Meat substitute product and method of manufacturing the same

Abstract

Methods of making meat substitute food products and meat substitute food products are provided. To improve texture, mouthfeel, and / or prevent or delay hardening of a protein matrix, a meat substitute food product is made using a high moisture protein texturization process using in an extruder under conditions resulting in a continuous protein fiber matrix structure containing a disruption, i) at least one protein matrix forming ingredient, such as an isolated protein or a concentrated protein, ii) a starch-containing grain selected from the group consisting of ii.a) steeped grains, ii.b) germinated grains, ii.c) malted grains, ii.d) sprouted grains, or ii.e) any combination of two, three, or four of these starch-containing grains as one of the ingredients, and iii) water or an aqueous liquid, the extrudate containing starch located at the disruption and not emulsified with the protein fiber matrix structure.

Description

TECHNICAL FIELD

[0001] The present invention relates to a meat substitute product and a method for manufacturing thereof. TECHNICAL BACKGROUND

[0002] In recent years, many people have turned to vegetarian or vegan diets, or at least increased the proportion of vegetables and vegetable products in their diet. While ecological concerns are the reason for some, it seems clear that vegetables and vegetable-made products should be a central part of a healthy diet. Many consumers find it difficult to ensure a daily intake of proteins using vegetables or vegetable-made products, and some find it time-consuming to prepare protein-containing foodstuffs for cooking or roasting.

[0003] There is therefore a market for the industrial production of vegetarian or vegan foodstuffs by extrusion cooking. Extrusion cooking is a continuous process that makes it possible to produce texturized proteins, which are unique products made by extrusion. Extrusion makes it possible to control functional properties such as density, rate and time of rehydration, shape, product appearance and mouthfeel.

[0004] For the extrusion of meat substitute products, also called meat analogues, or texturized vegetable products, a twin-screw extruder is generally used. There are mainly two extrusion cooking processes for preparing meat substitute products.

[0005] One type of meat substitute product is produced by low-moisture protein texturization extrusion. These products have a moisture content of 10% to 40% (moisture content during extrusion is 15% to 40%). They generally have a spongy texture and need to be rehydrated before consumption. These products are generally used as a ground meat substitute or as a meat extender in meat products, but they hardly mimic fibrous, whole muscle meat.

[0006] Another type of meat substitute product is manufactured by high-moisture protein texturization extrusion. These products have a moisture content of 40% to 80% (moisture content during extrusion is more than 40%). They generally resemble muscle foods more than meat substitute products manufactured by low-moisture texturization extrusion.

[0007] Meat substitute products are generally manufactured by mixing at least one protein base forming ingredient such as isolated or concentrated protein (often referred to as protein component), possibly starch-containing particles, possibly oil, and extruding the ingredients mixed into a slurry in an extruder configured to perform protein texturization extrusion.

[0008] In tests performed by the inventors with high moisture protein texturized extrusion we found that the mouthfeel of freshly extruded meat substitute products is generally very appealing. However, after a relatively short time (typically in the range of a few minutes, typically 5-10 minutes), when the meat substitute product cools down, the mouthfeel becomes unacceptable.

[0009] Currently, meat substitute products manufactured with high moisture protein texturized extrusion are generally sold deep-frozen. Alternatively, the meat substitute products are sold in a shredded or flaked form, so that the unacceptable mouthfeel becomes less apparent. SUMMARY

[0010] OBJECT OF THE INVENTION

[0011] It is the general object of the present invention to improve the texture and mouthfeel of meat substitute food products (i.e. meat imitations) and to further delay the structural hardening (firming up) of the protein matrix of meat substitute food products. It is a further object to improve the nutritional properties and the flavor of meat substitute food products (i.e. meat imitations). All these objects or at least some of these objects can be achieved by the method according to claim 1 and the meat substitute food product according to the parallel claim 22.

[0012] Dependent claims describe advantageous embodiments of the method and the meat substitute product.

[0013] ADVANTAGES OF THE INVENTION

[0014] In the method of manufacturing a meat substitute food product,

[0015] i. at least one protein matrix forming ingredient, such as isolated protein or concentrated protein,

[0016] ii. starch-containing grains selected from the group consisting of ii.a) steeped grains, ii.b) germinated grains, ii.c) malted grains, ii.d) sprouted grains, or ii.e) any combination of two, three or four of these starch-containing grains as one of the ingredients, and

[0017] iii. water or an aqueous liquid

[0018] a. is fed to an extruder suitable for high moisture protein texturized extrusion; and

[0019] b. is extruded in the extruder.

[0020] The extrusion is performed under conditions resulting in a continuous protein fibre matrix structure comprising a site of disruption, the extrudate comprising starch located at the site of disruption and not emulsified with the protein fibre matrix structure. The starch located at the site of disruption, not emulsified, can contribute to the long lasting improved mouthfeel.

[0021] The use of the method can improve the texture and mouthfeel of the meat substitute product. It can also prevent or delay the structural hardening (densification) of the protein matrix.

[0022] Furthermore, the use of the method can improve the nutritional profile and flavour of the meat substitute product. Although it is known that sprouted grains have an improved nutritional profile [see "5 Sprouted Seeds for Human Health" in reference 14], the sprouted grains are used in powder form if not consumed as is or used in a beverage fermentation process. The use of the method, starch containing grains selected from the group consisting of: ii.a) steeped grains, ii.b) germinated grains, ii.c) malt, ii.d) sprouted grains, or ii.e) any combination of two, three or four of these starch containing grains can be higher than starch containing grains that have not been steeped, germinated, malted or sprouted, while having advantages in terms of mouthfeel, texture profile and protein matrix hardening profile. The inventors are not aware that these ingredients would be used as starting material for protein texturised extrusion to make a meat substitute product.

[0023] Preferably, the extrusion is performed with a high moisture protein texturised extrusion method in which the starch containing grains are gelatinised and the protein forming the protein matrix is melted. This makes the structure of the meat substitute food product more compact, which in turn can contribute to improved texture profile and mouthfeel and / or delay or prevent the hardening of the protein matrix.

[0024] Preferably, in the extrusion, the starch containing grains are gelatinised before they are substantially pulverised by the extruder screw to create a substantially linearly oriented continuous protein fibre matrix structure with some starch not emulsified with the protein fibre matrix structure. This can contribute to the long lasting improved mouthfeel.

