WO2025188915A1 - Plant based whole cuts organized on different lengthscales - Google Patents

Abstract

The present disclosure describes methods and formulations to produce a plant-based meat analog, resembling whole cuts of meat, especially beefsteak. To achieve the most similar texture to real steak, the composition and structure of the plant¬ based steak are engineered on two lengthscales, where the smaller lengthscale comprises inclusions that may or may not be anisotropic and the larger lengthscale comprises strands that are semi-aligned on average along a principal axis. The degree of alignment of the strands is controlled to incorporate some randomness in order to improve the natural perception of the product and to increase its chewiness. Additionally, in some embodiments, some proteins within the composition are intentionally kept raw to create a setting (firming) effect of the product in the final cooking step by the end user.

Description

PLANT BASED WHOLE CUTS ORGANIZED ON DIFFERENT LENGTHSCALES

FIELD OF THE INVENTION

[0001] The present disclosure generally relates to plant-based meat analogs, such as analogs of whole cuts, including analogs of beefsteak. The present disclosure further relates to methods of preparing such plant-based meat analogs.

BACKGROUND OF THE INVENTION

[0002] In recent years, plant-based diets and lifestyle have gained in popularity. This rise in popularity is motivated by a number of concerns related to meat consumption, such as the negative environmental impact of livestock, health concerns related to meat consumption, and ethical concerns about animal welfare in industrial farming practices and slaughter.

[0003] While some consumers may readily agree with these concerns around meat consumption, they may still enjoy the taste, texture, flavor and aroma of meat, including the combined sensory perception of high-quality whole cuts of meat, for example, beefsteak. In addition, such meat dishes have societal cache and are often associated with celebratory events and luxurious experiences. Replicating the sensory experience of high value whole cuts with a plant-based product is a unique challenge that relies on replicating both the cooking experience and the complete sensory experience while eating the steak. While burgers and sausages are structurally and visually homogenous at the centimeter (cm) level, whole cuts are intact pieces of animal tissue and therefore are anisotropic, inhomogeneous, and possess a complex structure all the way from a molecular, nanometer (nm) lengthscale to the cm lengthscale. This complexity may include the arrangement of different tissues, like muscle tissue, fat (adipose) tissue and connective tissue; the orientation and alignment of the fiber-like structure of muscle tissue; and, more generally, the sub-structures within each tissue. An example of sub-structures in muscle tissue would be muscle fibers, enclosed by connective tissue (endomysium) and grouped into bundles by additional connective tissue (perimysium). Because of this structural complexity at multiple lengthscales, a realistic plant-based steak is sometimes considered the Holy Grail of plant-based meat analogs. In addition, the ability to reproduce these complex structures economically on an industrial production scale is itself a significant challenge. SUMMARY OF THE INVENTION

[0004] The present disclosure provides plant-based meat analogs, such as those resembling whole cuts of meat, including beefsteak. In a related aspect, provided herein are methods for producing the plant-base meat analogs, including replicating details like anisotropy, chewiness, juiciness, and/or the cooking experience of real steak.

[0005] To achieve a similar texture to real steak, the composition and structure of a plant-based steak can be engineered on at least two lengthscales, where the smaller lengthscale comprises inclusions that may or may not be anisotropic and the larger lengthscale comprises strands that are aligned on average along a principal axis. The degree of alignment of the strands can be varied to incorporate some randomness (referred to herein as “semi-aligned”) in order to improve the natural perception of the product and to increase its chewiness. Additionally, some proteins within the composition may be kept raw to create a setting (firming) effect of the product in the final cooking step by an end user.

[0006] For a steak-like texture, the following steps may be included:

• Selection of the best inclusions; these inclusions may or may not be fibrous. Often, these inclusions come from shredding or mincing textured proteins to the desired size and morphology

• Producing a firm muscle phase, in which these small inclusions are embedded in a dough- like matrix together with additional selected plant protein powders, binders, colors, flavors, and/or liquids. This dough is processed into strands resembling bundles of muscle fibers by, for example: o Sheeting the dough o Heating the sheets, wherein heating time and temperature can be adjusted to achieve the desired toughness based on the properties of the chosen protein powders and binders. o Cutting the sheets into strands, where the strands size and cross-section can be changed to produce textures that best mimic the desired cut and type of animal steak.

• Soaking of the strands in flavored and colored liquid (plant-based meat juice analog)

• Arranging the strands ‘semi-aligned’ on average along a principal axis to get both, anisotropy but also cohesion orthogonal to the main orientation of the strands • Binding together the soaked strands by a binder, for example a protein powder that is at least partially raw and still functional like vital wheat gluten. This binder acts as connective tissue analog between the strands, and firms up during the final cooking by the end user.

• Incorporating a fat emulsion (fat phase) that mimics adipose tissue.

• Forming the whole cut analog into a roast or other formation, for example by rolling, followed by slicing the roast into fillets.

• The final cooking step is done by the end user and sets the raw or partially raw binders to the desired doneness level.

[0007] In some embodiments, provided herein is a method of making a plant-based meat analog, comprising: (a) forming a dough-like wet protein mixture comprising i) texturized protein (TP), and ii) one or more of a plant protein powder, a binder, a coloring agent, a flavor agent and/or a spice, a liquid, and a leavening agent, wherein the TP has been processed into particles or fibers ranging from about 10 micrometer (pm) in diameter to about 1 mm in diameter; (b) processing the wet protein mixture to form fiber-like strands ranging from about 0.1 mm to about 4 mm, wherein the strands are flexible; (c) aggregating the strands into a desired alignment; and (d) stabilizing the strands to form a coherent strand mass resembling meat in appearance and texture. In some embodiments, the TP has been produced by (i) low moisture extrusion or high moisture extrusion, or similar methods involving temperatures above 100°C, pressures above ambient pressure, and/or shear. In some embodiments, the TP has been produced by any one of the following methods: shear cell technology, freeze alignment, wet spinning, electrospinning, and/or fermentation to produce microbial biomass.

[0008] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the liquid is water, vegetable oil, or a mix of the two.

[0009] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the stabilization is achieved through one or more of the following: rolling and pressing, compacting by subjecting to vacuum, chemical crosslinking, enzymatic crosslinking, heating, and cooling.

[0010] In some embodiments, according to any of the methods of making a plant-based meat analog described above, processing the wet protein mixture to form fiber-like strands comprises sheeting the wet protein mixture to form a coherent workable protein dough sheet followed by cutting the protein dough sheet to form strands. In some embodiments, the protein dough sheet is set prior to cutting. In some embodiments, the strands are set after cutting. In some embodiments, the setting is by heating to a temperature that allows for protein coagulation. In some embodiments, the setting is by means of cross-linking other than heat coagulation, optionally wherein the cross-linking is chemical crosslinking, and/or enzymatic crosslinking.

[0011] In some embodiments, according to any of the methods of making a plant-based meat analog described above, processing the wet protein mixture to form fiber-like strands comprises extruding the wet protein mixture into strands. In some embodiments, the method comprises extruding the wet protein mixture into strands followed by setting the extruded strands. In some embodiments, the setting is by heating to a temperature that allows for protein coagulation. In some embodiments, the setting is by means of cross-linking other than heat coagulation, optionally wherein the cross-linking is chemical crosslinking, and/or enzymatic crosslinking. In some embodiments, the method comprises extruding and heat-setting the wet protein mixture into strands in a single step through a heated nozzle or extrusion die.

[0012] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the TP is processed prior to adding to the wet protein mixture such that isotropic inclusions of the TP are formed in the wet protein mixture. In some embodiments, the TP is minced into particles prior to adding to the wet protein mixture. In some embodiments, the TP particles are arranged in a random fashion.

[0013] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the TP is processed prior to adding to the wet protein mixture such that anisotropic inclusions of the TP are formed in the wet protein mixture. In some embodiments, the TP is shredded into fibers prior to adding to the wet protein mixture. In some embodiments, the TP fibers are semi-aligned along a common axis. In some embodiments, the TP fibers are arranged in a random fashion.

[0014] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the TP is from one or more of the following: wheat gluten, soy protein isolate, soy protein concentrate, soy flour, pea protein isolate, pea protein concentrate, pea flour, faba bean protein isolate, faba bean protein concentrate, faba bean flour, chickpea protein isolate, chickpea protein concentrate, chickpea flour, an isolate, concentrate, and/or flour from a cereal grain, and optionally commercial food starch(es) selected from wheat starch, corn starch, tapioca starch, potato starch, rice starch, or any suitable, commercially available starch. In some embodiments, the majority of the protein in the TP is from (a) wheat gluten, optionally wherein the TP is a fibrous product from the ProTerra family by MGP ingredients (for example ProTerra 1100, ProTerra 1200, ProTerra 1350), or (b) a blend of wheat gluten and one or more of soy protein isolate, soy protein concentrate, and soy flour, optionally wherein the TP is a fibrous product from the SuproMax family by Solae/IFF (for example SuproMax 5010, SuproMax 5050). In some embodiments, the TP comprises a granular or flaky TP, optionally wherein the granular or flaky TP is the textured soy flour Purelynature 165-118 (ADM), textured soy protein concentrate Arcon T 158-171 (ADM), Response 4310, Response 4380, Response 4400, Response 4410, Response 4438 (Solae/IFF), or textured pea protein TPP70, or TPP80 (Puris).

[0015] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the wet protein mixture comprises (a) water; (b) salt; (c) flavoring and/or spices;

(d) vegetable oil; (e) TP; (f) a binder, optionally wherein the binder is (i) a fiber from citrus peel, citrus pulp, carrot, pea hull, inner cell walls of pea, apple pomace, Psyllium hull, oat fiber, and/or flaxseed, or (ii) a hydrocolloid, optionally wherein the hydrocolloid is xanthan gum, guar gum, alginate, high methoxyl and low methoxyl pectin, sodium alginate, different types of carrageenan, methylcellulose, and/or hydroxypropyl methylcellulose; (g) vital wheat gluten; (h) wheat flour, optionally wherein the wheat flour is all-purpose wheat flour; and/or (i) a leavener, optionally wherein the leavener is baking powder or baking soda. In some embodiments, the wet protein mixture comprises (a) water from about 40% to about 60% by weight; (b) salt from about 0% to about 1.5% by weight; (c) flavoring from about 0% to about 8% by weight; (d) vegetable oil from about 0% to about 15% by weight; (e) TP from about 5% to about 35% by weight; (f) a binder from about 0% to about 6% by weight; (g) vital wheat gluten from about 8% to about 30% by weight; (h) wheat flour from about 0% to about 5% by weight; (i) baking powder or baking soda from about 0.1% to about 5% by weight; and/or (j) a spice blend from about 0% to about 5% by weight.

