaminase is obtained as ACTIVA TI or ACTIVA STG-TI, which consists of 1 wt% transglutaminase and 99 wt% maltodextrin, and contains no animal-derived ingredients.

In one embodiment, an amount of transglutaminase may be determined by its enzymatic activity, in terms of units (U). The following method can be exemplified as the method for measuring the enzymatic activity of transglutaminase. Here, transglutaminase is allowed to act on benzyloxycarbonyl-L-glutaminylglycine and hydroxylamine as substrates in a Tris buffer solution with a pH 6.0 at 37 °C to generate hydroxamic acid. Hydroxamic acid is made into an iron complex in the presence of trichloroacetic acid, then absorbance is measured at 525 nm, the amount of hydroxamic acid is obtained by using a calibration curve, and the enzymatic activity is calculated. The amount of the transglutaminase enzyme that produces 1 μmol of hydroxamic acid in 1 minute at 37 °C and a pH of 6.0 is defined as 1 U (1 unit).

In one embodiment, the transglutaminase is present in the plant protein-based food at a concentration in terms of U per mass preparation that is at least 1.0 × 10^-11 U/g, at least 1.0 × 10^-10 U/g, at least 1.0 × 10^-9 U/g, at least 1.0 × 10^-8 U/g, at least 1.0 × 10^-7 U/g, at least 1.0 × 10^-6 U/g, at least 1.0 × 10^-5 U/g, at least 1.0 × 10^-4 U/g, at least 1.0 × 10^-3 U/g, at least 1.0 × 10^-2 U/g, at least 5.0 × 10^-2 U/g, at least 0.1 U/g, at least 1 U/g, at least 10 U/g, at least 50 U/g, at least 100 U/g, at least 300 U/g, at least 500 U/g, at least 1,000 U/g, and/or at most 1,250 U/g, at most 1,000 U/g, at most 500 U/g, at most 300 U/g, at most 100 U/g, at most 50 U/g, at most 10 U/g, at most 1 U/g, at most 0.1 U/g, at most 5.0 × 10^-2 U/g, at most 1.0 × 10^-2 U/g, at most 1.0 × 10^-3 U/g, at most 1.0 × 10^-4 U/g, at most 1.0 × 10^-5 U/g, at most 1.0 × 10^-6 U/g, at most 1.0 × 10^-7 U/g, at most 1.0 × 10^-8 U/g, at most 1.0 × 10^-9 U/g.

Transglutaminase reactivity

In one embodiment, a plant protein ingredient may be determined to have high or low reactivity towards transglutaminase by determining the gel strength of a hydrogel reaction product formed by reacting the plant protein ingredient with the transglutaminase.

For instance, a hydrogel reaction product may be formed by mixing a composition comprising 0.001 to 0.1 wt%, preferably 0.005 to 0.05 wt%, or about 0.01 wt% transglutaminase, 0.90 to 0.999 wt%, preferably about 0.99 wt% starch or starch derivative, plant protein ingredient to supply 15 to 25 wt%, preferably 17 to 20 wt%, or about 18 wt% protein, and water for the remaining weight percentage, each weight percentage relative to a total weight of the first composition. Preferably, the transglutaminase and the starch or starch derivative are obtained mixed together, for instance, as ACTIVA TI being 1 wt% transglutaminase and 99 wt% maltodextrin.

Then composition is allowed to react for 15 min to 2 h, 0.5 to 1.5 h, or about 1 h at a temperature in a range of 35 to 60 °C, preferably 45 to 55 °C, or about 50 °C to form the hydrogel reaction product. The compositions may be subjected to one or more deaeration steps, for instance, in a vacuum bag and/or at a pressure in a range of 10 to 380 mm Hg, 12 to 200 mm Hg, 15 to 100 mm Hg, 20 to 75 mm Hg, or 25 to 40 mm Hg.

A protein is considered to have high reactivity with transglutaminase if it produces a hydrogel having a certain amount of gel strength to resist deformation. A low reactive protein produces a hydrogel with a lower gel strength and a lower ability to resist deformation. In one embodiment, the maximum force withstood by a hydrogel reaction product of the transglutaminase and a highly reactive plant protein ingredient is 3 to 10 times, or preferably 3.5 to 9, 4 to 8, 4 to 6, 4 to 5, or 4.3 to 5 times greater than the respective maximum force withstood by a hydrogel reaction product of the transglutaminase and the low reactive plant-based protein. In a related embodiment, this maximum force may be understood as a gel strength. In one embodiment, the gel strength may be measured with a ball probe, for instance, a 20 mm diameter ball probe, or a probe, weight, or press having other dimensions or other geometries. The reactivity of a protein may also be compared by observing gel strength with and without transglutaminase being used in the hydrogel.

In alternative embodiments, a strength, viscosity, elasticity, or some other measure of structural integrity or degree of crosslinking may be measured of a gel, solid, or slurry produced from a reaction product between transglutaminase and a plant protein ingredient in order to compare reactivities and to evaluate and/or distinguish a highly reactive plant-based protein and a low reactive plant-based protein. In an alternative embodiment, the reaction product may be characterized by spectroscopy to determine reactivity.

Without being bound to theory, it is believed that within the same plant species, a highly reactive plant protein ingredient has a tertiary protein structure that is less disturbed and closer to its native tertiary structure than a low reactive plant protein ingredient. This difference may arise from the highly reactive plant protein ingredient being produced with fewer processing steps and/or less extreme processing conditions than low reactive plant protein ingredient. For instance, the highly reactive plant protein ingredient may be produced with less intense heating conditions, such as no heating or heating at a maximum temperature at least 10, 20, 30, 40, or 50 °C lower than the maximum temperature used in the production of the low reactive plant protein ingredient. In other embodiments, less intense processing conditions may arise with using solutions and/or solvents with different concentrations or pH levels. Alternatively, different plant varieties within the same species may produce protein ingredients with different reactivities. Differences in reactivities across different plant species may additionally arise from differences in the biochemistry and compositions of the plant protein ingredients.

In one embodiment, comparing the gel strength or other physical aspects of hydrogel reaction products obtained across a variety of plant protein ingredients, both across and within plant species, offers a way to screen transglutaminase reactivities. More highly reactive protein ingredients may be selected for use with transglutaminase in a plant-based protein food, or a blend of highly reactive and low reactive plant protein ingredients may be used to balance different textural qualities.

Other ingredients

In one embodiment, the plant protein-based food further comprises at least one other ingredient selected from consisting of water, an oil, a fat, table salt (sodium chloride), and a seasoning or flavoring other than table salt. In one embodiment the plant protein-based food further comprises water at a concentration of 30 to 75 wt%, 35 to 70 wt%, 37 to 67 wt%, 40 to 65 wt%, 42 to 62 wt%, 45 to 60 wt%, 47 to 58 wt%, 50 to 57 wt%, 52 to 55 wt%, 53 to 54 wt%, or about 48 wt%, or about 54 wt%, or about 57 wt%, or about 58 wt% relative to a total weight of the plant protein-based food.