[0025] Preferably, in the extrusion, the starch containing grains are gelatinised and the protein forming the protein matrix is melted:

[0026] i. before the gelatinised starch containing grains form an emulsion with the protein forming the protein matrix, and / or

[0027] ii. before the gelatinised starch forms a complete barrier preventing the formation of a continuous protein fibre cross-linked matrix.

[0028] This can contribute to the long lasting improved mouthfeel.

[0029] Preferably, after said extrusion, at least 10.5% of said starch is washable starch,

[0030] i. when the protein content of said extrudate is more than 55 wt% but less than 70 wt%, at least 10.5% of said starch is washable starch,

[0031] ii. when the protein content of said extrudate is at least 70 wt% but less than 90 wt%, at least 15% of said starch is washable starch,

[0032] iii. when the protein content of said extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of said starch is washable starch,

[0033] wherein the wt% is based on a dry basis.

[0034] These percentages of washable starch can contribute to obtaining a long-lasting improved mouthfeel.

[0035] Preferably, said protein matrix structure comprises disruptions, wherein some of said disruptions are in the form of cavities, the walls of which are at least partially coated with a cluster of gelatinized starch, said cluster of gelatinized starch being formed from starch, preferably from soluble starch or washable starch. Cavities with walls at least partially coated with a cluster of gelatinized starch formed from starch can contribute to obtaining a long-lasting improved mouthfeel.

[0036] In practice, from a commercially available material point of view, said at least one protein matrix forming ingredient can be at least one isolate protein and / or at least one concentrate protein or comprise at least one isolate protein and / or at least one concentrate protein.

[0037] Preferably, said starch-containing grain has an average or median microparticle volume of at least 0.125 mm 3 , preferably at least 1 mm 3 , most preferably at least 6 mm 3 .

[0038] Preferably, said extrusion is performed:

[0039] a. water or an aqueous liquid is fed into said extruder;

[0040] b. the mixture is heated in said extruder to gelatinize said starch-containing grain;

[0041] c. after starch gelatinization is reached, the mixture is further heated in said extruder to melt at least one protein matrix forming ingredient;

[0042] d. extruding the mixture through an extrusion die at a temperature of 70°C to 100°C.

[0043] In this way carrying out the extrusion, the inventors have managed to produce a meat substitute food product which has a long lasting improved mouthfeel.

[0044] Preferably,

[0045] e. heating step b. is carried out with impact heating, such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw, preferably:

[0046] i. such that the starch gelatinization takes place between 0s and 18s, preferably between Is and 15s, after the water / aqueous liquid dosing step a; and / or

[0047] ii. before the starch-containing grains are ground to a particle volume of less than 5000 pm 3 by the extruder screw, and preferably before the starch-containing grains are ground to a particle volume of less than 1000 pm 3 by the extruder screw; and / or

[0048] f. heating step c. is carried out with impact heating, such that the melting temperature of the protein of the protein matrix forming ingredient is reached, preferably a temperature of 140°C to 200°C, and / or such that protein melting takes place between Is and 40s, preferably between 10s and 30s, after the water / aqueous liquid dosing step a; and / or

[0049] g. after the heating step c. the extrusion of the mixture is continued for more than 5s, preferably more than 10s, at a temperature which is not higher than the temperature of the heating step c., preferably at a temperature of 90°C to the temperature in heating step c.

[0050] The starch-containing grains can be treated before being fed into the extruder, such that the starch is at least partially gelatinized before being fed into the extruder.

[0051] Preferably, water or an aqueous liquid is fed into the extruder at an elevated temperature, preferably the temperature of the water is higher than 60°C, more preferably higher than 65°C, most preferably higher than 75°C.

[0052] The starch-containing grains can be selected from, comprise or consist of one or more of the following: oat, barley, rye, wheat, rice, corn, lentil, chickpea, mung bean, haricot bean, pea, quinoa, pigeon pea, sorghum, buckwheat.

[0053] Preferably, the starch-containing grain consists of whole grains or the starch- containing grain comprises whole grains. The hull (e.g. bran layer) of the whole grains can prevent or slow down powderization of the grains in the extruder, thereby preventing some starch from forming an emulsion with the protein matrix. This can contribute to a long-lasting improved mouthfeel.

[0054] The starch-containing grain can consist of or comprise mechanically processed starch- containing grains, such as in particular one or more of the following: flakes (e.g. pressed, rolled or flaked), steel-cut grains, dehulled pearled grains, ground grains, dehulled but not pearled grains. The mechanically processed starch-containing grains can prevent or slow down powderization of the grains in the extruder, thereby preventing some starch from forming an emulsion with the protein matrix. This can contribute to a long-lasting improved mouthfeel.

[0055] Preferably, in the method, in addition to

[0056] i. at least one protein matrix-forming ingredient, such as for example isolated protein or concentrated protein;

[0057] ii. a starch-containing grain selected from the group consisting of: ii.a) malted grains, ii.b) germinated grains, ii.c) malt, ii.d) sprouted grains, or ii.e) any combination of two, three or four of these starch-containing grains as one of the ingredients, and

[0058] iii. water or a water-containing liquid

[0059] Also

[0060] iv. flour and / or bran and / or starch and / or fiber can be used.

[0061] This can bring benefits such as for example an improved nutritional value of the meat substitute product. Furthermore, these materials are widely commercially available. They can also control hardening to some extent. Different protein sources with different protein contents can have different combined effects with starch, flour and fiber.

[0062] The flour can comprise, consist of or be selected from at least one of the following: oat, barley, rye, wheat, rice, corn, lentil, chickpea, mung bean, haricot bean, pea, quinoa, pigeon pea, sorghum, buckwheat, potato, sweet potato, lupin, any mixture thereof.

[0063] The bran can comprise, consist, or consist essentially of at least one of the following: oat bran, barley bran, rye bran, wheat bran, rice bran, corn bran, millet bran, any mixture thereof.

[0064] The starch can comprise, consist, or consist essentially of at least one of the following: oat starch, barley starch, rye starch, wheat starch, rice starch, corn starch, lentil starch, chickpea starch, mung bean starch, broad bean starch, pea starch, quinoa starch, pigeon pea starch, sorghum starch, buckwheat starch, potato starch, sweet potato starch, lotus root starch, any mixture thereof.

[0065] The fiber can comprise, consist, or consist essentially of at least one of the following: oat fiber, barley fiber, rye fiber, wheat fiber, rice fiber, corn fiber, lentil fiber, chickpea fiber, mung bean fiber, broad bean fiber, pea fiber, quinoa fiber, pigeon pea fiber, sorghum fiber, buckwheat fiber, potato fiber, sweet potato fiber, lupin fiber, apple fiber, any mixture thereof.