[0016] In some embodiments, according to any of the methods of making a plant-based meat analog described above, i) the vegetable oil is selected from canola oil, soybean oil, sunflower oil, safflower oil or any other vegetable oil liquid at room temperature; ii) the TP is one of the fibrous TP products described above; and/or iii) the binder is Psyllium hull fiber. In some embodiments, the TP comprises one of the granular or flaky TPs described above, and the sum of the fibrous TP and the granular or flaky TP is about 5% to about 35% of the wet protein mixture by weight. [0017] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the fibrous TP is from soy protein, wheat gluten, and wheat starch.

[0018] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the fibrous TP is SuproMax 5050 or SuproMax 5010.

[0019] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the wet protein mixture further comprises a coloring agent and/or potato protein isolate. In some embodiments, the wet protein mixture comprises (i) a coloring agent from about 0.1% to about 5% by weight; and/or (ii) potato protein isolate from about 1% to about 5% by weight, optionally wherein the potato protein isolate is rich in native patatin, further optionally wherein the potato protein isolate is Solanic 200 (Avebe).

[0020] In some embodiments, according to any of the methods of making a plant-based meat analog described above, forming the dough-like wet protein mixture comprises: (a) soaking the TP in a mixture containing the water, salt, flavoring, vegetable oil and optionally color for a sufficient amount of time to allow for absorption of the liquid phase, thereby forming soaked textured proteins; (b) grinding the soaked textured proteins in such a manner as to result in small anisotropic fibers with the majority having a diameter between about 10 pm and about 1 mm, thereby forming ground textured proteins; and (c) combining the ground textured proteins with any remaining dry ingredients in such a manner as to result in a cohesive, extensible dough-like mass, thereby forming the dough-like wet protein mixture.

[0021] In some embodiments, according to any of the methods of making a plant-based meat analog described above, processing the wet protein mixture to form fiber-like strands comprises sheeting the wet protein mixture to form a coherent workable protein dough sheet followed by cutting the protein dough sheet to form strands, and wherein the protein dough sheet is from about 0.1 mm to about 4 mm in thickness. In some embodiments, the protein dough sheet is heated at a sufficient temperature and for a sufficient duration to allow for setting of the dough prior to cutting to form strands. In some embodiments, the protein dough sheet is cut to form strands from about 0.1 mm to about 4 mm wide.

[0022] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the method further comprises soaking the strands in a first portion of meat juice analog for a sufficient duration to allow for the meat juice analog to be at least mostly absorbed, optionally wherein the weight of the first portion of meat juice analog is from about 10% to about 80% of the weight of the strands. In some embodiments, the method further comprises coating the strands with a binder mix, optionally wherein the weight of the binder mix is from about 5% to about 50% of the weight of the strands. In some embodiments, the binder mix comprises a protein powder.

[0023] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the protein powder is at least partially undenatured (raw). In some embodiments, the protein powder is capable of being crosslinked and increasing in firmness during a final cooking step of the meat analog at about 60°C to about 95 °C carried out by an end user. In some embodiments, the end user can control the doneness level by cooking to a final temperature between about 60°C and about 95°C, optionally wherein the final temperature is between about 74°C and about 95°C for food safety.

[0024] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the binder mix comprises at least 50% vital wheat gluten. In some embodiments, the binder mix comprises vital wheat gluten, optionally wherein the binder mix further comprises a spice blend, further optionally wherein the vital wheat gluten is present in the binder mix from about 90% to about 100% by weight and/or the spice blend is present in the binder mix from about 0% to about 10% by weight.

[0025] In some embodiments, according to any of the methods of making a plant-based meat analog described above, aggregating the strands into a desired alignment comprises aggregating the strands such that they are semi-aligned along a common axis. In some embodiments, the degree of alignment of the strands is between 0.9965 and 0.7500.

[0026] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the method further comprises adding a second portion of meat juice analog to the aggregated strands, optionally wherein the weight of the second portion of meat juice analog is from about 10% to about 30% of the weight of the strands. In some embodiments, the meat juice analog comprises (a) water, optionally wherein the water is from about 85% to about 99% by weight; (b) flavoring, optionally wherein the flavoring is from about 0% to about 8% by weight; (c) salt, optionally wherein the salt is from about 0% to about 1.5% by weight. In some embodiments, the meat juice analog further comprises baking soda or another mildly alkaline, food-safe ingredient, from about 0% to about 5% by weight to neutralize the pH. In some embodiments, the meat juice analog further comprises a coloring agent, optionally wherein the coloring agent is from about 0.1% to about 10% by weight.

[0027] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the method further comprises applying an adipose tissue analog to the strands, optionally wherein the weight of the adipose tissue analog is from about 0% to about 35% of the weight of the strands.

[0028] In some embodiments, according to any of the methods of making a plant-based meat analog described above, stabilizing the strands to form a coherent strand mass comprises rolling up and pressing together the aggregate strands, followed by compacting by subjecting to vacuum.

[0029] In some embodiments, according to any of the methods of making a plant-based meat analog described above, a first portion of the adipose tissue analog is applied during aggregation of the strands, and a second portion of the adipose tissue analog is applied to an outside surface of the coherent strand mass.

[0030] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the adipose tissue analog comprises (a) a lipid component (optionally wherein the lipid component is vegetable oil and/or vegetable fat), optionally wherein the lipid component is from about 5% to about 40% by weight; (b) vital wheat gluten, optionally wherein the vital wheat gluten is from about 0% to about 10% by weight; (c) flavoring, optionally wherein the flavoring is from about 0% to about 6% by weight; (d) salt, optionally wherein the salt is from about 0% to about 1% by weight; (e) methylcellulose, optionally wherein the methylcellulose is from about 0.5% to about 4% by weight; and/or (f) water, optionally wherein the water is from about 35% to about 60% by weight. In some embodiments, the lipid component comprises a vegetable fat that is at least partly solid at 15-20°C, optionally wherein the lipid component further comprises liquid vegetable oil that is fully liquid at 15-20°C. In some embodiments, the solid vegetable fat is selected from cocoa butter, coconut oil, palm oil, palm kernel oil, shea butter, sal butter and/or hydrogenated fats. In some embodiments, the solid vegetable fat is shortening or margarine. In some embodiments, the liquid vegetable oil is selected from canola oil, soybean oil, sunflower oil, safflower oil, and/or any other suitable plant-based oil.

[0031] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the methylcellulose is a type that gels below 50°C and forms gels that are on the very firm end of the spectrum achievable with methylcellulose, measured with suitable texture methods, including TPA. In some embodiments, the methylcellulose is Wellence Vegeform 183 (IFF/Dupont).

[0032] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the method further comprises preparing the adipose tissue analog by (a) dispersing the dry ingredients of the adipose tissue analog in the vegetable oil and/or vegetable fat while heating at a temperature sufficient to melt the vegetable fat; (b) removing the oil mixture from heat; and (c) adding the water as a mix of ice and liquid with mixing to form a stable emulsion.

[0033] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the method further comprises cutting the coherent strand mass into individual fillets, resembling fillets of an animal based whole-cut.

[0034] In some embodiments, according to any of the methods of making a plant-based meat analog described above, there is no plane, whose normal vector is orthogonal to the average strand alignment in the whole-cut meat analog that the strands do not cross.

[0035] In some embodiments, according to any of the methods of making a plant-based meat analog described above, the tension in the whole-cut meat analog along a direction orthogonal to the average strand alignment is at least about 10% of the tension in the whole-cut meat analog along the average strand alignment. In some embodiments, the tension in the whole-cut meat analog along a direction orthogonal to the average strand alignment is no greater than about 90% of the tension in the whole-cut meat analog along the average strand alignment.

[0036] Also provided herein is a plant-based meat analog prepared according to any of the methods described above. For example, in some embodiments, provided herein is a plant-based meat analog comprising strands made from a wet protein mixture that range from about 0.1 mm to about 4 mm, the strands comprising i) texturized protein (TP), and ii) one or more of a plant protein powder, a binder, a coloring agent, a flavor agent and/or a spice, a liquid, and a leavening agent, the strands being aggregated into a desired alignment and stabilized to form a coherent strand mass resembling meat in appearance and texture.

BRIEF DESCRIPTION OF THE DRAWINGS

[0037] FIGS. 1A and IB depict an exemplary eye round roast. Butcher shop beef round (Giant

Supermarket house brand). FIG. 1A: surface, FIG. IB: cross-section. There is an element of randomness in both the length of the visual fiber bundles before being disrupted by layers of fat or connective tissue, and the orientation of the various fiber bundles relative to each other. This randomness makes the appearance “natural”.

[0038] FIGS. 2A-2F show examples of 3d printed meat analogs (see Example 1 and Example 2). FIG. 2A shows firm muscle phase separated by soft fat phase, fully aligned in parallel. FIG. 2B shows firm muscle phase separated by soft fat phase. Every second layer angled at 10 degrees to create crosslinks between overlapping muscle phase strands. FIG. 2C shows core-shell nozzle arrangement. FIG. 2D shows coaxial printing of core (firm muscle phase) and shell (alginate phase). 2% Ca-lactate in firm muscle phase. Alginate shell gelled by calcium ions diffusing from core to shell. FIG. 2E shows coaxial printing of core (firm muscle phase) and shell (alginate phase). 1% Ca- lactate in firm muscle phase. Alginate shell insufficiently gelled by calcium ions. FIG. 2F shows coaxial printing of core (firm muscle phase) and shell (alginate phase). No Ca-lactate in firm muscle phase. Alginate shell not gelled by calcium ions, mushy sample.

[0039] FIGS. 3A-3C show examples of full-sized roast analogs (sliced). FIG 3A shows a full-sized roast with aligned small lengthscale fibrous inclusions, but no large lengthscale strands (steak 2.0, example 3); FIGS. 3B and 3C show full-sized roast analogs with semi-aligned strands (FIG. 3B: example 4, FIG. 3C: example 6). The clearest visual difference is from strands. In FIG. 3A, there are no strands, only embedded small lengthscale fibers, upon breaking the steak looks too random (burger-like); in FIG. 3B, the cut strands are cut too large (about 3 mm), while in FIG. 3C, the strands were cut to 2 mm. Additionally, small lengthscale inclusions (fibers) and juiciness were improved in FIG. 3C (not visible).

[0040] FIGS. 4A and 4B show steps in a process for roast analog formation. FIG. 4A shows semialigned strands before rolling up into roast (as in examples 4 and 6), FIG. 4B shows a rolled-up roast. The strands in FIG. 4 A are aligned on average left to right (direction of arrow). A natural looking surface of the roast results as shown in FIG. 4B after rolling from front to back (2^nd arrow). The slices look realistically aligned and natural (see FIGS. 3B and 3C).

