CN117794382A

CN117794382A - Method for producing a cookable fibrous meat analogue by directional freezing

Info

Publication number: CN117794382A
Application number: CN202280049919.2A
Authority: CN
Inventors: 克里斯托弗·布赖森; 克里斯托弗·马克·格雷格森; 德里克·鲁索; 奥克·德·弗里斯; 拉斐尔·德·埃诺
Original assignee: New School Food Co
Current assignee: New School Food Co
Priority date: 2021-05-21
Filing date: 2022-05-20
Publication date: 2024-03-29

Abstract

The present disclosure provides a method for producing a "cookable", fibrous meat analogue using directional freezing. The method includes subjecting an ingestible hydrocolloid to directional freezing for inducing the formation of elongated ice crystals, wherein the elongated ice crystals are aligned in a given direction in the directional frozen hydrocolloid. The elongated ice crystals are then removed and replaced with protein and any other additives, such as supplements, located in the aligned channels that initially contain aligned ice crystals. Once the desired protein loading is achieved, the protein loaded hydrocolloid is subjected to conditions suitable to induce gelation of some of the proteins to form a protein gel in the aligned elongate channels.

Description

Method for producing a cookable fibrous meat analogue by directional freezing

Cross Reference to Related Applications

The present application claims priority from U.S. patent application Ser. No. 17/326,567, filed on even 21 at 5 at 2021 (now published as US11,241,024), and U.S. patent application Ser. No. 17/666930 filed on even 8 at 2 at 2022, both of which are incorporated herein by reference in their entireties.

Technical Field

The present disclosure relates to a method of producing a "cookable", fibrous meat analogue using directional freezing of hydrogels.

Background

Due to the growth of plant-based alternatives, the global meat industry of $ 1.4 trillion is experiencing unprecedented confusion, estimated to reach $ 1400 billions (approximately 10% of the global meat market) in the next 10 years. Plant-based alternatives to meat and fish are becoming a need for many consumers who are striving to address the ethical issues of eating animal-based protein products, such as pure vegetarian, etc. Consumers who are allergic to meat or fish to various foods are also looking for plant-based alternatives. The growth in this industry is expected to last for decades; by 2050, the global meat industry would need to grow 69% to meet population growth. This would be particularly challenging because animal-based protein production is significantly more resource intensive in terms of water resource usage, land usage, and GHG emissions than plant-based proteins. In addition, 90% of wild fish on earth have been classified as over-caught or caught at maximum capacity. This means that the opportunities for plant-based foods only increase over time, driven by increasing pressures from customers, special interest groups/NGOs and governments. Due to the increasing shortage of resources of the earth, it is of best interest in every country to have ready-made alternatives to animal-based protein foods from the point of view of food safety.

Another impetus for the transition from animal-based proteins to plant-based protein substitutes is driven by an increasing outbreak of animal disease known to be transmitted to humans. These diseases are known as zoonotic diseases (or zoonotic), which are caused by bacteria transmitted from animals to humans, and those poor agricultural practices in the world lead to particular problems in areas of intimate contact between animals and humans. There are two ways of infection of animals to humans, most commonly viral infection between humans in intimate contact with the animals, due to the transmission of virus from animal to human through the air, and the other due to ingestion and consumption of the animal by the human. The impact of these diseases on the economy of countries that rely heavily on animal-based agriculture can be devastating, let alone the death of humans that these animal-based diseases cause when they begin to spread to humans.

Of these diseases transmitted to humans by eating beef, most notably Bovine Spongiform Encephalopathy (BSE), which was first identified in 1985 in british cattle. It is associated with human variant Creutzfeldt-Jakob disease (Creutzfeldt-Jakob disease) and is the food of infected meat that results in transmission to humans. In the uk, nearly two hundred (200) people die from this disease and result in millions of cattle being slaughtered. BSE is a neurological disorder caused by a rare transmission factor called prion and appears in cattle beginning in the 70 s of the 20 th century. The main reasons are identified as animal feeds, including Meat and Bone Meal (MBM) from infected or infected cattle. Over 100,000 cattle were confirmed to be infected. The use of MBM was banned in 1988, but it has been associated with a rare disease known as creutzfeldt-jakob disease (CJD) in humans. The version associated with BSE is named variant CJD and may appear on consumers eating infected meat many years later, in many cases fatal.

Similarly, ingestion of fish by humans can also lead to disease. Zoonotic diseases associated with fish exposure are mainly bacterial infections. These include Mycobacterium (Mycobacterium), erysiphe (Erysipelothrix), campylobacter (Campylobacter), aeromonas (Aeromonas), vibrio (Vibrio), edwardsiella (Edwardsiella), escherichia (Escherichia), salmonella (Salmonella), klebsiella (Klebsiella) and Streptococcus iniae (Streptococcus iniae). Although these infections do not always make fish look ill, they can cause serious illness in humans. The impact of global warming on the growth of zoonotic disease in animals and fish has not been quantified or understood and it is reasonable to consider that an increase in ocean and ambient temperature can lead to an outbreak of more zoonotic disease. Thus, there is an increasing drive to find plant-based alternatives to meat and/or fish from safer protein sources that do not require as much resources as are currently required for animal-based protein sources.

Unfortunately, plant-based meat substitutes currently on the market have proven difficult to appeal to mass market consumers because it is difficult to adequately mimic the taste, texture, nutritional profile, and cooking behavior of real meat. These products sometimes use basic formulations and untested methods, which highlight the lack of development investment for manufacturers. Consumer research indicates that consumers rely primarily on 3 criteria in making decisions to purchase meat and replace meat products: price, taste and convenience. Thus, if a plant-based meat substitute does not have a truly meat-comparable taste, is more expensive, and/or is less convenient to prepare, it may only appeal to the small population of vegetarians and pure vegetarians.

In the alternative protein industry, almost all efforts have focused on mimicking alternatives to beef, chicken and pork, with little effort directed to alternative plant-based seafood. In 2019, plant-based seafood was only 1% of the total sales of plant-based meat replacement, and only 0.07% of the total sales of seafood (dollars of 1.05 billion). By 2030, the plant-based seafood market can reach 10% of the entire seafood market, which would be worth up to $200 billion as the entire plant-based meat market would be expected to reach 10% of the global meat sales.

There are methods using directional freezing as a method for producing a fiber similar to a muscle fiber. In us patent No. 4,423,083, a method of producing fibers by combining protein and hydrocolloid followed by freezing is described. After thawing, the fibers are reinforced with chelating agents, preserving the fiber structure. In the methods disclosed herein, directional freezing of the hydrocolloid gel is performed first, and the second step is immersing in the protein solution without the need for a chelating agent, thereby avoiding the need for a chelating agent. This very advantageously provides flexibility in controlling fiber formation prior to the addition of other components such as proteins and other supplements as disclosed herein.

Furthermore, U.S. patent No. 4,423,083 discloses that it is necessary to slice the frozen material before immersing in the chelating solution to produce a fiber bundle having a maximum thickness of 8mm, thereby maintaining the fiber characteristics of the innermost portion of the sample. In the present disclosure, no sectioning is required, the fibrous structure can be preserved without chelating agents, and the sample can be of any size or shape.

Disclosure of Invention

In one aspect, a method for producing a fibrous meat analogue is provided, comprising: a) Preparing an ingestible biopolymer gel, solution or dispersion comprised of one or more ingestible proteins and/or hydrocolloids and water; b) Subjecting the biopolymer gel, solution or dispersion to directional freezing, inducing the formation of aligned elongated ice crystals to form a directionally frozen biopolymer gel, solution or dispersion having aligned elongated channels in which aligned elongated ice crystals are located; c) Replacing the aligned elongated ice crystals with an ingestible protein and/or hydrocolloid to produce an infused gel; and d) subjecting the injected gel to suitable conditions to gel at least some of the ingestible proteins and/or hydrocolloids to produce gelled proteins and/or hydrocolloids within the aligned channels, thereby forming a fibrous food product.

In one embodiment, the ingestible biopolymer gel, solution or dispersion is: a hydrocolloid gel, solution or dispersion comprising one or more different ingestible hydrocolloids and water; a protein gel, solution or dispersion comprising one or more different ingestible proteins and water; or a complex gel, solution or dispersion comprising one or more different ingestible hydrocolloids and one or more different ingestible proteins and water.

In one embodiment, the ingestible proteins and/or hydrocolloids of steps a) and c) are the same or different. In one embodiment, the ingestible hydrocolloid comprises one or more of conventional and/or recombinant gelatin, agar, alginate, curdlan, kappa-carrageenan, kappa 2-carrageenan and iota-carrageenan, red algae gum, starch, modified starch, seaweed extract, dextrin, konjac glucomannan, methylcellulose, pectin, gellan gum, xanthan gum, guar gum, locust bean gum, gum arabic, tara gum, or polysaccharide.

In one embodiment, the method comprises: a) Preparing an ingestible hydrocolloid gel consisting of one or more ingestible hydrocolloids and water; b) Subjecting the ingestible hydrocolloid gel to directional freezing, inducing the formation of aligned elongate ice crystals to form a directional frozen hydrocolloid gel having aligned elongate channels in which the aligned elongate ice crystals are located; c) Replacing the aligned elongated ice crystals with an ingestible protein to produce a protein infused hydrocolloid gel; and d) subjecting the protein-infused hydrocolloid gel to suitable conditions to gel at least some of the ingestible proteins, thereby producing gelled proteins within the aligned channels.

In one embodiment, the method comprises a) preparing an ingestible protein gel comprised of one or more first ingestible proteins and water; b) Subjecting the ingestible protein gel to directional freezing, inducing the formation of aligned elongated ice crystals to form a directional frozen protein gel having aligned channels in which the aligned elongated ice crystals are located; c) Replacing the aligned elongated ice crystals with a second ingestible protein to produce a protein gel infused with the protein; and d) subjecting the protein gel injected with the protein to suitable conditions to gel at least some of the second ingestible protein, thereby producing a gelled protein in the aligned channels.

In one embodiment, the ingestable protein comprises a gellable protein, a non-gellable protein, or a combination thereof. In one embodiment, the ingestable protein comprises a culture protein; animal proteins, such as recombinant animal proteins; a plant protein; bacterial proteins; fungal proteins, such as yeast proteins; algae proteins; or a combination thereof. In one embodiment, the ingestible protein is mammalian whey protein, casein or caseinate; any one or any combination of soy protein, potato protein, rubisco protein, duckweed protein, rice protein, almond protein, egg protein, oat protein, linseed protein, euglena protein, schizochytrium limacinum protein, mung bean protein, pea protein, recombinant mammalian whey, cultured mammalian whey, recombinant egg albumin, cultured egg albumin, recombinant gelatin or collagen, cultured gelatin or collagen, canola protein, lupin protein, broad bean protein, wheat protein, lentil protein, amaranth protein, peanut protein, moringa seed protein, pumpkin seed protein, chickpea protein, sunflower seed protein, safflower seed protein, mustard seed protein, chlorella protein, and spirulina protein.

In one embodiment, step c) comprises: thawing the directionally frozen biopolymer gel, solution or dispersion by immersion in a solvent containing the ingestible protein and/or hydrocolloid of step c), said solvent having a temperature suitable for thawing ice crystals; freeze-drying the directionally frozen biopolymer gel, solution or dispersion to remove substantially all water, and then immersing the dried gel in a solution containing the ingestible protein and/or hydrocolloid of step c); evaporating the ice crystals and then immersing the dried gel in a solution containing the ingestible protein and/or hydrocolloid of step c); or placing one end of the injected gel under vacuum to extract ice crystals and pulling the ingestible protein and/or hydrocolloid of step c) from the other end of the injected gel into the aligned elongate channels. In one embodiment, step c) comprises thawing the directionally frozen biopolymer gel, solution or dispersion; and wherein the method comprises a plurality of cycles of directional freezing and thawing. In one embodiment, the method further comprises controlling the diameter of the aligned elongate channels by controlling a temperature gradient across the material so as to vary the speed of the directional freezing process, and wherein the diameter of the gelled protein in the aligned elongate channels is proportional to the diameter of the aligned elongate channels. In one embodiment, the diameters of the aligned elongate channels are controlled to obtain elongate gelled proteins having diameters in the range of about 20 to about 500 microns.

In one embodiment, the biopolymer gel, solution or dispersion is a solution or dispersion and the method comprises (i) prior to step c), gelling the solution or dispersion by further subjecting the directionally frozen solution or dispersion to suitable conditions, or (ii) prior to step c), inducing gelling of the solution or dispersion by immersion into a suitable solution. In one embodiment, the biopolymer gel, solution or dispersion is a solution or dispersion and the method comprises gelling the solution or dispersion while directionally freezing. In one embodiment, the biopolymer solution or dispersion comprises a first hydrocolloid and a second hydrocolloid, and wherein the method comprises gelling the first hydrocolloid, subjecting the biopolymer to directional freezing, subsequently gelling the second hydrocolloid, and replacing the ice crystals with an ingestible protein.

In one embodiment, suitable conditions include: heat treating the injected gel, and wherein the ingestable protein comprises at least a heat-gellable protein; penetrating a salt or ion into the injected gel, the salt being selected to induce gelation of the at least some of the ingestible protein and/or hydrocolloid; adjusting the pH of the injected gel to a value suitable to cause gelation of the at least some of the ingestible proteins and/or hydrocolloids; penetrating a solution containing a cross-linking agent into the injected gel, the cross-linking agent being selected to induce gelation of the at least some of the ingestible protein and/or hydrocolloid; subjecting the injected gel to pressure treatment to induce gelation of the at least some of the ingestible proteins and/or hydrocolloids; and/or irradiating the injected gel with radiation of a suitable wavelength and intensity to induce cross-linking of the proteins, thereby inducing gelation of the at least some of the ingestible proteins.

In one embodiment, the osmotic salt comprises contacting the injected gel with a salt solution of sufficient concentration to allow gelation of the at least some of the ingestible protein and/or hydrocolloid; and wherein the salt is any one of sodium (Na), potassium (K), calcium (Ca) and magnesium (Mg) sulfate, citrate, chloride, carbonate, ascorbate, acetate, sorbate, lactate, tartrate, gluconate and phosphate, and any combination thereof. In one embodiment, adjusting the pH of the injected gel comprises adding a food safe pH adjuster comprising acetic acid, hydrochloric acid, ascorbic acid, malic acid, formic acid, lactic acid, tartaric acid, citric acid, gluconic acid, glucono-delta lactone, sodium hydroxide, potassium hydroxide, calcium hydroxide, or a combination thereof. In one embodiment, the crosslinking agent is a chemical crosslinking agent comprising glutaraldehyde, tannic acid, genipin (genipin), liquid smoke (liquid smoke), or a combination thereof. In one embodiment, the cross-linking agent is an enzyme-based cross-linking agent comprising transglutaminase (EC 2.3.2.13), sortase a (EC 3.4.22.70), tyrosinase (EC 1.14.18.1), laccase (EC 1.10.3.2), peroxidase (EC 1.11.1. X), lysyl oxidase (EC 1.4.3.13), amine oxidase (EC 1.4.3.6), or a combination thereof.

In one embodiment, the ingestible biopolymer gel, solution or dispersion has multiple layers. In one embodiment, the alternating layers are made of the same or different biopolymers or biopolymer blends. In one embodiment, the method further comprises producing a layer that mimics the skin layer of meat or fish by producing a mixture of agar and an alginate solution with an alginate-oil emulsion and gelling the mixture to produce a skin layer. In one embodiment, the method further comprises producing a plurality of protein infused biopolymer gels each having a preselected thickness, and preparing interstitial layers made of a material selected to mimic connective tissue of meat and/or fish. In one embodiment, a plurality of protein infused biopolymer gels are stacked and adhered to the interstitial layer. In one embodiment, the interstitial layer comprises a material selected to mimic connective tissue of meat and/or fish, the material comprising any one or a combination of proteins, hydrocolloids, oil-in-water emulsions, solid particles, fats, and oil gels. In one embodiment, the solid particles comprise any one or a combination of titanium dioxide, protein, calcium carbonate, starch, solid fat crystals, and algae.