[0066] The starch-containing cereal grain can be selected such that the steeping treated grain (ii.a) is only used in combination with the germination treated grain (ii.b) and / or malt (ii.c) and / or germinated grain (ii.d), i.e. option ii.a) can be excluded such that it is not selected alone in the method.

[0067] Preferably, the extrusion step is performed with an extrusion die having a length of more than 300 mm, preferably more than 1000 mm.

[0068] Preferably, some of the non-emulsified starch can be soluble starch, preferably the starch that is not bound to the protein matrix is determined to be soluble starch; preferably the compressibility is controlled by varying the extrusion parameters such that the ratio of the amount of soluble starch to the total amount of starch in the extruded meat substitute product is 3 to 10 wt.%, and / or the soluble starch content is 0.03 to 1.1 wt.%. This can contribute to a long-lasting improved mouthfeel.

[0069] A meat substitute food product, which is an extrudate manufactured according to the method of the first aspect of the invention or which comprises an extrudate manufactured according to the method of the first aspect of the invention.

[0070] Preferably, the extrudate comprises starch, at least 5.1%, preferably at least 5.2% of which is soluble starch. This can contribute to a long-lasting improved mouthfeel.

[0071] The extrusion can comprise disruptions in the matrix structure, such that some of the disruptions are in the form of cavities, the walls of which are at least partially coated with clusters of gelatinized starch, the clusters of gelatinized starch being formed from a leachable starch that does not emulsify with the matrix structure. This can help to obtain a long-lasting improved mouthfeel.

[0072] The clusters of starch can comprise a leachable starch that is leachable in water at a temperature of 50°C. A leachable starch can help to obtain an improved mouthfeel.

[0073] Preferably, the matrix structure has disruptions, and some of the disruptions in the matrix structure are preferably in the form of cavities, the walls of which are at least partially coated with clusters of gelatinized starch, the clusters of gelatinized starch being formed from a starch, preferably from a soluble starch. Such cavities can help to obtain a long-lasting improved mouthfeel.

[0074] Preferably, the meat substitute product is in the form of chunks, chops, nuggets, fillets, steaks, or doner meat-like slices, or in the form of doner kebab-like layer-wise stratification layers in a yoghurt or a vegan yoghurt and a spice. BRIEF DESCRIPTION OF DRAWINGS

[0075] The meat substitute product and the method of manufacturing the meat substitute product will be described in more detail below with reference to the accompanying drawings, in which:

[0076] Figure 1 is a photograph of sample no. 5, 7 and 8;

[0077] Figure 2A is a Micro-CT scan image taken after sample no. 5 was soaked in water at 60°C for 24 hours and air dried;

[0078] Figure 2B is a Micro-CT scan image taken after sample no. 8 was soaked in water at 60°C for 24 hours and air dried. This sample was cut in the same way as Figure 2A

[0079] Figure 3 illustrates the relationship observed between the solubility of the starch and the compression force needed to compress the meat substitute product (exponential curve fit to the measured points);

[0080] Figure 4 ​Particle weight distribution of extruded material affected by ingredient composition and extrusion heating temperature profile is shown for experiments 1-6;

[0081] Figure 5 Results of compression testing of dry (un-soaked) steel cut oats versus soaked steel cut oats (soaked in hot water) are shown;

[0082] Figure 6A and 6B are microscope images (10x magnification) taken from a sample of sample 2;

[0083] Figure 6C and 6D are microscope images (10x magnification) taken from a sample of sample 2;

[0084] Figure 6E and 6F are microscope images (10x magnification) taken from a sample of sample 6;

[0085] Figure 6G and 6H are microscope images (20x magnification) taken from a sample of sample 6;

[0086] Figure 7A are microscope images taken from a sample of washable starch washed out of sample 2 with water at 50°C;

[0087] Figure 7B are microscope images taken from a sample of washable starch washed out of sample 2 with water at 50°C;

[0088] Figure 8 are examples of food made from the meat substitute product (sample 2) after shredding into pieces;

[0089] Figure 9 are examples of food made from the meat substitute product (sample 2) after shredding into pieces, marinating the pieces (left side), breading the extruded product, breading the extruded product and deep frying in oil (right side);

[0090] Figure 10 Gelation of pea protein affected by heating temperature is shown;

[0091] Figure 11 Analysis methods for cutting force and compression force are illustrated;

[0092] Figure 12A and B illustrate the schematic arrangement of the extrusion process;

[0093] Figure 13 Quantitative analysis methods for soluble starch and washable starch are illustrated;

[0094] Figure 14A Starch coated on the inner surface of the cavity of the extruded product is shown;

[0095] Figure 14B The inner surface of the cavity of the extruded product as observed by iodine staining is shown;

[0096] Figure 14C The inner surface of the cavity of the extruded product as observed by iodine staining is shown;

[0097] Figure 14D and Figure 14E The inner surface of the cavity of the extruded product as observed by iodine staining is shown; and

[0098] Figure 15 Photos of sample 2 are shown before (top photo) and after (bottom two photos) expansion.

[0099] In all figures, like reference numerals refer to like parts. DETAILED DESCRIPTION

[0100] Previous work - not published so far - is described in detail

[0101] I: State of the art and objectives

[0102] Cooked chicken drumstick meat has a different mouthfeel than cooked chicken breast meat. The difference in mouthfeel relates, inter alia, to tenderness. Cooked chicken breast meat usually requires a relatively high compression force at 40% compression, which indicates that cooked chicken breast meat usually has a relatively low compressibility.

[0103] As described in the introductory part, the inventors have been working on meat replacement products manufactured with high-moisture protein texturized extrusion. Figure 12A An extruder 12 configured to implement a conventional high-moisture protein texturized extrusion process is schematically shown. In the extruder 12, ingredients in powder form are mixed in a mixer 121, which is connected with a supply line 122 leading to an inlet hopper 123. The extruder 12 has a liquid feed line 124 connected with a conventional tap water supply (tap water usually has a temperature not higher than room temperature or, for example, 30°C) (preferably via a valve 130 and a collection tank 131 to achieve a constant water volume flow). The extruder 12 has a long cooling die 125. The extrusion is implemented with two extruder screws 126, hence the name “twin-screw extruder”.

[0104] The focus of the research has been to improve the mouthfeel and find ways to produce a meat substitute made with high moisture protein texturized extrusion that has a suitable high compressibility and chewiness so that the mouthfeel is as close as possible to that of a cooked chicken drumstick. Furthermore, to optimize the mouthfeel, the meat substitute should have a long continuous fibrous protein matrix structure.