[0041] FIGS. 5A and 5B show the firm phase of example 6. FIG. 5 A shows embedded fibers (small lengthscale inclusions) marked (visible after bending of firm phase sheet), FIG. 5B shows pores marked (a stack of firm phase sheets was rolled up, individual sheets are about 2 mm high).

[0042] FIGS. 6A and 6B show a schematic drawing of aligned (FIG. 6A) vs. randomly embedded (FIG. 6B) small fibrous inclusions in strands. Aligned as in FIG. 6A is expected to provide maximum strength in fiber direction, but weakness if pulled against fiber direction, thus maximum anisotropy. Randomly embedded small fibrous inclusions provide more cohesion in any direction. This latter situation was found to be desirable: anisotropy of the steaks is generated by the semialigned strands, chewiness by the randomly embedded fibers.

DETAILED DESCRIPTION OF THE INVENTION

[0043] The present disclosure describes plant-based meat analogs and methods of producing the same. This includes meat analogs that resemble to some degree whole cuts of meat, such as beefsteak, in terms of appearance, texture (presence of anisotropy, chewiness, juiciness), visible structure, taste, flavor, and/or aroma. The possibility that these meat analogs undergo changes similar to real steak during the final cooking step is included; for example, a color change from red to brown, a texture change towards firmer in compression, the release of roasted aroma notes, the disappearance of white fat tissue, and/or the release of liquid fat.

[0044] Unless described otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. Whenever appropriate, terms used in the singular will also include the plural and vice versa. All patents, applications, published applications, and other publications are incorporated by reference in their entirety.

[0045] As used herein, the singular terms “a,” “an,” and “the” include the plural reference unless the context clearly indicates otherwise.

[0046] As explained above, the present disclosure has many advantages over conventional methods and products. Some existing attempts to reproduce whole cuts include making of a protein dough sheet, cutting it into strands, and aggregating those. While this approach can generate aligned strands, further sub-structures and the opportunity to use those sub-structures to fine time texture are missing. In addition, this technology is not tailored to plant-based products, but instead is designed to be used with protein ingredients derived from animals, like egg white and gelatin.

[0047] Other existing attempts include forming monolayers from essentially parallelly aligned elongate texturized vegetable protein (TVP) strands to produce, e.g., an alternative whole muscle cut. While texture of TVP or protein from high moisture extrusion (HME) can be adjusted in the extrusion process, the possibilities to design finer sub-structures by extrusion are limited.

Additionally, the technology produces a product with perfect alignment of unidirectional continuous fibers arranged in a periodic layered structure. This results in a visual appearance that does not resemble a whole-cut from an animal. In a whole-cut meat piece, there is an element of randomness in both the length of the visual fiber bundles before being disrupted by layers of fat or connective tissue, and the orientation of the various fiber bundles relative to each other. This randomness makes the appearance ‘natural’ see FIGS. 1A and IB), as is the case with the current disclosure. [0048] When it comes to sub-structures, other existing attempts include digitally printed strands, which contain bundles of axially aligned elongated textured protein fibers. While this creates substructures similar to real meat, directly mimicking real muscle structure with plant-based proteins and ingredients is likely not the optimal way to achieve a plant-based steak with comparable texture and organoleptic perception to an animal steak. For example, muscle fiber is known to be hierarchical and fibrous over multiple lengthscales, with the fibers having a high degree of alignment. However, it is not clear that aligned fibrous substructures is necessary or even desirable in a plant-based steak.

[0049] An additional aspect of the current disclosure concerns the heating protocol. In an ideal steak analog, tough, muscle fiber-like strands would be present already before the final cooking (pan searing) step by the end user (consumer or restaurant chef). Additionally, this steak would undergo a change in texture during the final cooking step similar to real meat when cooked from raw, becoming firmer in compression and drier with increasing cook time. In existing attempts, there is sometimes a final heating (stabilizing) step in the main production process, and additional optional heating steps during sheeting, and after cutting but before final stabilization, to toughen the fibers. Some existing work also can include an optional final thermal treatment after the multilayer product has been produced, and indirectly, the use of TVP or HME protein indicates that heating was used during the extrusion process when making these texturized materials. The current disclosure instead tailors heating conditions in the process to the properties of the used plant protein types, and by focusing on both, raw and cooked texture of the steak, providing an advantage over the existing attempts.

Analogs of tissue and other components

[0050] In order to mimic the texture and broader organoleptic properties of animal whole cuts, plant-based analogs for the various animal tissues can be used. These include muscle tissue engineered on two lengthscales (as described in detail below under ‘Muscle phase analogs and their arrangement into a roast’), fat (adipose) tissue, and connective tissue. Additionally, a plant-based analog for meat juice can be included to provide juiciness, color and/or flavor. In some aspects, a muscle tissue analog as described herein has anisotropy through orientation of strands and/or inclusions.

[0051] In some aspects, a connective tissue analog as described herein includes connecting or binding individual muscle analog strands together. This can be achieved through vital wheat gluten, as outlined in examples 4 and 6, but also a range of other proteins and/or hydrocolloids, for example soy protein isolate, pea protein isolate, sodium alginate plus calcium salts and others. Cohesion of proteins used as connective tissue analog can be improved by cross-linking enzymes like transglutaminase. In an additional aspect, the binder can be a protein powder that is at least partially raw and still functional, and firms up during the final cooking by the end user, allowing some control of the doneness level. This is the case, for example, for vital wheat gluten.

[0052] Adipose tissue aims at mimicking the white marbling inside and on the surface of raw animal steak; the fat release that occurs when cooking animal steak; and the transition of the raw white adipose tissue to a final, translucent, cooked adipose tissue. A wide range of lipids can be used, including solid plant based fats and/or liquid plant based oils, for example coconut oil, cocoa butter, palm oil, palm kernel oil, shea butter, sal butter, hydrogenated fats (solid), canola oil, soybean oil, sunflower oil, and/or safflower oil (liquid), optionally also including pre-made shortening or margarine. Such fats and oils can be mixed to create the desired firmness and melting properties of the resulting mixture. Adipose tissues can include other components besides fats and oils, for example proteins and hydrocolloids. Adipose tissue analogues can be emulsions, and proteins, hydrocolloids and/or polar lipids can contribute to stabilize these emulsions. Hydrocolloids (for example, methylcellulose) and/or proteins (for example, vital wheat gluten, soy protein, pea protein or potato protein) can also help create non-lipid structures that remain in the meat analog after the fat has rendered in the final cooking step.

[0053] The properties of the plant-based muscle analog, the plant-based adipose tissue, and the plant-based connective tissue may be tuned independently to best mimic different cuts and types of animal whole cuts. In addition, the composition and proportion of the different plant-based tissue analogs within a plant-based steak may also be adjusted to best mimic different cuts and types of animal whole cuts. As an example, in a plant-based steak, the muscle phase analog can be about 40- 45% by weight, connective tissue analog (dry) about 15%, adipose tissue analog about 10-20%, with the rest being meat juice analog. Furthermore, the adipose tissue can be varied, for example between 0 and 30% by weight, to imitate leaner versus fattier cuts.

Muscle phase analogs and their arrangement into a roast

[0054] In some embodiments of the meat analogs described herein, firm muscle phase strands are the largest elongated units, with diameters of less than one mm to several mm, and may contain inclusions as part of their sub-structure. In some embodiments, these small inclusions are embedded in a dough-like matrix together with additional selected plant protein powders, binders, colors, flavors and spices, liquids (water, liquid oils), leaveners and/or pH adjusters. In some embodiments, this dough-like matrix is heat-set and processed into strands. In some embodiments, the heating process takes place at ambient pressure. The dough may also be set via other means than heat, for example, by cross-linking with an enzyme reaction. The dough-like matrix may be processed into strands before or after heat-setting; for example, by sheeting of the raw dough to a suitable thickness (for example between 0.1 to 4 mm), heating it and then slicing the heat-set sheets into strands (for example, 0.1 mm to 4 mm wide), by extruding the raw dough into strands (for example, with a cross-sectional lengthscale of 0.1 mm to 4 mm) and then heat-setting the extruded strands, or by extruding and heat-setting the raw dough into strands in a single step through a heated nozzle or extrusion die. A wide range of heating conditions is possible for heat setting. In principle, only temperatures above coagulation/gelation point of the proteins used are required, and if desired also high enough to reach a microbiological kill step, e.g., above 74°C (165°F). For heating following sheeting, oven temperature settings may be higher, for example 135°C (275°F), 160°C (320°F), 177°C (350°F), or 260°C (500°F) for any time that leads to the required setting temperature inside the sheet, for example 7 min, 8 min, 12 min, 20 min, or longer as needed. Furthermore, for those skilled in the art it is clear that the required time and temperature settings also interact with other oven conditions like presence/absence of forced convection, or air speed in forced convection, and that such settings can be different between household ovens and commercial ovens or different oven models. The listed examples in this disclosure give some examples of suitable time/temperature combinations.

[0055] The firm muscle phase strands can be further soaked in a plant-based meat juice analog to increase juiciness, boost flavor and modify color. It can be favorable to create porosity in the strands to increase the absorption of liquid. Methods to create porosity may include leaveners releasing carbon dioxide like baking powder or baking soda, steam created during heating, physical incorporation and expansion of gases through pressure and vacuum cycles, or adding of supercritical carbon dioxide. These gases (carbon dioxide, steam etc.) may be trapped as bubbles in the matrix, followed by setting of the matrix. This is known to those skilled in the art from producing leavened baked goods (see also example 5). In some embodiments, after soaking the optionally porous strands in a plant-based meat juice, those strands are arranged ‘semi-aligned’ with respect to a principal axis together with a binder. “Semi-aligned” as used herein means that not all strands are exactly parallel to the principal axis, but that instead an element of randomness is included, for example through arranging the strands such that not all of the strands are aligned along a principal axis. The strand orientation is measured with respect to the principal axis and can take values of -180 degrees to 180 degrees. In some embodiments, the average orientation of all the strands will be 0 degrees, indicating that the principal axis represents the average orientation of all the strands. The semialigned nature of the strands is quantified by the degree of alignment (doa). For a set of strand orientations having an average orientation of 0, the doa is calculated by taking the average of cos(2X) over all the strands, where cos() represents the cosine function and X represents the orientation of a strand. Arranging all the strands exactly parallel to the principal axis results in a doa of 1.0000. In some embodiments, the semi-aligned nature of strands results in a doa between 0.9965 and 0.7500. Semi-aligned strands were found to be favorable for appearance, cohesion orthogonal to the main direction of the strands, and overall meat-like chewiness. In some embodiments, additional orthogonal cohesion is achieved by a connective tissue analog (binder) described herein, for example vital wheat gluten. Additionally, an adipose tissue analog can be incorporated between the strands to generate a marbling effect. Soaked strands, binder and fat can be formed into a ‘roast’, simulating whole muscles (similar to a ribeye roll subprimal or a tenderloin subprimal), and then sliced into individual steaks or ‘fillets’. The roast can be formed and the shape stabilized by rolling, pressing, and/ or otherwise compacting the semi-aligned strands, binder, and adipose tissue along a direction perpendicular to the principal axis, for example, to yield a final cylindrically-shaped roast where the principal axis and the cylinder axis are parallel. The term “stabilization” is used in the present disclosure to describe any technique that mechanically stabilizes the final shape of the roast. Besides the described rolling and/or pressing, compacting and thus stabilization can be also achieved in some embodiments by placing the roast in a vessel (e.g., a plastic bag) and subjecting the vessel to vacuum (e.g., -0.97 bar relative to ambient pressure), so that gas pockets are removed and the vessel exerts pressure from all sides on the roast. Chemical and/or enzymatic crosslinking can contribute to the stabilization (e.g., by crosslinking the proteins via transglutaminase). Additionally, controlled heating can induce crosslinking and thus stabilization, while cooling (for example to 5 °C) can solidify the fats added to adipose tissue and/or other tissue analogs and thus stabilize the shape and structure of the roast. Inclusions from texturized proteins (TP)