In one embodiment, the method further comprises: i) Adding a pigment during the preparation of the biopolymer gel, solution or dispersion; or ii) replacing the aligned elongated ice crystals with a mixture of a second ingestible protein and/or hydrocolloid and a pigment; wherein the pigment comprises carotenoids, beta-carotene, astaxanthin, lycopene, carmine, anthocyanidins, betalains, hemoglobin, myoglobin, beet juice extract, carthamin, lutein, curcumin, capsanthin, norbixin, anthocyanin, curcuminoids, turmeric root powder, phycocyanin, melanoidin, or combinations thereof.

In another aspect, a method for producing a fibrous meat analogue is provided, comprising: subjecting the ingestible biopolymer gel to directional freezing, inducing the formation of aligned elongated ice crystals to form a directionally frozen biopolymer gel having aligned channels in which the aligned elongated ice crystals are located; thawing the directionally frozen biopolymer gel having aligned channels by immersing the frozen biopolymer in a solution containing at least one ingestible soluble thermal gelling protein, thereby thawing and replacing the aligned elongated ice crystals with the at least one ingestible soluble thermal gelling protein at a temperature below the gelling temperature of the soluble thermal gelling protein to produce a protein infused biopolymer gel; and heating the protein infused biopolymer gel at a temperature above the gelation temperature of the at least one ingestible soluble thermogelling protein to produce protein fibers, thereby forming a fibrous meat analog food product.

In another aspect, there is provided a fibrous meat analogue food product produced by the method as described herein.

Many additional features and combinations thereof with respect to the embodiments described herein will be apparent to those of skill in the art upon reading this disclosure.

A further understanding of the functionality and advantageous aspects of the present disclosure may be realized by reference to the following detailed description and the attached drawings.

Drawings

Embodiments will now be described, by way of example only, with reference to the accompanying drawings, in which:

fig. 1 shows a schematic representation of an isotropic hydrogel subjected to directional freezing, wherein the hydrogel in contact with a pre-cooled substrate begins to freeze to form ice crystals, which grow in a direction perpendicular to the plane of the substrate, and these aligned crystals grow away from the cooled substrate until the whole substance is present in the form of elongated ice crystals surrounded by now concentrated hydrogel, as disclosed in Yokoyama, f, achife, e.c., momoa, j, shimamura, k, and Monobe, k, 1990. Morphology of optically anisotropic agarose water gels prepared by directional freezing (Morphology of optically anisotropic agarose hydrogel prepared by directional freezing), colloid and polymer science (Colloid and Polymer Science), 268 (6), pages 552 to 558.

Figures 2A and 2B show polarized light micrographs of 2A) transverse and 2B) longitudinal cross-sections of oriented frozen/thawed agarose gels, as disclosed in Yokoyama, f., achife, e.c., momoda, j., shimamura, k., and Monobe, k.,1990. Morphology of optically anisotropic agarose gels prepared by oriented freezing, colloid and polymer science, 268 (6), pages 552 to 558.

FIG. 3A is a photomicrograph showing a side view of the elongate channel in an agar-alginate hybrid gel after directional freezing using an optical microscope.

FIG. 3B is a photomicrograph showing a cross-sectional view of the elongate channel in the agar-alginate hybrid gel after directional freezing using an optical microscope.

Fig. 3C is a scanning electron microscope image showing a cross-sectional view of an elongated structure in an agar gel after directional freezing and subsequent freeze-drying. Ice in the elongate channels in this image is removed by a freeze drying process.

Fig. 4A is an optical photograph showing a cross-section of a layered directionally frozen gel in which alternating layers are fibrous or non-fibrous.

Fig. 4B is an optical photograph showing a cross-section of another layered directionally frozen gel, wherein alternating layers are fibrous or non-fibrous, similar to fig. 4A.

Fig. 4C is an optical photograph showing a side view of another layered directionally frozen gel, wherein alternating layers are fibrous or non-fibrous, similar to fig. 4A.

Fig. 5A is an optical photograph showing 3 wt% agar containing 0.15 wt% pigment in a salmon mold. The right panel shows the final result after adding myodiaphragm (myomamma).

Fig. 5B is an optical photograph showing sarcomere-diaphragmatic gel sectioned and directionally frozen at-15 ℃.

Fig. 5C is an optical photograph showing a sarcomere-diaphragmatic gel after directional freezing.

Fig. 6 is an optical photograph showing a stacked 3 wt% agar sarcomere gel, wherein the myodiaphragm layer is made of 5 wt% mung bean and 3 wt% agar, with an alginate-seaweed based epidermis covered thereon.

Fig. 7A is an optical photograph showing the appearance during frying of agar, alginate and agar-alginate composite gels.

FIG. 7B is an optical photograph showing the structure of alginate, agar and agar-alginate composite gel after frying.

Fig. 7C is an optical photograph showing the appearance of a 12 wt% potato protein gel after directional freezing.

Fig. 8 is an optical photograph showing the fibrous appearance of a 15 wt% canola-potato protein sodium alginate gel after being directionally frozen and cooked, with the left drawing showing it intact and the right drawing pulled apart.

FIG. 9 is an optical photograph showing a 3 wt% agar, 0.75 wt% alginate mix gel containing a 1:1 blend of canola to potato protein and a pigment after directional freezing. The left hand sample was heated to 55 ℃ (resulting in a "raw" appearance) and the right hand sample was subjected to subsequent heat treatment in the frying pan (showing the appearance after cooking).

Fig. 10 is a photograph showing a 0.75 wt% alginate, 7.5 wt% whey, 3 wt% potato protein complex gel after frying and directed freezing.

Fig. 11 is an optical photograph showing a side view of another layered, directionally frozen gel, wherein alternating layers are fibrous or non-fibrous, similar to fig. 4A, and the product is raw/uncooked and translucent.

Detailed Description

Various embodiments and aspects of the disclosure will be described with reference to details discussed below. The following description and drawings are illustrative of the disclosure and are not to be construed as limiting the disclosure. Numerous specific details are described to provide a thorough understanding of various embodiments of the disclosure. However, in some instances, well-known or conventional details are not described in order to provide a concise discussion of embodiments of the present disclosure.

As used herein, the terms "comprise" and "comprising" are to be construed as inclusive and open-ended, rather than exclusive. In particular, the terms "comprises" and "comprising," and variations thereof, when used in the specification and claims, are intended to include the specified features, steps or components. These terms should not be interpreted to exclude the presence of other features, steps or components.

As used herein, the term "exemplary" means "serving as an example, instance, or illustration," and should not be construed as preferred or advantageous over other configurations disclosed herein.

As used herein, the terms "about" and "approximately" are meant to encompass variations that may exist in the upper and lower limits of a numerical range, such as variations in properties, parameters, and dimensions. In one non-limiting example, the terms "about" and "approximately" mean plus or minus 10% or less.

As used herein, the phrase "fibrous meat analogue" refers to food analogues that mimic food products characterized by a fibrous structure, including fish and meat (beef, lamb, pork, chicken, etc.).

As used herein, "protein" includes natural proteins and recombinant proteins.

As used herein, the word "protein denaturation" means the alteration of the structure of a protein from its natural state. This may be accomplished, for example, by breaking some of the intramolecular bonds (e.g., hydrogen bonds) in the protein molecule. Cleavage of these bonds, for example due to heat treatment, means that the highly ordered protein structure is altered from its natural or natural state. This process may involve exposing hydrophobic side groups, which are typically buried in the center of the protein molecule, and transferring or creating intermolecular disulfide bonds. This may lead to the formation of protein aggregates.

As used herein, the phrase "protein gel" means a three-dimensional viscoelastic network of water-immobilized proteins. This can be accomplished, for example, by heating the protein solution to a temperature above the denaturation temperature of the protein under solvent conditions (e.g., ionic strength and pH) that favor formation of a continuous network. Another possible approach is to create a fluid suspension or dispersion of proteins or protein aggregates and, via changing solvent conditions (e.g., ionic strength or pH), form a protein gel due to a decrease in repulsive forces between proteins or protein aggregates.

As used herein, the phrases "protein gelling" and "gelling of a protein" and "gelled protein" mean the process of producing a protein gel, as defined previously, e.g., via heat treatment of a protein solution. By "gelled protein" is meant a volume of protein that undergoes a gelling process.

As used herein, the term "biopolymer" means an organic molecule, such as proteins and polysaccharides, that is composed of repeating monomers and is produced by or biocompatible with a living organism.

As used herein, the term "diameter" represents the cross-sectional width of an individual channel or fiber, but does not necessarily mean a circular cross-sectional shape.

As used herein, elongated ice crystals refer to phase separated ice domains (ice domains) having a high aspect ratio.

As used herein, the term "hydrogel" means a three-dimensional network of hydrophilic biopolymer molecules that can immobilize large amounts of water.

As used herein, the phrase "protein fiber" means an elongated protein gel having a high aspect ratio in a size range similar to muscle fibers found in meat or fish.

As used herein, the phrase "fibrous" means containing biopolymer fibers, such as protein and/or polysaccharide fibers.

The present disclosure provides a method for producing a "cookable", fibrous meat analogue by employing directional freezing.

In some embodiments, the present disclosure provides a method for producing a "cookable," fibrous meat analogue by employing directional freezing.

The present disclosure provides a two-step process for producing a fibrous meat analogue by directional freezing of a biopolymer gel, solution or dispersion, such as a hydrocolloid gel, a protein gel or a complex gel of hydrocolloids and/or proteins. The first step of the method is directed freezing of the biopolymer gel, solution or dispersion. This method results in the formation of meat-like or fish-like muscle fibers and a change in the texture of the biopolymer gel, solution or dispersion gel due to the formation of ice crystals that align the hydrogel fibers. The second step involves replacing ice crystals with ingestible proteins and/or hydrocolloids to form an infused gel. In some embodiments, the protein and/or hydrocolloid of the first step is the same as or different from the protein and/or hydrocolloid of the second step.

In some embodiments, the first step is directed chilled hydrocolloid gel and the second step is to replace ice crystals with an ingestible protein. In some embodiments, the first step is to orient the frozen hydrocolloid gel and the second step is to replace the ice crystals with ingestible hydrocolloid. In some embodiments, the first step is to orient the frozen hydrocolloid gel and the second step is to replace the ice crystals with the ingestible protein and hydrocolloid. In some embodiments, the first step is to orient the frozen protein gel and the second step is to replace the ice crystals with an ingestible hydrocolloid. In some embodiments, the first step is to orient the frozen protein gel and the second step is to replace the ice crystals with an ingestible protein. In some embodiments, the first step is to orient the frozen protein gel and the second step is to replace the ice crystals with an ingestible protein and hydrocolloid. In some embodiments, the first step is directed freezing of the composite hydrocolloid gel and the second step is replacing ice crystals with an ingestible protein. In some embodiments, the first step is directed freezing of the composite hydrocolloid gel and the second step is replacing ice crystals with ingestible hydrocolloid. In some embodiments, the first step is directed freezing of the composite hydrocolloid gel and the second step is replacing ice crystals with ingestible proteins and hydrocolloids.

In some embodiments, injecting the gel comprises soaking the hydrogel in the protein solution at a preselected temperature for a specific time such that the aligned ice crystals are replaced with soluble proteins in the organized hydrogel. Subsequent heating of the injected hydrogel results in gelation. Preferably, hydrocolloids with melting temperatures above the gelation temperature of the protein are used to maintain the size, structure and fibrosis of the product.

In one embodiment, the method includes subjecting an ingestible polysaccharide-containing hydrogel of a selected size and shape to directional freezing to induce formation of elongated ice crystals, wherein the elongated ice crystals are aligned in a given direction in the ingestible hydrogel to form an organized hydrogel containing ice crystals. Subsequently, the organized hydrogel is immersed in a solution containing the ingestible soluble protein at a preselected temperature such that when the ice crystals melt, the ingestible thermal gelling proteins diffuse into the organized hydrogel, replacing the melted ice crystals. The organized hydrogel is immersed in a solution containing the ingestible, thermal gelling protein for a selected period of time as required to achieve a desired protein loading. The protein-permeable hydrogel is then heat treated at a temperature sufficient to induce gelation and fiber formation within the hydrogel for use in producing a cookable fibrous meat analog food product. An exemplary product is a salmon side analog product.

In another embodiment, the method includes subjecting an ingestible hydrocolloid gel consisting of one or more different ingestible hydrocolloids and water to directional freezing to induce the formation of elongated ice crystals, wherein the elongated ice crystals are aligned in a given direction in the ingestible hydrocolloid gel. The aligned elongated ice crystals are then replaced with an ingestible protein to produce a protein infused hydrocolloid gel that is subsequently subjected to gelation to produce a food product.

Directional freezing

The process of directional freezing involves freezing a material by controlling the direction of water freezing. The step of directionally freezing the hydrogel is performed by placing the hydrogel of a selected size and shape in contact with a pre-cooled substrate to induce formation of ice crystals extending through the fibrous hydrogel structure in a direction perpendicular to the pre-cooled substrate, and wherein the pre-cooled substrate is cooled to a temperature in the range of about-2 ℃ to about-196 ℃.

Referring to fig. 1 and 2A and 2B, the main concept of directional freezing of hydrogels is shown. Here, a hydrogel of selected mass and shape is placed in contact with a cold substrate, whereupon elongated ice crystals begin to form perpendicular to the freezing front. This forces the biopolymer chains of the hydrogel to align perpendicular to the pre-cooled substrate to form an aligned fibrous gel structure, which also results in the formation of aligned anisotropic elongated ice crystals that are separated from each other by aligned fiber bundles, wherein the elongated ice crystals are aligned in a given direction to form a organized hydrogel containing aligned ice crystals. As can be seen from fig. 3A, 3B and 3C, once the aligned elongated ice crystals are removed or moved from the hydrocolloid they will leave aligned elongated channels. This is a freeze concentration effect in which water and polysaccharide gradually phase separate from the initial solution as the elongated ice crystals form, the biopolymer chains of the hydrocolloid being pushed together into a higher concentration of smaller volume.

In some embodiments, this method produces a fibrous texture that mimics the typical muscle fiber structure found in many fish species, such as but not limited to salmon, trout, tuna, and cod, to name a few. Although it is not even limited thereto; we can also use it to make other analogues of food products characterized by a fibrous texture, such as but not limited to steak or chicken fillet.

In some embodiments, upon thawing the organized hydrogel in the presence of an aqueous solution or dispersion containing a substance (such as, but not limited to, an ingestible soluble protein capable of diffusing into the product), the substance diffuses into the organized product to replace the thawed ice crystals as the aligned ice crystals melt.

Directional freezing mould

For each different type of fibrous meat analog food produced, whether they be fish, poultry, pork, veal, beef, etc., a directional freezing mold for a particular product can be produced. There are several variable parameters for each particular type of mold, which may vary depending on the commercial product to be sold. In particular, the shape of each mold may be adjusted to mimic the shape of the food product being produced. For fish, the shape may reflect the shape of the whole fish, or it may reflect the shape of the fillet rather than the whole fish. Salmon side has a unique shape, and the mold may reflect this unique shape. The depth of the mold may vary depending on the desired thickness of the final product. Similarly, many steak cuts have different characteristics, which can be reflected in the mold, for example, a T-shaped steak has characteristic "T" shaped vertebrae, and the shape of the mold can be shaped to reflect this. The size of the mold may be made to truly reflect the typical size of the meat chunk. Since the thickness of these cuts ranges from a few inches to a few inches, this can be reflected in the depth of the mold.

In the case of salmon, the size and shape of the mould may be substantially the same as the size and shape of the salmon side produced, wherein a single substrate may be used, which substrate is moulded such that when filled with hydrogel the final product will have substantially the same size and shape as the salmon side. Alternatively, the whole fish may be produced by sizing and shaping the top and bottom molds such that when the top and bottom are connected to the inner hydrogel, the inner size and shape mimics the whole fish.