In one embodiment, the plant protein-based food further comprises 2 to 35 wt%, 4 to 32 wt%, 5 to 30 wt%, 6 to 28 wt%, 7 to 25 wt%, 8 to 24 wt%, 9 to 23 wt%, 10 to 22 wt%, 11 to 21 wt%, 12 to 20 wt%, 13 to 19 wt%, 14 to 18 wt%, 15 to 17 w%, 16 to 21 wt%, or about 10 wt%, or about 11 wt%, or about 20 wt% oil and/or fat relative to a total weight of the plant protein-based food. In one embodiment, the one or more oils and/or fats is selected from the group consisting of sunflower seed oil, canola oil, corn oil, soybean oil, rapeseed oil, olive oil, perilla oil, hempseed oil, sesame oil, groundnut oil, linseed oil, rice bran oil, peanut oil, grape seed oil, safflower oil, cottonseed oil, sunflower oil, tiger nut oil, vegetable oil, coconut oil, palm oil, avocado oil, cocoa butter, shea butter, algal oil, or some other edible oil or fat. Preferably the one or more oils and/or fats is selected from the group consisting of sunflower oil, coconut oil, canola oil, algal oil, and soybean oil. The oil or fat may be added in an encapsulated form. In one embodiment, the oil and/or fat is treated with a hydrogenation process. The algal oil may be present at a weight percentage in a range of 5 to 40 wt%, 8 to 35 wt%, 9 to 30 wt%, 10 to 25 wt%, 10 to 20 wt%, 11 to 18 wt%, 12 to 15 wt%, or about 13 wt% relative to a total weight of all oils and/or fats in the plant protein-based food. In one embodiment two or more oils may be used, for instance, algal oil may be used with sunflower oil, or coconut oil may be used with sunflower oil. In one embodiment, coconut oil and sunflower oil are used together, where the coconut oil is present at a weight percentage in a range of 45 to 80 wt%, 50 to 60 wt%, or 65 to 75 wt% relative to a combined mass of the coconut oil and sunflower oil.

In one embodiment, the plant protein-based food further comprises table salt at a concentration of 0.1 to 1.5 wt%, 0.2 to 1.4 wt%, 0.3 to 1.3 wt%, 0.4 to 1.2 wt%, 0.5 to 1.1 wt%, 0.6 to 1 wt%, 0.7 to 0.9%, 0.8 to 1 wt%, or about 0.5 wt%, about 0.9 wt%, or about 1.3 wt%, relative to a total weight of the plant protein-based food. In one embodiment the table salt may be in the form of sea salt or a rock salt, such as Himalayan salt.

In one embodiment, the plant protein-based food further comprises one or more seasonings and/or flavorings other than table salt at a combined or individual weight percentage in a range of 0.1 to 12 wt%, 0.2 to 11 wt%, 0.5 to 10 wt%, 0.8 to 9 wt%, 1 to 8 wt%, 1.2 to 7 wt%, 1.5 to 6 wt%, 1.8 to 5 wt%, 2 to 4 wt%, 2.2 to 3.8 wt%, 2.5 to 3.5 wt%, 2.7 to 3.2 wt%, 2 to 3 wt%, or about 2.1 wt%, or about 2.6 wt%, or about 3.6 wt%, or about 5.1 wt%, relative to a total weight of the plant protein-based food. In one embodiment, the plant protein-based food comprises a seasoning or flavoring other than sea salt at an individual weight percentage in a range of 0.01 to 0.1 wt%, 0.05 to 0.09 wt%, 0.06 to 0.08 wt%, or 0.07 to 0.09 wt% relative to a total weight of the plant protein-based food.

In one embodiment, a seasoning or flavoring may be an amino acid seasoning, a nucleotide seasoning, or a yeast derivative. An amino acid seasoning includes but is not limited to monosodium glutamate, disodium glutamate, monopotassium glutamate, a glycine salt, glutamic acid, calcium glutamate, monoammonium glutamate, magnesium glutamate, and mixtures thereof. A nucleotide seasoning includes but is not limited to inosine monophosphate, inosinic acid, disodium guanylate, guanosine monophosphate, dipotassium guanylate, calcium guanylate, disodium inosinate, and mixtures thereof. A yeast derivative includes but is not limited to yeast extract, yeast autolysate, and nutritional yeast, as well as preparations such as Yeastock(trademark) HR-Pd, and Savorboost(trademark) UM.

In one embodiment, a seasoning or flavoring may include, but is not limited to, onion, garlic, black pepper, vinegar powder, chili powder, paprika, oregano, bay leaf, cinnamon, cumin, coriander, clove, cayenne pepper, cumin, cilantro, parsley, coriander seed, cocoa powder, and mixtures thereof.

In one embodiment, the flavoring or seasoning may be some other herb, including but not limited to angelica, basil (holy, Thai, or lemon basil), bay leaf, Indian bay leaf (tejpat), boldo, borage, chervil, chives (garlic or Chinese chives), cicely, culantro, cress, curry leaf, dill, epazote, hemp, hoja santa houttuynia cordata (giap ca), hyssop, jimbu, kinh gioi (Vietnamese balm), kkaennip, lavender, lemon balm, lemon grass, lemon myrtle, lemon verbena, limnophila aromatica (rice-paddy herb), lovage, marjoram, mint, mugwort, mitsuba, parsley, perilla, rosemary, rue, sage, savory, sansho leaf, shiso, sorrel, tarragon, thyme, woodruff, and mixtures thereof.

In one embodiment, the flavoring or seasoning may be some other spice, including but not limited to aonori, ajwain, alligator pepper, allspice, amchoor, anise, asafoetida, peppercorn (black/green/white), Brazilian pepper, camphor, caraway, cardamom, cassia, celery powder, celery seed, charoli, chenpi, chia, chili (chili powder, chipotle, crushed red pepper, jalapeno, New Mexico, tabasco), citrus zest, cubeb, deulkkae, dill / dill seed, fennel, fenugreek, fingerroot, galangal, ginger, aromatic ginger, golpar, grains of paradise, grains of selim, horseradish, Japanese pricklyash, juniper berry, kokum, korarima, dried lime, liquorice, litsea cubeba, long pepper, mango-ginger, mastic, mahleb, mustard (black, brown, white), nigella, njangsa, nutmeg, Peruvian pepper, pomegranate seed, poppy seed, radhuni, rose, saffron, sarsaparilla, sassafras, sesame, shiso, Sichuan pepper (huajiao), sumac, tamarind, Tasmanian pepper, tonka bean, turmeric, uzazi, vanilla, voatsiperifery, wasabi, yuzu zest, zedoary, zereshk, and mixtures thereof.

