Animal-free food substitutes

Abstract

A method for producing a substitute food product, such as a dairy product such as cheese or yogurt, or an egg substitute product, using an amylase enzyme, which may be at least partially denatured and may be present in an amount of 3 g / 100 g protein.

Description

[Technical Field]

[0001] The present invention relates to a method for producing a food substitute, such as a dairy product (such as cheese or yogurt), or an egg replacer composition. The invention further extends to a food substitute, such as a cheese replacer or egg replacer product, made using the method of the invention, and to compositions used to produce the food replacer. [Background technology]

[0002] Protein is an important nutritional requirement and forms an important component of the diet of humans and animals. Proteins can be of animal origin (e.g., meat, fish, milk, eggs, etc.) or can be sourced from plants, e.g., legumes, or microorganisms.

[0003] Cheese is typically made from milk or milk-derived ingredients. The sustainability of cheese made from milk or milk-derived ingredients has been called into question, as milk production is a resource-intensive process that has a negative impact on the environment. In fact, cheese is the third most unsustainable animal food in the world in terms of greenhouse gas emissions per kg of product.

[0004] Therefore, significant efforts have been made to produce dairy substitutes, i.e., dairy substitutes that do not use milk or other animal-derived ingredients, such as vegan-sourced cheese-like dairy substitutes and other dairy substitutes (including milk and yogurt substitutes). However, to date, the use of vegan ingredients, such as plant proteins, has resulted in products that lack the organoleptic properties of cheese. Furthermore, the nutritional quality of these products is also low due to the lower protein content and / or nutritional value of plant proteins compared to animal-derived proteins. The lack of similarity of the organoleptic properties and / or lower nutritional quality of plant-based substitutes to dairy products is one of the main reasons why these plant-based products have not slowed down dairy cheese consumption.

[0005] Eggs, particularly those from birds such as chickens, while high in protein, contain components that may be allergenic or unhealthy for certain groups of people. However, eggs possess several desirable functional properties that allow them to be used in the preparation of a wide variety of egg-based foods with pleasing taste / flavor and texture, including emulsions, batters, doughs, baked goods, and prepared foods. Eggs are useful in these products because they contribute to processes such as gelling / thickening / coagulation, foaming / leavening, emulsification, and water binding. As such, eggs are an essential ingredient in many commonly consumed foods, and are believed to impart or contribute to size, texture, emulsification, shape, taste / flavor, cooking stability, processing resistance, and shelf life.

[0006] There has been growing interest in vegan egg substitutes. Known prior art egg substitutes are based on, for example, mung bean protein isolate, chickpea flour, pea protein, or lupin bean protein isolate. Companies producing such egg substitutes include Greenforce, Just Egg, Rewe Bio, and Plant-B. Aquafaba from commercially available canned chickpeas can also be used as a potential egg substitute; the Institute of Food Science and Technology, October 24, 2019, discusses recipe optimization and storage stability of aquafaba in vegan mayonnaise. Similarly, WO2022124988 discloses an egg yolk substitute.

[0007] There is a need for egg replacers that provide similar nutritional content, similar functionality, taste, flavor, and organoleptic properties, but with fewer or no undesirable properties. It is an object of the present invention to provide improved egg replacers.

[0008] Therefore, there is a need to provide animal-free food substitutes, including dairy and egg substitutes, particularly substitutes that mimic the organoleptic properties of the corresponding animal-based foods. Summary of the Invention

[0009] The inventors have found that modified amylases can be used in products as ingredients, for example, to the extent of a minimum of 3% (w / w protein) in the food substitute. Optionally, amylases can be present as the main protein component of the food substitute. Amylases allow for the formation of products with suitable physical characteristics as well as acceptable organoleptic properties. In egg replacers (intended to be combined with other ingredients before cooking), the enzymes can be denatured as part of the cooking process itself.

[0010] Optionally, the amylase is present in an amount of from about 0.01 g / L to about 1000 g / L, inclusive, typically from 10 g / L to 450 g / L, inclusive, and more specifically from 60 g / L to 350 g / L, inclusive.

[0011] The inventors have discovered that food substitutes can be formed by using amylase as an ingredient in a food substitute. When referring to an enzyme present as an "ingredient," it is meant that the enzyme is not present as a biocatalyst to degrade other biomolecules or as a dietary supplement, but rather is used for its structural or nutritional properties. Thus, the amylase is subjected to a processing step that converts the amylase into at least one converted protein with a structural function. Because the enzyme is not used as a biocatalyst, it can be conveniently denatured, optionally during cooking of the food itself. Optionally, the amylase is heat-treated before or during cooking to make it suitable for human consumption. The term "amylase" is used herein to refer to a wild-type amylase enzyme or a variant thereof having an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of the wild-type amylase enzyme. Optionally, the variant has 80% or more sequence identity to the wild-type amylase enzyme, for example, 85% or more, 90% or more, 95% or more, or 98% or more sequence identity to the amino acid sequence of the wild-type amylase enzyme.

[0012] More specifically, the inventors have found that by using at least one amylase, it is possible to obtain a curdled or emulsified composition to produce an animal-free substitute food product that resembles the organoleptic or other desirable properties of a comparable food product, e.g., a traditional dairy product such as cheese. Optionally, the amylase is denatured by heat treatment, which further ensures suitability for human consumption.

[0013] At least one amylase can be treated to partially or substantially lose its catalytic function before or during the food product formation process. Such treatment can be carried out by hydrolysis (pH-based hydrolysis or enzymatic hydrolysis), heat treatment, or mechanical shearing to partially or completely denature the catalytic activity of the amylase. The at least partially denatured amylase is suitable for use as a structural ingredient for producing food substitutes. This treatment can be carried out by enzymatic hydrolysis using, for example, a protease, and / or heat treatment.

[0014] The present invention therefore extends to dairy substitutes in which the major protein component is at least one partially or fully denatured amylase.

[0015] In accordance with another aspect of the present invention, there are provided amylases that, when converted according to an aspect of the present invention, result in protein components that have desirable properties, by which is meant properties that may be useful in producing food-like or substitute foods, such as egg replacers.

[0016] In a further aspect, the present invention provides a process for forming a substitute egg product, comprising mixing amylase with water to form a mixture and homogenizing the mixture with fat or oil, wherein the amylase provides at least 5g / 100g of protein in the product. Optionally, the amylase is mixed with water and a carbohydrate source, such as seed mucilage or wheat flour. Fibers such as flaxseed fiber, hydrocolloids, modified starches, or water-soluble fibers such as beta-glucan, xanthan gum, and pectin can be used instead of seed mucilage. Optionally, fat or oil can be added to the mixture and homogenized to form the substitute egg product. Optionally, other flavors or colorants can be added, as described below with respect to the composition.

[0017] Thus, the present invention also extends to egg substitute products whose major protein component is at least one partially or fully denatured amylase.

[0018] The inventors have found that the use of amylases, which may be partially or fully denatured, can provide improved structural or nutritional value within the animal-free food substitutes of the present invention. [Brief explanation of the drawings]

[0019] [Figure 1] FIG. 1 shows an imitation cheese product made as described in the Examples. [Figure 2] FIG. 2 shows the substitute cream cheese product made as described in the Examples. [Figure 3] FIG. 3 shows a feta cheese substitute product made as described in the Examples. [Figure 4] FIG. 4 shows a substitute jogurt product made as described in the Examples. [Figure 5] Amplitude sweeps of alpha amylase (top), glucoamylase (middle), and yogurt (bottom). [Figure 6]Figure 6 shows imitation fresh cheese products made as described in the Examples. Fresh cheese made with alpha amylase (left) and glucoamylase (right). [Figure 7] Figure 7 shows egg replacer products made as described in the Examples: Egg replacer based on alpha amylase (left) and glucoamylase (right). [Figure 8] FIG. 8 shows the gelation of glucoamylase (product A) at different pH values ​​and salt concentrations. [Figure 9] Figure 9 shows the gelation of glucoamylase (Product B) at different pH values ​​and salt concentrations. Top: no salt, bottom: 1% NaCl (w / w). From left to right: pH 5.0, 5.5, 6.6. [Figure 10] Figure 10 shows the change in storage and loss modulus of heat-induced gels based on purified lactase (β-APC) and alpha-amylase (a-APC) gels as a function of temperature at pH 6 (a and b) and pH 4.8 (c and d). Detailed Description of the Invention

[0020] Dairy milk and eggs form an important part of the Western diet. Generally, most animal-free foods rely heavily on the inclusion of plant proteins. The process of separating plant proteins from associated carbohydrates and other molecules is inefficient and expensive. Therefore, known animal-free egg replacer compositions made with plant proteins inherently suffer from poor performance due to co-extracted compounds that adversely affect taste, odor, and texture. The proteins used in these products are structural in nature.

[0021] Typically, cheese products are made using animal milk or proteins and fats extracted from plants. To date, most animal-free dairy alternatives rely heavily on the inclusion of plant proteins. The process of separating plant proteins from associated carbohydrates and other molecules is inefficient and expensive. Therefore, animal-free cheese alternatives (for example) made with plant proteins inherently suffer from poor performance due to co-extracted compounds that adversely affect taste, odor, and texture. Surprisingly, the present applicant has discovered that amylases, commonly used in the animal and human food industries (but previously employed only as biocatalysts to break down carbohydrates or as dietary supplements, not for their structural or nutritional properties), can be used as a primary raw material to create alternative foods such as dairy alternatives. It was surprising that amylases can serve the same structural purpose as animal- or plant-based globular proteins. Optionally, amylases can be processed to make them suitable for human consumption, for example, by heat treatment. Specifically, amylase is heat treated to produce a cheese-like gel, emulsion, or curd, which can be further processed to produce a cheese substitute or cheese-like product, specifically a vegan or animal-free cheese substitute.