In addition to parameters of shape, size and depth of the mold, another parameter is surface topography. For foods with unique surface features that are non-planar or planar, the mold can be produced to reflect the non-planar topography such that the non-planar surface is visible when packaged and the product looks very realistic. It should be appreciated that the die may incorporate mechanical design features that may be incorporated into the die to more easily control the variation in channel diameter to control fiber diameter. A non-limiting example is by having sharp points on the inner surface of the mold to seed ice crystal nucleation (and thus their number and size). Another method may be to vary the rate at which the hydrocolloid sample/mould descends into the cooling bath.

While a single pre-cooled plate may be used for the directional freezing process, it should be understood that two (2) plates may be used, one below the hydrocolloid (or protein) mass and one on top of the hydrocolloid (or protein) mass.

Physiologically compatible hydrogels and hydrocolloids

In some embodiments, the method for producing a cookable fibrous meat analog involves directional freezing of an ingestible hydrocolloid gel or protein gel suitable for food products. The hydrocolloid gel may be, but is not limited to, a polysaccharide hydrogel, conventional gelatin, recombinant gelatin, or a combination of both. Hydrocolloids may be naturally occurring, they may be recombinant, or they may be laboratory grown or cultured, or they may be chemically or enzymatically modified.

In some embodiments, the method for producing a cookable fibrous meat analogue uses an ingestible polysaccharide-containing hydrogel. Hydrogels are composed of a network of crosslinked polymer chains that are generally hydrophilic. Interactions between polymer chains cause cross-linking and result in the formation of a three-dimensional network, entrapping aqueous liquids in the semi-solid structure. Crosslinking between biopolymers may be chemical or physical and includes, but is not limited to, hydrogen bonding, hydrophobic or ionic interactions, and chain entanglement. These crosslinks are strong enough that the integrity of the hydrogel network is maintained and the polymer does not readily dissolve back into solution. Hydrogels are characterized by having a highly absorbent natural or synthetic polymer network, and they can easily contain more than 90% water.

There are many types of polysaccharide hydrogels. Non-limiting examples of such hydrogels include carrageenans, a family of natural linear sulfated polysaccharides extracted from red edible seaweed, and exhibit high potency for strong binding to food proteins. Carrageenans are large, highly flexible molecules that form a coiled helical structure that imparts them the ability to form a variety of different gels at room temperature, and therefore they are widely used in the food industry, particularly as stabilizers and thickeners.

These carrageenans typically contain from about 15% to about 40% by weight of sulfate content, which produces anionic polysaccharides. They are classified into three different categories based on their sulfate content. Kappa-carrageenan (K-carrageenan) has one sulfate group per disaccharide, iota-carrageenan (I-carrageenan) has two sulfate groups, and lambda-carrageenan (L-carrageenan) has three sulfate groups. K-carrageenan is characterized in that it forms a strong and hard gel in the presence of potassium ions and reacts with dairy proteins, while I-carrageenan forms a soft gel in the presence of calcium ions and, finally, L-carrageenan does not gel but can be used to thicken dairy products. Carrageenan is a high molecular weight polysaccharide and is made predominantly of alternating 3-linked b-D-galactose-pyranose (G-units) and 4-linked a-D-galactopyranose (D-units) or 4-linked 3, 6-anhydro-a-D-galactopyranose (DA-units), forming disaccharide repeating units of carrageenan.

Another type of ingestible hydrogel includes agar hydrogels, which are gelatinous materials obtained from red algae, and are a mixture of two components, a heterogeneous mixture of the linear polysaccharide agarose and smaller molecules called agaropectins. It forms a support structure in the cell wall of certain species of algae and is released upon boiling. These algae are called agarophytes (agarophytes) and belong to the phylum rhodophyta (red algae).

Agar hydrogels have been used as food ingredients, for example as thickeners for vegetarian substitutes for gelatin, soups, ice cream, fruit preserves, and the like. Agar hydrogels are also used in other physiological applications such as appetite suppressants and laxatives, to name a few. The gelling agent in agar is an unbranched polysaccharide isolated from the cell walls of various red algae. Those skilled in the art will know that such ingredients as agar and carrageenan are widely used in the food industry.

Thus, non-limiting examples of ingestible hydrocolloid gels include agar, fermentation-derived gelatin, alginates, curdlan, carrageenan selected from the group consisting of kappa-carrageenan, kappa 2-carrageenan, and iota-carrageenan, furcellaran, starches (including modified starches and dextrins), konjac glucomannan, gellan, and combinations comprising xanthan gum, guar gum, locust bean gum, and tara gum.

Proteins

In the method of the present invention, the protein incorporated into the hydrocolloid or protein gel is not limited to naturally occurring proteins. For example, recombinant proteins suitable for use in food products, cultured (laboratory cultured) proteins, chemically or enzymatically modified proteins may be used. The protein may be an animal protein or a recombinant animal protein. The protein may also be any one or a combination of a plant-based protein, a bacterial-based protein, a fungal-based protein and an algae-based protein. As an example, the fungal-based protein may include yeast. The algae-based protein may be any one or a combination of macroalgae and microalgae.

Non-limiting examples of ingestible proteins are any one or any combination of whey protein, soy protein, potato protein, rubisco protein, duckweed protein, rice protein, almond protein, oat protein, linseed protein, euglena protein, schizochytrium limacinum protein, mung bean protein, pea protein, recombinant whey, cultured whey, recombinant egg albumin, cultured egg albumin, recombinant gelatin or collagen, cultured gelatin or collagen, canola protein, lupin protein, faba protein, wheat protein, lentil protein, amaranth protein, peanut protein, moringa seed protein, pumpkin seed protein, chickpea protein, sunflower seed protein, safflower seed protein, mustard seed protein, chlorella protein, and spirulina protein.

Whey Protein Isolate (WPI) is a dietary supplement and a food ingredient, which is produced by separating ingredients from whey. Whey proteins are mixtures of proteins and some of them gel very well, while others in the mixture (if any) do not gel very well. Whey is a milk byproduct of cheese making that can be processed to produce three different forms of whey protein, including whey isolates, whey concentrates, and whey hydrolysates. The differences between these protein forms are related to the composition of the product, in particular the protein content. Whey isolates contain the highest amount of protein and may be lactose-free, carbohydrate-free, fat-free and cholesterol-free.

These proteins are characterized by high bioavailability and rapid absorption into the body and contain high concentrations of Branched Chain Amino Acids (BCAAs) which are highly concentrated in muscle tissue and serve to stimulate protein synthesis in addition to providing energy to the working muscle.

While the food products of the present invention include the use of WPI in the examples of the present disclosure, those skilled in the art will appreciate that many other plant-based proteins that provide excellent thermal gelling proteins can be used and readily identified by those skilled in the art. Non-limiting examples include soy protein, potato protein isolate, rubisco protein, mung bean protein, and pea protein. To be effective in thermal gelation, the protein will have the following properties: solubility (> 85%), viscosity (preferably low at room temperature and high at >50 ℃), denaturation temperature (about 45 to about 85 ℃) and gel strength standard (G' >100 pascals).

The ingestible soluble proteins are preferably natural thermal gelling proteins and when these proteins are used, the ingestible polysaccharide-containing hydrogels and the ingestible thermal gelling proteins are selected such that the hydrogels have a melting temperature above the gelling temperature of the proteins to maintain the size, structure and fibrosis of the fibrous meat analogue food product.

In some embodiments, the ingestible soluble protein is an ingestible non-thermal gelling protein, in which case a thermally induced trigger is included to trigger gelling upon an increase in temperature. The trigger induces gelation of the otherwise non-gelled protein. The trigger may be pre-mixed with the protein or hydrogel. The thermally induced trigger may be any one or a combination of salt, enzyme or pH adjuster. For example, salts, pH modifiers or enzymes are microencapsulated within a meltable coating that is triggered by heating. The microencapsulated material may be in any one of the phases. Non-limiting examples of pH adjusting agents include glucono-delta-lactone. Non-limiting examples of enzyme-based triggers include transglutaminases. Non-limiting examples of salt-based triggers include calcium phosphate.

Whether or not the protein is thermally gelled, the protein-impregnated hydrogel product is heated such that the internal temperature rises to between 50 ℃ and 100 ℃ to induce gelation of the protein. This can be accomplished using a technique that uses higher temperatures (oven, grill, frying pan, roaster, to name a few). The purpose of this heating step is to produce a product that undergoes a transition upon heating, resulting in a change in color and/or texture (preferably both) similar to that possessed by conventional fish or meat.

The solution immersed in the frozen (or thawed) hydrogel may contain only 100% of the heat-gelling protein, but may also contain a mixture of heat-gelling protein and non-heat-gelling protein or protein hydrolysate. The concentration of the thermogelling protein may be less than the total protein content. For example, in a 15 wt% total protein solution, 5 wt% may be a heat gelling protein, the remainder being non-heat gelling proteins. It should be noted that these amounts are non-limiting.

In one embodiment, the concentration of the ingestible soluble total protein in the aqueous solution or dispersion is in the range of about 1 to about 35 wt.%. More preferably, the concentration of the ingestible soluble protein in the aqueous solution or dispersion is in the range of about 10 to about 30 wt.%. More preferably, the concentration of the ingestible soluble protein in the aqueous solution or dispersion is in the range of about 15 to about 25 wt.%.

Controlling protein gel diameter

The method of the present invention allows the diameter of the aligned elongate channels to be controlled by controlling the relative temperature between the cooling surface in contact with the ingestible hydrogel and the surface air temperature above the surface in order to vary the speed of the directional freezing process. The diameter of the protein gel in the aligned channels will be proportional to the diameter of the aligned elongate channels. Faster freezing typically results in thinner ice crystal channels. Controlling this phenomenon is the rate of ice crystal nucleation, which is faster at lower temperatures. If the nucleation rate is faster, then the result is more ice cores (and thus more ice crystals), so each individual ice crystal is thinner.

Ice nucleation will be most important during the first few seconds of the freezing process and at the colder surfaces where nucleation occurs. At deeper depths in the hydrogel, these crystals will grow with the freezing front, and ice nucleation may not be so important. Thus, controlling the crystal thickness is the nucleation rate at the first frozen surface, which is controlled by the amount of supercooling, and in turn, by the temperature of the frozen surface and the rate at which the temperature is reduced. In the case of thinner ice crystals, once the ice crystals are moved, they leave correspondingly thinner aligned channels that when filled with protein, when gelled with some of the gellable protein, result in thinner protein fibers.

In a preferred embodiment, the diameters of the aligned elongate channels are controlled such that the diameter of the protein gel in the channels is in the range of about 20 to about 200 microns, thereby allowing the diameter of the protein gel to be in the same range. For example, the protein fiber in salmon has a diameter of about 100 microns. The diameter of the protein fibers in meats such as beef, chicken, pork, etc., for chicken, typically ranges from about 30 to 50 microns, and for beef, from 20 to 85 microns.

Method for replacing elongated ice crystals with proteins and other additives

There are several methods that can be used to replace elongated ice crystals with proteins and/or other ingredients, such as flavors, dietary supplements, and the like. In one embodiment, replacing the aligned elongated ice crystals with the ingestible protein comprises thawing the directionally frozen hydrocolloid gel by immersion in a solvent containing the ingestible protein, the solvent having a temperature suitable for thawing the ice crystals replaced with the ingestible protein to produce a protein infused hydrocolloid gel. In this embodiment, thawing the directionally frozen hydrocolloid gel comprises adjusting the temperature of the solvent comprising the ingestible gellable protein to a range from the melting point of water to the melting point of the hydrocolloid gel, and replacing the aligned elongated ice crystals with the ingestible protein comprises adjusting the temperature of the solvent comprising the ingestible protein to a range from the freezing point of the solvent comprising the protein solution to the gelation onset temperature of at least one of the ingestible proteins. Typically, thawing the directionally frozen hydrocolloid gel is performed at a temperature in the range of between about 0 ℃ and about 85 ℃ depending on the type of hydrocolloid, and similarly, replacing aligned elongated ice crystals with an ingestible protein may be performed at a temperature in the range of between about 0 ℃ and about 45 ℃.

The solvent containing the protein and/or other ingredients may be an aqueous solution, or alternatively it may be a non-aqueous solvent suitable for the food product in which the ingestible protein is soluble. The non-aqueous solvent may be any one or combination of acetic acid, formic acid, ethanol, methanol, propanol, and mixtures thereof with water. When the solvent is an aqueous solution in which the ingestible protein is soluble, this solution may be maintained at a temperature between about 1 ℃ and about 99 ℃. It may also be heated to a higher temperature in the range of between about 99 ℃ and about 130 ℃ and subjected to a pressure in the range of about 0 to 1.7b bar in a self-pressurizing closed vessel.

In another embodiment, a method of replacing aligned elongated ice crystals with an ingestible protein comprises subjecting an oriented frozen hydrocolloid gel to conditions suitable for sublimating the elongated ice crystals in the presence of the ingestible gellable protein. This may include subjecting the directionally frozen hydrocolloid gel to a vacuum, causing sublimation of ice, and immersing the sublimated hydrocolloid gel into a solution containing the ingestible protein, thereby injecting the solution containing the ingestible protein into the sublimated hydrocolloid gel.

In another embodiment, a method of replacing aligned elongated ice crystals with an ingestible protein comprises freeze-drying a directionally frozen hydrocolloid gel to remove substantially all water, and then immersing the dried gel in a solution containing the ingestible protein.

In another embodiment, a method of replacing aligned elongated ice crystals with an ingestible protein comprises subjecting a directionally frozen hydrocolloid gel to conditions suitable to cause evaporation of ice in the presence of a solution containing the ingestible protein to remove substantially all of the ice.

Thus, it will be appreciated that there are many ways to move ice crystals using phase changes, such as sublimation (e.g. freeze drying) or solid to liquid phase transition (melting), evaporation of ice crystals, and furthermore any type of physical movement of ice crystals may be used.

Conditions suitable for producing protein gels in aligned channels

Proteins can be classified into gellable proteins and non-gellable proteins. In the methods of the invention, non-gellable proteins can be mixed with gellable proteins to provide increased protein content. The types of gellable proteins include thermally gellable proteins, wherein heating to the gelation temperature of a particular protein results in the formation of a protein gel. However, there are other methods of gelling proteins, and thus the proteins used in the methods and food analog products of the present invention are not limited to non-heat-gellable proteins.

Thus, once the protein is loaded into the hydrocolloid or protein gel, if the protein is not thermogellable, several methods of inducing gellation of the gellable protein can be used. In one embodiment, the conditions suitable for gelling at least some of the ingestible proteins may include penetration of a salt into the hydrocolloid gel of the injected protein, wherein the salt is selected to induce gelling of at least some of the ingestible proteins, thereby producing a protein gel. Such salts may be penetrated into the protein-infused hydrocolloid gel by injecting a saline solution into the protein-infused hydrocolloid gel or protein gel.

Alternatively, the salt may be allowed to permeate into the protein-infused gel by adding the salt as a crystalline solid to the surface of the protein-infused hydrocolloid gel, and then the salt is dissolved and diffused into the protein-infused gel by any available water present in the protein-infused hydrocolloid gel.

Alternatively, the salt may be infiltrated into the protein-infused gel by immersing the protein-infused gel in a concentrated salt solution that diffuses into the protein-infused hydrocolloid gel. The salt may be any one of sulfate, citrate, ascorbate, acetate, gluconate, and phosphate of sodium (Na), potassium (K), calcium (Ca), and magnesium (Mg), and any combination thereof.

Another method of inducing gelation of an infused protein includes adjusting the pH of the hydrocolloid gel of the infused protein to a value suitable to cause gelation of the at least some of the ingestible proteins. The pH can be adjusted by adding pH adjusters suitable for food (liquid form, solution form) or by adding soluble pH adjusters (solid form). The modifier may be any one or combination of acetic acid, hydrochloric acid, ascorbic acid, malic acid, formic acid, tartaric acid, citric acid, glucose-delta lactose, sodium hydroxide, potassium hydroxide and calcium hydroxide.