In one embodiment, the flavoring or seasoning may be a blend of herbs and/or spices, including but not limited to berbere, curry powder, garam marsala, harissa, five-spice powder, herbes de Provence, hung liu, jerk spice, Old Bay(registered), and za’atar.

In one embodiment, the plant protein-based food may comprise other natural flavoring, artificial flavoring, protein hydrolysates, protein partial decomposition products, leghemoglobin, alkaline agents (pH adjustor) such as sodium carbonate, potassium carbonate, and calcined calcium, chelating agents such as gluconate and citrate, oxidation or reduction agents such as sodium ascorbate, glutathione, and cysteine, vinegar, other food additives such as alginic acid, brine, oil and fats, a coloring agent (including but not limited to beet juice and/or leghemoglobin), acidulants, rosemary extract, tocopherol, bread crumbs, soy sauce, flaxseed, psyllium seed husk, lecithin, wheat fiber, corn fiber, barley fiber, rye fiber, oat fiber, rice fiber, cellulose powder fiber, sugar beet fiber, inulin, soy fiber, pea fiber, citrus fiber, cottonseed fiber, mustard fiber, and leguminous fiber, calcium chloride, potassium chloride, magnesium chloride, as well as sulfate salts of calcium, potassium, and magnesium, and mixtures thereof; sugar or sweetening agents such as dextrose, sucrose, fructose, maltitol, sorbitol, lactose, trehalose, and the like. The plant protein-based food may comprise artificial coloring or natural coloring, such as caramel color, beet juice, turmeric, lycopene, leghemoglobin, or annatto extract.

In an alternative embodiment, the plant protein-based food may comprise one or more gums (also called hydrocolloids), such as xanthan gum (e.g., SATIAXANE CX 90, available from Cargill), guar gum, ghatti gum, karaya gum, tragacanth gum, carrageenan, psyllium seed gum, tamarind seed gum, alginates including salts of alginic acid (e.g., sodium alginate, potassium alginate, ammonium alginate, calcium alginate, etc.) and modified alginates (e.g., propylene glycol alginates, with specific mention being made to KIMILOID products such as KIMILOID HV, available from Kimica Corp.), locust bean gum, agar, tapioca, gelatin, pectin, gum arabic (acacia), including mixtures or blends of gums such as CARAGUM available from TIC gums; said gums being used alone or optionally in combination with a gelling agent such as calcium acetate, calcium citrate, calcium gluconate, calcium glycerol phosphate, mono-, di-, and tri-calcium phosphate, calcium sulfate, calcium carbonate, calcium lactate, and calcium phosphite.

In an alternative embodiment, the plant protein-based food may comprise one or more modified cellulose polymers such as methylcellulose (MC), ethyl cellulose (EC), hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxyethyl methyl cellulose (HEMC), ethyl hydroxyethyl cellulose (EHEC), methyl ethyl hydroxyethyl cellulose (MEHEC), hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (CMC), polyanionic cellulose (PAC), as well as modified versions thereof, for example, hydrophobically modified versions of any of the above.

In one embodiment, the plant protein-based food may comprise a cereal grain flour, starch, a grain flour not of a cereal grain, a milk-derived protein, or an egg-derived protein. Examples of grain flour (both cereal grains and others) include, but are not limited to, rice flour (including those derived from long grain, medium grain, or short grain rice, and glutinous (e.g. mochi-gome) or non-glutinous (e.g. uruchi-mai)), wheat flour, corn flour, potato flour, sweet potato starch, high amylose starch, tapioca flour (derived from cassava), amaranth flour, corn starch, waxy corn starch, arrowroot flour, sorghum flour (Jowar), soybean flour, adzuki flour, teff flour, chia seed flour, barley flour, pure buckwheat flour, gram flour (derived from chick-peas), millet flour (e.g., Chinese millet, Japanese millet, proso millet, etc.), oat flour, quinoa flour, almond flour, coconut flour, chestnut flour, tiger nut flour, and garbanzo flour, as well as combinations thereof. Examples of starch may include, but are not limited to, cornstarch including waxy corns starch, tapioca starch, arrowroot starch, wheat starch, rice starch including glutinous or non-glutinous rice starch, potato starch, sweet potato starch, sago starch, and mung bean starch, as well as modified or pre-gelatinized versions of such starches, and mixtures thereof. The starch may be a processed starch, and examples of the processed starch include oxidized starch obtained by oxidizing the above-mentioned starch, starch obtained by treating the above-mentioned starch with acid, crosslinked starch such as phosphoric acid-crosslinked starch obtained by phosphoric acid crosslinking the above-mentioned starch, and the like, starch obtained by processing the above-mentioned starch with fat, soy flour, etc., esterified starch obtained by esterifying the above-mentioned starch into phosphate ester, acetic acid ester, or the like, etherified starch obtained by etherification of the above-mentioned starch into hydroxypropylether or the like, starch obtained by a combination of these processes, and the like.

Examples of a milk-derived or egg-derived protein in may include, but are not limited to, albumin powder, defatted powdered milk, whey protein, whey protein concentrate, whey protein isolate, casein, cheese, cheese product, dried whole egg powder (e.g., lyophilized whole eggs), egg yolk powder (e.g., lyophilized egg yolks), and egg white powder (e.g., lyophilized egg whites). In one embodiment, water may be added as an ingredient.

In one embodiment, the plant protein-based food comprises whole, ground, minced, sprouted, germinated, malted, dried, and/or pulverized grains, legumes, or seeds such as those previously described. In another embodiment, the plant protein-based food may comprise vegetables or legumes such as onion, carrot, leafy greens, peppers, potato, whole lentils, split peas, soybeans, whole chickpeas, whole pinto beans, whole black beans, water chestnut, chili, and the like.

In one embodiment, the plant protein-based food may comprise one or more vitamins or minerals, including but not limited to zinc gluconate, iron, thiamine hydrochloride (vitamin B₁), niacin, pyridoxine hydrochloride (vitamin B₆), riboflavin (vitamin B₂), vitamin D₂, and vitamin B₁₂. In one embodiment, the plant protein-based food may be supplemented with docosahexaenoic acid (DHA, an omega-3 fatty acid), for instance, from algal oil.