[0105] On the market, there are meat substitutes made with high moisture protein texturized extrusion that are sold shredded or torn into pieces and that have a mouthfeel that is somewhat comparable to that of a cooked chicken breast fillet when the meat substitute is cooled after extrusion. Table I shows certain data for selected existing meat substitutes compared to tofu, chicken breast fillet and chicken drumstick.

[0106] Table I: Physical properties of selected meat substitutes on the market

[0107]

[0108] [ is a registered trademark of Food for Progress Scandinavia Ab in at least the European Union, the United States, New Zealand, Switzerland, Australia, the Islands and Norway. The product "the chunk" has the ingredients water, soy protein (23%) and salt.

[0109] None of these products that the inventors were able to test resembled a cooked chicken drumstick, which is more tender, more compressible and has a more flexible structure than a cooked chicken breast fillet.

[0110] Due to the long continuous fibrous protein matrix structure of a cooked chicken drumstick, it has a chewy mouthfeel that is comparable to that of a chicken breast fillet.

[0111] In tests that the inventors have performed with high moisture protein texturized extrusion, we have found that the mouthfeel of a fresh extruded meat substitute made with high moisture protein texturized extrusion is generally very appealing.

[0112] However, after a relatively short time, generally in the range of a few minutes, often 5 to 10 minutes, the mouthfeel becomes unacceptable when the meat substitute cools. This unacceptable mouthfeel is caused by the meat substitute losing its tenderness, becoming less compressible, and the structure of the meat substitute becoming less flexible.

[0113] Currently, most meat substitutes made with high moisture protein texturized extrusion are sold deep-frozen. After thawing, these products will have a mouthfeel that is comparable to that of a cooked chicken breast fillet, but not comparable to that of a cooked chicken drumstick.

[0114] To improve the mouthfeel of meat substitute products manufactured with low moisture extruded protein texturization, it is known to add microparticles to the extrudate, such as starch; flour; soluble and insoluble polymeric fibers such as pea fiber, cellulose, agar, xanthan gum (e.g., in U.S. Patent Application Publication 2016 / 0205985 Al); insoluble salts such as gypsum (e.g., in U.S. Patent 5,922,392); and fat to disrupt protein fibers to tenderize the extruded product to produce meat substitute products (e.g., in U.S. Patent Application Publication 2016 / 0205985 Al).

[0115] However, these compounds are mostly small in size (less than 100 pm in each dimension) prior to extrusion, or will be broken into small parts (less than 100 pm in each dimension) during the extrusion process. In practice, they will all be homogenized by the extruder screw, and emulsified with the protein material that covers them.

[0116] Tolstoguzov [Reference 1] has investigated and described in detail different types of emulsions in protein extrusion, including polysaccharide emulsions in protein. Tolstoguzov found that the emulsion system extruded under protein texturization extrusion conditions is different from the typical water-in-water or oil-in-water emulsions that exist at temperatures below 140°C. The protein-in-polysaccharide emulsion can be considered as an emulsion of a polysaccharide melt in a protein melt. During the meat substitute product manufacturing process, i.e., in the high moisture protein texturization extrusion process, protein is the main ingredient. Typically, protein constitutes 50% to 100% of the weight of the extruded raw material on a dry basis. Usually, plant proteins suitable for such extrusion processes can be melted in the extruder at heating temperatures of 140°C to 200°C. Therefore, protein can form the continuous phase.

[0117] Thus, the microparticles disclosed in US 2016 / 0205985 Al and 5,922,392 will be dispersed in the protein and form the dispersed phase. The dispersed microparticles are stably trapped or embedded within the continuous phase, evenly distributed throughout the continuous phase, and have a small particle size.

[0118] The spinneretless spinning effect in extrusion leads to the formation of anisotropic (fibrous or lamellar) structures of the heterogeneous liquid system in the flow.

[0119] In the final stage of the extrusion process, the shape of the emulsion, the liquid filaments and the anisotropic structure are fixed by the rapid gelation of the protein phase, which is shorter than the lifetime of the liquid filaments. After this, if the protein layer or protein matrix structure covering the dispersed particles is not broken open, the dispersed particles remain uniformly dispersed, firmly embedded and almost inseparable from the protein matrix by mechanical forces (e.g. centrifugation, gravity) or by extraction (e.g. water washing, water extraction).

[0120] When producing meat substitute products with protein texturized extrusion, the known method of including particles in the extrusion is known to tenderize the extruded product to some extent, especially when the extruded product is freshly produced and before cooling and storage overnight. The particles can break the protein fibers by being located in between the protein fibers or between adjacent protein fibers.

[0121] The addition of such particles also dilutes the concentration (proportion) of protein in the ingredients used for the extrusion, which protein forms the protein fiber matrix and contributes to the strength of the extruded product. In this way, the addition of particles can soften the extruded product, especially when the product is fresh and warm before cooling at a cool temperature (e.g. 0°C to 6°C) and storage overnight. In low moisture protein texturized extrusion for the production of meat substitute products (e.g. with a moisture content of the material between 15% and 40% during the extrusion process), the extruded product is mostly largely expanded and contains a large number of air bubbles between the protein fibers. The expansion and air bubbles are due to the large amount of water evaporation that occurs when the extruded material, just after leaving the extrusion die, is at a high temperature (e.g. above 100°C). In such a case, the broken protein fibers are further separated by the air bubbles and are fixed in a position far away from each other. Therefore, in low moisture protein texturized extrusion for the production of meat substitute products, the breaking effect from these particles can be attractive to some extent.

[0122] However, in high moisture protein texturized extrusion for the production of meat substitute products (e.g. with a moisture content of the material between 40% and 80% during the extrusion process), the expansion of the extruded material is much less and contains much less air bubbles that are uniformly distributed between the protein fibers to break their cross-linking between adjacent fibers.

[0123] Akdogan [Reference 2] found that the reduced level of expansion in high moisture protein texturized extrusion is caused by the increased concentration of water during the extrusion process. More specifically, extrusion with higher moisture content has a different distribution of shear (the shear forces present are typically less in high moisture protein texturized extrusion), mixing, mechanical heat (the mechanical heat dissipation is typically less in high moisture protein texturized extrusion) and convective heat. Due to the greatly reduced melt viscosity and the reduced pressure build-up in the extruder barrel, the viscous dissipation of energy in the extruder barrel is much less in extrusion with higher moisture content. The pressure along the die is greatly reduced and this is therefore partly responsible for the minimal to non-existent expansion at the die. The extruded material is cooled during the high moisture protein texturized extrusion process with a long cooling die and therefore the evaporation of moisture is greatly reduced. It is also known from the background art that when the starch content of the extruded material is low, and when the starch gelatinization level is low, the level of expansion of the extruded material leaving the extrusion die will be low. The low viscosity of the extruded material associated with the higher moisture content also causes it to be unable to some extent to maintain (sustain) a stable expansion without collapsing into a dense mass.