[0022] It has been found that the gels, emulsions or curds formed can also be used to form other dairy substitute products such as yogurt, and other food substitutes such as egg substitute products.

[0023] Surprisingly, the present applicant has discovered that globular enzymes, particularly amylases, commonly used in the animal and human food industries (but previously employed only as biocatalysts to degrade other biomolecules or as dietary supplements, and not for their structural properties), can be treated or manipulated to produce animal-free egg replacer compositions, egg-based foods, or egg substitute foods, such as egg white replacers. It was therefore surprising that amylases, as native egg proteins, can provide the necessary structure to form egg-like foods, egg white replacers, or egg replacer compositions for use in egg-based foods, or compositions for egg-based foods. Specifically, amylases can be heat-treated to produce egg-like gel or emulsion products that can be used to form egg replacer compositions (optionally vegan or animal-free) or egg-like products, such as scrambled eggs (optionally vegan or animal-free).

[0024] To successfully deliver an egg substitute food product, the technological properties of the protein are crucial to provide the characteristics of the final product, as is the ability to produce the protein in sufficient quantities in a sustainable manner. At the same time, the protein needs to provide similar health (nutritional) benefits as egg protein.

[0025] Egg white contains the functionally important proteins ovalbumin (54%), ovotransferrin (12%), ovomucoid (11%), ovomucin (3.5%), and lysozyme (3.5%). However, producing all of these proteins using genetic engineering and precision fermentation to mimic animal-free egg substitutes is extremely difficult and costly. Therefore, it is important to find a single protein that provides one or more of the technological, nutritional, and / or organoleptic properties of egg proteins and delivers the egg experience to consumers.

[0026] Amylase has the ability to gel (coagulate) when heated.

[0027] Biocatalytic enzymes such as amylases are used almost exclusively for their precisely defined catalytic function. In contrast, in the present invention, amylases are used to provide the main protein component (i.e., the main protein nutrition) and improved structure to food substitutes such as cheese substitute or egg substitute products. Advantageously, the amylases used herein are already generally regarded as safe ("GRAS") due to their widespread use in the food industry.

[0028] According to a first aspect of the present invention, there is provided a method for producing a food substitute, the method comprising the steps of: i. providing at least one amylase ii. subjecting at least one amylase to one or more processes to at least partially denature the amylase or to provide the amylase in an amount of at least 3 g / 100 g protein in the product. iii. incorporating the amylase into a composition to form a food substitute.

[0029] The term "amylase" is used herein to refer to a wild-type amylase enzyme or a variant thereof having an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of the wild-type amylase enzyme. Optionally, the variant has 80% or more sequence identity to the wild-type amylase enzyme, e.g., 85% or more, 90% or more, 95% or more, or 98% or more sequence identity to the amino acid sequence of the wild-type amylase enzyme.

[0030] Optionally, the amylase used in the present invention is recombinantly produced and expressed under controlled conditions using a heterologous host cell, e.g., a genetically modified organism (GMO), such as a microorganism (GMO). Optionally, the amylase is naturally expressed from the host cell. The host cell may be a fungus, e.g., Aspergillus oryzae or Aspergillus niger. Optionally, the amylase is naturally produced by a plant. Optionally, the amylase is naturally produced by a microorganism. Optionally, the amylase can be provided in the form of a crude fermentate or a purified fermentate.

[0031] The amylases used in the present invention can be engineered to form gels, emulsions, or curds with a neutral flavor and texture and mouthfeel that can be optimized by modifying processing conditions well known to those skilled in the art. Amylases can be commercially available, including alpha or beta amylases, such as alpha amylase and glucoamylase (amyloglucosidase). The availability of amylases provides an opportunity to improve the suboptimal economics involved in producing alternative foods, such as producing dairy substitutes from recombinantly produced milk proteins.

[0032] Alpha-amylase (1,4-α-d-glucan glucohydrolase) is an endo-acting glucanase belonging to glycoside hydrolase family 13 (GH13). This enzyme catalyzes the cleavage of α-1,4-glycosidic bonds in starch and other related polysaccharides, producing oligosaccharides of various lengths, including α-constituent dextrins and α-limit dextrins, which constitute branched oligosaccharides. β-amylase (EC 3.2.1.2) is an exo-acting enzyme that cleaves the α-(1→4) bond from the non-reducing end of the polymer chain, releasing the disaccharide maltose.

[0033] The present invention is based on the recognition that amylases can be used to produce compositions that exhibit similar taste, aroma, and mouthfeel to equivalent foods, e.g., dairy products such as cheese made using animal milk ("traditional milk" or "traditional dairy" when animal milk is used to make the product such as cheese. Accordingly, the present invention provides compositions useful for producing these substitute foods, methods of making the compositions, and kits containing the amylases, compositions, curds, and mixtures.

[0034] The present invention includes the use of amylase enzymes produced by precision fermentation, which convert a catalytically active protein (i.e., an enzyme) into a protein that provides a structural function. The present invention also provides proteins with structural functions produced using the methods of the present invention. Thus, the present invention extends to amylase enzymes that have been processed according to the methods of the present invention and at least partially modified to make them suitable for inclusion in food substitutes as a structural building block of a food product, i.e., suitable for inclusion in a food product as a structural building block of a food product.

[0035] The substitute food products or compositions of the present invention have similar taste, texture, aroma, and nutritional value compared to comparable products or compositions made using animal products.

[0036] For example, the dairy substitute products or compositions of the present invention have similar taste, mouthfeel, aroma, and nutritional value compared to comparable products or compositions made using animal milk.

[0037] Additionally, the inventors have found that it is possible to obtain an egg replacer composition ("egg replacer") that can be used with other ingredients to form an egg-based food or egg-based food composition. The egg replacer composition can be formed using at least one amylase enzyme. The formed egg replacer composition is preferably an animal-free product. The formed egg replacer composition preferably exhibits the organoleptic properties or other desirable properties of avian eggs.

[0038] Thus, the present invention extends to egg-based food products, i.e. food products in ready-to-eat form, comprising the egg replacer composition of the present invention.

[0039] The present invention also extends to egg-based food compositions, i.e., compositions for making food requiring cooking into a ready-to-eat form, comprising the egg replacer composition of the present invention. Thus, the present invention provides a food or food composition comprising the egg replacer composition of the present invention together with at least one other ingredient to form an egg-based food or egg-based food composition.

[0040] In one embodiment of the invention, an alpha amylase glucosidase is used. In one embodiment of the invention, a glucosidase, specifically amyloglucosidase, commonly known as glucoamylase, is used. In one embodiment of the invention, an alpha amylase and a glucoamylase are used.

[0041] Optionally, the food substitute, e.g., egg replacer composition / product, comprises an amylase enzyme, wherein the amylase provides a minimum level of 3% (protein w / w), optionally at least 5% (protein w / w), optionally at least 10% (protein w / w), for example at least 20% (protein w / w) of the protein of the food product. Optionally, the amylase can be the major protein component of the food product.

[0042] The term "major protein component" refers to an ingredient that provides the majority of the protein content in a product of the present invention compared to other ingredients or the sum of the other ingredients. For example, a "major protein component" can provide 50% or more (w / w) of the protein content in the product, e.g., at least 80% (w / w), e.g., at least 90% (w / w) of the protein content in the product.

[0043] The term "dairy substitute" refers to a composition similar to or equivalent to dairy milk or a product made using dairy milk components. In particular, dairy substitutes are similar to or equivalent to dairy milk or a product made using dairy milk components in one or more respects, such as taste, aroma, flavor, mouthfeel, organoleptic attributes, or color. Thus, as used herein, the term "dairy substitute" refers to a product that resembles traditional dairy products but is made using proteins produced in a process that does not involve mammals. For example, the proteins used may be plant-based proteins, microbially expressed proteins, or, more preferably, recombinantly expressed proteins, e.g., proteins expressed using heterologous microbial host cells. The terms "synthetic dairy product" or "synthetic cheese product" can be used interchangeably with the term "dairy substitute."

[0044] The term "animal milk" refers to milk obtained by lactation from female mammals such as cows, goats, sheep, camels, and humans.

[0045] The term "isolated" protein or "isolated" enzyme refers to a protein or enzyme that has been substantially separated from other cellular components that naturally accompany the native protein or enzyme within a natural host cell.

[0046] The term "animal-free" refers to components that are not obtained or produced from animals, but excludes recombinantly produced proteins, even if the recombinantly produced protein has the same amino acid sequence as its wild-type counterpart.

[0047] "Heat treated" means that the amylase is heat treated to a temperature sufficient to reduce the possibility of bacterial contamination, thereby rendering the amylase suitable for use in foodstuffs. A "heat treated" amylase enzyme may be at least "partially denatured."

[0048] By "partially denatured" is meant that the protein structure of the amylase has been irreversibly altered such that its native catalytic activity is impaired by at least 50%, preferably at least 75%, relative to the activity of the intact amylase.

[0049] By "totally denatured" or "completely denatured" is meant that the protein structure of the amylase has been irreversibly altered and its catalytic activity has been reduced by 10% or less, e.g., 5% or less. Optionally, all catalytic activity of the amylase has been lost.