In one embodiment, one or more of the above mentioned ingredients may be present in the plant protein-based food at a weight percentage of at least 0.0001 wt%, at least 0.001 wt%, at least 0.01 wt%, at least 0.05 wt%, at least 0.1 wt%, at least 0.25 wt%, at least 0.5 wt%, at least 0.75 wt%, at least 1 wt%, at least 1.5 wt%, at least 2 wt%, at least 2.5 wt%, at least 3 wt%, at least 4 wt%, at least 5 wt%, at least 6 wt%, at least 7 wt%, at least 8 wt% at least 9 wt%, at least 10 wt%, at least 12 wt%, at least 15 wt%, at least 20 wt%, at least 30 wt%, at least 40 wt%, at least 50 wt%, at least 60 wt%, at least 70 wt%, at least 80 wt%, and/or at most 0.001 wt%, at most 0.01 wt%, at most 0.05 wt%, at most 0.1 wt%, at most 0.25 wt%, at most 0.5 wt%, at most 0.75 wt%, at most 1 wt%, at most 1.5 wt%, at most 2 wt%, at most 2.5 wt%, at most 3 wt%, at most 4 wt%, at most 5 wt%, at most 6 wt%, at most 7 wt%, at most 8 wt% at most 9 wt%, at most 10 wt%, at most 12 wt%, at most 15 wt%, at most 20 wt%, at most 30 wt%, at most 40 wt%, at most 50 wt%, at most 60 wt%, at most 70 wt%, at most 80 wt%, at most 90 wt%, at most 95 wt%.

In one embodiment, the plant protein-based food contains less than 20 wt%, less than 15 wt%, less than 12 wt%, less than 10 wt%, less than 8 wt%, less than 5 wt% carbohydrates relative to a total weight of the plant protein-based food.

In one embodiment, the plant protein-based food is free or substantially free of one or more of methylcellulose, carrageenan, casein, egg white, glucose oxidase, phospholipase, grain flour, gluten, cellulose, gelatin, and collagen.

In one embodiment, the plant protein-based food is free or substantially free of gluten. In one embodiment, the plant protein-based food is free or substantially free of soy. In a further embodiment, the plant protein-based food is free or substantially free of both soy and gluten.

In one embodiment, the plant protein-based food is free or substantially free of methylcellulose. In one embodiment, the plant protein-based food is free or substantially free of enzymes other than transglutaminase.

In one embodiment, the plant protein-based food is free or substantially free of meat and meat-derived ingredients such as gelatin and collagen, and the plant protein-based food may be considered vegetarian, lacto-vegetarian, ovo-vegetarian, lacto-ovo vegetarian, and/or vegan. In a further embodiment, the plant protein-based food is free or substantially free of egg, egg white, and egg-derived ingredients. In one embodiment, the plant protein-based food is free or substantially free of meat and meat-derived ingredients as well as dairy and dairy-derived ingredients such as whey and casein. In one embodiment, the plant protein-based food is free or substantially free of animal-derived ingredients. Here, the plant protein-based food may be considered vegetarian and/or vegan.

Application

A method of making one of the above plant protein-based foods is described. This method comprises mixing the texturized vegetable protein, plant protein ingredient, transglutaminase, and water to produce a first mixture. Preferably the TVP is rehydrated by stirring in a mixture with the water, preferably cold water, and then the transglutaminase and plant protein ingredient is added and mixed. Additional ingredients may also be added and mixed. This mixing may be for a time in a range of 15 s to 5 min, preferably 30 s to 2 min, 45 s to 90 s, preferably 60 s. This produces a first mixture, which is preferably shaped into one or more loaves or patties, or any other shape as described throughout this disclosure. The first mixture is maintained at a temperature in a range of 1 to 15 °C, preferably 2 to 10 °C, 3 to 8 °C, or about 5 °C for a time in a range of 15 min to 72 h, preferably 30 min to 48 h, 45 min to 36 h, 50 min to 18 h, 55 min to 4 h, or 60 min. After the maintaining, the mixture may be frozen or cooked. In one embodiment the first mixture after the maintaining is cooked in a range of 100 to 250 °C, preferably 150 to 200 °C, 170 to 190 °C, or about 180 °C for a time in a range of 1 to 20 min, 2 to 15 min, 3 to 10 min, 4 to 8 min, or about 5 min. The plant protein-based food may be cooked on a skillet, flame grill, charcoal grill, convection oven, conventional oven, rotisserie, deep fryer, solar oven, pressure cooker, pressure fryer, steamer, or microwave oven. The plant protein-based food kept in a freezer may be cooked from frozen or allowed to thaw before cooking.

In one embodiment, the plant protein-based food is a pork, a lamb, or a seafood mimetic. The pork mimetic may be a chorizo, sausage, bratwurst, kielbasa, pepperoni, salami, andouille, or some other ground pork product. The seafood mimetic may be calamari, a fish, an octopus, or a crustacean mimetic including but not limited to lobster, shrimp, crawfish, and crab. In one embodiment, plant protein-based food may be in the form of a patty, burger patty, steak, cutlet, fillet, slice, flake, loaf, nugget, chunk, cube, ring, kabob, ground meat, crumble, ball, drumstick, meatball, jerky, shreds, stick, lollipop, tail (i.e., a lobster tail), claw (i.e., a crustacean claw), a shrimp shape, a fish shape, any other shape typical of a surimi product, or may be stuffed in a casing similar to a sausage, or may be filled into a stuffed food such as a potsticker, dumpling, roll, or bun. Other fish products include fish stick, fish sausage, kamaboko, chikuwa, satsuma-age (deep-fried fish cake), etc.) The shapes of the plant protein-based food may have an individual mass in a range of 5 to 1,000 g, 10 to 800 g, 20 to 500 g, 50 to 200 g, 80 to 150 g, or about 100 g. In one embodiment the plant protein-based food may be pressed into a shape, for instance, pressed into a mold. In one embodiment, a liquid mixture that forms the plant protein-based food may be extruded or filled into a mold where the plant protein-based food solidifies within. In another embodiment, the plant protein-based food may be formed into a block, slab, or other large shape from which one or more forms of the final shape may be cut or carved out.

According to an additional aspect, the present invention discloses a method of making a plant protein-based food, for instance, the plant protein-based food described previously in above embodiments or any variation thereof.

In one embodiment, the plant protein ingredient comprises at least one first plant-based protein concentrate or isolate and at least one second plant-based protein concentrate or isolate. In one embodiment, the at least one first plant-based protein concentrate or isolate is a protein concentrate or isolate of soy protein, pea protein, fava bean protein, and/or navy bean protein. In one embodiment, the at least one second plant-based protein concentrate or isolate is chickpea protein concentrate or isolate and/or pea protein concentrate or isolate. In one embodiment, the protein of the at least one first plant-based protein concentrate or isolate is derived from a different plant source species than the protein of the at least one second plant-based protein concentrate or isolate. In another embodiment, the at least one first and second plant-based protein concentrates or isolates are from the same or similar pulse source but are produced differently. In one embodiment, the at least one third plant-based protein concentrate or isolate is derived from soy, pea, chickpea, fava bean, lentil, and/or dry bean.