[0056] In some embodiments of the meat analogs described herein, much of the texture of the muscle phase analog and of the overall steak is determined by the inclusions embedded in the strands (see example 5). Inclusions are small (10 pm lengthscale to 1 mm lengthscale) elements of plant proteins or other plant-based material that act as texture modifiers. These inclusions can be produced by shredding or mincing hydrated texturized proteins. Isotropic inclusions can be produced by mincing the texturized proteins, while anisotropic, fibrous inclusions can be produced by shredding inherently fibrous texturized proteins (e.g. SuproMax 5050) using a blunted blade or a typical kitchen processor with the blade spinning in reverse, so that the material is hit primarily by the blunt backward side of the blade and more tom than minced. The inclusions are, on average, smaller in diameter than the firm phase strands.

[0057] Textured or texturized proteins are plant proteins or mixtures of plant proteins that have been processed in ways known to the art to produce products with the common names, “textured vegetable protein”, “texturized protein”, “structured vegetable protein”, “structured plant protein”, “extrudate”, “low-moisture extrudate”, “high-moisture extrudate”. Common examples of processes for texturization include low moisture extrusion (10-40% water, producing expanded dry products requiring rehydration), high moisture extrusion (40-70% water, requiring controlled cooling to prevent water flashing to steam, and producing anisotropic fibrous products). Common principles of low and high moisture extrusion include high temperature (e.g. >130°C), pressures clearly above ambient pressure, and high mechanical shear, to fully denature the plant proteins and then set them in the desired structure.

[0058] Other methods to produce texturized proteins and inclusions may include shear cell technology (Krintiras, Georgios A., et al. "On the use of the Couette Cell technology for large scale production of textured soy-based meat replacers." Journal of Food Engineering 169 (2016): 205- 213), freeze alignment, wet spinning and electrospinning, fermentation based production of microbial (e.g., fungal) biomass, or any other methods known in the art to produce plant-based edible texturized materials, and grinding them into inclusions of the appropriate shape and size scale.

[0059] Raw materials for texturized proteins may include wheat gluten, soy protein isolate, soy protein concentrate, soy flour, pea protein isolate, pea protein concentrate, pea flour, faba bean protein isolate, faba bean protein concentrate, faba bean flour, chickpea protein isolate, chickpea protein concentrate, chickpea flour, as well as protein isolates, concentrates and flours from other cereal grains like barley, rye, oats, and so on. Texturized proteins may be made from a single, or a combination of multiple of these raw materials, and may also include additional ingredients, for example food starches (e.g., wheat starch, corn starch, tapioca starch, potato starch, rice starch). Commercial examples include texturized soy flour (e.g., granular TVP Purelynature 165-118 by ADM), texturized soy protein concentrate (e.g., Arcon T 158-171 flakes by ADM, Response 4310, Response 4380, Response 4400, Response 4410, Response 4438 by Solae/IFF), fibrous texturized wheat (gluten) proteins (e.g., ProTerra 1100, 1200 and 1350 by MGP Ingredients), texturized pea proteins (e.g., ProTerra 2200 and 2350 by MGP Ingredients, TPP70 and TTP80 by Puris), and mixtures of soy and wheat resulting in expanded, fibrous texturized proteins or structured vegetable proteins (e.g., SuproMax 5010, SuproMax 5050 by Solae/IFF from soy protein isolate, wheat gluten, wheat starch).

Protein powders, binders, color, flavors

[0060] In some embodiments of the meat analogs described herein, plant protein powders include vital wheat gluten, soy protein isolate, soy protein concentrate, soy flour, pea protein isolate, pea protein concentrate, pea flour, faba bean protein isolate, faba bean protein concentrate, faba bean flour, chickpea protein isolate, chickpea protein concentrate, chickpea flour, protein isolates, concentrates and flours from other cereal grains like barley, rye, oats, protein isolates and concentrates from potatoes, pseudocereals (amaranth, quinoa, buckwheat), protein isolates and concentrates from leaves (e.g., RuBisCO), protein isolates from microorganisms (e.g., yeasts, bacteria, fungi). Examples include vital wheat gluten (e.g. Gem of the West by Manildra), potato protein isolate (e.g., Solanic 200 by Avebe) and pea protein isolate (e.g., Vitessence Pulse 1853 by Ingredion). Plant protein powders can differ widely in their solubility, denaturation temperature, gelation, isoelectric point, purity, off flavors, cost and commercial availability. Relevant examples for the current disclosure include potato protein isolate and vital wheat gluten. The commercially available potato protein isolate Solanic 200 by Avebe is rich in the patatin fraction of potato protein, which is soluble in water and coagulates, at neutral pH, in the 50-60°C range, forming a strong gel (Creusot, Nathalie, et al. “Rheological properties of patatin gels compared with p-lactoglobulin, ovalbumin, and glycinin.” Journal of the Science of Food and Agriculture 91.2 (2011): 253-261; Alting, A. C., et al. “Potato proteins.” Handbook of food proteins. Woodhead Publishing, 2011. 316- 334). Vital wheat gluten is predominantly the gliadin and glutenin fraction of wheat, isolated and dried gently enough that functionality is maintained. Gluten is largely insoluble in water, and instead aggregates into a viscoelastic, dough-like mass, which ultimately firms up (sets) upon heating. Because gluten proteins cover a wide range of molecular weights and physicochemical properties, there is not one single specific setting temperature, but generally, setting occurs higher than for patatin, with major changes in functionality occurring between 55 and 75 °C, major solubility changes in SDS solutions at 90°C and gliadins even requiring temperatures above 100°C for polymerization (Schofield, J. D., et al. “The effect of heat on wheat gluten and the involvement of sulphydryl-disulphide interchange reactions.” Journal of Cereal Science 1.4 (1983): 241-253; Lavelli, Vera, Nicoletta Guerrieri, and Paolo Cerletti. “Controlled reduction study of modifications induced by gradual heating in gluten proteins.” Journal of Agricultural and Food Chemistry 44.9 (1996): 2549-2555; Singh, H., and F. MacRitchie. “Changes in proteins induced by heating gluten dispersions at high temperature.” Journal of Cereal Science 39.2 (2004): 297-301).

[0061] The term “additional binders” is taken to mean ingredients other than plant protein powders that may be added to the matrix to improve cohesion and allow for sheeting and forming before heat setting. These can be natural and at least partly soluble fibers from, e.g., citrus peel, citrus pulp, carrot, pea hull, inner cell walls of pea, apple pomace, Psyllium hull, oat fiber, flaxseed.

Alternatively, isolated and potentially chemically modified hydrocolloids can be used, including, for example, xanthan gum, guar gum, alginate, high methoxyl and low methoxyl pectin, sodium alginate, different types of carrageenan, methylcellulose, and hydroxypropyl methylcellulose. Methylcellulose is unique because it forms strong, reversible, heat-induced gels (thermoreversible gelation), and because it is also an emulsifier. It is also used in some of the subsequent examples to emulsify and stabilize the fat phase (adipose tissue), together with some vital wheat gluten. Sodium alginate forms strong gels with divalent ions like calcium via so-called ‘egg-box junctions’. Calcium chloride reacts fastest, while calcium lactate reacts slower but has less off flavor issues (Lee, P., and M. A. Rogers. "Effect of calcium source and exposure-time on basic caviar spherification using sodium alginate." International Journal of Gastronomy and Food Science 1.2 (2012): 96-100). [0062] Colors include natural or synthetic food colors. Natural colors are sometimes preferred for labeling reasons and can be based on fruit and vegetable juice concentrates (e.g., red tones from beetroot, radish, orange carrot or purple carrot), brown tones from apple with other plant extracts for fine tuning. Those skilled in the art will aim to mix different colors to obtain the desired raw meat color (reddish-brown tones) at the given pH, and also consider heat stability to obtain a color change to brown upon heating, similar to real meat. For example, betacyanin in beetroot is known to be fairly instable upon heating, so that its reddish purple color may fade upon heating, while the anthocyanins in radish are more heat stable, and may be more suitable when a more permanent red color is needed to get the appearance of rare steak.

[0063] Flavors, for the purpose of this disclosure, include low notes (e.g., salt and umami), middle notes (including reaction flavors, generated through Maillard reaction, e.g., from cysteine, glutamate, and reducing sugars like ribose, or from more complex mixtures), and top notes (volatile components, for example volatile fat degradation products, smokey and grilled notes, or volatiles from spices). Yeast extracts and autolysates can provide both basic umami from glutamate, ribonucleotides, some peptides and also more specific desirable middle notes. Top notes, for example smokey, grilled and fatty notes can be purchased from specialized flavor houses. Furthermore, spices can be added as in the preparation of real meat, for example paprika powder, chili powder, black pepper, granulated onion, granulated garlic, dried rosemary, thyme, and/or oregano leaves. For the purpose of plant-based meat, flavors preferably avoid animal-based ingredients like animal fat.

Texture Analysis Methods

[0064] Food texture may be defined as physical properties of a food sensed by touch with either the hands or in mouth. Some aspects of it can be objectively measured by instrumental methods, using a texture analyzer. In the widest sense, a texture analyzer measures the relation between force, distance and time, using specific probes or fixtures interacting with the food. More practically, depending on the probe or fixture, the texture analyzer can measure food under conditions of, for example, compression, extension, cutting, bending and shearing. Tests may be conducted until fracture or to a certain, defined force or deformation. As a demonstration, example 7 shows tests with strips of firm muscle phase in tension comparing the way of alignment of fibrous inclusions. The test was conducted until fracture, and maximum force, distance at this force and area under the resulting curve recorded.