[0050] The term "lipid" refers to one or more molecules containing a fatty acyl group (e.g., a saturated or unsaturated acyl chain). For example, the term "lipid" includes fats, oils, phospholipids, free fatty acids, monoglycerides, diglycerides, and triglycerides. Non-limiting examples of lipids are described herein and include sunflower oil, coconut oil, rapeseed oil, palm oil, tributyrin, mono- and diglycerides, free fatty acids, and phospholipids, cultured oils or fats, or microbially produced or derived oils or fats. Non-animal fats or oils are also an option, and refer to plant, microbial, fungal, or recombinantly produced fats or oils. Additional examples of lipids are known in the art.

[0051] The term "sweetening agent" refers to a sugar (e.g., a monosaccharide, disaccharide, or polysaccharide) or an artificial sweetener (e.g., a low-molecular-weight artificial sweetener or a protein artificial sweetener) that, when added to a composition, causes the composition to taste sweet when ingested by a mammal, such as a human. Non-limiting examples of sweetening agents are described herein. Additional examples of sweetening agents are known in the art.

[0052] The term "ash" is known in dairy science and refers to one or more ions, elements, minerals, and / or compounds that can be found in animal milk. Non-limiting examples of ions, elements, minerals, and compounds found in animal milk and encompassed by the term "ash" are described herein. Additional ions, elements, minerals, and compounds found in animal milk are also known in the art. As used herein, the term "ash" refers to the residue remaining after a product is burned for analysis and includes minerals such as salts. In some embodiments, the minerals may be salts or ions or one or more of sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc, chloride, manganese, selenium, and iodine. The term "ash" also includes retinol, carotenes, and other vitamins (such as vitamin D, vitamin E, vitamin B12, thiamine, and riboflavin). As used herein, the term "ash" also includes anions. Examples of such anions include one or more of phosphate, citrate, sulfate, carbonate, and chloride.

[0053] The term "color-matching agent" or "colorant" refers to an agent added to a composition to adjust the color of the composition, for example, to make the color of the composition appear more similar to a comparable product made using animal milk or avian eggs. Non-limiting examples of color-matching agents or colorants include 3-carotene and annatto. Other examples of color-matching agents are known in the art. Optionally, the color-matching agent or colorant may be produced by or obtained from a plant.

[0054] Desirably, the substitute foods and compositions of the present invention exhibit the functional characteristics and organoleptic properties of the equivalent food-based product. In some embodiments, the substitute foods and compositions of the present invention have a similar nutritional profile as the equivalent conventional food, replicating one or more, if not all, of its core functionality.

[0055] The term "core functionality" refers to the sensory, chemical, biochemical, or mechanical characteristics of a traditional food product. The term "core functionality" includes, but is not limited to, flavor, taste, appearance, nutritional value, handling and mouthfeel, desirable density, structure, texture, elasticity, springiness, coagulation, binding, leavening, breathability, foaming, creaminess, and emulsifying properties.

[0056] The term "flavor" refers to the taste and aroma of food and drink.

[0057] As used herein, the term "majority," or variations thereof, is understood to mean, for example, a) in the context of fats and oils, the amount of a particular fat and oil composition relative to the total amount of fat and oil compositions; b) in the context of proteins, the amount of a particular protein composition (e.g., amylase) relative to the total amount of protein compositions (e.g., α-, β-, and κ-casein).

[0058] The terms "about," "approximately," or "similar" mean within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which may depend in part on how the value is measured or determined or the limitations of the measurement system. All ranges and amounts set forth below are approximate and should be understood as not intended to limit the invention. When ranges and numerical values ​​are used, they may be approximated to include statistical ranges or measurement error or variation. In some embodiments, for example, measurements may be plus or minus 10%.

[0059] According to one aspect of the present invention, there is provided a method for producing a food substitute, the method comprising the steps of providing at least one amylase and incorporating the at least one amylase into a composition to form a food substitute, and the amylase is subjected to one or more treatment steps to at least partially denature the amylase during formation of the food substitute.

[0060] In one embodiment, the amylase is denatured. Thus, the amylase gels or solidifies during denaturation. Optionally, the amylase is in a solution or suspension having a pH of about 5.0 to about 7.2 (e.g., about 6.2 to about 6.8) prior to gelation / solidification.

[0061] The step of at least partially denaturing the amylase may occur before or after the amylase is added to the other ingredients to form the composition.

[0062] Optionally, the at least partially denatured enzyme amylase comprises at least 3% by weight of the protein in the food substitute, such as at least 5% by weight, for example at least 7% (w / w protein), such as 7-10% (w / w protein), up to 10% (w / w protein).

[0063] Optionally, the amylase is the major protein component of the food substitute, i.e., comprises at least 10% (w / w protein) in the food substitute, such as 20% (w / w protein) or more, for example 20-50% (w / w protein), for example 30-50% (w / w protein). Specifically, the amylase may comprise 3-15% (w / w protein) in the dairy substitute, for example 5-12% (w / w protein), 5-10% (w / w protein) or 5-12% (w / w protein).

[0064] Optionally, the amylase is totally modified in the production of the food substitute.

[0065] Thus, the present invention further provides a substitute food composition comprising at least one amylase and one or more selected from lipids, proteins, sweeteners, and / or ash, wherein the amylase comprises at least 3% by weight of the protein in the substitute food composition. Optionally, the amylase content can be greater than 3%, such as at least 10% w / w protein, for example at least 20% w / w protein.

[0066] Optionally, the amylase is at least partially denatured in the food substitute.

[0067] Optionally, the food substitute is a dairy substitute, such as a cheese substitute or a yogurt substitute.

[0068] Optionally, the food substitute is an egg substitute product.

[0069] Optionally, the composition comprises one or more of the following components: lipid, protein, sweetener, color matching agent, and / or ash. Optionally, the composition comprises a fat or oil.

[0070] Compositions used to form food substitutes form a further aspect of the present invention.

[0071] In one aspect, the present invention provides an egg-like food product comprising an amylase enzyme providing a protein level of at least 3% (w / w protein), optionally at least 5% (w / w protein), optionally at least 10% (w / w protein), e.g., at least 20% (w / w protein). Optionally, the enzyme can be the major protein component of the food product. The egg-like food product has one or more properties selected from foaming, emulsifying, or binding properties similar to whole eggs obtained from natural (avian) eggs. The egg-like food product can, for example, be used in place of avian eggs, e.g., directly fried to produce scrambled egg-type products or omelets.

[0072] In one aspect, the present invention provides an egg white substitute having properties suitable for use in baked or cooked goods, the egg white substitute comprising an amylase enzyme, which provides a protein level of at least 3% (w / w protein), optionally at least 5% (w / w protein), optionally at least 10% (w / w protein), for example at least 20% (w / w protein). Optionally, the amylase enzyme can be the major protein component of the egg white substitute. The egg white-like substitute has one or more properties selected from foaming, emulsifying, or binding properties similar to egg whites obtained from natural (avian) eggs. The egg white substitute can be added to dough, for example, in place of avian eggs, to contribute to a foamy texture.

[0073] The egg replacer compositions of the present invention may also be prepared as spray-dried or dehydrated compositions and may be in the form of a powder that can be reconstituted with water and / or other liquids. Egg replacer composition products and egg-based foods can have a natural egg-like texture achieved by combining proteins with other modifiers, such as texture modifiers (e.g., mucilages such as flaxseed extract or carbohydrate compounds such as dextrose) and taste modifiers (e.g., yeast extract or salt). Salt added as a taste modifier can include conventional edible salts such as sodium chloride and potassium chloride, as well as salts such as kala namak, also known as Indian black salt, or Hawaiian black lava salt.

[0074] The egg white-like emulsion product can have a natural egg white-like texture obtained by combining the protein with other modifiers, such as mucilages, e.g., flaxseed extract or flaxseed flour, carbohydrates (e.g., glucose), yeast extract, or texture and taste modifiers such as the addition of salts, which can include traditional edible salts such as sodium chloride and potassium chloride, as well as salts such as kala namak, also known as Indian black salt, or Hawaiian black lava salt.

[0075] Another taste modifier is yeast, which provides a versatile food product, particularly an egg replacer or egg white-like emulsion product, suitable for culinary applications where eggs, particularly egg whites, are commonly used.

[0076] The amylase enzymes of the present invention can be commercially produced either by microbial fermentation, purification from non-animal or non-mammalian sources, or by recombinant expression in microbial host cells (generally microbial or plant cells).

[0077] Optionally, the amylase can be mixed or blended with one or more vegetable proteins to form the compositions of the present invention. Because purified or recombinant proteins are generally more expensive to produce than traditional foods, it may be desirable to include one or more vegetable proteins to improve the economics of producing food substitutes using purified or recombinant proteins.

[0078] <Dairy substitute products> In one embodiment of the present invention, a method for producing a dairy substitute product is provided, the method comprising the steps of providing at least one amylase and incorporating the at least one amylase into a composition to form a dairy substitute product, such as cheese, yogurt or other dairy-based product, and subjecting the amylase to one or more treatment steps to at least partially denature the amylase during formation of the dairy substitute product.

[0079] Optionally, the amylase comprises at least 3% by weight of the protein in the dairy replacer, such as at least 5% by weight, for example at least 7% (w / w protein) of the dairy replacer or at least 10% (w / w protein) of the dairy replacer.

[0080] Optionally, amylase is the major protein component of the dairy substitute.

[0081] Optionally, the amylase is totally denatured.