This method involves mixing texturized vegetable protein, one or more plant protein concentrates or isolates, transglutaminase, table salt, and one or more seasonings and/or flavorings other than table salt to form a first mixture of dry ingredients. Water is mixed with the first mixture to form a second mixture, and this mixing may take 10 s to 5 min, 30 s to 3 min, preferably 1 to 2 min. Oil and/or fat is then mixed with the second mixture to form a third mixture, and this mixing may take 15 s to 5 min, 20 to 90 s, preferably 30 to 60 s.

At this point the third mixture may be transferred to a vacuum bag and drawn under pressure to compress the mixture and remove air bubbles. For instance, the third mixture may be drawn under a pressure of 28 to 32 in. Hg for 2 to 15 s, or 3 to 10 s, and then sealed in the vacuum bag. Alternatively, the third mixture may be drawn under a pressure in a range of 10 to 380 mm Hg, 12 to 200 mm Hg, 15 to 100 mm Hg, 20 to 75 mm Hg, or 25 to 40 mm Hg for 2 to 15 s, or 3 to 10 s, and then sealed in the vacuum bag. Other means of removing air bubbles are possible, such as centrifugation, kneading, compressing under a press, or some other method of mechanical agitation. Alternatively, the third mixture may be poured or pressed into a slab or a mold.

The third mixture is then maintained at a temperature in a range of 1 to 65 °C, 2 to 55 °C, 3 to 40 °C, 3 to 30 °C, 3 to 25 °C, 3 to 20 °C, 3 to 15 °C, 3 to 10 °C, 3 to 6 °C, or about 4 °C, or 45 to 60 °C, 48 to 55 °C, 50 to 53 °C, or about 52 °C (125 °F) for a time in a range of 30 s to 72 h, 30 s to 48 h, 30 s to 36 h, 30 s to 24 h, 30 s to 18 h, 30 s to 12 h, 30 s to 6 h, 30 s to 4 h, 30 s to 2 h, 45 s to 105 min, 1 to 90 min, 10 to 80 min, 20 to 75 min, 30 to 70 min, 45 to 65 min, or about 60 min to produce the plant protein-based food.

In one embodiment, the method further involves cooling the third mixture and/or the plant protein-based food to a temperature in a range of -80 to 6 °C, -60 to 5 °C, -40 to 5 °C, -25 to 5 °C, -15 to 5 °C, -5 to 5 °C, 1 to 5 °C, or about 4 °C. For instance, the third mixture and/or the plant protein-based food may be placed in a refrigerator, ice bath, or freezer for a time in a range of 4 to 72 h, 6 to 48 h, 8 to 36 h, 12 to 24 h, 16 to 20 h, or about 18 h or about overnight to 72 hours.

In one embodiment, the method further involves heating the third mixture and/or the plant protein-based food at a temperature in a range of 80 to 120 °C, 90 to 110 °C, or about 100 °C, for instance by placing the vacuum bag containing the third mixture and/or the plant protein-based food into boiling water or exposed to heat or steam, in order to inactivate or denature the transglutaminase enzyme. This heating may be for a time in a range of 30 s to 10 min, 1 to 8 min, 2 to 5 min, or about 3 min.

In one embodiment, the method involves a combination of one or more steps of heating, cooling, and freezing the third mixture and/or the plant protein-based food. In one embodiment, the third mixture may be refrigerated, for instance at a temperature in a range of 1 to 5 °C or about 4 °C, for 4 to 20 h, or 8 to 18 h, and then frozen at a temperature of -80 to -15 °C indefinitely until needed for cooking or further preparation.

In one embodiment, the method involves forming the plant protein-based food into one or more different shapes as mentioned previously.

The plant protein-based food may be prepared for human or animal consumption. The plant protein-based food may be cooked, partially cooked, or frozen either in uncooked, partially cooked, or cooked state. Cooking may include frying either as sauteing or as deep-frying, pressure frying, baking, smoking, impingement cooking, steaming, pressure steaming flame grilling, charcoal grilling, solar cooking, radiating, drying, microwaving, and combinations thereof. In one embodiment, the plant protein-based food may be combined with another ingredient and then cooked, such as breading or battering and then fried or baked. In some embodiments, the plant protein-based food is used in cooked meals, including but not limited to soups, burritos, chilis, chowders, tacos, gyros, sandwiches, lasagnas, dumplings, bao, salads, pasta sauces, stews, kebabs, pizza toppings, spring rolls, potstickers, dumplings, and meat sticks. In some embodiments, the plant protein-based food is mixed with other protein products, including but not limited to other plant-derived products and/or animal meat. Critical aspects of the plant protein-based food include but are not limited to moisture level (both interior and exterior), concentration of TVP, protein concentration, protein type, and transglutaminase concentration.

The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventors to function well in the practice of the invention. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention

EXAMPLE 1
Plant Protein Ingredients

The plant protein ingredients used for reactivity analysis were generally obtained from commercial sources as indicated in Table 1.

Bambara groundnut flour, black-eyed pea flour, and pigeon pea flour were produced by grinding whole legumes into a flour using professional-grade Vitamix E310 Explorian(registered) Blender with a 32 oz. Vitamix Dry Grains container.

Protein concentrates of Bambara groundnut, black-eyed pea, and pigeon pea were produced by a two-stage process that separates the water-soluble proteins from the starch and then separates a water-insoluble fraction.

The water-soluble protein concentration uses the isoelectric point of water-soluble globulin proteins to separate from starch. First, a 1:10 slurry of pulse flour to water is mixed and stirred for a minimum of 1 hour to hydrate. The pH is adjusted to 4.5 with the addition of 2 M HCl while stirring. At this point, the pH adjusted slurry may be stored at 4 °C no longer than 24 hours. Sedimentation is allowed to occur for a minimum of 1 h at 4 °C, and the liquid is then centrifuged at 10,000 RPM for 15 min in a temperature-controlled centrifuge at 15 °C for 15 min. Sedimentation is allowed to occur again for at least 120 minutes at 4 °C. The supernatant is collected, and the remaining sediment is centrifuged to collect more supernatant.

The combined supernatant is adjusted to a pH of 7 using 2 M NaOH and dried in a vacuum oven at 50 °C. Alternatively, the combined supernatant may be adjusted to a pH of 10 with the 2 M NaOH, which could precipitate the proteins for collection by centrifugation.