[0065] A different, cutting type test was conducted with a rectangular blunt blade in compression, which penetrated the firm muscle phase sheets on a flat surface to 80% strain. The maximum force was extracted from the results.

[0066] A third type of test, texture profile analysis (TPA, also called two-bite test) was conducted with cylindrical cuts from slices of real steak and plant based steak analog for comparison of, among others, firmness (hardness), springiness, and cohesiveness. The sample was compressed to 60% strain with a plate larger than the sample on a flat surface. The plate returned to its start point before, after a waiting period of 5 sec, starting a second compression cycle. Firmness (hardness) is defined as the maximum peak force during the first compression cycle (first bite). Springiness describes how much the height of the food recovers between the end of the first bite and start of the second bite (with 0 being no recovery and 1 being ideal recovery). Cohesiveness is defined as the ratio of the positive force area during the second compression to that during the first compression. Other parameters that can be extracted from the TPA are, for example, adhesiveness, chewiness, and/or resilience.

[0067] Additional measurements related to sensory of plant-based steak are cooking yield (mass ratio after cooking to before cooking of the plant based steak), which can also be calculated as % loss (cooking loss). This method captures losses from evaporation and from release of fat and other substances into the pan.

[0068] Juiciness perception is complex, but in a simplified way can be measured by compressing a defined piece of plant-based steak, capturing the released liquid, and if desired, differentiating between aqueous phase and fat phase in the released liquid.

Visualization and structural measurements

[0069] Visualization techniques that enable three-dimensional structural measurements are common in the food industry. Techniques like Micro-CT, or X-ray CT can construct a 3D image of the internal structure by scanning a sample with Xrays. Practically speaking, the images can resolve local density differences down to lengthscales of 1 pm. These techniques enable quantifying the semi-aligned nature of the firm phase strands in the final roast or fillet.

EXAMPLES

[0070] The specifics of the disclosure are illustrated through a series of examples following the development of a plant-based steak designed to mimic high-value beef whole cuts such as a filet mignon or ribeye steak. High-level summary of the examples

[0071] Example 1 demonstrates that some crosslinking generated through angled 3d printing is desirable over perfect parallel alignment. While anisotropy is maximal without crosslinking, the cohesion and chewiness increase with crosslinks, and such samples feel more meat-like.

[0072] Example 2 shows that a Ca-gelled alginate shell around a protein core generated by 3d printing can imitate connective tissue surrounding a muscle strand. However, high and practically infeasible calcium concentrations are needed to gel the alginate and create some strength orthogonal to fiber direction. The principle of strength orthogonal to fiber direction is similar to the importance of crosslinks in example 1.

[0073] Example 3 shows that by embedding aligned smallscale fibrous inclusions from shredded textured protein in a full sized roast, a chewy product can be generated. However, in the absence of large strands, the product appears more burger or sausage like despite the aligned small fibers. The meat juice analog does not penetrate well in the fillets, because the fillets are very compact, limiting juiciness.

[0074] Example 4 demonstrates a clear visual improvement relative to example 3, when strands with embedded inclusions from minced textured proteins are made and roughly aligned. To better mimic a juicy, high-value whole cut, juiciness, chewiness and strand size should be further improved. The next goal is to identify the best suited inclusions to embed in the strands, soak the strands longer for more juiciness, and decrease size of the cross-section of the strands.

[0075] Example 5 compares different inclusions to embed in the strands to improve juiciness and chewiness of the strands. Among the candidates tested in this example, small fiber-like inclusions from shredding SuproMax 5050 yielded the best results. Preparing firm muscle phase strands with these inclusions resulted in firm and chewy, meat-like, fibrous texture. These strands absorbed considerable amounts of liquid (over 70% meat juice analog if swollen overnight, and if baking soda was added to create pores to physically hold more liquid).

[0076] Example 6 applies the improved strands from example 5 to produce a plant-based steak that has improved chewiness, juiciness, and a decreased strand size, all measured with respect to the plant-based steak in example 4.

[0077] Example 7 shows that aligning fibrous inclusions mainly weakened the firm phase orthogonal to fiber direction, while strength in fiber direction was similar to not aligned fibrous inclusions. We conclude that not aligning the embedded fibrous inclusions produces an acceptable texture and is simpler to do in production. The working hypothesis is that the roughly aligned strands in the steaks provide anisotropy, while embedded fibrous inclusions strengthen the firm phase strands, provide cohesion and thus chewiness. This is illustrated in FIGS. 6A and 6B.

[0078] Example 8 demonstrates that potato protein isolate (Solanic 200) can be omitted from the dough for the firm phase and replaced with extra vital wheat gluten. Because of the higher setting temperature of gluten relative to Solanic 200, the firm phase without potato protein needs to be heated longer. More generally, the texture of the firm phase can be further tailored by adjusting the heating conditions in the setting step to the properties of the proteins used. In an even wider sense, the steak in Table 6 involves a range of different heating conditions providing a hierarchy of firmness, where the firmness decreases as the lengthscale of the structure increases:

• The small fibrous inclusions are from shredded TVP, commercially made by extrusion (very firm and chewy, heating in extrusion » 100 °C).

• The large strands are baked in a convection oven until the sheet reaches 90-100°C, and these heating conditions can be tailored to the proteins used, and to achieve the desired toughness of the strands.

• The binder mix (essentially raw gluten) is not heated before cooking of the steak by the end user (target there 74°C core temperature), and is thus least set, least firm and allows to control the steak texture (‘doneness’) by cooking duration.

Example 1: A 3d printed meat analog, printed in different angles

[0079] This example demonstrates the effect of perfect alignment versus angled arrangements as a means of creating crosslinks. Firm phase strands (a cohesive mass high in protein and low in fat) are separated by soft phase strands higher in fat and lower in protein (Tables la and lb), thus separating the firm phase strands. 3d printing allows for near perfect control of strand arrangement (FIGS. 2A- 2F).

[0080] The aligned samples (FIG. 2A) felt highly anisotropic, the firm muscle phase strands provide stability in fiber direction, but easily separate when pulled against fiber direction. The mouth feel is more similar to “pasta and sauce” than meat.

[0081] An angled sample with the even layers printed at 90 degrees is the alternate extreme. This sample feels anisotropic along the layer direction but separates far less easily than the aligned sample when pulled in the plain of the layer. This is due to crosslinks created by the overlapping firm muscle phase strands. The mouth feel is much more “burger” like and firmer.

[0082] Angled samples printed at 5 degrees and 10 degrees (FIG. 2B) still feel anisotropic while being firmer and more cohesive and chewier than the aligned samples. The 5 degree and 10 degree sample are distinguishable in the mouth and in the hand, with the 5 degree sample being more anisotropic and the 10 degree sample being more firm and cross-linked. Both the 5 degree and 10 degree sample are more meat-like than the aligned sample and both the 5 degree and 10 degree sample feel more like a whole cut in the mouth than the 90 degree sample. In this example, the 5 degree sample has a doa of 0.9962, the 10 degree sample has a doa of 0.9848, and the 90 degree sample has a doa of 0.0000.

Table la: Firm muscle phase analog

[0083] Mix with handheld immersion blender

Table lb: Soft fat phase (adipose tissue analog)

[0084] Mix with handheld immersion blender (water, color, proteins first, add oil last until stable emulsion results)

Procedure [0085] Load both, muscle phase and fat phase in syringes, degas by centrifugation (5000 rpm, 9 min)

[0086] Configure Hyrel Engine SR Printer to hold 30 ml syringes and to trigger pressure pumps (Nordson EFD Ultimus V) on command. Load Geode onto the printer, with G7 and G8 commands set to trigger the pressure pumps. Mount syringes to the printer with 20 GA (0.58 mm) dispensing tips (Nordson EFD) on each syringe and connect the syringe containing the firm muscle phase to a pressure pump set to 43 psi and the syringe containing the soft fat phase to a pressure pump set to 17 psi. Affix aluminum foil to the buildplate with tape and run the Geode to print the sample. For an aligned sample, the Geode is designed to produce a sample where each layer consists of alternating parallel strands of the firm muscle phase and soft fat phase. In addition, each subsequent layer switches the material that is first printed. The final sample thus has a “striped” appearance when viewed along a plane whose normal vector is orthogonal to the parallel strands and has a “checkerboard” appearance when viewed along a plane whose normal vector is aligned with the parallel strands. As a concrete example, let the first 5 parallel strands in the odd layers have the composition: firm muscle phase, soft fat phase, firm muscle phase, soft fat phase, firm muscle phase. In this example the first 5 parallel strands in the even layers have the composition: soft fat phase, firm muscle phase, soft fat phase, firm muscle phase, soft fat phase. The size of the sample is thus determined by the length of the parallel strands (setting the length of the sample) the number of parallel strands and with diameter of the strands (setting the width of the sample) and the number of layers and height of each layer (setting the height of the sample). Once the print is finished, pick up printed samples on aluminum foil carefully, place the sample within a plastic frame, seal in vacuum bag, heat to set proteins (sous vide bath set at 74°C/165°F for 20 min). Conduct a sensory evaluation.

[0087] To produce an angled sample, let the print direction (corresponding to the orientation of the strands) in the first layer (and every odd layer) be 0 degrees. Then, set the print direction of the even layers to an angle. Within this framework, the “aligned” sample has the even layers set to 0 degrees.

Example 2: A 3d printed meat analog, with coaxial connective tissue analog from alginate gelled by calcium [0088] This example demonstrates the approach of imitating a muscle strand surrounded by connective tissue via coaxial (core-shell) printing (FIGS. 2C and 2D). The calcium ions in the core gelled the alginate in the outer shell by diffusing from core to shell. A translucent, firm, elastic ‘alginate hose’ resulted that connected the stacked strands, and separated the firm muscle phase cores, similar to connective tissue (endomysium) around muscle fibers. However, the calcium also caused a bitter off flavor and made the firm phase gritty. Reducing the calcium concentration was not successful: at 1% Ca-lactate (FIG. 2E), the grittiness and bitterness persisted, while alginate gelling was already clearly reduced. Without calcium, as expected, the alginate did not gel and the sample was mushy (FIG. 2F). This example demonstrated once again the importance of some strength orthogonal to fiber direction (via Ca-alginate gel), even though the major strength has to be in fiber direction to create anisotropy. The example also shows that closest imitation of the situation in real meat (connective tissue around muscle strand) does not automatically create the best texture.

Table 2a: Firm muscle phase analog (‘core’)

[0089] Mix with handheld immersion blender; variations included reducing Ca-lactate to 1% and 0.