[0082] The milk replacer composition (an intermediate product leading to a dairy replacer) may be curd. The curd can be used to produce further dairy replacer products. The dairy replacer products may be butter replacer, cheese, yogurt, custard, cream cheese, medium-hard cheese, hard cheese, pasta filata-type cheese, soft-ripened cheese, feta-style cheese, etc.

[0083] Where the dairy replacer is in the form of cheese or yogurt, the method may include maturing the dairy replacer. Thus, the dairy replacer may be a matured dairy replacer cheese or a matured dairy replacer yogurt.

[0084] Depending on the type of cheese, one or more bacteria or other microbial species may be employed in the cheese-making process for ripening or fermentation, producing fermentation products and by-products such as lactic acid, carbon dioxide, alcohol, aldehydes, and ketones.

[0085] Optionally, the method of making a dairy replacer according to the present invention may include adding one or more lipids, oils or fats to form a dairy replacer composition comprising amylase.

[0086] Optionally, the method of making a dairy replacer according to the present invention may include adding one or more of a carbohydrate, a sweetener, and an ash to form a dairy replacer composition comprising amylase.

[0087] The milk replacer composition can include amylase at a concentration of from about 0.01 g / L to about 1000 g / L, for example, from 10 g / L to 450 g / L, and more specifically, from 60 g / L to 350 g / L.

[0088] The amylase can be added in a purified form (e.g., at least 70%, 80%, 90%, 95%, 99% or higher purity), or it can be produced by microbial fermentation and provided in the form of a crude extract having a purity of 70% or less.

[0089] Dairy substitute products, such as cheese and yogurt (jogurt) products, can contain one or more different amylases, such as alpha amylase and glucoamylase.

[0090] The dairy replacer of the present invention can include one or more other proteins to form a dairy replacer composition containing amylase. The one or more proteins included in the composition can include one or more non-dairy proteins or dairy proteins. For example, soy protein or fava bean protein can be included. The dairy protein can generally be included as purified or recombinant dairy protein. For the avoidance of doubt, recombinant dairy proteins are considered "animal-free." The dairy protein can include one or more casein proteins. Alternatively, or in addition, the dairy protein can include one or more whey proteins. Optionally, the casein proteins and / or whey proteins can be recombinant proteins, produced, for example, by microbial fermentation. In some embodiments of these methods, the protein mixture can include one or more proteins selected from the group consisting of α-lactalbumin, β-lactoglobulin, α-S1-casein, α-S2-casein, lactoferrin, transferrin, and serum albumin.

[0091] The dairy substitute product or composition can include one or more lipids at a total concentration of about 0% to about 45% by weight, one or more flavor compounds at a total concentration of about 0.01% to about 6% by weight, one or more carbohydrates or sweeteners at a total concentration of about 0.1% to about 6% by weight, and ash at a total concentration of about 0.15% to about 1.5% by weight, with all ingredients totaling 100% by weight, the remainder being amylase and water. The lipid can be cultured lipid or microbially produced lipid.

[0092] Optionally, the one or more lipids are selected from the group consisting of sunflower oil, coconut oil, tributyrin, mono- and di-glycerides, free fatty acids, and phospholipids. Optionally, the dairy substitute comprises one or more of: sunflower oil at a final concentration of about 1% to about 28% by weight, coconut oil at a final concentration of about 0.5% to about 14% by weight, mono- and diglycerides at a final total concentration of about 0.05% to about 1.0%, about 0.08% to about 1.2% by weight, free fatty acids at a final total concentration of about 0.02% to about 0.28% by weight, and phospholipids at a final total concentration of about 0.02% to about 0.3% by weight.

[0093] Optionally, the free fatty acid comprises at least one fatty acid selected from the group consisting of butyric acid, caproic acid, caprylic acid, myristic acid, and capric acid. In some embodiments of any of the compositions described herein, the phospholipid is soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid, or rapeseed lecithin phospholipid. In some embodiments of any of the compositions described herein, the mono- and diglycerides are plant-derived mono- and diglycerides or bacterial-derived mono- and diglycerides.

[0094] Optionally, the flavor compounds include at least one flavor compound selected from the group consisting of delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2,3-pentanedione, gamma-undecalactone, and delta-undecalactone.

[0095] Optionally, the one or more sweeteners are sugars. The sugars can be selected from the group consisting of glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose. Alternatively, the one or more sweeteners are artificial sweeteners. For example, the artificial sweetener can be selected from the group consisting of stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lasititol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osalazine, pentaazine, sorbitol, thaumatin, xylitol, acesulfame potassium, advantame, alitame, aspartame, acesulfame, sodium cyclamate, darcin, glucin, neohesperidin dihirudochalcone, neotame, and P-4000.

[0096] Optionally, the ash comprises one or more of calcium, phosphorus, potassium, sodium, citrate, and chloride. In some embodiments of any of the compositions described herein, the ash comprises one or more (e.g., one, two, or three) of CaCl, KHPO, and sodium citrate. For example, CaCl can have a final concentration of about 0.05 g / L to about 0.2 g / L, KHPO can have a final concentration of about 0.2 g / L to about 0.4 g / L, and / or sodium citrate can have a final concentration of about 0.1 g / L to about 0.3 g / L.

[0097] Optionally, the dairy substitute includes one or more color-matching agents, such as, for example, beta-carotene or annatto.

[0098] <Egg substitute food> According to a further aspect of the present invention there is provided an egg replacer food product comprising at least one amylase enzyme, said at least one amylase enzyme providing at least 3% (w / w) of the protein content of the product (optionally being the major protein component of the egg replacer food product).

[0099] In another aspect, a method for producing an egg substitute food product is provided, the method comprising the steps of providing at least one amylase, subjecting the at least one amylase to one or more treatment steps to at least partially denature the amylase, and incorporating the at least partially denatured amylase into a composition to form the egg substitute food product.

[0100] The egg substitute food product can be an egg replacer composition (egg replacer), an egg-like food product, or an egg white replacer.

[0101] Optionally, the amylase enzyme is partially or totally denatured, or the amylase is denatured during cooking of the food itself or a food composition incorporating the food (e.g., when an egg replacer composition or egg white replacer is added to other ingredients to form a food product).

[0102] Optionally, the egg substitute food product contains one or more lipids. Optionally, the egg substitute food product further comprises water, a carbohydrate (such as dextrose, flaxseed powder or extract), a fat (such as coconut fat), and salt, and optionally a colorant (such as beta-carotene). The egg substitute food product can be produced by hydrating the amylase and carbohydrate, followed by homogenization and adding the fat and colorant. The emulsion can then be pasteurized. If the product is to be used in liquid form, the pasteurized liquid can be bottled and used in various applications. Alternatively, the liquid can be made into a powder, such as by freeze-drying.

[0103] Optionally, the egg substitute food product comprises one or more additional proteins (preferably non-egg proteins, more preferably vegetable proteins). Small amounts of additional protein can be provided by seed mucilage, for example flaxseed mucilage. However, the seed mucilage primarily comprises carbohydrates. Optionally, the egg substitute food product comprises one or more carbohydrates. Optionally, the egg substitute food product comprises one or more sweeteners. Optionally, the egg substitute food product comprises one or more colorants. Optionally, the egg substitute food product comprises one or more vitamins, such as yeast extract. Optionally, the egg substitute food product comprises one or more minerals or salts.

[0104] Optionally, the amylase is denatured by heat treatment, for example, heating at a temperature of 40° C. to 100° C. for a suitable time, such as 10 to 90 minutes. The amylase enzyme can be denatured during cooking of the final food composition to provide a cooked food ingredient.

[0105] The amylase may be an isolated (purified) enzyme.

[0106] The amylase may be a commercially produced enzyme, optionally produced by a recombinant (eg, recombinant microbial) process, or purified from a non-animal source.

[0107] Optionally, the amylase used in the products of the invention is recombinantly produced, i.e., the gene encoding the enzyme protein sequence is expressed under controlled conditions. Any suitable host cell can be used, for example, a microorganism such as yeast or bacteria. Optionally, the gene encoding the enzyme is expressed using a heterologous host cell. Alternatively, the gene encoding the amylase can be expressed in a homologous host cell, optionally engineered to exhibit greater than normal levels of expression, for example, by manipulation of the native promoter and / or enhancer sequences.

[0108] The amylases used in the present invention can be engineered to form gels, emulsions, or egg replacers that have a neutral flavor and a texture and mouthfeel that can be optimized by modifying processing conditions known to those skilled in the art. Many of these amylases are commercially available on the market. The availability of these enzymes provides an opportunity to improve the cost economics associated with making food substitutes from recombinantly produced food proteins.

[0109] Optionally, the amylase can be combined with other ingredients to form an egg substitute food product. Accordingly, the present invention provides a method of making an egg substitute food product, the method comprising providing an amylase and adding one or more lipids, proteins, carbohydrates, sweeteners, colorants, vitamins, minerals, and / or salts to the amylase enzyme to form the egg substitute food product. Optionally, the amylase comprises at least 3% of the protein content of the egg substitute food product (optionally at least 5% w / w protein, e.g., at least 10% w / w protein, e.g., at least 20% w / w protein). Optionally, the amylase enzyme can be the majority protein component of the food product.

[0110] When the egg substitute food product is an egg replacer composition, the composition can form an egg-based food product such as a pancake, a baked good (biscuit, cake, bread, etc.), or a mixture thereof.