[0124] The difference in moisture content during extrusion also causes a change in the main contributing protein-protein forces that stabilize the protein matrix. Lin et al. [Reference 3] found that under high moisture extrusion (e.g. when the moisture content during extrusion is between 40% and 80%), a large portion of the proteins are connected and stabilized by hydrogen bonds, while disulfide bonds and hydrophobic interactions are not the main stabilizing forces of the protein matrix. Conversely, under low moisture extrusion (e.g. when the moisture content during extrusion is between 30% and 40%), the main important protein matrix stabilizing forces are disulfide bonds and hydrophobic bonds. After extrusion, the hydrogen bonds in the protein matrix can significantly contribute to further increasing the gel strength (firmness) of the extruded product during the cooling process. It is well known and disclosed by Sun and Arntfield [Reference 4] that low temperatures for storage (e.g. 0°C to 6°C) and the cooling process after protein gel formation can favor extensive and increasing formation of hydrogen bonds. Furthermore, it is well known that during the cooling process after starch is heated and gelatinized in water, the starch gel strength also increases mostly and substantially, because a large number of hydrogen bonds are created between the starch molecules during cooling. Starch retrogradation can occur after starch gelatinization. Longer storage time periods will result in further formation of hydrogen bonds and thus in a further tightening (firming) of the structure and a lower water holding capacity. Therefore, starch gelatinization and retrogradation are another factor that contributes to the problem of the texture firming and loss of attractive mouthfeel of the meat substitute product produced by high moisture protein texturized extrusion in the methods known in the background art.

[0125] In the context of baked bread, the adverse effect of retrogradation on the bread crumb texture is well known: retrogradation significantly causes the bread crumb to age and increase in firmness during storage.

[0126] Hydrogen bonds are short-range chemical bonds, which means that the cross-linking associated with hydrogen bonds primarily occurs between adjacent compounds (e.g., protein-protein, protein-starch, starch-starch) that are in close or direct contact with each other. Starch of the amylose type has a high capacity to form starch-starch hydrogen bonds because it has many hydroxyl groups and linear polymer chains in its molecular structure. Starch before gelatinization cannot form a gel in water because the starch is embedded in the starch granule structure and thus insoluble. Starch gelatinization can occur more extensively in high moisture extrusion processes than in low moisture extrusion. During high moisture protein texturization extrusion, starch is heated sufficiently to leach out into water by heat and shear forces, causing the leached amylose molecules to line up linearly and in close contact with each other.

[0127] Because the extruded product from high moisture protein texturization extrusion has a higher density (less expansion, higher density) and more protein-protein cross-linking force formation of the hydrogen bond type than the product from low moisture extrusion, the addition of particulates (e.g., starch powder, insoluble salt, fiber, fat, etc.) can hardly disrupt the protein-protein cross-linking or interaction forces that occur extensively during the cooling phase and after extrusion as much as in low moisture extrusion. Therefore, these extruded products with or without the addition of particulates are still subject to the very serious problems of structural hardening (densification) and loss of acceptable mouthfeel (e.g., compressibility) during cooling and storage. More specifically, the particulates are easily homogenized, covered, and emulsified by the protein matrix during the extrusion process or shortly after being extruded with the protein material. The particulates then cannot provide a large enough disruptive force or barrier effect between protein fibers, but can only provide a limited disruptive area around each individual particulate point without extending. More seriously, when starch is added in the form of starch powder (including or not including modified starch or pregelatinized starch) or grain flour powder, they are also homogenized, covered, and emulsified by the protein matrix shortly after being extruded with the protein material. The emulsified starch is then heated and gelatinized. The starch remains as small particulates throughout the extrusion process and in the final product. So the starch can hardly provide a large enough disruptive force or barrier effect between protein fibers, but can only provide a limited disruptive area around each individual particulate point without extending. The protein matrix around the starch particulates can continue to densify, forming protein-protein interaction forces, such as more hydrogen bonds, after extrusion. In addition, the starch, after being sheared, gelatinized, distributed within (between) linearly arranged protein fibers, and linearly arranged, becomes extremely susceptible to starch gelatinization, retrogradation, hardening, drying, and starch-protein interaction with hydrogen bond formation potential. In this way, the extruded product is subject to the very serious problems of structural hardening (densification) and loss of acceptable mouthfeel (e.g., compressibility) during cooling and storage.

[0128] II: Processing machine (extruder system) implementing the tests described in the following examples

[0129] Figure 12B An extruder 13 configured to implement the high-moisture protein texturized extrusion process according to the methods described in the present application is shown. The extruder 13 implements the technical features required for the new process.

[0130] In the new process, the mechanically processed starch-containing cereal grains are mixed with starch-containing cereal grains in powder form (preferably flour), at least one (preferably vegetable or dairy) isolated protein / at least one (preferably vegetable or dairy) concentrated protein / a mixture of at least one such isolated protein and at least one such concentrated protein, possibly oil and possibly spices and any other ingredients in a mixer 121 and fed into the extruder 13 through a feed line 122, for example through an inlet hopper 123. The extruder 13 has a liquid feed line 124 connected to a water heating element 14 configured to provide heated water (so that the heated water is much higher in temperature than tap water, for example, having a temperature of at least 50°C) and preferably configured to provide water having a stable temperature (for this purpose, the heating element 14 preferably has a pump 132 and a heater tank 133, and the heater tank 133 preferably has a water heating element and a temperature probe). The extruder 13 also contains a long cooling die 125. The pump 132 can be controlled so that the water fed to the tank 131 always has the target temperature, and the pump 130 can feed water to the extruder 13 at the target flow rate (for example, how many kilograms of water per hour). If the tap water is directly connected to the tank 131 and an attempt is made to heat the water in the tank 131, it will be more difficult to accurately control the temperature of the water.

[0131] In the following examples, the experiments carried out by the inventors are described in more detail.

[0132] III: First experiments (Examples 1 and 2)

[0133] In the following, also throughout the description of the composition of the samples in the other experiments and tests, the percentages of the ingredients are given in weight % on a dry basis.