Another method of inducing gelation of an infused protein includes penetrating a solution containing an enzyme-based cross-linking agent into a hydrocolloid gel of the infused protein, the enzyme-based cross-linking agent being selected to induce gelation of the at least some of the ingestible proteins. The enzyme cross-linking agent may include any one or a combination of transglutaminase, transglutaminase (EC 2.3.2.13), sortase a (EC 3.4.22.70), tyrosinase (EC 1.14.18.1), laccase (EC 1.10.3.2), peroxidase (EC 1.11.1. X), lysyl oxidase (EC 1.4.3.13), and amine oxidase (EC 1.4.3.6).

Another method of inducing gelation of injected proteins involves pressure treatment of the hydrocolloid gel of the injected proteins to induce gelation of at least some of the proteins. In this method, a protein-infused hydrocolloid gel food product is sealed and placed in a rigid sealed compartment containing a liquid, and the liquid is pressurized.

Another method of inducing gelation of an injected protein includes penetrating a solution containing a chemical cross-linking agent into a hydrocolloid gel of the injected protein, the chemical cross-linking agent being selected to induce gelation of the at least some of the ingestible proteins. Non-limiting examples of chemical cross-linking agents are any one or combination of glutaraldehyde, tannic acid, genipin, and liquid smoke.

Another method of inducing gelation of the injected protein involves irradiating the hydrocolloid gel of the injected protein with radiation of a suitable wavelength and intensity to induce cross-linking of the protein, thereby inducing gelation.

It will be appreciated that any combination of the above methods for gelling gellable proteins may be used if a mixture of thermally and non-thermally gellable proteins is loaded into aligned channels of a hydrocolloid or protein gel.

Thawed directionally frozen hydrocolloid

In embodiments using thawing in a solution containing protein and any other desired additives, when the organized hydrogel is thawed in the presence of an aqueous solution or dispersion containing a substance, as the aligned ice crystals melt, the substance diffuses into the organized product to replace the melted ice crystals. When the thermal gelling proteins replace melted ice crystals, several situations may occur:

1) In the first case, the hydrogel infused with the protein is heated to a temperature above the gelation temperature of the protein for a period of time such that some, but not all, of the protein is denatured. This results in the formation of protein fibres but is sufficient to avoid leakage of the remaining protein from the hydrogel;

2) The second case is that the thawed hydrogel may be heated above the gelation temperature for a period of time such that most, if not all, of the protein is involved in the production of protein fibers;

3) The third case is that the heating to the gelation temperature does not take place, so that gelation does not occur, but rather an ingestible substance is introduced into the hydrogel, which substance acts to prevent leakage, wherein the protein-infused hydrogel is then sealed; and is also provided with

4) In the fourth case, not only can the temperature be heated to a temperature at which the protein is gelled to form protein fibers, but the temperature can be further raised to actually cook the food analog, producing a "precooked" food product, which is then packaged and sent to the end-user and does not require cooking.

The step of thawing the directionally frozen hydrogel may be performed first and once the ice crystals have melted and left behind with elongated aligned channels, the thawed hydrogel may be immersed in an aqueous protein solution, whereupon the protein flows into the vacated channels.

The step of immersing the protein-infiltrated organized hydrogel in a solution containing protein is performed at a preselected temperature in the range of about 0 ℃ to about 80 ℃ and preferably about 1 ℃ to about 7 ℃ and may be performed at a typical refrigerator temperature of 4 ℃ where the temperatures are selected such that the aligned ice crystals slowly melt and as they melt the protein, and if the thawing and infiltration steps are performed simultaneously by immersing the directionally frozen hydrogel in a liquid solution containing protein, any other additional ingredients will diffuse to and replace the melted ice crystals as the liquid will be above the freezing point of water.

If the thawing and permeation steps are performed separately, the frozen hydrogel is first thawed by placing in air or liquid at a temperature between the melting point of water (about 0 ℃) and the melting point of hydrocolloid (about 85 ℃). The permeation step may be performed at any temperature between the freezing point of the protein solution (about 0 ℃) and the onset of gelation temperature of the protein (about 45 ℃, but depending on the protein).

The amount of protein loading, as well as any other ingredients or supplements, is controlled by varying the selected period of time that the organized hydrogel is immersed in a solution containing the ingestible soluble protein and other ingredients, so as to vary the amount of protein and other ingredients loaded into the organized hydrogel in a time-dependent manner. The amount of protein loading may also be varied by the concentration of protein in the immersion liquid and the ratio between the weight or volume of the chilled hydrogel and the immersion liquid in which it is placed. The upper limit on the amount of protein that may be present is the solubility limit of the protein.

In embodiments where the ingestible polysaccharide-containing hydrogel is a K-carrageenan hydrogel, specific ions may be included in the solution containing the ingestible soluble protein or inside the hydrogel (or both), for reducing swelling and shrinkage in a concentration-dependent manner and increasing the hardness of the K-carrageenan gel compared to the hardness in the absence of ions, and for maintaining the formation of fibers of the fibrous hydrogel in the directionally frozen K-carrageenan gel upon prolonged storage.

Ratio of ingestible protein to hydrocolloid

In fibrous meat analog foods, in embodiments, the protein is present in a range of about 5 wt% to about 35 wt% and the hydrocolloid is present in a range of about 0.2 wt% to about 10 wt%, such that the ratio of protein to hydrocolloid ranges from about 35:0.2 to about 5:10, or 175 to 0.5, (50 to 17500%).

In a more preferred embodiment, the protein is present in a range of about 10 wt% to about 30 wt% and the hydrocolloid is present in a range of about 0.5 wt% to about 8 wt%, such that the ratio of protein to hydrocolloid is in a range of about 30:0.5 to about 10:8, or 60 to 1.25, (125 to 6000%).

In the most preferred embodiment, the protein is present in a range of about 10 wt% to about 20 wt% and the hydrocolloid is present in a range of about 1 wt% to about 5 wt% such that the ratio of protein to hydrocolloid ranges from about 20:1 to about 10:5, or from about 20 to about 2, (200 to 2000%).

Ratio of gellable to non-gellable protein

In some embodiments, the ingestible gellable protein is a mixture of ingestible gellable proteins, at least some, but not all of which are thermally gellable. In fibrous meat analogue foods, the total amount of ingestible protein may be present in the range of about 5 wt% to about 35 wt%, and the hydrocolloid may be present in the range of about 0.2 wt% to about 10 wt%. In a more preferred embodiment, in a fibrous meat analogue diet, the total amount of ingestible protein is present in the range of about 10 wt% to about 30 wt%, and the hydrocolloid is present in the range of about 0.5 wt% to about 8 wt%. In the most preferred embodiment, in a fibrous meat analog diet, the ingestible protein is present in the range of about 10 wt% to about 20 wt%, and the hydrocolloid is present in the range of about 1 wt% to about 5 wt%.

For a mixture of thermally gellable and non-thermally gellable proteins, the minimum amount of thermally gellable protein that yields a good gel is about 5 wt% and the maximum is 35 wt%, and wherein the maximum total protein is about 35 wt%, then the maximum non-thermally gellable protein is about 30 wt%, and thus the ratio of non-gelling to gelling is about 30:5 to about 0:35, equal to 6 to 0, i.e. up to 6.

For a mixture of thermally gellable and non-thermally gellable proteins, the intermediate amount of thermally gellable protein yielding good gel is about 8 wt% and the maximum is 25 wt%, and the maximum total protein amount is 35 wt%, then the maximum non-thermally gellable protein is 27 wt%, such that the ratio of non-gelling to gelling is 27:8 to 0:25=about 3.5 to 0, i.e. up to 3.5.

For a mixture of thermogellable and non-thermogellable proteins, the commercially viable minimum amount of thermogellable protein that yields a good gel is about 10 wt% and the maximum amount is 20 wt%, and the maximum total protein present is 30 wt%, the maximum non-thermogellable protein is 20 wt%, such that the ratio of non-thermogellable protein to thermogellable protein is 20:10 to 0:20=2 to 0, i.e. up to 2.

When the hydrogel is a K-carrageenan hydrogel, it preferably has a concentration range of about 0.1% to about 15% by weight. Similarly, when the hydrogel is an agar hydrogel, it preferably has a concentration in the range of about 0.1% to about 15% by weight. This results in a modulus of about 100 to 5000 pascals. Non-limiting examples of additional ingredients or supplements include any one or combination of flavoring agents, emulsifying agents, preserving agents, coloring agents, and texture improving agents. Additional supplements may include emulsions of any one or combination of omega-3, omega-6, omega-9 fatty acids. With respect to omega-3 supplements, a preferred mode is to use omega-3 fatty acids, such as, but not limited to, triglycerides, which are mainly in the form of fatty acid esters. Examples of ingestible supplements include water-soluble vitamins including ascorbic acid (vitamin C), thiamine, riboflavin, niacin, vitamin B ₆ (pyridoxine, pyridoxal and pyridoxamine), folic acid, vitamin B ₁₂ Biotin and pantothenic acid. Water insoluble vitamins, including any one or combination of vitamins A, D, E and K, may also be included. Ingestible minerals may be included including any one or combination of iron, magnesium, manganese, zinc, and calcium. Other ingestible supplementsThe extender includes an antioxidant such as, but not limited to, tocopherol.

The heat treated proteins (and optionally other supplements or additives) that penetrate into the organized hydrogel may be packaged and stored at temperatures ranging from about 4 ℃ to about 7 ℃, but they may be stored over a wider temperature range.

It will be appreciated that different proteins may penetrate into the thawed hydrogel. For example, water-soluble heat-induced gelling proteins from non-animal sources with or without water-soluble non-gelling proteins may be used. The water-soluble heat-induced protein may be a variety of vegetable proteins, such as canola, rubisco (various sources such as duckweed/lentil), potato, or animal proteins expressed in a non-animal host, such as gelatin, beta-lactoglobulin, or ovalbumin. The water-soluble non-gelling proteins may be various types of hydrolyzed proteins, including hydrolyzed proteins from legumes, wheat, or algae. Soluble thermogelling non-protein polymers such as hydroxypropyl methylcellulose, methylcellulose or gelatin may also be added.

Precooked food

Heating the protein-containing fibrous meat analog food to a temperature in the range of about 50 ℃ to about 60 ℃ is advantageous because it provides adequate gelation that prevents significant water leakage that occurs after protein injection, while maintaining the "green" appearance of the product. Such products may then be packaged and shipped to a store, such as a grocery store, that sells the products. In this form, once purchased, the consumer will cook at a temperature above 60 ℃, where the cooking temperature depends on the particular type of food product. In order to obtain the desired meat or fish analogue they will produce different products with different heating profiles using different proteins/ingredients, as each type of meat has a different texture, so the ingredients and processing methods need to be changed to properly imitate each food product so that the heating parameters can be changed to seal the moisture. Once in the hands of the consumer, the final cooking temperature will vary from food to food as different foods will have different proteins, protein levels, hydrocolloids, etc.

On the other hand, in another embodiment, it may be desirable to actually cook the product after production and sell it as a "precooked" meal. For example, where it is tactically not advisable to cook meals in the field, precooked and ready-to-eat food analogs can be readily sold to the military as ready-to-eat meals. Similar logic applies to disaster relief situations for meals requiring precooking.

Composite mixed hydrocolloid gel

The inventors have observed that the production of mixed hydrocolloid gels followed by directional freezing improves texture and thermal stability. Some hydrocolloids that produce hydrocolloid gels may be prone to disintegration upon cooking prior to protein gelation and/or may result in loss of fibrosis. Mixing two or more hydrocolloids together to produce a hydrocolloid gel may solve this problem.

The complex hydrocolloids will be described using a non-limiting example of complexes produced using alginate mixed with agar (and compared to pure alginate and agar), but it should be understood that due to the availability of many hydrocolloids, there are many possible complexes and these complexes are not limited to being produced from only two different hydrocolloids.

Example # 1-Complex hydrocolloid

Three samples were prepared, the first being pure agar, the second being pure alginate and the third being a complex formed by mixing alginate with agar. For the latter, the alginate is mixed with deionized water until dissolved, after which agar powder is added, and the mixture is heated to 85 ℃ and held at this temperature for about 20 minutes while stirring to dissolve the agar. After dissolution, the temperature was reduced to 60℃and CaCO was added ₃ Dispersing for about 20 minutes. The Gluconolactone (GDL) was dissolved in the mixture at 60 ℃ for 5 minutes while vigorously mixing (e.g., using a magnetic stirrer), and then the sample was placed in an acoustic water bath to remove bubbles. The resulting viscous composite was then added to a cold plate (cylindrical mold maintained at a temperature of about-15 ℃ to orient the frozen composite gel) at a temperature of about 50 ℃After full orientation freezing, the oriented frozen product was kept frozen at about-18 ℃ for about 24 hours, which was determined to improve the integrity of the fibrous structure.

The pure agar sample was about 2 wt% agar and the pure alginate sample was about 0.5 wt% alginate. The complex comprises about 2% by weight agar + about 0.5 to about 0.75% by weight sodium alginate. All samples contained CaCO ₃ (0.17 to about 0.225 wt%) and GDL (about 0.6 to about 0.8 wt%). The sample was then placed in a protein solution containing 12 wt% potato protein, 50mM NaCl and about 0.1 wt% pigment overnight at 4℃to melt the ice crystals and inject the protein solution into the directionally frozen structure. The next day, the samples were heated in a glass beaker placed in a water bath at 55 ℃ for 20 minutes.

The comparison of pure agar, pure alginate and agar/alginate complex gel after soaking the protein solution is as follows. Both agar and agar-alginate gels are relatively hard gels, whereas alginate gels are relatively soft. The potato protein loaded composite agar-alginate gel was slightly larger than the potato protein loaded agar gel, which appeared to shrink more than the former. After heat treatment at about 55 ℃, the samples were significantly lighter and stiffer. The protein loaded gel was placed in the same frying pan for the same duration to test frying performance at the same temperature (see fig. 7A). The agar-alginate mixed gel maintains its shape well when heated, while the agar gel and the alginate gel shrink significantly.

After frying, the samples were placed on an anvil and the difference between agar and agar-alginate mixtures was again observed (see fig. 7B): the protein-loaded composite agar-alginate gel retains its shape better than agar or alginate alone. Furthermore, in cross section, the protein-loaded composite agar-alginate gel shows a more attractive structure than the protein-loaded gel made with hydrocolloid alone. It is shown that the addition of alginate to agar results in a gel with better heat resistance than agar alone, resulting in a more stable structure when heated in a frying pan, while retaining a fibrous structure.

The directionally frozen agar-alginate mixed gel was imaged using an optical microscope (prior to loading with protein). The photograph shows the aligned elongate channels (sides) and cross section. In fig. 3A, it can be seen that the channels in the polymer gel are mostly uninterrupted, resulting in channels that are hundreds of microns long, and thus become templates for fibrous structures. The cross section (fig. 3B) shows channels having diameters of about 50 to about 200 microns, being slightly elongated, being intersected by high density "lamellae" of polysaccharide.

These results indicate that a composite or hybrid gel may advantageously improve certain product characteristics compared to a non-composite produced from a single hydrocolloid. It will be appreciated that the complex may be prepared from more than two starting components. It will be appreciated that the complexes of the invention are not limited to alginate and agar, and in fact many such combinations are possible.

Construction of larger/more complex macrostructures

More complex fibrous meat analogue foods are produced by creating complex macrostructures through a stacking process that produces multiple layers of hydrocolloid/mixed hydrocolloid/protein plus hydrocolloid gels, each layer separated by thinner interstitial layers, using protein/starch/hydrocolloid/oil-in-water (O/W) emulsions/solid particles (examples are titanium dioxide, protein, calcium carbonate, starch)/any combination of these, to simulate the connective tissue of meat/fish.

In this process, the desired number of hydrogels are prepared, which will be included in the stacked structure. These steps include preparing a caulking layer made of a material selected to mimic connective tissue of meat and/or fish, applying a caulking material to a surface of one of the first one of the hydrocolloid gels, placing a second hydrocolloid gel on top of the caulking layer, and repeating the above steps until a desired number of individual hydrocolloid gels have been stacked together. Typically, the thickness of the interstitial layers will be the same, but it should be understood that they need not all have the same thickness.