[0111] The present invention is based on the recognition that amylases, when denatured (partially or fully) or hydrolyzed, can be used to produce egg-based food compositions that exhibit similar taste, aroma, and mouthfeel to those products made using eggs ("traditional eggs" or "traditional egg-based foods," the latter being, for example, baked goods, pancakes, desserts, etc., which typically include eggs as an ingredient, generally as a binder. Accordingly, the present invention provides compositions, methods for making the compositions, and kits comprising the amylases, compositions, emulsions, and mixtures, that are useful for making these egg-based food compositions.

[0112] The egg substitute foods (egg replacer compositions, egg-like foods, egg white substitutes) provided herein have similar taste, texture, aroma, and nutritional value compared to equivalent products made using avian eggs.

[0113] The egg replacer food product can include at least one amylase enzyme at a concentration of about 30-98 g enzyme / 100 g total protein content, typically 55-98 g enzyme / 100 g total protein content, and more specifically 70-98 g enzyme / 100 g total protein content.

[0114] The amylase enzymes used may be in a purified form containing at least 40% by weight of protein (enzyme) per enzyme composition (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or more pure). This reflects the content of commercially available enzyme compositions, which may contain bulking agents such as maltodextrin. Alternatively, the amylase may be produced by microbial fermentation and provided in the form of a crude extract having a protein content of 70% or less (w / w). These percentages are relative to the total content of the enzyme composition.

[0115] The egg substitute food product of the present invention can include one or more other additional proteins to produce a protein mixture. The one or more additional proteins included in the protein mixture can include one or more enzymes or food proteins. The term "food protein" refers to any non-animal-derived protein that can be added to a food product. Optionally, the additional protein can be a purified protein or a recombinant protein obtained from a non-animal source. The additional protein can include one or more avian egg proteins, but can also include other enzymes. Alternatively, or in addition, the additional protein can include one or more recombinant milk proteins, such as casein protein or whey protein, optionally produced by microbial fermentation. Optionally, the protein mixture can include one or more additional proteins selected from the group consisting of α-lactalbumin, β-lactoglobulin, α-S1-casein, α-S2-casein, lactoferrin, transferrin, and serum albumin.

[0116] Optionally, the egg substitute food product (e.g., egg replacer composition) can comprise one or more lipids. The lipids can be in the form of a fat or oil. The lipids can be selected from the group consisting of sunflower oil, coconut oil, rapeseed oil, palm oil, tributyrin, mono- and diglycerides, free fatty acids, and phospholipids. The lipids can be cultured oils or fats, microbially produced or derived oils or fats. Optionally, the egg substitute food product (e.g., egg replacer composition) can comprise a carbohydrate. The carbohydrate can be derived from a plant. The carbohydrate can be a monosaccharide, a disaccharide, a polysaccharide, or a mixture thereof. The carbohydrate can be a starch, a gum, an edible fiber, a flour, or a mixture thereof. When the carbohydrate is in the form of starch, the starch can be selected from the group consisting of tapioca starch, corn starch, potato starch, wheat starch, waxy maize starch, modified starch, and mixtures thereof. The starch may comprise 0.1% to 50% by weight of the egg replacer. When the carbohydrate is in the form of a gum, the gum may be selected from the group consisting of arrowroot starch, xanthan gum, tara gum, guar gum, agar gum, locust bean gum, and gum arabic, and mixtures thereof. The gum may comprise 0.1% to 50% by weight of the egg replacer. The carbohydrate may be in the form of exopolysaccharides typically derived from microbial cultures, such as dextrans, fructans, galactans, xanthans, and their derivatives with molecular weights of 1 to 10,000 kDa.

[0117] In one embodiment of the present invention, the egg substitute food product (e.g., egg replacer composition) can contain mucilage-containing plant seeds, or their flour, fractionated flour, or mucilage extract, to which amylase enzyme has been added. Optionally, the mucilage can be flaxseed mucilage or aloe vera extract. Optionally, flaxseed flour or fractionated flour can be used. Optionally, the mucilage is based on chia seeds, psyllium husk, aquafaba extract, chickpea water, hemp seeds, wheat germ, etc. Instead of flaxseed mucilage, fibers such as flaxseed fiber, hydrocolloids, modified starches, or water-soluble fibers such as beta-glucan, xanthan gum, pectin, etc. can be used.

[0118] Optionally, beta-carotene is added to the egg replacer food product (e.g., egg replacer composition) as a natural coloring agent. However, other yellow, orange, or red coloring agents, natural extracts, or artificial coloring agents may also be used, such as turmeric, capsicum extract, capsicum L, tomato extract, lycopene, carrot extract, annatto, safflower, etc. Optionally, one or more flavor compounds may be added to the egg replacer food product (e.g., egg replacer composition). The flavoring may be selected from the group consisting of fully or partially degraded beta-lactoglobulin, extracts containing fully or partially degraded beta-lactoglobulin (e.g., biomass extracts containing such compounds produced using precision fermentation), delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2,3-pentanedione, gamma-undecalactone, delta-undecalactone, sulfur-containing flavor compounds (e.g., 2-acetylthiazoline, dimethyl sulfide, allyl isothiocyanate, diallyl thiosulfinate, diallyl disulfide, etc.), and hydrogen sulfide. The flavoring compounds may be natural or artificial. Optionally, one or more sweeteners may be added to the egg substitute food product (e.g., egg replacer composition). The one or more sweeteners may be sugars. Optionally, the sugars are selected from the group consisting of glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose. The sweetener may also have a neutral taste, such as maltodextrin. Optionally, one or more sweeteners may be artificial sweeteners. Optionally, the egg substitute food product (e.g., egg replacer composition) may include one or more salts. Exemplary salts include one or more of calcium salts, phosphorus salts, potassium salts, sodium salts, citrate salts, and chloride salts.

[0119] Optionally, the egg replacer composition has a pH of about 4.6 to about 8.0, such as about 6.2 to about 7.2 (eg, about 6.2 to about 6.8).

[0120] The egg replacer compositions or egg white substitutes of the present invention can be used in baking or cooking as binding agents in frozen desserts, bakery mixes, meringues, coatings, batters, cakes, pancakes, tarts, confections, panades for use in e.g., croquettes, and as egg substitutes in alcoholic beverages such as whiskey sours, pisco sours, etc. The present invention also extends to baked or cooked products (i.e., "egg-based foods") made using the egg replacers or egg white substitutes of the present invention.

[0121] In one embodiment of the present invention, alpha amylase and / or glucoamylase are used to form the egg substitute food products (eg, egg replacer compositions) of the present invention.

[0122] The term "egg replacer composition" (or "egg substitute") refers to a composition formed using avian eggs as a primary component, e.g., a product similar to beaten eggs. Accordingly, as used herein, the term "egg replacer composition" refers to a product similar to traditional egg products, but made with proteins produced by a process that does not involve mammals. For example, the proteins used may be plant-based proteins, or more preferably, recombinantly expressed proteins, e.g., proteins expressed using microbial host cells. Optionally, the "egg replacer composition" of the present invention is similar, equivalent, or substantially identical to a corresponding product formed using avian eggs, particularly with respect to one or more of the following: taste, aroma, flavor, mouthfeel, organoleptic attributes, color, etc. Optionally, the "egg replacer composition" of the present invention is made with proteins produced by a process that does not involve animals.

[0123] An "egg replacer composition" of the present invention refers to a composition comprising, in addition to other ingredients, at least partially denatured amylase. The egg replacer composition is an intermediate composition for addition to an egg-based food product or composition.

[0124] The term "egg white substitute" (or "egg white substitute") refers to a product similar to the egg white (but not the yolk) or composition formed using avian eggs, without other ingredients. Thus, as used herein, the term "egg white substitute" refers to a product that resembles avian egg white, but is made with a protein produced by a process that does not involve mammals. For example, the protein used may be a plant-derived protein, or more preferably, a recombinantly expressed protein, e.g., a protein expressed using a microbial host cell. Optionally, the "egg white substitute" of the present invention is similar, equivalent, or substantially identical to a corresponding product formed using avian egg white, particularly with respect to one or more of the taste, aroma, flavor, mouthfeel, organoleptic attributes, color, etc. Optionally, the "egg white substitute" of the present invention is made with a protein produced by a process that does not involve animals.

[0125] The term "egg-based food product" refers to a food product made with the egg replacer composition or egg white replacer of the present invention in a form suitable for consumption or sale to consumers. Optionally, these products can be referred to as "composite foods." Optionally, these products can be cooked or baked by the consumer at home, or can be in the form of a kit, such as, for example, a cake mix, which requires the addition of other ingredients (e.g., fat, oil, milk, or water). The term "egg-based food composition" refers to a composition for making a food product that includes the egg replacer composition or egg white replacer of the present invention. This composition can include all the ingredients of the final food product, and can optionally be in a "ready-to-cook" or partially baked format, or can be in the form of a kit intended for the addition of other ingredients to form the final food product.

[0126] "Bird eggs" refers to eggs laid by birds such as chickens, ducks, and quails.

[0127] <Egg substitute composition (egg substitute)> Optionally, the amylase enzyme can be mixed or blended with one or more plant products, such as vegetable proteins or oils, to form the egg replacer compositions of the present invention. Because recombinant proteins are generally more expensive to produce than traditional foods, it may be desirable to include more plant products to improve the economics of making the egg replacer composition and any food products containing it from purified or recombinant proteins.

[0128] The egg replacer composition can include one or more lipids in a final total concentration of about 0% to about 30% by weight, one or more flavor compounds in a final total concentration of about 0.01% to about 4% by weight, one or more carbohydrates or sweeteners in a final total concentration of about 0.1% to about 60% by weight, and salt in a final total concentration of about 0.15% to about 1.5% by weight.