The water-insoluble protein concentration uses 70% ethanol solution to separate the water-insoluble proteins from the starch of the sedimentation pellet produced from the above water-soluble protein concentration. Here, a 1:10 slurry of the sedimentation pellet to ≧70% ethanol solution is mixed for at least one hour and stored for no longer than 24 hours at 4 °C. Sedimentation is allowed to occur for at least one hour at 4 °C, or the slurry is centrifuged for 15 minutes at 10,000 RPM (and at 15 °C if possible). The supernatant is collected and dried in a vacuum oven at 50 °C, producing a visually distinguishable top layer of protein. The top layer of protein is scraped off and collected, and the bottom layer may be resuspended in a 1:10 slurry with ethanol solution to repeat the above process.

The collected proteins fractions from both the water-soluble and water-soluble may be blended together before or after drying. The protein concentration may be determined by Kjeldahl nitrogen determination assay.

Plant Protein Ingredient Reactivity Analysis

The transglutaminase activities of plant protein ingredients and resulting structures were evaluated by a Texture Analyzer (TSM Pro - Food Technology Corp.) with a 500 N load cell and a 20 mm ball probe.

The plant protein ingredient as a dry powder was blended with water and ACTIVA STG-TI (1 wt% transglutaminase in 99 wt% maltodextrin) to produce a slurry with a total weight of 500 g. The amount of protein ingredient was chosen so that the slurry comprises 18 wt% protein relative to the total weight of the slurry. Since the protein sources vary in dry weight protein concentration, different amounts were added to provide the 18 wt% protein concentration. ACTIVA STG-TI was added to a concentration of 1 wt% relative to the total weight of the slurry, with water added as the remainder to provide the 500 g total mass of the slurry. A transglutaminase-free control for each protein ingredient was made with water replacing the 1 wt% ACTIVA STG-TI. An exception to these formulations was used in the case of the unmodified flours of black-eyed pea, Bambara groundnut, and pigeon pea. In these formulations the volume of water was doubled to reduce the high viscosities of the slurries to a workable level. This also reduced the overall protein content. The compositions of the samples for black-eyed pea, Bambara groundnut, and pigeon pea flours, with and without transglutaminase, are shown in Table 2.

The resulting slurry was mixed until smooth or hydrated depending on viscosity build and then placed into a 16” × 20” (40.6 × 50.8 cm) vacuum bag. The bag was sealed, and a vacuum was drawn to -0.98 atm or 27 mm Hg to produce a deaerated slurry. This slurry was pushed to the bottom of the sealed bag. A corner of the bag was cut off diagonally, and the dearated slurry was dispensed through the cut-off corner into a second vacuum bag measuring 8” × 12” (20.3 × 30.5 cm). Sealing marks were made on both sides of the second vacuum bag at 6” (15.2 cm) from the bottom. A vacuum was drawn on the bag to -0.98 atm or 27 mm Hg, and the second vacuum bag was sealed near the top. The dearated slurry within was pushed to the bottom of the bag, and the bag was heat sealed on the sealing marks. This second vacuum bag was placed in a 50 °C water bath for 60 minutes. The second vacuum bag was immediately placed in a 4 °C cooler overnight (14 to 18 hours) while maintaining the vacuum to produce a hydrogel reaction product.

This hydrogel reaction product was observed for resulting gel structure while in the bag. Where sufficient structure existed, the bag was opened and the hydrogel reaction product was cut into 1” (2.5 cm) cubes and evaluated using the TSM Pro Texture Analyzer and 20 mm ball probe, with average maximum force being determined for each gel. The maximum force relates to the maximum pressure from the ball probe that the gel can withstand before losing cohesive structure. Table 1 shows each plant protein ingredient tested. The dry basis weight protein content of each plant protein ingredient is also shown, with the weight percentage of the plant protein ingredient used in the hydrogel to obtain 18 wt% protein concentration. These are related by the equation:

The results of the force for each hydrogel reaction product formed with or without transglutaminase (TG) are included; however, where the reaction product remained a liquid or a very weak gel, a force measurement could not be obtained.

EXAMPLE 2
Evaluation of Plant Based Protein Patties

Protein patties were made using different ratios of plant protein ingredients, transglutaminase source (as ACTIVA STG-TI, Ajinomoto Co., Inc., 100 U/g), texturized soy protein, and water. The texturized pea protein (as a type of texturized vegetable protein, or TVP) was obtained as MGP PROTERRA 2100 granules from MGP Ingredients. The plant protein ingredients used in the patties have been tested for transglutaminase reactivity in Table 1 above. These plant protein ingredients include Bambara groundnut flour, Bambara groundnut protein concentrate, black-eyed pea flour, black-eyed pea concentrate (the four preceding ingredients were produced by Ajinomoto Health & Nutrition North America, Itasca, IL), chickpea protein concentrate (obtained from Greenboy), fava bean protein concentrate (Vitessence Prista(registered) P 360 fava bean from Ingredion), mung bean protein concentrate (obtained from Greenboy), pea protein concentrate (Vitessence Pulse(registered) 1853 pea protein from Ingredion), pinto bean flour (Honest Origins), lupin protein concentrate (CK Ingredients), pigeon pea flour and pigeon pea protein concentrate (both produced by Ajinomoto Health & Nutrition North America), and red lentil protein concentrate (Atura).

The protein contents of the above protein ingredients are indicated in Table 1 except for the black-eyed pea concentrate, the pigeon pea concentrate, and the Bambara protein concentrate. The black-eyed pea concentrate used in the patties had a protein content of 66.8 wt%; the pigeon pea protein concentrate had a protein content of 51.9 wt%, and the Bambara protein concentrate had a protein content of 65.2 wt%.

Of the additional materials, sunflower seed oil was obtained as Girasole from Zucchi; potato starch was obtained as Eliane 100 from Ingredion; and methylcellulose was obtained as (Metolose MCE-100TS from Shin-Etsu Chemical Co., Ltd.)

Table 3 shows the recipes (R1 to R8) used for the protein patties with (CR1 to CR4) as control recipes. CR1 is a notable control as it uses methylcellulose (MC) as the binding ingredient in place of transglutaminase (TG). The amounts of ingredients were chosen to form patties of 100 g each. The plant protein ingredients were tested at three main levels of concentration: “upper protein” included the plant protein ingredient in a range of 40 to 50 wt%; “ideal protein” is in a range of 1 to 3 wt%; and “below lower protein” is in a range of 0.1 to 0.5 wt%, each relative to a total mass of the patty before cooking. The transglutaminase source (ACTIVA STG-TI, 1 wt% transglutaminase) was tested at five levels of concentration: “lower TG” included the transglutaminase source at concentrations in a range of 0.0010 to 0.0050 wt%; “ideal TG” is in a range of 0.08 to 0.20 wt%; “middle TG” is in a range of 0.25 to 1 wt%; “above ideal TG” is in a range of 1.1 to 2.0 wt%; and “upper TG” is in a range of 8 to 10 wt%, each relative to a total mass of the patty before cooking. It is important to note that all patties included TVP; patties indicated as “no protein” have no plant protein ingredient added but still contain TVP.