[0134] With Examples 1 and 2, we show exemplary parameters of the manufacturing process (ingredients, impact heating) and their effect on the quality of the resulting meat substitute product (for example in terms of certain physical properties, for example compressibility, firmness, swelling, cavity structure).

[0135] The mechanically processed starch-containing cereal grains comprise or consist of one or more of the following: flakes (for example pressed, rolled or flaked), steel-cut grains, hull-less polished grains, crushed grains, hull-less but unpolished grains.

[0136] Mechanically processed starch-containing cereal grains comprise or consist of one or more of the following: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, haricot beans, peas, quinoa, pigeon peas, sorghum, buckwheat.

[0137] However, the following ingredients were excluded from the mechanically processed starch-containing cereal grains : hulled but unpolished oat grains, hulled but unpolished rye grains, hulled but unpolished barley grains and hulled but unpolished corn grains.

[0138] While other extruder configurations can be used, the extruder 13 used to carry out the experiments was a twin-screw co-rotating extruder with a screw 126 having a diameter of between 30 mm and 50 mm. The extruder 13 had a screw chamber 138 around the screw 126. The screw chamber 138 in the configuration used had 6 zones (although other numbers of zones are possible) which can be numbered from the side where the solid ingredients are fed into the extruder and extrusion starts as zone 1 to zone 6. Thus, there is an inlet aperture 139 for feeding the solid ingredients (for example in zone 1). Zones 2, 3, 4, 5 and 6 are all equipped with heating, cooling and temperature detection elements which preferably allow the temperature of each zone to be individually controlled between, for example, 10 °C and 220 °C. In addition, there is an inlet aperture 140 for feeding liquid into the extruder 13 to be extruded with the solid ingredients (for example in zone 2).

[0139] At typical screw rotation speeds (for example between 150 rpm and 300 rpm), the material can be passed through the screw chamber 138 in about 45 s to 75 s. The inventors set up the liquid feed line 124 and heating elements 14 to feed water at different temperatures from 5 °C to 99 °C, for example, in some cases, heated water was fed into the tank 131 and the heated water was pumped into the extruder 13 by the pump 130 of the liquid feeder. Tests were carried out to stop the extruder and remove the screw after continuously running the extrusion of dry oat flakes without water and it was observed that at a screw grinding time (for example calculated from the conveying distance) of 5-15 s, at about zone 2, the oat flakes were mostly (more than 90%) and essentially powdered into flour-like particles which were significantly smaller than their original size (for example their size was less than 200 μιη).

[0140] Differently, the conventional liquid feed line 124 was connected to regular tap water and tap water at a temperature of between 5 °C and 25 °C was fed into the extruder (as shown in Figure 12A The feed speed (for example kg / h) of the solid ingredients and the liquid could be individually controlled.

[0141] After the last zone (e.g. zone 6), there is a long cooling die 125 connected to the extruder 13, which also has heating, cooling and temperature detection elements. The long cooling die 125 is longer than 300 mm, preferably its length is between 300 mm and 5000 mm, most preferably between 1000 mm and 3000 mm. Between the last zone (e.g. zone 6) and the cooling die 125, there are pressure detection sensors and temperature detection sensors. Furthermore, after the long cooling die 125 a cutter can be connected.

[0142] The person skilled in the art has sufficient knowledge from the background art to know how to adjust or select the screw 126 diameter, the screw 126 speed, the cooling die 125 length and shape, the cutter type and cutting speed depending on different types of custom requirements such as production stability, production speed, product size and shape.

[0143] Example 1 (Sample Nos. 1, 2, 3, 4) - Effect of ingredients on the textural properties of the extruded product.

[0144] The inventors prepared 4 samples (No. 1, No. 2, No. 3, No. 4) which were extruded with the extruder 13 shown in Fig. 1 using high moisture protein texturization. Figure 12B The extruder 13 shown in Fig. 1 was used for extruding the samples with high moisture protein texturization.

[0145] No. 1 sample contains 90 wt% pea protein, 5 wt% oat flour, 4 wt% fiber, wherein further ingredients (e.g. salt, spices, yeast extract, oil, oat malt extract, starch-free cereal grains - e.g. sunflower seeds) are added.

[0146] No. 2 sample contains 90 wt% pea protein, 5 wt% steel-cut oats, 4 wt% fiber, wherein further ingredients (e.g. salt, spices, yeast extract, oil, oat malt extract, starch-free cereal grains - e.g. sunflower seeds) are added.

[0147] No. 3 sample contains 62 wt% pea protein, 20 wt% oat flour, 10 wt% fiber, wherein further ingredients (e.g. salt, spices, yeast extract, oil, oat malt extract, starch-free cereal grains - e.g. sunflower seeds) are added.

[0148] No. 4 sample contains 62 wt% pea protein, 1 wt% steel-cut oats, 19 wt% oat flour, 10 wt% fiber, wherein further ingredients (e.g. salt, spices, yeast extract, oil, oat malt extract, starch-free cereal grains - e.g. sunflower seeds) are added.

[0149] After production, the No. 1, No. 2, No. 3, No. 4 samples were cooled and stored overnight. The next day their mechanical properties were measured to investigate the texture. The measurement results are shown in Table II.

[0150] The results in Table II show that samples 1 and 3, produced from ingredients comprising starch-containing flour (oat flour), have a hard and rubbery texture and high resistance to cylindrical compression.

[0151] The results in Table II also show that samples 2 and 4, in which the starch-containing flour (oat flour) is replaced or partially replaced by starch-containing cereal grains (steel-cut oats), are more flexible and compressible than samples 1 and 3.

[0152] The thickness cooking expansion of sample 2 (265%) is much higher than that of sample 1 (143%) after cooking in water at 110°C in a high pressure cooker (e.g. in a pressure cooker). This difference is caused only by the change in the starch-containing ingredient (from flour to steel-cut grains). Other conditions such as extrusion parameters are kept the same; and the ingredients have the same chemical (nutritional) composition.

[0153] Table II. Texture of samples 1, 2, 3, 4

[0154]

[0155] • As the protein in Example 1 we use pea protein isolate. It can be replaced at least partially by pea protein concentrate, or by any other protein isolate or concentrate (e.g. fava, soy, chickpea, wheat gluten, oat), dairy (milk or whey) protein, or a mixture of at least one of these. The results are comparable.

[0156] • The cereal grains used in Example 1 are steel-cut oats. They can be replaced by any of the above-mentioned (note that the above-mentioned excluded species) Mechanically processed starch-containing cereal grains are excluded) species mentioned above. The results are comparable.