[0129] Optionally, the egg replacer composition can contain one or more lipids in the form of oil or fat. Optionally, the lipid can be selected from the group consisting of sunflower oil, coconut oil, rapeseed oil, palm oil, tributyrin, mono- and di-glycerides, free fatty acids, and phospholipids. Additionally or alternatively, the lipid can be cultured lipid or microbially produced lipid. Such lipids, including phospholipids, can be produced by sophisticated fermentation techniques. Optionally, the lipid is a free fatty acid selected from the group consisting of butyric acid, caproic acid, caprylic acid, and capric acid. Optionally, the lipid is a phospholipid, such as soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid, or rapeseed lecithin phospholipid. Optionally, the lipid can be a monoglyceride or diglyceride, which is a plant-derived monoglyceride or a bacterial-derived monoglyceride or diglyceride.

[0130] As described above, the egg replacer composition may comprise a carbohydrate. The carbohydrate may be derived from a plant. The carbohydrate may be a monosaccharide, a disaccharide, a polysaccharide, or a mixture thereof. The carbohydrate may be a starch, a gum, an exopolysaccharide, an edible fiber, a flour, or a mixture thereof. When in the form of starch, the starch may be selected from the group consisting of tapioca starch, corn starch, potato starch, wheat starch, waxy maize starch, modified starch, and mixtures thereof. The starch may comprise 0.1% to 50% by weight of the egg replacer. When the carbohydrate is in the form of a gum or exopolysaccharide, the gum may be selected from the group consisting of arrowroot starch, tara gum, guar gum, agar gum, locust bean gum, and gum arabic, and mixtures thereof. The exopolysaccharide may be selected from the group consisting of dextran, fructan, galactan, xanthan, and mixtures thereof. The gum may comprise 0.1% to 50% by weight of the egg replacer. Optionally, one or more sweeteners may be added to the egg replacer composition. The one or more sweeteners may be sugars. Optionally, the sugars are selected from the group consisting of glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose. The sweetener may have a neutral taste, such as maltodextrin. Optionally, the one or more sweeteners are artificial sweeteners. Optionally, the artificial sweetener is selected from the group consisting of stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osladin, pentasine, sorbitol, thaumatin, xylitol, acesulfame potassium, advantame, alitame, aspartame acesulfame, sodium cyclamate, darcin, glucin, neohesperidin dihirudochalcone, neotame, and P-4000. Optionally, the composition includes one or more coloring agents, such as beta-carotene.

[0131] As described above, the egg replacer composition can include one or more salts. Optionally, the egg replacer composition can include one or more salts selected from calcium salts, phosphorus salts, potassium salts, sodium salts, citrate salts, and chloride salts. Optionally, the salts used include one or more (e.g., one, two, or three) of CaCl, KHPO, and sodium citrate, KCO, and Mg(CHO). Optionally, the egg replacer includes CaCl2 at a final concentration of about 0.05 g / L to about 0.2 g / L, and / or KH2PO4 at a final concentration of about 0.2 g / L to about 0.4 g / L, and / or sodium citrate at a final concentration of about 0.1 g / L to about 0.3 g / L, and / or K2CO3 at a final concentration of about 0.1 g / L to about 0.5 g / L, and / or Mg3(C6H5O7)2 at a final concentration of about 0.01 g / L to about 0.3 g / L.

[0132] Optionally, one or more bacterial or other microbial species are employed in the process of forming the egg replacer composition for ripening or fermentation, depending on the desired end use of the egg replacer composition, to produce fermentation products and by-products such as lactic acid, carbon dioxide, alcohol, aldehydes and ketones.

[0133] The amylase enzymes used in the present invention may be commercially available enzymes produced by recombinant microbial processes or may be enzymes purified from non-animal or non-mammalian sources.

[0134] In one embodiment, the egg replacer combines amylase with potato protein, flaxseed fiber, sunflower oil, sunflower lecithin, salt, coloring, and water.

[0135] <Egg white substitute> Egg white substitutes are made as described above for egg replacer compositions, but generally do not include colorants such as β-carotene, and include, for example, amylase, water, flaxseed mucilage or flaxseed fiber, and fat (e.g., coconut oil or sunflower oil).

[0136] The egg replacer composition and / or egg white replacer can each be combined with other food ingredients to form a foodstuff that typically comprises avian eggs.

[0137] <Egg-like food> The egg-like food product is prepared as described above for the egg replacer composition and generally includes a colorant such as β-carotene, for example, amylase, water, flaxseed mucilage or flaxseed fiber, a fat (e.g., coconut oil or sunflower oil), a colorant such as β-carotene, and optionally dextrose.

[0138] In a further aspect, the present invention provides a process for forming an egg substitute product, comprising mixing an amylase enzyme with water to form a mixture and homogenizing the mixture with a fat or oil, wherein the amylase enzyme provides at least 5 g / 100 g of protein in the product. Optionally, the amylase enzyme is mixed with water and a carbohydrate source, such as a seed mucilage or wheat flour. One suitable seed mucilage is flaxseed mucilage. Optionally, a fat or oil can be added to the mixture and homogenized to form the egg substitute product. Suitable fats and oils are as described above, including, for example, vegetable oil, sunflower oil, and coconut oil. Optionally, other flavors, sweeteners, ash, salt, and / or colorants can be added, as described above with respect to the composition. These additional components can be added before or after homogenization, as convenient.

[0139] The invention can be further defined by reference to the following clauses. 1. According to one aspect of the present invention, there is provided a method for producing a dairy substitute product, the method comprising the steps of: providing an enzyme having a catalytic function; subjecting the enzyme to one or more treatment steps to convert the enzyme into a converted protein having a structural function; and incorporating the converted protein into a dairy substitute product. 2. The method according to item 1, wherein the enzyme is treated to substantially eliminate its catalytic function. 3. The method of claim 1 or 2, wherein at least one enzyme is a globular enzyme. 4. The method according to any one of items 1 to 3, wherein the enzyme is selected from the group consisting of lactase, glucosidase, cellulase, amylase, invertase, protease, xylanase, glutenase, phytase, lipase, gelatinase, glucose oxidase, transglutaminase, pectinase, beta-amylase, pullulanase, naringinase, limoninase, aminopeptidase, laccase, tyrosinase, cutinase, superoxide dismutase, endoglycosidase, glycosyltransferase, glucose isomerase, amidase, lignin peroxidase, invertase, and kinase. 5. The method according to paragraph 4, wherein the enzyme is selected from the group consisting of beta-galactosidase, alpha-glucosidase, cellulase, alpha-amylase, nattokinase, glucoamylase, and invertase. 6. The method according to any one of items 1 to 5, wherein a curd or emulsifying composition is obtained from such a spherical enzyme, and such a curd or emulsifying composition is suitable for producing an animal-free product that resembles one or more desirable properties of a dairy product. 7. The method of any one of 1 to 6, wherein the enzyme is treated by a method of hydrolysis (pH-based hydrolysis or enzymatic hydrolysis), heat treatment, or mechanical shear to at least partially or completely denature the enzyme into a structural protein building block or component. 8. The method according to any one of 1 to 5, wherein the enzyme is heat-treated at 40 to 100°C. 9. The method according to paragraph 8, wherein the heat treatment is carried out for between 1 minute and 500 minutes, typically between 5 minutes and 90 minutes, and most typically between 10 minutes and 30 minutes. 10. Non-milk protein components with desirable properties. 11. The non-dairy protein component of paragraph 10, which is a non-milk protein that has been processed to convert it into a protein with structural functionality suitable for use in the production of a dairy substitute product, such as a cheese substitute product. 12. A dairy substitute produced by treating at least one enzyme having catalytic function to make said enzyme suitable for the structural purpose of the dairy substitute. 13. Dairy substitutes according to paragraph 12, which are milk substitutes, milk substitutes, butter, cheese, yogurt, custard, cream cheese, medium-hard cheese, hard cheese, pasta filata-type cheese, soft-ripened cheese and other dairy substitutes. 14. The dairy substitute product according to paragraph 13, which, when in the form of a cheese substitute product or yogurt substitute product, is a matured cheese substitute product or a matured yogurt substitute product. 15. The alternative dairy product according to any one of items 12 to 14, wherein the protein processed according to the method of the present invention is present at a concentration of about 0.01 g / L to about 1000 g / L, generally 10 g / L to 450 g / L, more specifically 60 g / L to 350 g / L.