Example #2 lamellar hydrocolloid gels

The gel is formed by pouring alternating layers of agar and agar-protein mixtures. Two solutions were prepared separately, heated to 85 ℃ for 15 minutes, and then cooled to 70 ℃. The solution is poured into a container to form a layer of about 0.25cm to about 2 cm. Between pouring each layer, the temperature of the previous layer was allowed to cool for about 60 seconds for the thin layer and 120 seconds for the thicker layer, thereby increasing the viscosity of the solution and preventing the layers from mixing as poured. In this case, the optimum temperature for each layer was found to be about 42 ℃.

The resulting layer structure was left overnight at 4 ℃ to complete the gelation of the interstitial layer. Subsequently, the macrostructures are directionally frozen at once, followed by subsequent injection of the directionally frozen macrostructures into the protein, such as, but not limited to, thawing the directionally frozen stack in a protein solution as previously described. The protein is dispersed in all of the hydrogel layers in the stack, although the interstitial layers have different formulations.

Non-limiting examples of hydrocolloid gel layers include the use of alternating hydrocolloid gel layers; one produces sarcomere (muscle fiber) and the other produces muscle compartment (white interstitial connective tissue).

After directional freezing, these layers may have alternating fibrous/non-fibrous features throughout the structure, such as by incorporating particulate matter (e.g., protein particles) into the interstitial layer, which adversely affects the ability to form fibers (as seen in fig. 4A and 4B). Thus, the fiber properties of the stacked product can be controlled. In the resulting stacked product, the various gel layers adhere to each other (as shown in fig. 4C), but fracture to each other under stress before fracture inside the fiber layers, thereby simulating a "sheetlike" texture.

A variation of this method may involve the use of a mold (see fig. 5A, 5B and 5C), possibly 3D printed, that mimics the visual shape of a whole meat or fish, with a space/divider in the location of the connective tissue/fat layer. The hydrocolloid gel is added to the mould, followed by setting the gel, after which it is removed from the mould. Once removed, the space or gap in the superstructure where the connective tissue/fat layer separator is located is filled in the space with connective tissue formulation in liquid form and a single macroscopic structure is created as the connective tissue forms a gel to solidify. This macrostructure is then directionally frozen and subsequently injected with protein, as by any of the methods described previously. Various additives may be added, such as adding lipids and other flavor components via the interstitial layer, possibly via an oil-in-water (O/W) emulsion and/or through the main gel layer.

Method for producing meat or fish analogues with different texture by directional freezing of protein gel

Using protein gels (rather than hydrocolloid gels) and subsequently directionally freezing them as described previously, various food analog products with different textures can be produced. This still produced protein fibers, but with different textures obtained using hydrocolloid gels. All other steps of producing the final food product are the same as the hydrocolloid gels discussed above.

Example # 3-textured food analog product

The heat-set gel (a protein gel made by heating a solution of a heat-gellable protein) was produced by heating a solution of 12 wt.% protein, 50mm nacl, ph7 for about 30 minutes at about 80 ℃. The sample was cooled to about 4 ℃ in a refrigerator. The protein gel was directionally frozen until it was completely frozen, after which it was left at room temperature for thawing. After thawing, a fibrous/lamellar structure is observed (as shown in fig. 7C). Proteins that may be used to produce the protein gel include, but are not limited to, any one or combination of whey protein, soy protein, potato protein, rubisco protein, duckweed protein, rice protein, almond protein, egg protein, oat protein, linseed protein, euglena protein, schizochytrium, mung bean protein, pea protein, recombinant mammalian whey, cultured mammalian whey, recombinant egg albumin, cultured egg albumin, recombinant gelatin or collagen, cultured gelatin or collagen, canola protein, lupin protein, faba protein, wheat protein, hyacinth bean protein, amaranth protein, peanut protein, moringa seed protein, pumpkin seed protein, chickpea protein, sunflower seed protein, safflower seed protein, mustard seed protein, chlorella protein, and spirulina protein.

Directionally freezing a solution of a single biopolymer or mixture of biopolymers and injecting it with the single biopolymer or mixture of biopolymers

Those skilled in the art will appreciate that the first biopolymer solution may be a hydrocolloid solution, a protein solution, or a mixed protein hydrocolloid solution. The second biopolymer solution may be a hydrocolloid solution, a protein solution or a mixed protein hydrocolloid solution.

Method for producing meat or fish analogues with different texture by directional freezing of a solution containing both protein and hydrocolloid

A complex of hydrocolloid and protein can be produced and subjected to directional freezing. The resulting composite food analog forms a more fibrous texture than either a protein gel infused with pure protein or a hydrocolloid gel infused with protein. Oriented freezing may be followed by replacement of aligned elongated ice crystals with protein using any of the methods described previously.

Example # 4-hydrocolloid and protein Complex 1

In this example, canola and potato proteins, sodium alginate, caCO are used ₃ And GDL to produce gels. A 20 wt% protein solution was prepared and stored overnight at 4 ℃. Sodium alginate was dissolved in the solution at a concentration of 1 wt%. CaCO with equivalent weight of 15mM ₃ Disperse in solution for 20 minutes, then dissolve GDL equivalent to 30mM for 5 minutes. The mixture was then degassed in an ultrasonic bath for 5 minutes and poured into moulds on cold plates at-15 ℃. Once completely frozen, the samples were stored at-18 ℃ for 24 hours and then thawed at 4 ℃ for 24 hours. The thawed fibrous food analog product is then cooked, producing a visual chicken-like texture (fig. 8).

Directional freezing of a gel of a single biopolymer or mixture of biopolymers and injection thereof with the single biopolymer or mixture of biopolymers

Those skilled in the art will appreciate that the biopolymer gel may be a hydrocolloid gel, a protein gel, or a mixed protein hydrocolloid gel. The biopolymer solution may be a hydrocolloid solution, a protein solution or a mixed protein hydrocolloid solution.

Method for producing meat or fish analogues with different texture by directional freezing of a gel containing both protein and hydrocolloid

A complex of hydrocolloid and protein can be produced and subjected to directional freezing. The resulting composite food analog forms a fibrous texture different from that of a protein gel infused with pure protein or a hydrocolloid gel infused with protein. Oriented freezing may be followed by replacement of aligned elongated ice crystals with protein using any of the methods described previously.

Example # 5-hydrocolloid and protein Complex 2

In this example, the gel was produced by mixing a 20 wt% hydrolyzed rice protein solution with a 4 wt% agar solution at a ratio of 1:1 at 85 ℃. The homogeneous mixture was cooled, gelled, and frozen in a directed manner. The gel was then placed in a protein solution containing 12 wt% potato protein, 50mM NaCl and about 0.1 wt% pigment overnight at 4℃to melt the ice crystals and infuse the protein solution into the directionally frozen structure. The next day, the sample was heated in a glass beaker placed in a water bath at about 55 ℃ for about 20 minutes. The result is a product with a fibrous structure.

Production of skin layers

After the production of the fibrous meat analogue food, an algae-hydrocolloid composite film similar to animal/fish skin can be produced by immersing algae (e.g., laver) sheets in an alginate solution or an alginate-oil emulsion (see fig. 6), laminating it onto the fibrous meat analogue food, gelling the alginate solution, and then partially drying the resulting gel. A non-limiting method for gelling the mixture is to immerse it in a 2 wt% calcium chloride solution for about two (2) minutes. The skinned product may be packaged with the skin dry or wet. In either case, over time, the epidermis will become moist due to the balancing action.

The above examples of skin layers produced from algae and mixtures of alginate solutions or alginate-oil emulsions are merely illustrative and not limiting. The epidermis layer may be made of a variety of materials including vegetable proteins, carrageenan, furcellaran and konjak to give a few examples.

Color change

As shown in fig. 9, when the composition of the protein solution is optimized for color, there is a significant color and opacity change before and after heat treatment (i.e., from "as-grown" to "after cooking"), thereby improving the actual appearance of a piece of meat-like material undergoing cooking. In this case, the appearance before cooking is shown in the left figure, and after frying in a pan, the fibrous gel undergoes shrinkage and increases in brightness, as shown in the right figure. Fig. 10 shows that this improves the real appearance of a piece of meat-like material during frying in a pan. Fig. 11 shows the product in an uncooked state, so it is translucent. The change in color and opacity occurs at least due to partial denaturation and subsequent aggregation of the protein. The generation of aggregates greater than the visible wavelength results in scattering of light, resulting in a more opaque appearance. Such colors may undergo chemical changes during the cooking process, further resulting in a change in appearance and/or a change in opacity.

Food coloring

In order to obtain a fibrous meat analogue food product having a final color that looks like the meat product it simulates, various food colors may be mixed with the protein such that this mixture replaces the elongated ice crystals. For example, to make salmon analogs with the typical pink color of raw salmon, pigments including, but not limited to, carotenoids, astaxanthin, lycopene, carmine, anthocyanins, carthamin, lutein, curcumin, capsanthin, norbixin, curcuminoids, phycocyanins, melanoids, and betalains may be used. For beef, the following pigments may be used, but are not limited to hemoglobin, myoglobin, anthocyanin, pomegranate juice extract, beet juice extract, and betalains.

Raw fibrous meat analogue

Although the production of a fibrous meat analogue food has been described, involving a step of gelling at least some of the proteins, it will be appreciated that a "raw fibrous meat analogue" may be produced for transport, wherein no proteins are gelled in the production of protein-infused hydrocolloids. This method involves preparing an ingestible hydrocolloid gel comprised of one or more different ingestible hydrocolloids, then directionally freezing it, inducing the formation of aligned elongate ice crystals to form a directionally frozen hydrocolloid gel having aligned elongate channels in which the aligned elongate ice crystals are located. The ice crystals are then replaced with an ingestible protein to produce a protein infused hydrocolloid gel. After the protein-infused hydrocolloid gel is produced, an ingestible substance is introduced into the protein-infused hydrocolloid gel, the ingestible substance acting to prevent leakage of the protein from the protein-infused hydrocolloid gel to produce a raw fibrous meat analogue food product. The raw fibrous meat analogue food is then packaged for shipment.

Non-limiting examples of ingestible substances that are incorporated into protein-infused hydrocolloid gels to prevent protein leakage include pH, salts, heat treatments, chemical crosslinking, enzymatic crosslinking, injection of gelling hydrocolloids such as sodium alginate, curdlan, methylcellulose, or application of hydrocolloid coatings such as calcium-gelled alginate solutions.

The step of replacing aligned elongated ice crystals with an ingestible protein to produce a protein infused hydrocolloid gel and the step of adding an ingestible substance to the protein infused hydrocolloid gel to prevent protein leakage may be performed using the same steps described above for preparing analogs with some of the gelled proteins.

The end consumer will then cook the product for consumption, or alternatively, the fibrous meat analogue food product may be designed to be eaten raw. Non-limiting examples of such raw fibrous meat analog foods include, but are not limited to, sushi or other raw seafood products.

Biopolymer solutions and dispersions

In some embodiments of producing fibrous meat analogs described herein, the biopolymer solution and dispersion are gelled separately from the protein. In some embodiments, the biopolymer solution and dispersion are gelled prior to replacement of ice crystals with ingestible proteins and/or hydrocolloids. In one embodiment, the gelation of the biopolymer solution and dispersion is accomplished by the same suitable conditions as the gelation of the ingestable protein.

In some embodiments, the biopolymer solution and dispersion are gelled under the same type or step as directional freezing. In one example, alginate is dissolved in water at room temperature and then agar is added. To completely dissolve the agar, this solution was heated to 85 ℃. This heated solution was then partially cooled to 60℃at which point CaCO was added ₃ GDL solution. Adding CaCO ₃ The partially cooled solution of/GDL is poured onto the frozen surface in the mold to fully orient the freezing. The directionally frozen samples were placed in an immersion liquid and kept in the mold at 4 ℃ overnight.

In some embodiments, the biopolymer solution and dispersion comprise a mixed hydrocolloid solution in which one hydrocolloid is gelled before the second hydrocolloid. In one embodiment, the first hydrocolloid is gelled using a gelling agent that gels only the first hydrocolloid, and then the frozen biopolymer is oriented. Subsequently, the ingestible protein is thawed in a gelling agent that gels the second hydrocolloid prior to injection. In one example, agar and alginate powders are dry mixed at various concentrations and added to water to prepare an agar-alginate solution. This solution was heated to 90 ℃ and poured into a mold and allowed to set. At this stage, the agar was gelled. The agar-alginate gel was placed on a frozen surface and allowed to freeze in orientation. Placing the frozen sample into a freezer <CaCl at 7 ℃ C.) ₂ Solution [ ]>0.1 wt%) and stored overnight in an ice bath, allowing the alginate to gel.

The specific embodiments described above have been shown by way of example, and it should be understood that these embodiments may be subject to various modifications and alternatives. It should be further understood that the claims are not intended to be limited to the particular forms disclosed, but rather to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure.

Example A

1. A method for producing a fibrous meat analogue comprising: subjecting an ingestible biopolymer gel, such as a polysaccharide hydrogel, to directional freezing, inducing the formation of aligned elongate ice crystals to form a directionally frozen biopolymer gel (frozen polysaccharide hydrogel) having aligned channels in which the aligned elongate ice crystals are located; thawing a directionally frozen biopolymer gel (frozen ingestible polysaccharide hydrogel) having aligned channels by immersing the frozen biopolymer (frozen ingestible polysaccharide hydrogel) in a solution containing at least one ingestible soluble thermogelling protein, thereby thawing and replacing the aligned elongated ice crystals with the at least one ingestible soluble thermogelling protein at a temperature below the gelation temperature of the soluble thermogelling protein to produce a protein infused biopolymer gel (protein infused polysaccharide hydrogel), wherein the protein loading varies based on the immersion time; and heating the protein infused biopolymer gel (protein infused ingestible polysaccharide hydrogel) at a temperature above the gelation temperature of the at least one ingestible soluble thermogelling protein to produce protein fibers, thereby forming a fibrous meat analog food product.

2. The method of embodiment 1, wherein the at least one ingestible soluble thermogelling protein comprises: i) An ingestible soluble thermogelling protein, wherein the concentration of the ingestible soluble thermogelling protein in a solution comprising at least one ingestible soluble thermogelling protein is in the range of about 0.5% to about 30%; or ii) a mixture of ingestible soluble thermogelling proteins and non-thermogelling proteins.

3. The method of embodiment 1, wherein the ingestible biopolymer gel (ingestible polysaccharide hydrogel) has a melting temperature above the gelation temperature of the ingestible soluble thermal gelation protein.

4. The method of embodiment 1, wherein the ingestible soluble thermogelling protein is any one or a combination of Whey Protein Isolate (WPI), soy protein, potato protein isolate, rubisco protein, mung bean protein, and pea protein.

5. The method of embodiment 1 wherein the solution containing at least one ingestible soluble thermal gelling protein further comprises an ingestible non-thermal gelling protein and a thermally induced trigger to trigger gelling of the ingestible non-thermal gelling protein upon an increase in temperature.

6. The method of embodiment 1, wherein the step of heat treating the biopolymer gel of the ingestible soluble thermogelling infusion protein (polysaccharide hydrogel) is performed at a solution temperature in the range of about 40 ℃ to about 150 ℃.

7. The method of embodiment 5, wherein the thermally induced trigger comprises: salts, enzymes, pH modifiers or combinations thereof.

8. The method of embodiment 5, wherein the thermally induced trigger is an enzyme microencapsulated within a meltable coating.

9. The method of embodiment 1, wherein the ingestible biopolymer gel (ingestible polysaccharide hydrogel) is selected from the group consisting of: agar, fermentation-derived gelatin, alginates, curdlan, kappa-carrageenan, kappa 2-carrageenan and iota-carrageenan, furcellaran, starch, modified starch, dextrin, konjac glucomannan, gellan gum, and combinations of xanthan gum, guar gum, locust bean gum and tara gum.

10. The method of embodiment 1, wherein the solution comprising at least one ingestible soluble thermogelling protein comprises: an aqueous solution or an aqueous dispersion.

11. The method of embodiment 1, wherein the at least one ingestible soluble thermogelling protein is a mixture comprising: ingestible thermo-gelling proteins and ingestible non-thermo-gelling proteins; and wherein the solution containing at least one ingestible soluble thermogelling protein comprises from about 15 to about 25 wt% protein.