[0157] • In this example the mechanically processed starch-containing cereal grains are soaked in hot water before extrusion. The soaking is performed by gently mixing the grains with hot water (e.g. 90°C) 1:2, and then keeping at a warm temperature (e.g. 75°C) for 2 hours. After soaking the grains absorb all the water and become softer and larger.

[0158] • The flour in Example 1 is oat flour. It can be replaced by barley flour, wheat flour, rice flour, pea flour, chickpea flour, fava flour, lentil flour, etc. and mixtures thereof. The results are comparable.

[0159] • The fiber in Example 1 is pea fiber. It can be replaced with oat fiber, oat bran, potato fiber, fava bean fiber, etc., and mixtures thereof. The results are comparable.

[0160] • The other ingredients in Example 1 include all of the following: salt, spices, yeast extract, oil, oat malt extract, starch-free grains (e.g., sunflower seeds), etc. Some of these can be omitted or replaced with other desired ingredients.

[0161] • The resistance to cutting with a sharp blade is measured as the cutting force in Example 1. The measurement is performed with the texture analyzer described above.

[0162] • The resistance to compression with a cylinder is measured as the compression force in Example 1. The measurement is performed with the texture analyzer described above.

[0163] • The texture observations are analyzed from the texture observation notes by a panel of experts who perform sensory evaluations as in Example 1 in Table II.

[0164] Extrusion parameters used in Example 1:

[0165] (1) Liquid feed: hot water (e.g., with an elevated temperature of 65°C);

[0166] (2) The moisture content of the slurry (the material being extruded) during the extrusion process is about 50%. Depending on the desired properties of the extruded product (e.g., moisture content, color, etc.) and changes in ingredients (e.g., different proteins can have different melting requirements, different starches can have different pasting requirements), the moisture content of the slurry can be adjusted between 40% and 80%;

[0167] (3) Extruder heating profile: Impingement heating profile with temperatures 80-125-160-145-130 (°C) in zones 2-3-4-5-6. Cooling die temperature is 90°C. Depending on changes in ingredients (e.g., different proteins can have different melting temperatures, different starches can have different pasting temperatures), the temperatures can be adjusted within the ranges described in the attached method claims;

[0168] (4) Production rate: about 18 kg of product per hour is produced. Pressure at the end of the screw: 1.0 mPa to 3.0 mPa.

[0169] (5) The extruded product is immediately immersed in water (e.g., 20°C) for 2 hours to cool and prevent drying after extrusion. The extruded product is then removed from the water. The sample is then analyzed for cutting force, compression force, texture observations, and thickness of the cooked expansion rate after 24 hours of storage in a cold room (e.g., 5°C).

[0170] N.A. stands for not analyzed.

[0171] The swelling in Example 1 represents the cooking swelling ratio of the thickness analyzed by the cooking test method, which will be described below. Unless otherwise stated, for example, "extrusion swelling ratio", "swelling" or "swelling ratio" in this application always refers to the cooking swelling ratio of the thickness.

[0172] In further experiments, the ingredients of Sample No. 1 (90% by weight of pea protein + 5% by weight of oat flour + 5% by weight of fiber, to which other ingredients are also added) were processed with different extrusion parameters, such as different liquid feed water temperatures (15°C - 90°C), extruder heating profiles ("impact heating" such as 80°C - 125°C - 160°C - 145°C - 130°C (in zones 2-3-4-5-6), "massive heating" such as 80°C - 125°C - 160°C - 160°C - 160°C, "slow heating" such as 40°C - 75°C - 100°C - 140°C - 165°C), all resulting in unacceptable products (similar to Sample No. 1) which had a hard and rubbery structure and mouthfeel, cutting forces between 500 g and 1100 g, compression forces between 18200 g and 44000 g, and cooking swelling ratios between 125% and 149%. The results of these experiments were not satisfactory. The mouthfeel was not comparable at all to that of a boiled chicken leg.

[0173] Unacceptable results similar to those of Sample No. 1 were also produced by replacing the oat flour with other starch-containing flours, such as oat starch, potato starch, rice starch, chickpea starch, wheat starch, pea starch, etc. The inventors carried out a large number of tests.

[0174] Unacceptable products similar to those of Sample No. 1 were also produced by replacing the oat flour with starch-free grains, such as sunflower seeds, peanut pieces, almond seed pieces, coconut granules, chia seeds.

[0175] Unacceptable products similar to those of Sample No. 1 were also produced by replacing the oat flour with starch-containing grains having an intact shell or an intact, thick and strong seed coat (also known as bran layer) or an intact hull, such as whole grain oat seeds, whole grain barley seeds, whole grain rye seeds.

[0176] However, the addition of 0% to 20% (preferably between 0% and 10%) of these granules (such as sunflower seeds, chia seeds, whole grain oat seeds) to the ingredients to partially replace the protein in an acceptable sample, such as Sample No. 2, does not adversely affect the quality of the extruded product.

[0177] The addition of additives such as calcium chloride, calcium carbonate, gypsum powder (calcium sulphate dihydrate), baking powder, psyllium, alginate, ascorbic acid, xanthan gum, agar agar, etc. to the ingredients of sample No. 1 did not produce the desired properties observed in an acceptable sample (e.g. sample No. 2).

[0178] However, it is still possible to add from 0% to 5% of some of these additives (e.g. baking powder, gypsum powder, ascorbic acid) to the other ingredients of an acceptable sample such as sample No. 2, as it does not have a serious detrimental effect on the quality of the extruded product (compression properties and mouthfeel).

[0179] Example 2 (Sample Nos. 5, 6, 7, 8, 9) - Effect of extrusion ingredients and extrusion heat distribution on the textural and swelling properties of the extruded product. Figure 12B

[0180] The inventors prepared 5 samples (No. 5, No. 6, No. 7, No. 8, No. 9) which were textured extruded with a high moisture protein using an extruder 13 as shown in Figure Figure 1

[0181] Sample No. 5 comprises 70% by weight of pea protein, 30% by weight of oat flour.

[0182] Sample No. 6 comprises 70% by weight of pea protein, 30% by weight of oat flour, like sample No. 5.

[0183] Sample No. 7 comprises 70% by weight of pea protein, 10% by weight of oat flakes, 20% by weight of oat flour.

[0184] Sample No. 8 comprises 70% by weight of pea protein, 10% by weight of oat flakes, 20% by weight of oat flour, like sample No. 7.