[0140] The invention can be further defined by reference to the following clauses. 1A. A method for producing an egg replacer, the method comprising the steps of: providing an enzyme having a catalytic function; subjecting the enzyme to one or more treatment steps to convert the enzyme into a converted protein having a structural function; and incorporating the converted protein into an egg replacer. 2A. The method of paragraph 1, wherein the enzyme is at least partially denatured to substantially eliminate catalytic function. 3A. The method of paragraph 1 or paragraph 2, wherein at least one enzyme is an isolated enzyme, particularly a globular enzyme. 4A. The method of any one of paragraphs 1 to 3, wherein the enzyme is selected from the group consisting of lactase, glucosidase, cellulase, amylase, invertase, protease, xylanase, glutenase, phytase, lipase, gelatinase, glucose oxidase, transglutaminase, pectinase, beta-amylase, pullulanase, naringinase, limoninase, aminopeptidase, laccase, tyrosinase, cutinase, superoxide dismutase, endoglycosidase, glycosyltransferase, glucose isomerase, amidase, lignin peroxidase, invertase, and kinase. 5A. The method of paragraph 4, wherein the enzyme is selected from the group consisting of beta-galactosidase, alpha-glucosidase, cellulase, alpha-amylase, nattokinase, glucoamylase, and invertase. 6A. The method of any one of 1 to 5, wherein an egg white-like emulsion composition is obtained from such a spherical enzyme, and such an egg white-like emulsion composition is suitable for producing an animal-free food product comprising one or more desirable properties of the food product. 7A. The method of any one of 1-6, wherein the enzyme is treated by a method of hydrolysis (pH-based hydrolysis or enzymatic hydrolysis), heat treatment, or mechanical shear to at least partially or completely denature the enzyme into a structural protein building block or component. 8A. The method according to any one of 1 to 5, wherein the enzyme is heat-treated at 40 to 100°C. 9A. The method of paragraph 8, wherein the heat treatment is carried out for 1 minute to 500 minutes, typically for 5 minutes to 90 minutes, and most typically for 10 minutes to 30 minutes. 10A. Enzymes that have been processed to convert them into proteins with structural functions suitable for use in the manufacture of food substitutes, such as egg replacers. 11A. A food substitute produced by treating at least one enzyme having catalytic function to render said enzyme suitable for structural purposes in the food substitute. 12A. The food substitute of Item 11, wherein the protein processed according to the method of the present invention is present at a concentration of about 0.01 g / L to about 1000 g / L, generally 10 g / L to 450 g / L, more specifically 60 g / L to 350 g / L. 13A. Egg replacers that contain a non-egg protein as the primary structural protein. 14A. The egg replacer of paragraph 13, wherein the non-egg protein comprises an isolated enzyme that is at least partially denatured. 15A. The egg replacer of paragraph 14, wherein the enzyme is an inactivated isolated enzyme or a completely denatured isolated enzyme. 16A. Egg white-like emulsion product containing a non-egg protein as the primary protein source. 17A. The egg white-like emulsion product of paragraph 16, wherein the non-egg protein is an isolated enzyme that is at least partially denatured. 18A. The egg white-like emulsion product of paragraph 16 or paragraph 17, wherein the non-egg protein component is an inactivated isolated enzyme or a fully denatured isolated enzyme.

[0141] Further features of the present invention will become apparent from the following description, given by way of example only, with reference to the accompanying figures and examples. [Example]

[0142] Example 1. Cheese substitute products using alpha amylase Alpha amylase was procured from Rajvi Biotech Pvt. Ltd. Raw materials: Enzyme details: Supplier: Enzyme Bioscience, Alpha Amylase [Aspergillus oryzae], Protein content: 71.4%. (Refers to "alpha amylase.")

[0143] [Table 1]

[0144] Processing steps: Dissolve the alpha amylase in water and allow to hydrate with all other dry ingredients in a beaker on a stir plate at room temperature for 15 minutes. Add the melted coconut oil and emulsify (Ultraturrax, shear level 10,000 rpm, room temperature). Adjust the pH to 6.2 with 1M NaOH. Ferment with lactic acid bacteria to pH 5.5 and acidify with lactic acid (30% w / w) to pH 4.5. Heat the solution in a water bath (79°C, 20 minutes). Cool to below 40°C and slowly mix the salt and cultures to mature. Press the curds by hand for 1 minute, then turn the cheese over and press for another minute. Allow to mature for 3 days at 12°C and store in the refrigerator.

[0145] Results and Discussion The formulation produced a feta / salad cheese-like structure and was slightly crumbly. The product odor and taste were neutral. The product color was off-white. The overall taste and appearance was similar to mild feta / salad cheese when cubed. The dairy substitute product is shown in Figure 1.

[0146] Example 2: Procedure for making a cream cheese substitute Raw materials: Enzyme details: Supplier: Enzyme Bioscience, Alpha Amylase [Aspergillus oryzae], Protein content: 71.4%. (Refers to "alpha amylase.")

[0147] [Table 2]

[0148] Processing steps: 1. Milk production 1. Weigh out the loss on drying (see recipe). 2. Hydrate for 30 minutes. 3. Add oil (liquid). 4. Protease: clotting time is 2 hours 2. Homogenization 1. Pre-homogenize by UT: 1 min, 12000 rpm 2. Homogenize using a panda. 1. Batch 1: Total pressure 130 bar (second stage 50 bar) 2. Batches 2-8: Standard (250 bar, second stage 50 bar) 3. Adjust pH to 4.5 with 30% w / w lactic acid 4. Solidify in foil in a water bath 1.1a: 74°C for 15 minutes 2.1b: 74℃ 20 minutes 3.1c&2-8: 70℃ 25 minutes 5. Cool the curd in foil and store at 6°C until the next day. 6. Analysis of Milk Curds 1. Place the curds in a beaker and place on a plate. 2. Take a photo. 3. Sensory: Texture, mouthfeel, appearance 4. pH Measurement

[0149] Results and Discussion: This method produced a cream cheese-like curd within a range of temperatures and times. The dairy substitute product is shown in Figure 2.

[0150] Example 3: Feta / Salad Cheese Substitute Raw materials: Enzyme details: Supplier: Enzyme Bioscience, Alpha Amylase [Aspergillus oryzae], Protein content: 71.4%. (Refers to "alpha amylase.")

[0151] [Table 3]

[0152] Processing steps: 1. Milk production 1. Measure out the dry ingredients (see recipe). 2. Hydrate for 30 minutes on a stir plate. 3. Add protease. 4. Add oil (liquid). 2. Homogenization 1. Pre-homogenize by UT: 1 min, 12000 rpm. 2. Homogenize using a panda. 1.250 / 50 bar 3.Coagulation 1. Place milk in foil. Place in a 2.74°C water bath for 25 minutes. 4. Smoothing 1. Cool the curd in the foil to room temperature. 2. Place the curds into the Kenwood mixer. 3. Shear at level 1.5 for 30 seconds. 5. Compression 1. Place cheese in a mold using cheesecloth. 2. Compress in a hydraulic press at 1.5 bar for 2 minutes. 3. Turn the cheese over and compress it again. 6. Analysis of cooled cheese 1. Place the curds in a beaker and place on a plate. 2. Take a photo. 3. Sensory analysis: texture, mouthfeel, appearance

[0153] Results and Discussion: The dairy substitute products are shown in Figure 3. The addition of protease resulted in a creamier texture and reduced dryness (see Figure 3, 2 vs. 1).

[0154] Example 4: Substitute Jogurt Products Hydrated alpha amylase and gluco-amylase paste can be converted into a substitute jogurt. Raw materials: Details of the enzymes used: Supplier: Breatec, glucoamylase [Aspergillus niger], Bakemyl AGSX (referring to "glucoamylase") with a protein content of 43%. Details of the enzyme used: Supplier: Enzyme Bioscience, Alpha Amylase [Aspergillus oryzae], Protein content: 71.4%. (Refers to "alpha amylase.")

[0155] [Table 4]

[0156] procedure: Grind with a mortar and pestle or other suitable method, if necessary. Hydrate the dry ingredients (alpha amylase or glucoamylase, salt, nutriose, flaxseed fiber) for 30 minutes and add the oil. Homogenize the mixture. Adjust the pH to 4.5. Heat in a water bath at 75°C for 20 minutes. · Smoothing. · Cool the curd and refrigerate for 12 to 48 hours before analysis.

[0157] Results and Discussion: Visual inspection: Texture similar to that of jogurt. The dairy substitute product is shown in Figure 4. Rheological measurements: Jogurt amplitude sweep, alpha amylase, glucoamylase. Milk-based Jogurt contains 3.5g fat, 4.6g carbohydrates, 3.7g protein, and 0.12g salt and is manufactured for Netto by Marken Discount AG & Co Kg. The purpose of an amplitude sweep is to show the deformation behavior of a sample in the non-destructive deformation range and to determine the upper limit of this range. The results of an amplitude sweep measurement are usually shown as a plot of strain (or shear stress) on the x-axis and storage modulus G' and loss modulus G'' on the y-axis. The amplitude sweep results (see Figure 5) show that the height and progression of the storage modulus of alpha amylase and glucoamylase are very similar to those of the Jaw Gurt sample.

[0158] Example 5 Cheese Substitute Product - Fresh Cheese Hydrated alpha-amylase and gluco-amylase paste were converted into a fresh cheese substitute. Raw materials: Details of the enzyme used: Supplier: Breatec, glucoamylase [Aspergillus niger], Bakemyl AGSX (referring to "glucoamylase") containing 43% protein. Details of the enzyme used: Supplier: Enzyme Bioscience, Alpha Amylase [Aspergillus oryzae], Protein content: 71.4%. (Refers to "alpha amylase.")

[0159] [Table 5]

[0160] Fresh Cheese Procedure Grind with a mortar and pestle or other suitable method, if necessary. Hydrate the dry ingredients (alpha amylase or glucoamylase, salt) for 30 minutes and add the fat. Homogenize the mixture. Adjust the pH to 4.5. Heat in a water bath at 80°C for 20 minutes. Compression (3 bar, 10 minutes) · Cool the curd and refrigerate for 12 to 48 hours before analysis.