12. The method of embodiment 1, wherein the concentration of the at least one ingestible, soluble, thermogelling protein in the solution is in the range of about 10 to about 30 wt%.

13. The method of embodiment 1, wherein the solution comprising at least one ingestible soluble thermogelling protein has a temperature of about 1 ℃ to about 60 ℃.

14. The method of embodiment 1, wherein the protein loading is further varied by varying the volume ratio of the ingestible biopolymer gel (ingestible polysaccharide hydrogel) and the solution comprising the at least one ingestible soluble thermogelling protein.

15. The method of embodiment 1, wherein the step of directionally freezing the ingestible biopolymer gel (ingestible polysaccharide hydrogel) is performed by placing the ingestible biopolymer gel (ingestible polysaccharide hydrogel) in contact with a pre-cooled substrate at a temperature of about-2 ℃ to about-196 ℃.

16. The method of embodiment 1, wherein the ingestible biopolymer gel (ingestible polysaccharide hydrogel) comprises a kappa-carrageenan hydrogel.

17. The method of embodiment 1, wherein the ingestible biopolymer gel (ingestible polysaccharide hydrogel) is a kappa-carrageenan hydrogel having a modulus in the range of about 100 to about 5000 pascals.

18. The method of embodiment 1, wherein the ingestible biopolymer gel (ingestible polysaccharide hydrogel) is an agar hydrogel having an agar concentration in the range of about 0.1% to about 15% by weight.

19. The method of embodiment 1, wherein the solution containing at least one ingestible soluble thermogelling protein further comprises an ingestible supplement that diffuses into the aligned channels.

20. The method of embodiment 10, wherein the aqueous solution or aqueous dispersion comprises: flavoring agents, emulsifying agents, preserving agents, pigments, pH adjusting agents, texture adjusting agents, or combinations thereof.

21. The method of embodiment 19 wherein the ingestible supplement comprises: emulsions of esters of omega-3, omega-6, omega-9 fatty acids, or combinations thereof.

22. The method of embodiment 19 wherein the ingestible supplement comprises: water-soluble vitamins including ascorbic acid (vitamin C), thiamine, riboflavin, niacin, and vitamin B ₆ (pyridoxine, pyridoxal and pyridoxamine), folic acid, vitamin B ₁₂ Biotin and pantothenic acid.

23. The method of embodiment 19, wherein the ingestible supplement comprises an ingestible mineral.

24. The method of embodiment 19, wherein the ingestible supplement comprises a water-insoluble vitamin.

25. The method of embodiment 19, wherein the ingestible supplement comprises an antioxidant.

26. The method of embodiment 1, wherein the step of subjecting the ingestible biopolymer gel (ingestible polysaccharide hydrogel) to directional freezing and inducing the formation of aligned elongated ice crystals further comprises: placing the ingestible biopolymer gel (ingestible polysaccharide hydrogel) in contact with a pre-cooled substrate for directional freezing in one direction; or placing the ingestible biopolymer gel (ingestible polysaccharide hydrogel) between two pre-cooled substrates, wherein the directional freezing is performed from opposite directions.

27. The method of embodiment 2, wherein the concentration of the ingestible, thermal gelling protein in the solution comprising at least one ingestible, soluble, thermal gelling protein in the mixture of thermal gelling proteins and non-thermal gelling proteins is in the range of about 2 to about 10 wt%, the remainder being non-thermal gelling proteins, to make up a total of 25 wt% of the protein mixture.

Example B

1. A method for producing a fibrous meat analogue comprising: preparing an ingestible hydrocolloid gel consisting of one or more different ingestible hydrocolloids and water; subjecting the ingestible hydrocolloid gel to directional freezing, inducing the formation of aligned elongate ice crystals to form a directional frozen hydrocolloid gel having aligned elongate channels in which the aligned elongate ice crystals are located; replacing the aligned elongated ice crystals with an ingestible protein to produce a protein infused hydrocolloid gel; and subjecting the protein-infused hydrocolloid gel to conditions suitable for gelling at least some of the ingestible proteins to produce a protein gel within the aligned channels, thereby forming a fibrous meat, poultry, or seafood analog food product.

2. The method of embodiment 1, wherein the hydrocolloid gel is a polysaccharide hydrogel.

3. The method of embodiment 1, wherein the hydrocolloid gel is conventional gelatin, recombinant gelatin, or a combination of both.

4. The method of embodiment 1, wherein the ingestible protein is a mixture of gellable and non-gellable proteins.

5. The method of embodiment 1, wherein the ingestible protein is a gellable protein.

6. The method of embodiment 1, wherein the ingestible protein is a mixture of ingestible and non-thermal gellable proteins, and wherein the conditions suitable for gelling the at least some of the ingestible, thermal gellable proteins comprise heating the protein-infused hydrocolloid gel to a temperature that enables gelling of the at least some of the thermal gellable proteins.

7. The method of embodiment 1, wherein the at least some of the ingestible proteins are ingestible, thermogelable proteins, and wherein the conditions suitable for gelling the ingestible, thermogelable proteins comprise heating the protein-infused hydrocolloid gel to induce denaturation of the at least some of the thermogelable proteins.

8. The method of embodiment 7, wherein the protein infused hydrocolloid gel is heat treated at a temperature in the range of about 40 ℃ to about 75 ℃ to induce gelation, wherein the temperature depends on the gelation temperature of the protein.

9. The method of embodiment 6, wherein the ingestible, thermogelable proteins comprise one or more different types of ingestible, thermogelable proteins, one or more types of non-thermogelable proteins, and one or more types of non-gelable proteins.

10. The method of embodiment 1, wherein the conditions suitable for gelling the at least some of the ingestible proteins comprise penetration of a salt into the hydrocolloid gel of the injected protein, the salt being selected to induce gelling of the at least some of the ingestible proteins.

11. The method of embodiment 10, wherein salt is infiltrated into the protein infused hydrocolloid gel by injecting a salt solution into the protein infused hydrocolloid gel.

12. The method of embodiment 10 wherein salt is infiltrated into the protein infused hydrocolloid gel by adding salt as a crystalline solid to the surface of the protein infused hydrocolloid gel, which is then dissolved and diffused into any available water present in the protein infused hydrocolloid gel.

13. The method of embodiment 10 wherein salt is infiltrated into the protein-infused hydrocolloid gel by contacting the protein-infused hydrocolloid gel with a concentrated salt solution that diffuses into the protein-infused hydrocolloid gel, whereby the concentration of salt solution is sufficient to allow gelation of the protein, and the desired concentration will depend on the type of protein and the type of salt.

14. The method of embodiment 10, wherein the salt is any one of sodium (Na), potassium (K), calcium (Ca), and magnesium (Mg) sulfate, citrate, ascorbate, acetate, sorbate, lactate, tartrate, gluconate, and phosphate, and any combination thereof.

15. The method of embodiment 1, wherein the conditions suitable for gelling the at least some of the ingestible proteins comprise adjusting the pH of the protein-infused hydrocolloid gel to a value suitable for causing gelling of the at least some of the ingestible proteins.

16. The method of embodiment 15, wherein the pH is adjusted by adding a pH adjuster suitable for food in liquid form, in solution form, or adding a soluble pH adjuster in solid form.

17. The method of embodiment 16, wherein the pH adjuster is any one or combination of acetic acid, hydrochloric acid, ascorbic acid, malic acid, formic acid, tartaric acid, citric acid, glucono-delta lactone, sodium hydroxide, potassium hydroxide, and calcium hydroxide.

18. The method of embodiment 1, wherein the conditions suitable for gelling the at least some of the ingestible proteins comprise penetrating a solution containing an enzyme-based cross-linking agent into the protein-infused hydrocolloid gel, wherein the enzyme-based cross-linking agent is selected to induce gelling of the at least some of the ingestible proteins.

19. The method of embodiment 18, wherein the enzyme cross-linking agent comprises any one or a combination of transglutaminase (EC 2.3.2.13), sortase a (EC 3.4.22.70), tyrosinase (EC 1.14.18.1), laccase (EC 1.10.3.2), peroxidase (EC 1.11.1. X), lysyl oxidase (EC 1.4.3.13), and amine oxidase (EC 1.4.3.6).

20. The method of embodiment 1, wherein the conditions suitable for gelling the at least some of the ingestible proteins comprise pressure treating the protein-infused hydrocolloid gel to induce gelling of the at least some of the ingestible proteins.

21. The method of embodiment 20 wherein the protein infused hydrocolloid gel food product is sealed and placed in a rigid sealed compartment containing a liquid and the liquid is pressurized.

22. The method of embodiment 1, wherein the conditions suitable for gelling the at least some of the ingestible proteins comprise penetrating a solution containing a chemical cross-linking agent into the protein-infused hydrocolloid gel, the chemical cross-linking agent being selected to induce gelling of the at least some of the ingestible proteins.

23. The method of embodiment 22, wherein the chemical cross-linking agent is any one or a combination of glutaraldehyde, tannic acid, genipin, and liquid smoke.

24. The method of embodiment 1, wherein the conditions suitable for gelling the at least some of the ingestible proteins comprise irradiating the protein-infused hydrocolloid gel with radiation of a suitable wavelength and intensity to induce cross-linking of proteins, thereby inducing gelling of the at least some of the ingestible proteins.

25. The method of embodiment 1 wherein replacing the aligned elongated ice crystals with the ingestible protein comprises thawing the directionally frozen hydrocolloid gel by immersion in a solvent containing the ingestible protein, the solvent having a temperature suitable for thawing ice crystals replaced by the ingestible protein to produce the protein infused hydrocolloid gel.

26. The method of embodiment 25, wherein thawing the directionally frozen hydrocolloid gel comprises adjusting the temperature of the solvent comprising the ingestible gellable protein to a range from the melting point of the solvent within the hydrocolloid gel to the melting point of the hydrocolloid gel, and wherein replacing the aligned elongated ice crystals with the ingestible protein comprises adjusting the temperature of the solvent comprising the ingestible protein to a range from the freezing point of the solvent comprising a protein solution to the gelation onset denaturation temperature of the at least some of the ingestible proteins.

27. The method of embodiment 26, wherein thawing the directionally frozen hydrocolloid gel is performed between about 0 ℃ and about 85 ℃.

28. The method of embodiment 26, wherein replacing the aligned elongated ice crystals with the ingestible protein occurs between about 0 ℃ to about 45 ℃.

29. The method of embodiment 25, wherein the solvent is a non-aqueous solvent suitable for a food in which the ingestible protein is soluble.

30. The method of embodiment 29, wherein the solvent is any one or combination of acetic acid, formic acid, ethanol, methanol, propanol, and mixtures thereof with water.

31. The method of embodiment 25, wherein the solvent is an aqueous solution and the ingestible protein is soluble or dispersible in the aqueous solution.

32. The method of embodiment 31, wherein the aqueous solution containing the ingestible protein is at a temperature between about 1 ℃ and about 99 ℃.

33. The method of embodiment 31 wherein the aqueous solution containing the ingestible protein is at a temperature between about 99 ℃ and about 130 ℃ and comprising subjecting the thawed hydrocolloid gel to a pressure in the range of about 0 to 1.7 bar in a self-pressurized closed container.

34. The method of embodiment 1, wherein replacing the aligned elongated ice crystals with the ingestible protein comprises subjecting the directionally frozen hydrocolloid gel to conditions suitable for sublimating the elongated ice crystals in the presence of the ingestible gellable protein.

35. The method of embodiment 34 wherein subjecting the directionally frozen hydrocolloid gel to conditions suitable for sublimating the elongated ice crystals comprises subjecting the directionally frozen hydrocolloid gel to vacuum, resulting in sublimation of ice, and immersing the sublimated hydrocolloid gel in a solution containing the ingestible protein, thereby injecting a solution containing the ingestible protein into the sublimated hydrocolloid gel.

36. The method of embodiment 1 wherein replacing the aligned elongated ice crystals with an ingestible protein comprises freeze-drying the directionally frozen hydrocolloid gel to remove substantially all water, and then immersing the dried gel in a solution containing the ingestible protein.

37. The method of embodiment 1 wherein replacing the aligned elongated ice crystals with an ingestible protein comprises subjecting the directionally frozen hydrocolloid gel to conditions suitable to cause evaporation of ice to remove substantially all ice, and then immersing the dried gel into a solution containing the ingestible protein.

38. The method of embodiment 1, wherein the ingestible protein is an animal protein.

39. The method of embodiment 35, wherein the animal protein comprises a recombinant animal protein.

40. The method of embodiment 1, wherein the ingestible protein is any one or a combination of a plant-based protein, a bacterial-based protein, a fungal-based protein, and an algae-based protein.

41. The method of embodiment 40 wherein the fungal-based protein comprises yeast.

42. The method of embodiment 40, wherein the algae is any one or a combination of macroalgae and microalgae.

43. The method of embodiment 1, wherein the ingestable gellable protein is any one or any combination of an animal based protein, a recombinant protein, a culture protein, a plant based protein, a bacterial based protein, a fungal based protein, and an algae based protein, all of which are suitable for use in food products.

44. The method of embodiment 1, wherein the ingestible protein is any one or any combination of whey protein, soy protein, potato protein, rubisco protein, duckweed protein, rice protein, almond protein, egg protein, oat protein, linseed protein, euglena protein, schizochytrium, mung bean protein, pea protein, recombinant mammalian whey, cultured mammalian whey, recombinant egg albumin, recombinant gelatin or collagen, cultured gelatin or collagen, canola protein, lupin protein, fava protein, wheat protein, lentil protein, amaranth protein, peanut protein, pepperylene protein, pumpkin seed protein, chickpea protein, sunflower seed protein, safflower seed protein, mustard seed protein, chlorella protein, and spirulina protein.

45. The method of embodiment 1, wherein in the fibrous meat analog food product the ingestible protein is present in a range of about 5 wt% to about 35 wt% and the hydrocolloid is present in a range of about 0.2 wt% to about 10 wt%.

46. The method of embodiment 1, wherein in the fibrous meat analog food product the ingestible protein is present in a range of about 10 wt% to about 30 wt% and the hydrocolloid is present in a range of about 0.5 wt% to about 8 wt%.

47. The method of embodiment 1, wherein in the fibrous meat analog food product the ingestible protein is present in a range of about 10 wt% to about 20 wt% and the hydrocolloid is present in a range of about 1 wt% to about 5 wt%.

48. The method of embodiment 6, wherein in the fibrous meat analog food product the ingestible protein is present in a range of about 5 wt% to about 35 wt% and the hydrocolloid is present in a range of about 0.2 wt% to about 10 wt%.

49. The method of embodiment 6, wherein in the fibrous meat analog food product the ingestible protein is present in a range of about 10 wt% to about 30 wt% and the hydrocolloid is present in a range of about 0.5 wt% to about 8 wt%.

50. The method of embodiment 6, wherein in the fibrous meat analog food product the ingestible protein is present in a range of about 10 wt% to about 20 wt% and the hydrocolloid is present in a range of about 1 wt% to about 5 wt%.

51. The method of embodiment 1, wherein the ingestible gellable protein is a mixture of ingestible thermogellable proteins and non-gellable proteins to provide an increased protein content.

52. The method of embodiment 51, wherein in the fibrous meat analog food product the total amount of the ingestible protein is present in a range of about 5 wt% to about 50 wt%, and the hydrocolloid is present in a range of about 0.2 wt% to about 10 wt%.

53. The method of embodiment 51, wherein in the fibrous meat analog food product the total amount of ingestible protein is present in the range of about 10 wt% to about 30 wt%, and the hydrocolloid is present in the range of about 0.5 wt% to about 8 wt%.

54. The method of embodiment 51, wherein in the fibrous meat analog food product the total amount of ingestible protein is present in the range of about 15 wt% to about 25 wt%, and the hydrocolloid is present in the range of about 1 wt% to about 5 wt%.

55. The method of example 51, wherein the maximum amount of protein present is 25% by weight, then the minimum amount of ingestible, thermally gellable protein in the mixture is 2% by weight, and the maximum amount of non-thermally gellable protein is 23% by weight.