Except for CR1, all recipes and control recipes of the plant protein patties involved the following procedure. The TVP was added to the water (as cold or ice-cold water) and rehydrated for 5 minutes in a mixer. The plant protein ingredient and ACTIVA STG-TI were added and stirred for 1 minute. The mixture was formed into 100 g burger patties using a patty mold and left to sit at 5 °C for 1 h. The patties were then cooked in an oven at 180 °C for 5 min.

For the methylcellulose sample, CR1, an emulsion curd was formed by mixing sunflower seed oil, potato starch (Eliane 100 Ingredion), water, and methylcellulose (Metolose MCE-100TS) at high shear using a stick blender. Separately, the TVP was added to an additional volume of water (as cold or ice-cold water) and rehydrated for 5 minutes in a mixer. The emulsion curd was added to the hydrated TVP in the mixture with an additional amount of methylcellulose and mixed for 1 minute. The mixture was formed into 100 g burger patties using a patty mold and left to sit at 5 °C for 1 h. The patties were then cooked in an oven at 180 °C for 5 min.

Tables 4 to 16 provide results of the different plant protein ingredients, as determined by an expert panel of five evaluators, except for the Bambara groundnut protein concentrate (Table 5), the black-eyed pea protein concentrate (Table 7), and the pigeon pea protein concentrate (Table 15), which each used an expert panel of four evaluators. On a scale of 1 to 3, with 1 being the least preferable, and 3 being the most preferable, the expert panel judged the cohesiveness of the patty after the chilling step and before the cooking, the firmness after cooking, and the moistness after cooking. The cohesion was evaluated by the formability and binding of the patty, and the patty’s resistance to crumbling. The firmness was considered as the patty’s firmness to the touch and when being eaten. The moistness after cooking was considered both as the moisture level on the outside by touch, and by the inside of the patties while being eaten. The values presented on the tables are the averages of the rankings from the expert panel. The description provided for “Pre-test” relates to the condition of the patties after being formed in the mold but before the 1 h chilling. The patties were generally compared with the methylcellulose control having preferred cohesion, firmness, and moistness. For some plant protein ingredients, the patties are ranked by similarity to the methylcellulose control.

Additional comments on the patties follow:

Chickpea protein concentrate

Results shown in Table 8. All variables showed some binding. CR3 had the most binding and cohesion even though there was no TG. R4 was comparable to CR3; however, the structure crumbled more easily. R8 and CR4 had the least binding and cohesion.

Mung bean protein concentrate

Results shown in Table 10. All patties with the exception of CR4 had binding. Samples R3, R4, R6, and CR3 had the most binding. CR4 had minimal to no binding-low practicability and usage. Mung beans had the closest attributes to methylcellulose overall compared to the other pulses.

Pea protein concentrate

Results shown in Table 11. Significant binding was observed in R3, R4, and R6. Binding was minimal in CR3 and R8 or not present in CR4. Patties R1 and CR2 were unable to form patties due to lack of water. No effect with the addition of water.

Pinto flour

Results shown in Table 12. All patties contained some sort of binding aside from CR4, which provided minimal to no binding. R1 and R2 were the most firm, but R3 had the most binding. No samples completely matched the methylcellulose control (CR1) in all aspects.

Lupin protein concentrate

Results shown in Table 13. Lupin protein has some binding; however, it is not a match for the methylcellulose control, CR1. Of the patties, R1 and CR2 have potential for other uses due to their floral notes as well as gum-like textures.

Red lentil protein concentrate

Results shown in Table 16. All patties that contained the lentil protein concentrate had some sort of binding. Samples with upper TG (R1, R3, R5) provided more binding and cohesion. However, no samples were judged to be a good methylcellulose replacer.

EXAMPLE 3
Plant-based Protein Lobster Mimetic
A lobster mimetic was prepared using ingredients at the concentrations listed in Table 17. Dry ingredients of TVP (ProTerra(registered) 1100 wheat TVP or CP-Xtura(trademark) 65 chickpea TVP), monopotassium glutamate, Yeastock(trademark) HR-Pd, table salt, Savorboost(trademark) F, lobster flavor, seafood flavor, Activa(registered) STG-TI transglutaminase preparation, dextrose, life’sDHA(registered) S10-P200 encapsulated algal oil, and protein concentrate (Supro(registered) EX 45 soy protein concentrate or InnovoPro(registered) 70 Chickpea protein concentrate) were first combined and blended thoroughly. Water was added and mixed for 1 minute, followed by the addition of sunflower oil and an additional 1 minute of mixing. The mixture was transferred to a large vacuum bag and vacuum sealed at a pressure of 29.5 in. Hg. The mixture was heated to 125 °F (51.7 °C) and held at that temperature for 1 hour. Next, the mixture was refrigerated overnight and then processed into different shapes, such as sticks.

EXAMPLE 4
Plant-based Protein Fish Mimetic

A fish mimetic was prepared using ingredients at the concentrations listed in Table 17. Dry ingredients of TVP (ProTerra(registered) 1100 wheat TVP or CP-Xtura(trademark) 65 chickpea TVP), monopotassium glutamate, Yeastock(trademark) HR-Pd, table salt, Savorboost(trademark) F, tuna flavor, Activa(registered) STG-TI transglutaminase preparation, dextrose, life’sDHA(registered) S10-P200 encapsulated algal oil, and protein concentrate (Supro(registered) EX 45 soy protein concentrate or InnovoPro(registered) 70 Chickpea protein concentrate) were first combined and blended thoroughly. Water was added and mixed for 1 minute, followed by the addition of sunflower oil and an additional 1 minute of mixing. The mixture was transferred to a large vacuum bag and vacuum sealed at a pressure of 29.5 in. Hg. The mixture was heated to 125 °F (51.7 °C) and held at that temperature for 1 hour. Next, the mixture was refrigerated overnight and then processed into different shapes, such as flakes.