[0161] Results and Discussion: The dairy substitute products are shown in Figure 6. Fresh cheese using alpha amylase (left) and glucoamylase (right). Rheological measurements: Amplitudes of commercial Almette fresh cheese and fresh cheese made with alpha amylase and glucoamylase. Almette, a milk-based fresh cheese, contains 25g fat, 3g carbohydrates, 5.6g protein, and 0.68g salt per 100g. It is manufactured by Hochland Deutschland GmbH. Ingredients list: Fresh cheese, salt, nitrogen The amplitude sweep data revealed that the height and progression of storage modulus of amylase and glucoamylase fresh cheese samples were very similar to those of Almette fresh cheese samples.

[0162] Example 6: Egg substitute Hydrated alpha amylase and gluco-amylase paste was converted into an egg replacer. Raw materials: Details of the enzyme used: Supplier: Breatec, glucoamylase [Aspergillus niger], Bakemyl AGSX containing 43% protein. (Refers to "glucoamylase"). Details of the enzyme used: Supplier: Enzyme Bioscience, Alpha Amylase [Aspergillus oryzae], Protein content: 71.4%. (Refers to "alpha amylase.")

[0163] procedure Grind with a mortar and pestle or other suitable method, if necessary. Hydrate the dry ingredients (alpha amylase or glucoamylase, salt, nutriose, flaxseed fiber, beta-carotene, flavoring) for 30 minutes and adjust the pH to 6.3. · Add oil / lecithin mix. Homogenize the mix. Stir-fry in a frying pan.

[0164] Results and Discussion: The egg replacer products are shown in Figure 7. Alpha amylase (left) and glucoamylase (right) based egg replacer.

[0165] Example 7 Glucoamylase functionality / gelling: Tap water was mixed with the enzyme preparation to obtain a solution containing 10% (w / w) protein. A) Glucoamylase [Aspergillus niger], powder, 43% protein (w / w, determined by Dumas method), Breatec B) Glucoamylase [Aspergillus niger], liquid solution, 25% protein (w / w, determined by Dumas method), WeissBio Tech

[0166] Each solution was divided into three tubes, and the pH was adjusted to 6.5, 5.5, and 5.0 with 80% (w / w) lactic acid or 1 M NaOH (w / w), respectively. Each sample was divided into two microtubes, one of which was supplemented with 1% (w / w) NaCl.

[0167] Processing steps: The sample was mixed. The sample was centrifuged (10 seconds, 13300 rpm). Incubate at room temperature for 30 minutes Heat treatment (80℃, 30 minutes) Cool in cold water Remove the sample from the microtube and analyze it visually.

[0168] Results and Discussion: All samples containing 10% (w / w) protein (glucoamylase product A or B) formed strong gels after incubation at 80°C. Gel strength was higher at pH 6.5 than at pH 5.0 and 5.5. The crumbly texture suggests suitability as a substitute for grated cheese, such as a Parmesan cheese substitute. (See Figures 8 and 9.)

[0169] Example 8: Thermally induced gelation - Analysis of gel structure Heat-induced gels made in water from alpha-amylase enzyme powder were compared with gels made with a lactase (beta-galactosidase) enzyme base. The resulting properties are influenced not only by the composition but also by the processing conditions. The gel properties are measured by rheological methods using small deformation oscillatory tests.

[0170] Standard Procedure: Use alpha amylase or purified lactase (beta galactosidase) enzymes. Details of enzymes used: 1. Source: Vita Actives, Lactase [Beta-D-Galactosidase] [Aspergillus oryzae] 100,000 ALU / g powder, protein content 44% according to the supplier. Contains lactase activity 100,000 ALU / g powder and 18-22% maltodextrin. The protein content of this preparation was increased to 60-70% by standard filtration procedures followed by spray drying (referred to as "purified lactase"). 2. Supplied by Angel Yeast Co., Alpha Amylase [Aspergillus oryzae] 55,000 U / g powder. The protein content of this preparation was 52.7% (referring to "alpha amylase").

[0171] If necessary, grind with a mortar and pestle or other suitable method. Standard procedures and recipes call for adding powder to liquid at a 6% protein weight / volume ratio (6g protein per 100ml total). Add coconut oil at a weight / volume ratio of 6% (6 grams of oil per 100 ml). Add salt (0.1 wt / vol sodium chloride and 1.39 wt / vol calcium chloride) Adjust the pH (to pH 4.8 or pH 6) Heat at a rate of 4 K / min to a final temperature of 80°C to denature the enzymes and induce coagulation, and measure the coagulation process with a rheometer. Cool the gel to room temperature in the rheometer at a rate of 10 K / min. · Analyze the rheological properties (gel properties) in a rheometer.

[0172] One common testing method for studying the rheological properties of protein gels is oscillatory testing using a rheometer. The basic principles of small deformation oscillatory testing can be explained using a two-plate model.

[0173] In a rheometer, by applying an oscillatory test, the viscoelastic behavior of a sample can be described using the storage modulus G' (unit: [Pa]). The G' value is a measure of the deformation energy stored in a material during shear processing. G' indicates the elastic behavior of the sample. High gel elasticity corresponds to a high elastic component (high G').

[0174] The data (see Figure 10) show the temperature dependence of G' and G'' for alpha amylase and lactase heated to 80°C. For both proteins, the G' curves exceed the G'' curves, indicating that solid behavior predominates over liquid behavior. The temperature at which G' increases reflects the heat-induced changes that lead to gel formation. For alpha amylase, G' increases almost linearly. The temperature at which the G' change occurs is between 47 and 50°C, while the G' of the lactase gel increases at 66°C. This may indicate that under the selected conditions, gel structure formation begins early but is much slower for alpha amylase gels than for lactase gels.

Claims

1. A method for producing a food substitute, comprising: i. providing at least one amylase enzyme; ii. subjecting at least one of said amylases to one or more treatment steps to at least partially denature said amylase or providing said amylase in an amount of at least 3 g / 100 g protein in the product; and iii. incorporating the amylase into a composition to form a food substitute.

2. 10. The method of claim 1, wherein the amylase is the primary protein component of the food substitute.

3. 3. The method of claim 1 or claim 2, wherein the amylase is at least partially denatured.

4. 4. The method of any one of claims 1 to 3, wherein the amylase is totally denatured within the food substitute.

5. The method according to any one of claims 1 to 4, wherein the food substitute is curdled milk.

6. 6. The method of any one of claims 1 to 5, wherein the amylase is denatured by hydrolysis, heat treatment, or mechanical shear.

7. 7. The method according to any one of claims 1 to 6, wherein the amylase is heat-treated by heating to a temperature of 40 to 100°C.

8. The method of claim 7, wherein the heat treatment is carried out for 10 to 30 minutes.

9. 1. A dairy substitute food composition comprising: at least one amylase, comprising one or more of lipids, proteins, sweeteners, and / or ash; A composition wherein said amylase comprises at least 3% by weight of the protein in said substitute food composition.

10. 1. An egg replacer food composition comprising an amylase enzyme present in the product in an amount of at least 3 g / 100 g of protein.

11. 11. The composition of claim 9 or claim 10, comprising a lipid.

12. The composition of claim 11 , wherein the lipid is coconut oil or a vegetable oil.

13. 13. The composition of any one of claims 9 to 12, which is a liquid composition and comprises the amylase in a concentration of 0.01 g / L to 1000 g / L.

14. 14. The composition of claim 13, comprising the amylase at a concentration of 60 g / L to 350 g / L.

15. The composition of any one of claims 9 to 14, wherein the amylase is the major protein component of the composition.

16. 16. The composition of any one of claims 9 to 15, wherein the amylase comprises at least 5% by weight of the composition.

17. 17. The composition of any one of claims 9 to 16, further comprising mucilage formed from soaked flaxseed.

18. 17. A dairy replacer formed using the composition of any one of claims 9 or 11-16, which is a curd replacer, milk replacer, butter, cheese, yogurt, custard, cream cheese, medium hard cheese, hard cheese, pasta filata type cheese, soft ripened cheese, and other dairy replacer products.

19. 20. The dairy substitute product of claim 18, which is a matured cheese substitute product or a matured yogurt substitute product.

20. 18. An egg substitute food product according to any one of claims 10 to 17 in the form of an egg white substitute.

21. A substitute egg-based food product comprising the egg replacer composition of any one of claims 10 to 17 together with one or more food grade ingredients.

22. 22. The egg based food substitute of claim 21, comprising the at least partially denatured amylase in a concentration of from 6g / 100g to 35g / 100g.

23. 23. The egg substitute based food product of claim 22 selected from a baked good, dessert, coating, batter, pancake, tart, confection, or panade.

24. The egg replacer composition according to any one of claims 10 to 17, together with one or more ingredients, An egg-based substitute food composition in the form of a ready-to-bake or partially baked food product.

25. The egg replacer composition according to any one of claims 10 to 17, together with one or more ingredients, An egg-based substitute food composition in the form of a food kit.

26. 1. A process for forming an egg replacer product, comprising: A process comprising mixing an amylase enzyme with water to form a mixture, and homogenizing said mixture with an oil or fat, wherein said enzyme provides at least 3 g of protein per 100 g of product.

27. 27. The process of claim 26, wherein the enzyme is combined with water and a carbohydrate to form a mixture.

28. 28. The process of claim 27, wherein the carbohydrate is a seed mucilage.

29. 29. The process of claim 28, wherein the seed mucilage is flaxseed mucilage.

30. 30. The method of any one of claims 26 to 29, wherein the oil or fat is vegetable oil, sunflower oil, or coconut oil.

31. 31. The method of any one of claims 26 to 30, wherein one or more flavor modifiers, sweeteners, ash, salt or colorants are added.

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