56. The method of example 51, wherein the maximum amount of protein present is 25 wt% and the intermediate minimum amount of ingestible, thermally gellable protein in the mixture is about 8 wt% and the maximum amount of non-thermally gellable protein is about 17 wt%.

57. The method of example 51, wherein the maximum amount of protein present is 25 wt% and the intermediate minimum amount of ingestible, thermally gellable protein in the mixture is about 10 wt% and the maximum amount of non-thermally gellable protein is about 15 wt%.

58. The method of embodiment 1, further comprising controlling the diameter of the aligned elongate channels by controlling a temperature gradient through the material so as to vary the speed of the directional freezing process, and wherein the diameter of the protein fibers is proportional to the diameter of the aligned elongate channels.

59. The method of embodiment 58 wherein the diameters of the aligned elongated channels are controlled to obtain protein fibers having diameters in the range of about 20 to about 200 microns.

60. The method of embodiment 1, wherein the ingestible hydrocolloid gel is a composite ingestible hydrocolloid gel composed of one or more different types of ingestible hydrocolloids.

61. The method of embodiment 60, wherein the two or more different types of hydrocolloids are at least one of a polysaccharide hydrocolloid, gelatin, or recombinant gelatin.

62. The method of embodiment 60, wherein the composite ingestible hydrocolloid gel is produced from a homogeneous mixture of the two or more different types of hydrocolloids.

63. The method of embodiment 60 wherein the composite ingestible hydrocolloid gel has a layered structure wherein alternating layers are made of different hydrocolloids or hydrocolloid blends.

64. The method of embodiment 61 wherein the composite ingestible hydrocolloid gel has a layered structure wherein alternating layers are made of the same hydrocolloid or hydrocolloid blend.

65. The method of embodiment 25, further comprising subjecting the ingestible hydrocolloid gel to multiple cycles of directional freezing and thawing in the absence of protein.

66. The method of embodiment 25, further comprising subjecting the ingestible hydrocolloid gel to multiple cycles of directional freezing and thawing in the presence of protein.

67. The method of embodiment 1, further comprising preparing a layer simulating the skin layer of meat or fish by preparing a mixture of agar and an alginate solution with an alginate-oil emulsion, gelling the mixture to prepare a skin layer, and partially drying the skin layer, followed by lamination of the skin layer onto a fibrous meat analogue food.

68. The method of embodiment 1, further comprising producing a plurality of protein infused hydrocolloid gels of preselected thickness, comprising the steps of: a) preparing a caulking layer made of a material selected to mimic connective tissue of meat and/or fish, applying the caulking material to a surface of one of the protein-infused hydrocolloid gels, b) placing the other protein-infused hydrocolloid gel on top of the caulking layer, and c) repeating steps a) and b) until a plurality of protein-infused hydrocolloid gels are stacked together.

69. The method of embodiment 68 wherein the interstitial layer of a material selected to mimic connective tissue of meat and/or fish comprises any one or combination of proteins, hydrocolloids, oil-in-water emulsions, solid particles, fats, and oil gels.

70. The method of embodiment 69, wherein the solid particles comprise any one or a combination of titanium dioxide, protein, calcium carbonate, and starch, solid fat crystals, and algae.

71. A method for producing a fibrous meat analogue comprising: subjecting the ingestible protein gel to directional freezing, inducing the formation of aligned elongated ice crystals to form a directional frozen protein gel having aligned channels in which the aligned elongated ice crystals are located; replacing the aligned elongated ice crystals with an ingestible protein to produce a protein gel infused with the protein; and subjecting the protein infused protein gel to conditions suitable for gelling at least some of the ingestible proteins to produce protein fibers in the aligned channels, thereby forming a fibrous meat analog food product.

72. The method of embodiment 71, wherein the ingestible protein is any one or any combination of whey protein, soy protein, potato protein, rubisco protein, duckweed protein, rice protein, almond protein, oat protein, linseed protein, euglena protein, schizochytrium limacinum protein, mung bean protein, pea protein, recombinant whey, cultured whey, recombinant egg albumin, cultured egg albumin, recombinant gelatin or collagen, cultured gelatin or collagen, canola protein, lupin protein, faba protein, wheat protein, hyacinth bean protein, amaranth protein, peanut protein, pepperylene seed protein, pumpkin seed protein, chickpea protein, sunflower seed protein, safflower seed protein, mustard seed protein, chlorella protein, and spirulina protein.

73. The method of embodiment 71, wherein the ingestible protein is made of the same ingestible protein that comprises the ingestible protein gel.

74. The method of embodiment 71 wherein the ingestible protein that replaces ice crystals comprises a mixture of gellable and non-gellable proteins.

75. The method of embodiment 71, wherein the ingestable protein comprises a mixture of gellable and non-gellable proteins.

76. The method of embodiment 71, wherein the ingestible protein gel is a complex ingestible protein gel comprising a mixture of different proteins.

77. The method of embodiment 71, further comprising subjecting the ingestible protein gel to multiple cycles of directed freezing and thawing in the absence of protein to increase the gel strength of the protein gel.

78. The method of embodiment 71, further comprising subjecting the ingestible protein gel to multiple cycles of directed freezing and thawing in the presence of a protein to increase the gel strength of the protein gel and the protein content of the protein gel.

79. A method for producing a fibrous meat analogue comprising: preparing a complex ingestible gel comprised of one or more different types of ingestible hydrocolloid and one or more different types of ingestible protein; subjecting the composite ingestible gel to directional freezing, inducing the formation of aligned elongate ice crystals to form a directionally frozen composite gel having aligned channels in which the aligned elongate ice crystals are located; replacing the aligned elongated ice crystals with any one or a combination of an ingestible protein, hydrocolloid, and complex to produce a complex ingestible gel infused with the protein; and subjecting the protein infused complex ingestible gel to conditions suitable for gelling at least some of the ingestible proteins to produce protein fibers in the aligned channels, thereby forming a fibrous meat analog food product.

80. The method of embodiment 71, further comprising subjecting the protein-infused complex ingestible gel to multiple cycles of directed freezing and thawing in the absence of protein.

81. The method of embodiment 71, further comprising subjecting the protein-infused, composite ingestible gel to multiple cycles of directed freezing and thawing in the presence of a protein to increase the gel strength of the protein-infused, composite ingestible gel and the protein content of the protein-infused, composite ingestible gel.

82. The method of embodiment 1 wherein the step of replacing the aligned elongated ice crystals with an ingestible protein to produce a protein infused hydrocolloid gel comprises replacing the aligned elongated ice crystals with a mixture of an ingestible protein and a hydrocolloid.

83. The method of embodiment 71 wherein the step of replacing the aligned elongated ice crystals with an ingestible protein to produce a protein gel infused with protein comprises replacing the aligned elongated ice crystals with a mixture of an ingestible protein and a hydrocolloid.

84. The method of embodiment 79 wherein the step of replacing the aligned elongated ice crystals with an ingestible protein to produce a protein-infused hydrocolloid gel comprises replacing the aligned elongated ice crystals with a mixture of an ingestible protein and a hydrocolloid.

85. The method of embodiment 1, wherein the step of preparing an ingestible hydrocolloid gel comprised of one or more different ingestible hydrocolloids comprises: adding a color during the preparation of the ingestible hydrocolloid gel, or replacing the aligned elongated ice crystals with a mixture of an ingestible protein and a color, and wherein the color is selected to impart a color to the fibrous meat analogue food product that is the actual color of the meat product of which the fibrous meat analogue food product is an analogue.

86. The method of embodiment 85, wherein the pigment is selected from the group consisting of: carotenoids, beta-carotene, astaxanthin, lycopene, carmine, anthocyanidins, betalains, haemoglobin, myoglobin, betalains extract, carthamin, lutein, curcumin, capsanthin, bilirubin, anthocyanidins, curcuminoids, phycocyanin and melanoidin.

87. The method of embodiment 67 wherein the step of producing a layer that mimics the skin layer of meat or fish comprises adding flavoring agents and coloring agents to the mixture to impart to the skin layer an appearance and taste that mimics an actual food that the fibrous meat analog food product mimics.

88. The method of embodiment 1, wherein subjecting a solution of a single or a set of hydrocolloids, or a solution of a single or a set of hydrocolloids and a single or a set of proteins to simultaneous directional freezing and gelling induces the formation of aligned elongated ice crystals to form a directional frozen gel having aligned elongated channels in which the aligned elongated ice crystals are located.

89. The method of embodiment 1 wherein the ice crystals are replaced with a solution or dispersion containing protein and hydrocolloid.

90. The method of embodiment 1 wherein the step of replacing the elongated ice crystals comprises replacing the elongated ice crystals with a mixture of proteins and pigments selected to impart a preselected color to the fibrous meat analogue food product.

91. The method of embodiment 1 wherein the step of preparing an ingestable prepares a mixture of the one or more different ingestible hydrocolloids with water and an ingestible protein, and wherein the step of subjecting the ingestible hydrocolloid gel to directional freezing comprises subjecting the ingestible hydrocolloid gel composed of one or more different ingestible hydrocolloids, water, and protein to directional freezing.

92. A fibrous meat analogue food product produced according to the method of example 1.

93. The fibrous meat analog food of embodiment 92, comprising packaging the fibrous meat analog food to form a food product that is transported to a consumer for cooking by the consumer.

94. A fibrous meat analogue food product produced according to the method of claim 71.

95. The fibrous meat analog food product of embodiment 94, comprising packaging the fibrous meat analog food product to form a food product that is transported to a consumer for cooking by the consumer.

96. A fibrous meat analogue food product produced according to the method of example 79.

97. The fibrous meat analog food product of embodiment 96, comprising packaging the fibrous meat analog food product to form a food product that is shipped to a consumer for cooking by the consumer.

98. A fibrous, edible, protein-enriched food analog product that can be cooked, the fibrous, edible, protein-enriched food analog product comprising an ingestible protein-infused hydrocolloid gel, wherein in the fibrous, edible, protein-enriched food analog product the protein is present in a range of about 2 wt% to about 50 wt% and the hydrocolloid is present in a range of about 0.2 wt% to about 10 wt%.

99. The product of embodiment 98, wherein in the fibrous, edible, protein-enriched food analog product, the ingestible protein is present in a range of about 10 wt% to about 30 wt%, and the hydrocolloid is present in a range of about 0.5 wt% to about 8 wt%.

100. The product of embodiment 98, wherein in the fibrous, edible, protein-enriched food analog product, the ingestible protein is present in a range of about 15 wt% to about 25 wt%, and the hydrocolloid is present in a range of about 1 wt% to about 5 wt%.

101. The product of embodiment 98 wherein the fibrous, edible, protein-enriched food analog product is any one of fibrous mammalian meat, poultry, or seafood analog food.

102. The product of embodiment 98, wherein the fibrous, edible, protein-enriched food analog product further comprises a skin layer formed from an ingestible ingredient selected to give an appearance and taste that mimics a food product that is emulated by the fibrous, edible, protein-enriched food analog product.

103. The product of embodiment 98 wherein the fibrous, edible, protein-enriched food analog product has a translucent appearance and is characterized by a transition from translucent to opaque when cooked.

104. A method for producing a fibrous meat analogue comprising: preparing an ingestible biopolymer gel or solution or dispersion of one or more ingestible proteins and/or hydrocolloids and water; subjecting the ingestible biopolymer gel or solution or dispersion to directional freezing, inducing the formation of aligned elongated ice crystals to form a directional frozen gel or solution having aligned elongated channels in which the aligned elongated ice crystals are located; replacing the aligned elongated ice crystals with ingestible proteins and/or hydrocolloids to produce a protein and/or hydrocolloid infused gel; and subjecting the ingestible protein and/or hydrocolloid infused gel to conditions suitable to gel at least some of the ingestible protein and/or hydrocolloid to produce a protein and/or hydrocolloid gel within the aligned channels, thereby forming the fibrous meat analogue comprising protein.

105. A method for producing a fibrous meat analogue food product comprising: preparing an ingestible hydrocolloid gel consisting of one or more different ingestible hydrocolloids; subjecting the ingestible hydrocolloid gel to directional freezing, inducing the formation of aligned elongate ice crystals to form a directional frozen hydrocolloid gel having aligned elongate channels in which the aligned elongate ice crystals are located; replacing the aligned elongated ice crystals with an ingestible protein to produce a protein infused hydrocolloid gel; exposing the protein-infused hydrocolloid gel to an agent for preventing leakage of protein from the protein-infused hydrocolloid gel; and packaging the fibrous meat analogue food for transportation.

106. The method of embodiment 105, wherein the agent for preventing protein leakage comprises any one or a combination of a pH adjuster, a salt, a heat treatment, a chemical cross-linking agent, an enzymatic cross-linking agent, injection of gelling hydrocolloid, and application of a hydrocolloid coating to the protein-injected hydrocolloid gel.

107. The method of embodiment 105 wherein the step of replacing the aligned elongated ice crystals with an ingestible protein comprises replacing the aligned elongated ice crystals with a mixture of an ingestible protein, a flavoring agent, and a coloring agent to impart the appearance and taste to the fibrous meat analog raw seafood product.

Claims

1. A method for producing a fibrous meat analogue comprising:

a) Preparing an ingestible biopolymer gel, solution or dispersion comprised of one or more ingestible proteins and/or hydrocolloids and water;

b) Subjecting the biopolymer gel, solution or dispersion to directional freezing, inducing the formation of aligned elongated ice crystals to form a directionally frozen biopolymer gel, solution or dispersion having aligned elongated channels in which the aligned elongated ice crystals are located;

c) Replacing the aligned elongated ice crystals with an ingestible protein and/or hydrocolloid to produce an infused gel; and

d) Subjecting the injected gel to suitable conditions to gel at least some of the ingestible proteins and/or hydrocolloids to produce gelled proteins and/or hydrocolloids within the aligned channels, thereby forming a fibrous food product.

2. The method of claim 1, wherein the ingestible biopolymer gel, solution, or dispersion is:

(i) A hydrocolloid gel, solution or dispersion comprising one or more different ingestible hydrocolloids and water;

(ii) A protein gel, solution or dispersion comprising one or more different ingestible proteins and water; or (b)

(iii) A composite gel, solution or dispersion comprising one or more different ingestible hydrocolloids and one or more different ingestible proteins and water.

3. The method according to claim 1 or 2, wherein the ingestible proteins and/or hydrocolloids of steps a) and c) are the same or different.

4. A method according to any one of claims 1 to 3, wherein the ingestible hydrocolloid comprises one or more of regular and/or recombinant gelatin, agar, alginate, curdlan, kappa-carrageenan, kappa-2-carrageenan and iota-carrageenan, red algae gum, starch, modified starch, seaweed extract, dextrin, konjac glucomannan, methylcellulose, pectin, gellan gum, xanthan gum, guar gum, locust bean gum, gum arabic, tara gum or polysaccharide.

5. The method according to claim 1, comprising:

a) Preparing an ingestible hydrocolloid gel consisting of one or more ingestible hydrocolloids and water;

b) Subjecting the ingestible hydrocolloid gel to directional freezing, inducing the formation of aligned elongate ice crystals to form a directional frozen hydrocolloid gel having aligned elongate channels in which the aligned elongate ice crystals are located;

c) Replacing the aligned elongated ice crystals with an ingestible protein to produce a protein infused hydrocolloid gel; and

d) Subjecting the protein-infused hydrocolloid gel to suitable conditions to gel at least some of the ingestible proteins, thereby producing gelled proteins within the aligned channels.

6. The method according to claim 1, comprising:

a) Preparing an ingestible protein gel comprised of one or more first ingestible proteins and water;

b) Subjecting the ingestible protein gel to directional freezing, inducing the formation of aligned elongated ice crystals to form a directional frozen protein gel having aligned channels in which the aligned elongated ice crystals are located;

c) Replacing the aligned elongated ice crystals with a second ingestible protein to produce a protein gel infused with protein; and

d) Subjecting the protein gel infused with protein to suitable conditions to gel at least some of the second ingestible proteins, thereby producing gelled proteins in the aligned channels.