Table 2b: Connective tissue analog (outer shell, alginate phase)

Table 2c: Meat juice analog

[0090] Firm phase analog and connective tissue analog were printed simultaneously through a coaxial needle (see FIG. 2C; Rame-Hart, Succasunna, NJ, USA, 100-10-COAXIAL-2016, core diameter 0.584 mm, shell diameter 1.19 mm; shortened to 15 mm needle length to reduce pressure). A parallel stack of this coaxial arrangement was printed using a Hyrel Engine SR 3D Printer configured to hold two syringes connected to pressure pumps that can be triggered by the 3D Printer. The printing pressure was 66 psi for core and 69 psi for outer shell (connective tissue). The sample was printed using Geode designed to produce an aligned sample (see example 1) and then heat-set (sous vide bath set at 74°C/165°F for 20 min). The sample was then soaked in imitation meat juice analog for 20 min under vacuum, seared and tasted.

Example 3: full sized roast with aligned small scale fibrous inclusions (steak 2.0)

[0091] This is an example of a full-sized roast (FIG. 3A). Fibrous inclusions are created through shredding of texturized proteins (mostly texturized wheat protein) and these inclusions are embedded in a matrix, in which vital wheat gluten, supported by potato protein isolate (patatin rich fraction Solanic 200), provide the heat setting.

[0092] The firm phase analog dough was printed through a large, 6 mm, nozzle to provide basic alignment of the fibrous inclusions. This diameter is large enough to not clog from the fibrous inclusions. The embedded fibrous inclusions do provide a chewy bite, chewier than the previous, 3d printed samples. However, the individual printed 6 mm strands merge back together when forming the roast. Anisotropy only originates from the aligned embedded fibers, large lengthscale strands are absent. The appearance of the product is more burger or sausage like, rather than fibrous in the sense of a whole-cut (FIG. 3A). Other than aligning embedded fibers, the print process allows for distribution of fat in the firm matrix. A clear flaw is that the meat juice analog does not penetrate well into the steak slices / fillets because of the compact structure. Consequently, the product is not very juicy, and we are unable to tune the flavor and color of the product using the meat juice analog.

Table 3a: Firm muscle phase analog

[0093] Soak ProTerra in excess water for 20-40 min, then squeeze out excess water. For coarse ground, grind for 20 sec in Thermomix TM5 on level 6 with the blades spinning in reverse. For fine ground, grind for 20 sec in Thermomix TM5 on level 10 with blades spinning in reverse. Mix canola oil, colors, salt, sodium bicarbonate and water. Premix all remaining dry ingredients and mix them with ground textured proteins and liquids in a KitchenAid mixer with batter blade for 1 min at lowest speed, followed by 2 min at medium speed. Table 3b: Adipose tissue analog

[0094] Melt cocoa butter, mix everything in Kitchen blender.

Table 3c: Meat juice analog

Assembly:

[0095] Sample printed using a custom 3D printer configured to print material through custom printheads. Material is loaded into custom 2 kg hoppers and extruded using a computer-controlled piston whose feed rate is linked to the diameter of the nozzle and the speed of the printhead. In this example, the diameter of the final outlet nozzle was 6 mm, the diameter of the coextrusion inlet (for the adipose tissue analog) was 2 mm, and the printhead has a linear max speed of 20 cm/sec. The ratio of firm phase dough to adipose tissue analog in the extruded material was 3.57:1.00. The Geode was configured to print the material layer-by-layer in a continuous serpentine pattern, forming a block of material 12 cm long by 20 cm wide by 3.2 cm high. The material was printed onto a standard sheet pan lined with aluminum foil placed on the printing platform. [0096] Once the material was printed, the pan was removed from under the printer, the material was rolled into a cylinder 120 mm long, and the cylinder was vacuum sealed in a plastic bag. The sealed cylinder was heated in a water bath held at 85°C/185°F for 140 minutes, stabilizing the cylinder.

[0097] Post heating: The cylinder was sliced into fillets (2 cm thick). To maximize meat juice penetration, each fillet was first soaked in meat juice analog for 2 hr, and then placed in a vacuum bag with 5% additional meat juice analog and vacuum sealed.

Example 4: Process for making a full sized meat roast with semi aligned strands

[0098] This example demonstrates the making of strands with embedded texturized soy (granular TVP 165-118) with a smaller amount of texturized wheat protein. The ‘semi alignment’ is achieved as shown in FIG. 4A. The resulting steaks show clear visible strands in the 3 mm range (FIG. 3B). However, the steaks are fairly dry because the meat juice analog has insufficient time to penetrate into the strands. Because of the high level of granular texturized soy flour (TVP 165-118) and fine grinding, after the initial bite the steak breaks down quickly, meaning it lacks chewiness relative to a high-value whole-cut. Finally, the 3 mm strands appear too large for a high-value whole cut.

Table 4a: Firm muscle phase analog [0099] Mix water, color, salt, flavoring, canola oil and soak both textured proteins in it for 30-40 min. Grind the soaked textured proteins in a Thermomix TM5 at level 8 for 30 sec. Premix all remaining dry ingredients and mix them with ground textured proteins in a stand mixer equipped with batter blade for 1 min at lowest speed, followed by 2 min at medium speed. Sheet to a thickness of 2 mm, and heat for 8 min at 135°C (275°F) in a commercial convection oven. Cut the sheets with a deli slicer to 3 mm wide strands.

Table 4b: Adipose tissue analog

[0100] Disperse all dry materials in oil and cocoa butter while heating to 40°C to melt cocoa butter (Thermomix TM5, level 5, 40°C). Turn off heat, add water as 50/50 ice/water mix and emulsify at level 5 until a stable emulsion with final temperature of about 10°C has formed.

Table 4c: Meat juice analog

[0101] Mix/dissolve all ingredients until homogeneous. Table 4d: Binder mix

[0102] Premix.

Table 4e: Assembly

[0103] To assemble the steak, follow the proportions in Table 4e. Strands are soaked in excess meat juice analog for 2 min and then drained in a colander (the meat juice analog 1 levels in Table 4e are typical values retained by the strands mainly at their surface). Place the strands in excess binder mix and move around to achieve uniform coating, followed by sieving to remove excess binder mix (the binder mix amounts in Table 4e are typical values). Roughly align the strands (FIG. 4A), add the second meat juice analog portion to make the gluten in the binder mix sticky, apply 2/3 of the adipose tissue analog in small pieces, and roll up into a roast (FIG. 4B). Apply the remaining adipose tissue analog on the outside of the roast. Wrap in parchment paper, place in vacuum bag, and seal under vacuum to further compress and stabilize the roast. Refrigerate for at least 1 hr. Slice the roast in 1 inch slices.

Example 5: Comparison of different commercial TVP types

[0104] This example demonstrates the improvement of chewiness and juiciness by selecting the most suitable texturized protein, and measuring the liquid absorption of the resulting strands. A suitable texturized protein contributes good texture when loaded into the strands, even if a large amount of liquid is absorbed. Comparison of texturized proteins

[0105] Evaluation of fully hydrated texturized proteins: hydrated in excess water and then excess water squeezed out:

• Textured wheat protein (ProTerra 1350): very rubbery (too rubbery for realistic muscle meat bite)

• Textured soy flour (ADM TVP 165-118) or textured soy concentrate (Arcon 171-158): soft, crumbly (more like burger or minced meat)

• Textured SuproMax 5050: fibrous, shreds into meat-like fibers when ground in Thermomix on reverse. Most suitable for meat-like bite

Applying of SuproMax5050 in firm muscle phase

[0106] Strands were prepared with shredded SuproMax 5050 fibers and a small amount of textured soy flour to modulate texture slightly, with and without added baking soda.

[0107] We concluded that this firm phase with SuproMax5050 has the potential to produce a meat analog that is both, fibrous and juicy, if prolonged swelling in meat juice analog is allowed. Baking soda can help increase the moisture absorption via pores (Table 5b). Overnight swelling of these strands prepared with baking soda yields the highest absorption and still decent texture. For practical feasibility, we recommend to soak overnight in the future and if required limit the amount of meat juice analog to not overhydrate. This will give the best penetration of the meat juice into the core of the strands.

Table 5a: Firm muscle phase analog with SuproMax 5050

[0108] Mix water, color, salt, flavoring, canola oil and soak both textured proteins in the mixed liquid for 1 hr. Grind the soaked textured proteins in a Thermomix TM5 at level 6 with blades spinning in reverse for 50 sec. Premix all remaining dry ingredients and mix them with ground textured proteins in a stand mixer equipped with batter blade for 1 min at lowest speed, followed by 2 min at medium speed. Sheet to a thickness of 2 mm, and heat in a household convection oven for 12 min at 177°C (350°F). Cut the sheets with a deli slicer to 3 mm wide strands. The strands were firm, chewy, leathery and resembled beef jerky. They were allowed to swell at refrigeration temperature for different times in excess meat juice analog and after removal from excess meat juice analog weighed and texture evaluated.

[0109] A second treatment of firm muscle phase was prepared exactly the same way, except that 0.15% baking soda was added with the dry ingredients to promote porosity by reacting with the acidic ingredients (e.g., colors).

Table 5b: Effect of different soaking times at refrigeration temperatures on absorption and texture of strands Example 6: A juicier, semi aligned meat analog with secondary not aligned embedded fibrous inclusions

[0110] This example incorporates the learnings on swelling time and fiber type from example 5, and applies them in a full size roast with semi aligned strands similar to example 4. The strands have fibrous inclusions from SuproMax 5050 embedded (FIG. 5A), use baking powder for porosity (FIG. 5B) and are swollen overnight with a large, but limited amount of meat juice analog (50% based on strand weight) - if more juice is absorbed, the strands get too wet and absorb too much binder mix (gluten). The combination of improved embedded fibrous inclusions and more meat juice analog results in improved chewiness and juiciness, while anisotropy results from the semi aligned arrangement of the strands. The strand width was reduced from 3 mm to 2 mm for better visuals (FIG. 3C). The adipose tissue analog was made with coconut oil instead cocoa butter (reduces white chocolate-like off flavor).

Table 6a: Firm muscle phase analog

[0111] Mix water, color, salt, flavoring, canola oil and soak both textured proteins in it for 70 min. Grind the soaked textured proteins in a Thermomix TM5 at level 5 with blades spinning in reverse for 70 sec. Premix all remaining dry ingredients and mix them with ground textured proteins in a stand mixer equipped with batter blade for 1 min at lowest speed, followed by 2 min at medium speed. Sheet to a thickness of 2 mm, and heat in a commercial convection oven for 7 min at 160°C (320°F). Cut the sheets with a deli slicer to 2 mm wide strands.