EXAMPLE 5
Plant-based Protein Lamb Mimetic

A lamb mimetic was prepared using ingredients at the concentrations listed in Table 18. Dry ingredients TVP pea granules (eg. ProTerra(trademark) 2100C), monopotassium glutamate, Plant Answer(registered) Mask B natural flavor, Savorboost(trademark) UM, other natural flavor, Savorboost(trademark) F, garlic powder, salt, Activa(registered) STG-TI transglutaminase preparation, fava protein concentrate or isolate (eg. Ingredion Vitessence(registered) Prista P360), and soy protein concentrate or isolate (eg. Supro(registered) EX 45), were mixed and blended thoroughly. Water with dissolved caramel color was added and mixed for 1 to 2 minutes. The sunflower oil, coconut oil, and additional natural flavor were then added and mixed for 30 to 60 seconds. The mixture was next transferred to a large vacuum bag and vacuum sealed with 9 seconds of contact to form the lamb mimetic mixture. The lamb mimetic mixture was formed into 55 g balls and refrigerated covered overnight. The balls were then frozen to retain shape and vacuum sealed at 4.5 seconds of contact for protection from exposure to oxygen and humidity fluctuations during storage at 4 °C or -20 to 0 °C.

EXAMPLE 6
Plant-based Protein Chorizo Mimetic

A chorizo mimetic was prepared using ingredients at the concentrations listed in Table 18, with the ingredients of the chorizo dry seasoning blend provided in Table 19. Dry ingredients TVP pea granules (eg. ProTerra(trademark) 2100C), the chorizo dry seasoning blend, salt, Activa(registered) STG-TI transglutaminase preparation, and Supro(registered) EX 45 soy protein concentrate were mixed and blended thoroughly. Water was added and mixed for 1 to 2 minutes. The sunflower oil, coconut oil, additional natural flavor, and annatto extract were then added and mixed for 30 to 60 seconds. The mixture was next transferred to a large vacuum bag and vacuum sealed with 9 seconds of contact to form the chorizo mimetic mixture. The chorizo mimetic mixture was formed into 100 g patties or 1000 g sheets having a thickness of 9.5 mm (3/8”), and incubated in a 50 °C water bath or steam oven for 60 to 120 min. The chorizo preparation was then frozen, or browned and frozen.

This application claims the benefit of priority from U.S. Provisional Application No. 63/346,683 filed May 27, 2022; U.S. Provisional Application No. 63/352,781 filed June 16, 2022; U.S. Provisional Application No. 63/359,530 filed July 8, 2022; and U.S. Provisional Application No. 63/492,293 filed March 27, 2023; the entire contents of all the above applications are incorporated herein by reference in their entirety.

Claims

A plant protein-based food, comprising:
15 to 85 wt% texturized vegetable protein;
0.35 to 45 wt% plant protein ingredient; and
0.0005 to 0.05 wt% transglutaminase, each weight percent relative to a total weight of the texturized vegetable protein, the plant protein ingredient, and the transglutaminase,
wherein the plant protein ingredient is at least one selected from the group consisting of a flour, a protein concentrate, or a protein isolate.
The plant protein-based food of claim 1, which is substantially free of one or more of methylcellulose, carrageenan, casein, egg white, glucose oxidase, phospholipase, cereal grain flour, gluten, cellulose, gelatin, and collagen.
The plant protein-based food of claims 1 or 2, which is substantially free of animal-derived ingredients.
The plant protein-based food of any one of the above claims, wherein the plant protein ingredient is derived from at least one selected from the group consisting of Bambara groundnut, black-eyed pea, dry bean, chickpea, fava bean, lentil, lupin, pea, pigeon pea, soybean, and wheat.
The plant protein-based food of any one of the above claims, wherein the dry bean is at least one selected from the group consisting of mung bean, navy bean, and pinto bean.
The plant protein-based food of any one of the above claims, wherein the plant protein ingredient is derived from at least one selected from the group consisting of Bambara groundnut, black-eyed pea, chickpea, fava bean, mung bean, pea, pinto bean, lupin, pigeon pea, and red lentil.
The plant protein-based food of any one of the above claims, wherein the plant protein ingredient comprises 19 to 100 wt% protein relative to a total weight of the protein ingredient.
The plant protein-based food of any one of the above claims, wherein the plant protein ingredient comprises 80 to 100 wt% protein relative to a total weight of the protein ingredient.
The plant protein-based food of any one of the above claims, wherein the texturized vegetable protein is made from at least one selected from the group consisting of wheat, soy, pea, dry bean, broad bean, chickpea, cowpea, pigeon pea, lentil, Bambara groundnut, and lupin.
The plant protein-based food of any one of the above claims, wherein the texturized vegetable protein is made from pea.
The plant protein-based food of any one of the above claims, further comprising 30 to 75 wt% water relative to a total weight of the plant protein-based food.
The plant protein-based food of any one of the above claims, further comprising 2 to 35 wt% of one or more oils and/or fats relative to a total weight of the plant protein-based food.
The plant protein-based food of claim 12, wherein the one or more oils and/or fats is selected from the group consisting of sunflower seed oil, canola oil, corn oil, vegetable oil, coconut oil, palm oil, and algal oil.
The plant protein-based food of any one of the above claims, which is substantially free of soy.
The plant protein-based food of any one of the above claims, which is a chorizo, pork, beef, chicken, sausage patty, sausage link, lamb, lobster, shrimp, crab, crustacean, calamari, or fish mimetic.
The plant protein-based food of any one of the above claims, wherein the plant protein-based food comprises:
15 to 85 wt% of at least one texturized vegetable protein;
0.35 to 45 wt% of one or more plant protein ingredients;
0.0005 to 0.01 wt% transglutaminase;
0.3 to 1.5 wt% table salt;
0.5 to 12 wt% of one or more seasonings and/or flavorings other than table salt;
30 to 75 wt% water; and
2 to 35 wt% of one or more oils and/or fats, each weight percent relative to a total weight of the plant protein-based food.
A method of making the plant protein-based food of claim 16, the method comprising:
mixing at least one texturized vegetable protein, one or more plant protein ingredients, transglutaminase, table salt, and one or more seasonings and/or flavorings other than table salt to form a first mixture;
mixing water with the first mixture to form a second mixture;
mixing oil and/or fat with the second mixture to form a third mixture; and
maintaining the third mixture at a temperature in a range of 1 to 65 °C for a time in a range of 30 s to 72 h to produce the plant protein-based food.
The method of claim 17, further comprising, before the maintaining, subjecting the third mixture to a vacuum to remove air bubbles.
The method of claims 17 or 18, further comprising cooling the plant protein-based food to a temperature in a range of -80 to 6 °C.
A method of making the plant protein-based food of any one of claims 11, 14, and 15, the method comprising:
mixing the texturized vegetable protein, plant protein ingredient, transglutaminase, and water to produce a first mixture; and
maintaining the first mixture at a temperature in a range of 1 to 15 °C for a time in a range of 15 min to 72 h.
The method of claim 20, further comprising shaping the first mixture between the mixing and the maintaining.
The method of claim 20 or 21, further comprising, after the maintaining, cooking the first mixture at a temperature in a range of 100 to 250 °C for a time in a range of 1 to 20 minutes.