7. The method of any one of claims 1-6, wherein the ingestable protein comprises a gellable protein, a non-gellable protein, or a combination thereof.

8. The method of claim 7, wherein the ingestible protein comprises a culture protein; animal proteins, such as recombinant animal proteins; a plant protein; bacterial proteins; fungal proteins, such as yeast proteins; algae proteins; or a combination thereof.

9. The method of claim 8, wherein the ingestible protein is mammalian whey protein, casein, or caseinate; any one or any combination of soy protein, potato protein, rubisco protein, duckweed protein, rice protein, almond protein, egg protein, oat protein, linseed protein, euglena protein, schizochytrium limacinum protein, mung bean protein, pea protein, recombinant mammalian whey, cultured mammalian whey, recombinant egg albumin, cultured egg albumin, recombinant gelatin or collagen, cultured gelatin or collagen, canola protein, lupin protein, broad bean protein, wheat protein, lentil protein, amaranth protein, peanut protein, moringa seed protein, pumpkin seed protein, chickpea protein, sunflower seed protein, safflower seed protein, mustard seed protein, chlorella protein, and spirulina protein.

10. The method according to any one of claims 1 to 9, wherein step c) comprises:

(i) Thawing said directionally frozen biopolymer gel, solution or dispersion by immersion in a solvent containing said ingestible protein and/or hydrocolloid of step c), said solvent having a temperature suitable for thawing said ice crystals;

(ii) Freeze-drying the directionally frozen biopolymer gel, solution or dispersion to remove substantially all water, and then immersing the dried gel into a solution containing the ingestible protein and/or hydrocolloid of step c);

(iii) Evaporating the ice crystals and then immersing the dried gel into a solution containing the ingestible protein and/or hydrocolloid of step c); or (b)

(iv) Placing one end of the injected gel under vacuum to extract the ice crystals and pulling the ingestible protein and/or hydrocolloid of step c) from the other end of the injected gel into the aligned elongate channels.

11. The method of any one of claims 1 to 9, wherein step c) comprises thawing the directionally frozen biopolymer gel, solution or dispersion; and wherein the method comprises a plurality of cycles of directional freezing and thawing.

12. The method of any one of claims 1 to 11, further comprising controlling the diameter of the aligned elongate channels by controlling a temperature gradient through the material so as to vary the speed of the directional freezing process, and wherein the diameter of gelled protein in the aligned elongate channels is proportional to the diameter of the aligned elongate channels.

13. The method of claim 12, wherein the diameters of the aligned elongate channels are controlled to result in elongate gelled proteins having diameters in the range of about 20 to about 500 microns.

14. The method of any one of claims 1 to 13, wherein the biopolymer gel, solution or dispersion is a solution or dispersion, and the method comprises:

(i) Prior to step c), gelling the directionally frozen solution or dispersion by further subjecting said solution or dispersion to suitable conditions; or (b)

(ii) Prior to step c), gelation of the solution or dispersion is induced by immersion in a suitable solution.

15. The method of any one of claims 1 to 13, wherein the biopolymer gel, solution or dispersion is a solution or dispersion, and the method comprises gelling the solution or dispersion while directional freezing.

16. The method of claim 14 or 15, wherein the biopolymer solution or dispersion comprises a first hydrocolloid and a second hydrocolloid, and wherein the method comprises gelling the first hydrocolloid, subjecting the biopolymer to directional freezing, subsequently gelling the second hydrocolloid, and replacing the ice crystals with an ingestible protein.

17. The method of any one of claims 1 to 16, wherein the suitable conditions comprise:

(i) Thermally treating the injected gel, and wherein the ingestible protein comprises at least a thermally gellable protein;

(ii) Penetrating a salt or ion into the injected gel, the salt selected to induce gelation of the at least some of the ingestible proteins and/or hydrocolloids;

(iii) Adjusting the pH of the injected gel to a value suitable to cause gelation of the at least some of the ingestible proteins and/or hydrocolloids;

(iv) Penetrating a solution containing a cross-linking agent into the injected gel, the cross-linking agent being selected to induce gelation of the at least some of the ingestible protein and/or hydrocolloid;

(v) Subjecting the injected gel to pressure treatment to induce gelation of the at least some of the ingestible proteins and/or hydrocolloids; and/or

(vi) Irradiating the injected gel with radiation of a suitable wavelength and intensity to induce cross-linking of the proteins, thereby inducing gelation of the at least some of the ingestible proteins.

18. The method of claim 17, wherein penetrating the salt comprises contacting the injected gel with a salt solution of sufficient concentration to allow gelation of the at least some of the ingestible protein and/or hydrocolloid; and wherein the salt is any one of sodium (Na), potassium (K), calcium (Ca) and magnesium (Mg) sulfate, citrate, chloride, carbonate, ascorbate, acetate, sorbate, lactate, tartrate, gluconate and phosphate, and any combination thereof.

19. The method of claim 17, wherein adjusting the pH of the infused gel comprises adding a food-safe pH adjuster comprising acetic acid, hydrochloric acid, ascorbic acid, malic acid, formic acid, lactic acid, tartaric acid, citric acid, gluconic acid, glucono-delta lactone, sodium hydroxide, potassium hydroxide, calcium hydroxide, or a combination thereof.

20. The method of claim 17, wherein the cross-linking agent is a chemical cross-linking agent comprising glutaraldehyde, tannic acid, genipin, liquid smoke, or a combination thereof.

21. The method of claim 17, wherein the cross-linking agent is an enzyme-based cross-linking agent comprising transglutaminase (EC 2.3.2.13), sortase a (EC 3.4.22.70), tyrosinase (EC 1.14.18.1), laccase (EC 1.10.3.2), peroxidase (EC 1.11.1. X), lysyl oxidase (EC 1.4.3.13), amine oxidase (EC 1.4.3.6), or a combination thereof.

22. The method of any one of claims 1 to 21, wherein the ingestible biopolymer gel, solution, or dispersion has multiple layers.

23. The method of claim 22, wherein alternating layers are made of the same or different biopolymers or biopolymer mixtures.

24. The method of claim 22 or 23, further comprising producing a layer that mimics the skin layer of meat or fish by producing a mixture of agar and an alginate solution with an alginate-oil emulsion and gelling the mixture to produce a skin layer.

25. The method of any one of claims 1 to 24, further comprising producing a plurality of protein infused biopolymer gels each having a preselected thickness, and preparing interstitial layers made of a material selected to mimic connective tissue of meat and/or fish.

26. The method of claim 25, wherein the plurality of protein infused biopolymer gels are stacked and adhered to the interstitial layer.

27. The method of claim 25 or 26, wherein the interstitial layer comprises a material selected to mimic connective tissue of meat and/or fish, the material comprising any one or a combination of proteins, hydrocolloids, oil-in-water emulsions, solid particles, fats, and oil gels.

28. The method of claim 27, wherein the solid particles comprise any one or a combination of titanium dioxide, protein, calcium carbonate, starch, solid fat crystals, and algae.

29. The method of any one of claims 1 to 28, the method further comprising:

i) Adding a pigment during the preparation of the biopolymer gel, solution or dispersion; or (b)

ii) replacing the aligned elongate ice crystals with a second ingestible protein and/or hydrocolloid and a mixture of said pigments;

wherein the pigment comprises carotenoids, beta-carotene, astaxanthin, lycopene, carmine, anthocyanidins, betalains, hemoglobin, myoglobin, beet juice extract, carthamin, lutein, curcumin, capsanthin, norbixin, anthocyanidins, curcuminoids, turmeric root powder, phycocyanin, melanoidin, or combinations thereof.

30. A method for producing a fibrous meat analogue comprising:

subjecting the ingestible biopolymer gel to directional freezing, inducing the formation of aligned elongated ice crystals to form a directionally frozen biopolymer gel having aligned channels in which the aligned elongated ice crystals are located;

thawing said oriented frozen biopolymer gel having said aligned channels by immersing the frozen biopolymer in a solution containing at least one ingestible soluble thermal gelling protein, thereby thawing and replacing said aligned elongated ice crystals with said at least one ingestible soluble thermal gelling protein at a temperature below the gelling temperature of said soluble thermal gelling protein to produce a protein infused biopolymer gel; and

Heating the protein infused biopolymer gel at a temperature above the gelation temperature of the at least one ingestible soluble thermogelling protein to produce protein fibers, thereby forming a fibrous meat analog food product.

31. The method of claim 30, wherein the ingestible biopolymer gel is:

a) A hydrocolloid gel comprising one or more different ingestible hydrocolloids and water;

b) A protein gel comprising one or more different ingestible proteins and water; or (b)

c) A complex gel comprising i) at least two different ingestible hydrocolloids, ii) at least two different ingestible proteins, or iii) one or more different ingestible hydrocolloids and one or more different ingestible proteins and water.

32. The method of claim 30 or 31, wherein the biopolymer gel is comprised of one or more of polysaccharides, conventional and/or recombinant gelatin, agar, fermentation-derived gelatin, alginates, curdlan, kappa-carrageenan, kappa-2-carrageenan and iota-carrageenan, furcellaran, starch, modified starch, dextrin, konjac glucomannan, pectin, methylcellulose, gellan gum, xanthan gum, guar gum, locust bean gum, gum arabic and tara gum.

33. The method of any one of claims 30 to 32, comprising varying the immersion time of the frozen biopolymer to control protein loading.

34. The method of any one of claims 17 and 30 to 33, wherein the at least one ingestible soluble thermogelling protein comprises: i) An ingestible, soluble, thermogelling protein, wherein the concentration of the ingestible, soluble, thermogelling protein in the solution comprising at least one ingestible, soluble, thermogelling protein is in the range of about 0.5% to about 30%; or ii) a mixture of ingestible soluble thermogelling proteins and non-thermogelling proteins.

35. The method of claim 34, wherein the ingestible biopolymer gel has a melting temperature above the gelation temperature of the ingestible soluble thermal gelation protein.

36. The method of any one of claims 10 and 30-35, wherein thawing the directionally frozen hydrocolloid gel comprises adjusting a temperature of a solvent containing the ingestible gellable protein to a range from a melting point of the solvent within the hydrocolloid gel to a melting point of the hydrocolloid gel, and wherein replacing the aligned elongated ice crystals with the ingestible protein comprises adjusting a temperature of a solvent containing the ingestible protein to a range from a freezing point of a solvent containing the protein solution to a gelation onset denaturation temperature of the at least some of the ingestible proteins.

37. The method of claim 36, wherein thawing the directionally frozen hydrocolloid gel is performed at between about 0 ℃ and about 85 ℃, preferably between about 0 ℃ and about 45 ℃.

38. The method of any one of claims 17 and 30-37, wherein the solution containing at least one ingestible soluble thermal gelling protein further comprises an ingestible non-thermal gelling protein and a thermally induced trigger to trigger gelling of the ingestible non-thermal gelling protein upon an increase in temperature.

39. The method of claim 38, wherein the thermally induced trigger comprises a salt, an enzyme, a pH adjuster, or a combination thereof.

40. The method of claim 38, wherein the thermally induced trigger is an enzyme microencapsulated within a meltable coating.

41. The method of any one of claims 30 to 40, wherein the solution containing at least one ingestible soluble thermogelling protein comprises: an aqueous solution or an aqueous dispersion.

42. The method of any one of claims 10 and 30-41, wherein the solution containing the at least one ingestible soluble thermogelling protein has a temperature of about 1 ℃ to about 60 ℃.

43. The method of claim 33, wherein protein loading is further varied by varying the volume ratio of the ingestible biopolymer gel and the solution containing the at least one ingestible soluble thermogelling protein.

44. The method of any one of claims 30 to 43, wherein the step of directionally freezing the ingestible biopolymer gel is performed by placing the ingestible biopolymer gel in contact with a pre-cooled substrate at a temperature of about-2 ℃ to about-196 ℃.

45. The method of any one of claims 30 to 44, wherein the ingestible biopolymer gel is a kappa-carrageenan hydrogel having a modulus in the range of about 100 to about 5000 pascals.

46. The method of any one of claims 30 to 44, wherein the ingestible biopolymer gel is an agar hydrogel having an agar concentration in the range of about 0.1% to about 15% by weight.

47. The method of any one of claims 30 to 46, wherein the solution containing at least one ingestible soluble thermogelling protein further comprises an ingestible supplement that diffuses into the aligned channels, wherein the ingestible supplement comprises: a water-soluble vitamin which is a compound of formula (I), Comprises ascorbic acid (vitamin C), thiamine, riboflavin, nicotinic acid, and vitamin B ₆ (pyridoxine, pyridoxal and pyridoxamine), folic acid, vitamin B ₁₂ Biotin, pantothenic acid, and combinations thereof; or an emulsion of omega-3, omega-6, omega-9 fatty acid esters, or combinations thereof; an ingestible mineral; and/or antioxidants.

48. The method of claim 41, wherein the aqueous solution or dispersion comprises: flavoring agents, emulsifying agents, preserving agents, pigments, pH adjusting agents, texture adjusting agents, or combinations thereof.

49. The method of any one of claims 30 to 48, wherein the step of subjecting the ingestible biopolymer gel to directional freezing and inducing the formation of aligned elongated ice crystals further comprises: placing the ingestible biopolymer gel in contact with a pre-cooled substrate for directional freezing in one direction; or placing the ingestible biopolymer gel between two pre-cooled substrates, wherein the directional freezing is performed from opposite directions.

50. The method of claim 34, wherein the concentration of the ingestible, thermal gelling protein in the solution containing at least one ingestible, soluble, thermal gelling protein in the mixture of thermal gelling proteins and non-thermal gelling proteins is in the range of about 2 to about 10 wt%, preferably about 2 to about 15%, more preferably about 2 to about 20%, with the remainder being non-thermal gelling proteins, to make up a total 25 wt% protein mixture.

51. The method of any one of claims 17 and 30-50, wherein the step of heat treating the biopolymer gel of the ingestible soluble thermogelling injection protein is performed at a solution temperature in the range of about 40 ℃ to about 150 ℃.

52. The method of any one of claims 1 to 51, further exposing the infused gel to an agent for preventing leakage of protein from the infused gel, wherein the agent comprises any one or a combination of a pH adjuster, a salt, a heat treatment, a chemical cross-linking agent, an enzymatic cross-linking agent, injection of gelled hydrocolloid, and application of a hydrocolloid coating to the protein infused hydrocolloid gel.

53. The method of any one of claims 1 to 52, wherein the step of replacing the aligned elongated ice crystals further comprises replacing with a flavoring and/or coloring agent.

54. A fibrous meat analogue food product produced by the method of any one of claims 1 to 53.

55. The food product of claim 54 which is a fibrous mammalian meat, poultry or seafood analog food.

56. The food product of claim 54 or 55, wherein the ingestible protein is present in the food product in a range of about 2 wt% to about 50 wt%, preferably about 5 wt% to about 35 wt%, preferably about 10 wt% to about 30 wt%, preferably about 15 wt% to about 25 wt%, preferably about 10 wt% to about 20 wt%.

57. The food product of any one of claims 54-56, wherein the hydrocolloid is present in the food product in a range of about 0.2 wt.% to about 10 wt.%, preferably about 0.5 wt.% to about 8 wt.%, preferably about 0.5 wt.% to about 5 wt.%.

58. The food product of any one of claims 54-57, wherein the ingestible protein comprises a mixture of ingestible, thermogelable proteins and non-gelable proteins to provide an increased protein content.

59. The food product of claim 58, wherein the maximum amount of protein present is 25 weight percent, then the minimum amount of ingestible, thermally gellable protein in the mixture is 2 weight percent, and the maximum amount of non-thermally gellable protein is 23 weight percent.

60. The food product of claim 58, wherein the maximum amount of protein present is 25% by weight and the intermediate minimum amount of ingestible, thermally gellable protein in the mixture is about 8% by weight and the maximum amount of non-thermally gellable protein is about 17% by weight.

61. The food product of claim 58, wherein the maximum amount of protein present is 25% by weight and the intermediate minimum amount of ingestible, thermally gellable protein in the mixture is about 10% by weight and the maximum amount of non-thermally gellable protein is about 15% by weight.