Table 6b: Adipose tissue analog

[0112] Disperse all dry materials in canola oil and coconut oil while heating to 40°C to melt coconut oil (Thermomix TM5, level 5, 40°C). Turn off heat, add water as 50/50 ice/water mix and emulsify at level 5 until a stable emulsion with final temperature of about 10°C has formed.

Table 6c: Meat juice analog

[0113] Mix/dissolve all ingredients until homogeneous.

Table 6d: Binder mix [0114] Premix.

Table 6e: Assembly

[0115] To assemble the steak, follow the proportions in Table 6e. Strands are soaked overnight in meat juice analog 1 in the refrigerator. (The strands should absorb almost all of the meat juice analog overnight). Place the soaked strands in excess binder mix and move around to achieve uniform coating, followed by sieving to remove excess binder mix (the binder mix amounts in Table 6e are typical values). Roughly align the strands (FIG. 4A), add the second portion of meat juice analog to make the gluten in the binder mix sticky, smear on 2/3 of the adipose tissue analog in small patches, and roll up into a roast (FIG. 4B). Apply the remaining adipose tissue analog on the outside of the roast. Wrap in parchment paper, place in vacuum bag, and seal under vacuum (target - 0.97 bar relative to ambient pressure) to further compress and stabilize the roast. Refrigerate for at least 1 hr. Slice the roast in 1-inch slices.

Example 7: Evaluating the need for alignment of the small lengthscale inclusions

[0116] Texture tests were conducted with a simplified firm phase (model formula), omitting ingredients with limited texture impact (Table 7a). Embedded fibrous inclusions (shredded SuproMax 5050) were either aligned or not aligned. These samples were compared by texture analysis (tension test to rupture, Table 7b). Results showed clear anisotropy for the aligned samples. Pulling in the direction of the embedded fibrous inclusions resulted in specifically longer distances at maximum force, somewhat higher maximum forces and consequently clearly higher toughness than pulling orthogonal to the fibrous inclusions. Samples where the fibrous inclusions were not aligned showed no relevant anisotropy. It is notable that not aligned samples reached similar values than the aligned samples in fiber direction. The data therefore suggest that alignment did not strengthen the sheet in direction of the fiber alignment, but rather weaken it in the orthogonal direction.

[0117] We conclude that embedded SuproMax fibrous inclusions have a strengthening effect on the firm phase whether aligned or not. As anisotropy of the steaks is achieved by aligned larger strands, strength and good cohesion of embedded fibrous inclusions is the relevant criteria, while the alignment of the fibrous inclusions can be ignored. This provides a more practical approach for production. See also illustration in FIGS. 6A and 6B.

Table 7a: Simplified model formula for firm muscle phase analog

[0118] Mix water, color, salt, canola oil and soak both textured proteins in the mixed liquid for 70 min. Grind the soaked textured proteins in a Thermomix TM5 at level 5 with blade spinning in reverse for 70 sec. Premix all remaining dry ingredients and mix them with ground textured proteins in a stand mixer equipped with batter blade for 1 min at lowest speed, followed by 2 min at medium speed. From here, different treatments were prepared (aligned vs. not aligned). Aligned: The dough was pressed through a syringe with 7 mm opening in parallel strands in sheeting direction. The shredded fibrous inclusions inside these strands are aligned by this procedure. Afterwards the strands were pressed together and then the mass sheeted and heated as the firm phase in example 5 above. The alignment direction of the sheet was marked for subsequent texture measurements. [0119] A control (no alignment) was prepared by sheeting and heating as before, but without the prior alignment step through the syringe. Intentionally, the control dough was turned 90 degree during sheeting to minimize any orientation imposed by sheeting. All heat-set dough sheets were then sealed in plastic bags and kept over night in the fridge. The next day, strips were cut out of the sheets (2 x 3 inches). The strips were cut out of the sheet either in fiber direction (3 inches side corresponds to fiber or sheeting direction), or against fiber direction (3 inches side was 90 degree to fiber or sheeting direction). In the case of the not aligned control, the strips were still cut along the main sheeting or against the main sheeting direction. The strips were then subjected to a tension test, using a texture analyzer (TA-XTplusC, Stable Micro Systems) with the following settings: test mode: tension, test speed: 5.0 mm/sec, distance 45.0 mm; Probe code TA-226 (Tug fixture set for pulling apart pizza crust bagels bread, Texture Technologies Corp.) Results were analyzed for maximum force, distance at this force, and area under the curve (‘toughness’) with the basic macro function.

Table 7b: Tension test of firm phase strips with embedded fibrous inclusions (aligned or not aligned)

³ Anisotropy index calculated as value in direction of fibers / value against direction of fibers (major sheeting direction in case of not aligned samples) ^b average ± standard deviation. 4 replicates

Example 8: tailoring the heating protocol to the protein composition

[0120] Two firm phase treatments were prepared similar as in example 6, with one treatment having the potato protein isolate omitted and replaced with extra gluten (Table 8). A fibrous shredded textured protein sourced from a Chinese manufacturer similar to SuproMax 5050 was used for inclusions. Both treatments yielded chewy firm phases. It was, however, necessary to heat the version without potato protein longer, because gluten sets at higher temperature than potato protein (Solanic 200).

[0121] The example shows that by longer heating, gluten setting can provide sufficient firmness without the contribution of potato protein.

[0122] If assembled into steaks similar as in example 6 above, both firm phase treatments produced steaks that were juicy, flavorful, chewy, and fibrous. The version without potato protein but longer heating was even a bit chewier.

[0123] We conclude that the texture of the firm phase can be further tailored by adjusting the heating conditions in the setting step to the properties of the proteins used.

Table 8: Firm muscle phase analog with and without potato protein

[0124] Mix water, color, salt, flavoring, canola oil and soak textured protein in it for 1 hr. Grind the soaked textured protein in a Thermomix TM5 at level 6 with blade spinning in reverse for 20 sec. Premix all remaining dry ingredients and mix them with ground textured proteins in a stand mixer equipped with batter blade for 1 min at lowest speed, followed by 2 min at medium speed. Sheet to a thickness of 2 mm, and heat in a household convection oven at 135°C (275°F). The potato protein treatment is baked for 12 min, the treatment without potato protein for 20 min. Cut into about 3 mm wide strands.

EQUIVALENTS AND INCORPORATION BY REFERENCE

[0125] While the invention has been particularly shown and described with reference to a preferred embodiment and various alternate embodiments, it will be understood by persons skilled in the relevant art that various changes in form and details can be made therein without departing from the spirit and scope of the invention.

[0126] All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry, patent application, or patent, was specifically and individually indicated incorporated by reference in its entirety, for all purposes.

[0127] Specifically, US provisional application no. 63/561,588, filed March 5, 2024, is incorporated herein by reference in its entirety for all purposes.

Claims

WHAT IS CLAIMED:

1. A method of making a plant-based meat analog, comprising: a) forming a dough-like wet protein mixture comprising i) texturized protein (TP), and ii) one or more of a plant protein powder, a binder, a coloring agent, a flavor agent and/or a spice, a liquid, and a leavening agent, wherein the TP has been processed into particles or fibers ranging from about 10 pm in diameter to about 1 mm in diameter; b) processing the wet protein mixture to form fiber-like strands ranging from about 0.1 mm to about 4 mm, wherein the strands are flexible; c) aggregating the strands into a desired alignment; and d) stabilizing the strands to form a coherent strand mass resembling meat in appearance and texture.

2. The method of claim 1, wherein the TP has been produced by (i) low moisture extrusion, (ii) high moisture extrusion, or (iii) a similar method involving temperatures above 100°C, pressures above ambient pressure, and/or shear.

3. The method of claim 1, wherein the TP has been produced by any one of the following methods: shear cell technology, freeze alignment, wet spinning, electrospinning, and/or fermentation to produce microbial biomass.

4. The method of any one of claims 1-3, wherein the liquid is water, vegetable oil, or a mix of the two.

5. The method of any one of claims 1-4, wherein the stabilization is achieved through one or more of the following: rolling and pressing, compacting by subjecting to vacuum, chemical crosslinking, enzymatic crosslinking, heating, and cooling.

6. The method of any one of claims 1-5, wherein processing the wet protein mixture to form fiberlike strands comprises sheeting the wet protein mixture to form a coherent workable protein dough sheet followed by cutting the protein dough sheet to form strands.

7. The method of claim 6, wherein the protein dough sheet is set prior to cutting.

8. The method of claim 6, wherein the strands are set after cutting.

9. The method of claim 7 or 8, wherein the setting is by heating to a temperature that allows for protein coagulation.

10. The method of claim 7 or 8, wherein the setting is by means of cross-linking other than heat coagulation, optionally wherein the cross-linking is chemical crosslinking, and/or enzymatic crosslinking.

11. The method of any one of claims 1-5, wherein processing the wet protein mixture to form fiberlike strands comprises extruding the wet protein mixture into strands.

12. The method of claim 11, comprising extruding the wet protein mixture into strands followed by setting the extruded strands.

13. The method of claim 12, wherein the setting is by heating to a temperature that allows for protein coagulation.

14. The method of claim 12, wherein the setting is by means of cross-linking other than heat coagulation, optionally wherein the cross-linking is chemical crosslinking, and/or enzymatic crosslinking.

15. The method of claim 11, comprising extruding and heat-setting the wet protein mixture into strands in a single step through a heated nozzle or extrusion die.

16. The method of any one of claims 1-15, wherein the TP is processed prior to adding to the wet protein mixture such that isotropic inclusions of the TP are formed in the wet protein mixture.

17. The method of claim 16, wherein the TP is minced into particles prior to adding to the wet protein mixture.

18. The method of claim 16 or 17, wherein the TP particles are arranged in a random fashion.

19. The method of any one of claims 1-15, wherein the TP is processed prior to adding to the wet protein mixture such that anisotropic inclusions of the TP are formed in the wet protein mixture.

20. The method of claim 19, wherein the TP is shredded into fibers prior to adding to the wet protein mixture.

21. The method of claim 19 or 20, wherein the TP fibers are semi-aligned along a common axis.

22. The method of claim 19 or 20, wherein the TP fibers are arranged in a random fashion.

23. The method of any one of claims 1-22, wherein the TP is from one or more of the following: wheat gluten, soy protein isolate, soy protein concentrate, soy flour, pea protein isolate, pea protein concentrate, pea flour, faba bean protein isolate, faba bean protein concentrate, faba bean flour, chickpea protein isolate, chickpea protein concentrate, chickpea flour, an isolate, concentrate, and/or flour from a cereal grain, and optionally commercial food starch(es) selected from wheat starch, com starch, tapioca starch, potato starch, rice starch, or any suitable, commercially available starch.

24. The method of claim 23, wherein the majority of the protein in the TP is from (a) wheat gluten, optionally wherein the TP is a fibrous product from the ProTerra family by MGP ingredients (for example ProTerra 1100, ProTerra 1200, ProTerra 1350), or (b) a blend of wheat gluten and one or more of soy protein isolate, soy protein concentrate, and soy flour, optionally wherein the TP is a fibrous product from the SuproMax family by Solae/IFF (for example SuproMax 5010, SuproMax 5050).

25. The method of claim 24, wherein the TP comprises a granular or flaky TP, optionally wherein the granular or flaky TP is the textured soy flour Purelynature 165-118 (ADM), textured soy protein concentrate Arcon T 158-171 (ADM), Response 4310, Response 4380, Response 4400, Response 4410, Response 4438 (Solae/IFF), or textured pea protein TPP70, or TPP80 (Puris).

26. The method of any one of claims 1-25, wherein the wet protein mixture comprises: a) water; b) salt; c) flavoring and/or spices; d) vegetable oil; e) TP; f) a binder, optionally wherein the binder is (i) a fiber from citrus peel, citrus pulp, carrot, pea hull, inner cell walls of pea, apple pomace, Psyllium hull, oat fiber, and/or flaxseed, or (ii) a hydrocolloid, optionally wherein the hydrocolloid is xanthan gum, guar gum, alginate, high methoxyl and low methoxyl pectin, sodium alginate, different types of carrageenan, methylcellulose, and/or hydroxypropyl methylcellulose; g) vital wheat gluten; h) wheat flour, optionally wherein the wheat flour is all purpose wheat flour; and/or i) a leavener, optionally wherein the leavener is baking powder or baking soda.

27. The method of claim 26, wherein the wet protein mixture comprises: a) water from about 40% to about 60% by weight; b) salt from about 0% to about 1.5% by weight; c) flavoring from about 0% to about 8% by weight; d) vegetable oil from about 0% to about 15% by weight; e) TP from about 5% to about 35% by weight; f) a binder from about 0% to about 6% by weight; g) vital wheat gluten from about 8% to about 30% by weight; h) wheat flour from about 0% to about 5% by weight; i) baking powder or baking soda from about 0.1% to about 5% by weight; and/or j) a spice blend from about 0% to about 5% by weight.

28. The method of claim 26 or 27, wherein i) the vegetable oil is selected from canola oil, soybean oil, sunflower oil, safflower oil or any other vegetable oil liquid at room temperature; ii) the TP is one of the fibrous TP products of claim 24; and/or iii) the binder is Psyllium hull fiber.

29. The method of claim 28, wherein the TP comprises one of the granular or flaky TPs of claim 25, and the sum of the fibrous TP and the granular or flaky TP is about 5% to about 35% of the wet protein mixture by weight.

30. The method of any of the claims 23-29, wherein the fibrous TP is from soy protein, wheat gluten, and wheat starch.

31. The method of any of the claims 23-30, wherein the fibrous TP is SuproMax 5050 or SuproMax 5010.

32. The method of any one of claims 26-31, wherein the wet protein mixture further comprises a coloring agent and/or potato protein isolate.

33. The method of claim 32, wherein the wet protein mixture comprises i) a coloring agent from about 0.1% to about 5% by weight; and/or ii) potato protein isolate from about 1% to about 5% by weight, optionally wherein the potato protein isolate is rich in native patatin, further optionally wherein the potato protein isolate is Solanic 200 (Avebe).

34. The method of any one of claims 26-33, wherein forming the dough-like wet protein mixture comprises: a) soaking the TP in a mixture containing the water, salt, flavoring, vegetable oil and optionally color for a sufficient amount of time to allow for absorption of the liquid phase, thereby forming soaked textured proteins; b) grinding the soaked textured proteins in such a manner as to result in small anisotropic fibers with the majority having a diameter between about 10 pm and about 1 mm, thereby forming ground textured proteins; and c) combining the ground textured proteins with any remaining dry ingredients in such a manner as to result in a cohesive, extensible dough-like mass, thereby forming the dough-like wet protein mixture.

35. The method of any one of claims 1-34, wherein processing the wet protein mixture to form fiberlike strands comprises sheeting the wet protein mixture to form a coherent workable protein dough sheet followed by cutting the protein dough sheet to form strands, and wherein the protein dough sheet is from about 0.1 mm to about 4 mm in thickness.

36. The method of claim 35, wherein the protein dough sheet is heated at a sufficient temperature and for a sufficient duration to allow for setting of the dough prior to cutting to form strands.

37. The method of claim 35 or 36, wherein the protein dough sheet is cut to form strands from about 0.1 mm to about 4 mm wide.

38. The method of any one of claims 1-37, further comprising soaking the strands in a first portion of meat juice analog for a sufficient duration to allow for the meat juice analog to be at least mostly absorbed, optionally wherein the weight of the first portion of meat juice analog is from about 10% to about 80% of the weight of the strands.

39. The method of claim 38, further comprising coating the strands with a binder mix, optionally wherein the weight of the binder mix is from about 5% to about 50% of the weight of the strands.

40. The method of claim 39, wherein the binder mix comprises a protein powder.

41. The method of claim 40, wherein the protein powder is at least partially undenatured.

42. The method of claim 41, wherein the protein powder is capable of being crosslinked and increasing in firmness during a final cooking step of the meat analog at about 60°C to about 95°C carried out by an end user.

43. The method of claim 42, wherein the end user can control the doneness level by cooking to a final temperature between about 60°C and about 95°C, optionally wherein the final temperature is between about 74°C and about 95°C.

44. The method of any one of claims 39-43, wherein the binder mix comprises at least 50% vital wheat gluten.

45. The method of claim 44, wherein the binder mix comprises vital wheat gluten, optionally wherein the binder mix further comprises a spice blend, further optionally wherein the vital wheat gluten is present in the binder mix from about 90% to about 100% by weight and/or the spice blend is present in the binder mix from about 0% to about 10% by weight.

46. The method of any one of claims 1-45, wherein aggregating the strands into a desired alignment comprises aggregating the strands such that they are semi-aligned along a common axis.

47. The method of claim 46, wherein the degree of alignment of the strands is between 0.9965 and 0.7500.

48. The method of any one of claims 1-47, further comprising adding a second portion of meat juice analog to the aggregated strands, optionally wherein the weight of the second portion of meat juice analog is from about 10% to about 30% of the weight of the strands.

49. The method of any one of claims 38-48, wherein the meat juice analog comprises: a) water, optionally wherein the water is from about 85% to about 99% by weight; b) flavoring, optionally wherein the flavoring is from about 0% to about 8% by weight; c) salt, optionally wherein the salt is from about 0% to about 1.5% by weight

50. The method of claim 49, wherein the meat juice analog further comprises baking soda or another mildly alkaline, food-safe ingredient, from about 0% to about 5% by weight to neutralize the pH.

51. The method of claim 49 or 50, wherein the meat juice analog further comprises a coloring agent, optionally wherein the coloring agent is from about 0.1% to about 10% by weight.

52. The method of any one of claims 1-51, further comprising applying an adipose tissue analog to the strands, optionally wherein the weight of the adipose tissue analog is from about 0% to about 35% of the weight of the strands.

53. The method of any one of claims 1-52, wherein stabilizing the strands to form a coherent strand mass comprises rolling up and pressing together the aggregate strands, followed by compacting by subjecting to vacuum.

54. The method claim 52 or 53, wherein a first portion of the adipose tissue analog is applied during aggregation of the strands, and a second portion of the adipose tissue analog is applied to an outside surface of the coherent strand mass.

55. The method of any one of claims 52-54, wherein the adipose tissue analog comprises: a) a lipid component (optionally wherein the lipid component is vegetable oil and/or vegetable fat), optionally wherein the lipid component is from about 5% to about 40% by weight; b) vital wheat gluten, optionally wherein the vital wheat gluten is from about 0% to about 10% by weight; c) flavoring, optionally wherein the flavoring is from about 0% to about 6% by weight; d) salt, optionally wherein the salt is from about 0% to about 1% by weight; e) methylcellulose, optionally wherein the methylcellulose is from about 0.5% to about 4% by weight; and/or f) water, optionally wherein the water is from about 35% to about 60% by weight.

56. The method of claim 55, wherein the lipid component comprises a vegetable fat that is at least partly solid at 15-20°C, optionally wherein the lipid component further comprises liquid vegetable oil that is fully liquid at 15-20°C.

57. The method of claim 56, wherein the solid vegetable fat is selected from cocoa butter, coconut oil, palm oil, palm kernel oil, shea butter, sal butter and/or hydrogenated fats.

58. The method of claim 56, wherein the solid vegetable fat is shortening or margarine.

59. The method of any one of claims 56-58, wherein the liquid vegetable oil is selected from canola oil, soybean oil, sunflower oil, safflower oil, and/or any other suitable plant-based oil.

60. The method of any one of claims 55-59, wherein the methylcellulose is a type that gels below 50°C and forms gels that are on the very firm end of the spectrum achievable with methylcellulose, measured with suitable texture methods, including TPA.

61. The method of claim 60, wherein the methylcellulose is Wellence Vegeform 183 (IFF/Dupont).

62. The method of any one of claims 55-61, further comprising preparing the adipose tissue analog by: a) dispersing the dry ingredients of the adipose tissue analog in the vegetable oil and/or vegetable fat while heating at a temperature sufficient to melt the vegetable fat b) removing the oil mixture from heat; and c) adding the water as a mix of ice and liquid with mixing to form a stable emulsion.

63. The method of any one of claims 1-62, further comprising cutting the coherent strand mass into individual fillets, resembling fillets of an animal based whole-cut.

64. The method of any one of claims 1-63, wherein there is no plane, whose normal vector is orthogonal to the average strand alignment in the whole-cut meat analog that the strands do not cross.

65. The method of any one of claims 1-64, wherein the tension in the whole-cut meat analog along a direction orthogonal to the average strand alignment is at least about 10% of the tension in the whole-cut meat analog along the average strand alignment.

66. The method of claim 65, wherein the tension in the whole-cut meat analog along a direction orthogonal to the average strand alignment is no greater than about 90% of the tension in the wholecut meat analog along the average strand alignment.

67. A plant-based meat analog prepared according to the method of any one of claims 1-66.

68. A plant-based meat analog comprising strands made from a wet protein mixture that range from about 0.1 mm to about 4 mm, the strands comprising i) texturized protein (TP), and ii) one or more of a plant protein powder, a binder, a coloring agent, a flavor agent and/or a spice, a liquid, and a leavening agent, the strands being aggregated into a desired alignment and stabilized to form a coherent strand mass resembling meat in appearance and texture.