[0185] Sample No. 9 comprises 70% by weight of pea protein, 20% by weight of oat flakes, 10% by weight of oat flour.

[0186] Samples No. 5, No. 6, No. 7, No. 8, No. 9 were cooled after production and stored overnight. Their mechanical properties were measured the next day to study the texture. The results of the measurements are shown in Table III.

[0187] Table III. Texture of samples No. 5, No. 6, No. 7, No. 8, No. 9

[0188]

[0189] ​Table III shows that extruded products containing oat flakes (sample 8 and 9) produced with an impulse heating temperature profile (hot water liquid feed used with a temperature profile 80°C-125°C-160°C-145°C-130°C in zones 2-3-4-5-6) have a more flexible and compressible texture which results in a very good mouth feel and is pleasant to eat. After cooking in water, they also have a high cooking expansion (189%-206%) which is consistent with their property of having a flexible and extensible structure and texture.

[0190] When the oat flakes are completely replaced by oat flour having the same chemical composition but a much smaller particle size (sample 6), the extruded product becomes hard, rubbery and has a lower cooking expansion (129%). The mouth feel is completely not comparable to that of the cooked chicken drumstick. The impulse heating extrusion conditions do not lead to a big difference between products containing no oat flakes (between sample 5 and 6).

[0191] When the oat flakes are used in extrusion conditions without impulse heating set (for example if the liquid feed water temperature is 25°C and the zone 2 temperature is set to 40°C), the product (sample 7) has a hard and rubbery texture and a low expansion (164%). The mouth feel is completely not comparable to that of the cooked chicken drumstick.

[0192] • The protein in example 2 is pea protein isolate. It can be replaced by other proteins in the manner explained in the context of example 1.

[0193] • The oat flakes in example 2 are used as mechanically processed starch-containing cereal grains. Other mechanically processed starch-containing cereal grains can be used in place of the oat flakes in the manner explained above and in the context of example 1. In particular, barley flakes, steel cut oats, steel cut barley, rice kernels, broken rice, polished barley, polished rye, polished wheat, etc. and mixtures thereof can be used. The results are comparable.

[0194] • In example 2, the mechanically processed starch-containing cereal grains are not soaked in hot water before extrusion.

[0195] • The flour in example 2 is oat flour. It can be replaced by barley flour, wheat flour, rice flour, pea flour, chickpea flour, lentil flour, quinoa, pigeon pea, sorghum, buckwheat, etc. or mixtures thereof. The results are comparable.

[0196] • The expansion in example 2 represents the cooking expansion rate of the thickness analyzed by the cooking test method which will be described below.

[0197] • The visible air cavities in example 2 represent the visible air cavities in the extruded product analyzed by the visual inspection method which will be described below.

[0198] • The texture observations in Example 2 represent texture property observation notes generated from expert panel sensory evaluation.

[0199] • Extrusion parameters:

[0200] (1) The moisture content of the slurry (material being extruded) during extrusion was about 50%;

[0201] (2) The extruded products were immediately immersed in water (20°C) for 2 hours after extrusion to cool and prevent drying. They were then removed from the water. The samples were then analyzed for texture observations, visible air cavities, thickness, and cooking swell after 24 hours of storage in a cold room (e.g., 5°C).

[0202] (3) Production rate: about 18 kg of product per hour was produced. The cooling die temperature was 90°C.

[0203] Examples of air cavities can be seen in Figure 1 and in Figure 2, sample #8.

[0204] IV: Results of the first experiment

[0205] Figure 1 are photographs of samples #5, #7, and #8 (from bottom to top) after being soaked in 60°C water for 24 hours: on the right, the samples were cut parallel to the fiber direction so that the fibers, length, and thickness of the samples are visible. On the left, the samples were cut across the fiber direction so that the cross-section (width and thickness) of the samples are visible. Sample #8 has significantly more visible air cavities than samples #7 and #5. The air bubbles in sample #8 are more evenly distributed in the protein fiber matrix, with a greater total volume and larger average size than the air bubbles in samples #5 and #7. Figure 1 There are white particulates in sample #7 of Example 2 that are intact oat flake particulates contained within the protein matrix. The contained particulates do not address the problem of the product being rubbery, hard, and difficult to compress. The visible particulates are not pulverized by the extruder, primarily because some very small fraction (e.g., less than 5%) of the particulates slip through the narrow gap between the screw and the screw chamber. They remain mostly intact throughout the extrusion and do not mix effectively with other ingredients. The degree to which these particulates are gelatinized is insufficient and much lower than other particulates that are effectively mixed by the screw (e.g., those that are pulverized in sample #7). At the end of the process, they are covered by other materials. They do not disrupt the formation of the overall protein fiber structure or the formation of inter-fiber interaction forces. These are consistent with the results in Example 1. Figure 2A These are corroborated by the microscopy (although no photographs are provided in this application) and texture (compressibility) study results.

[0206] Figure 2Bis an X-ray microtomography (Micro-CT) scan image taken after sample No. 5 was soaked in water at 60 °C for 24 hours and air dried. The sample was cut parallel to the fiber direction so that the fibers, length, and thickness of the sample are visible.

[0207] Figure 2A is an X-ray microtomography (Micro-CT) scan image taken after sample No. 8 was soaked in water at 60 °C for 24 hours and air dried. The sample was cut parallel to the fiber direction so that the fibers, length, and thickness of the sample are visible. Figure 2A The sample was cut in the same way as in the middle. Figure 2B and Figure 6A The difference between the two is clear, and it can be seen that sample No. 8 has more air bubbles (white inter-fiber black cavities) that are widely and uniformly distributed in the protein fiber matrix, with a larger total volume and larger average size than sample No. 5. In addition, sample No. 8 has a clear structure of long continuous fibers. The fibers of sample No. 8 are thinner and more uniform in thickness than the fibers of sample No. 5. Most of the fibers are parallel to each other. This indicates that although the protein fibers tend to stick to each other and form larger bundles or clumps, the protein fibers in sample No. 8 are well broken up and separated. The thinner fiber structure of sample No. 8 contributes to a good, chewy, and compressible texture that can approach that of a cooked chicken drumstick. The aggregated and layered structure of sample No. 5 gives it an unfavorable, hard, leathery, and rubbery texture.

[0208] Figure 6B is a microscope image of a sample taken from sample No. 2. The sample was stained with a protein dye (Thermo Scientific Pierce Coomassie Brilliant Blue R-250). The sample was observed with an optical microscope (Zeiss Axio Lab. Al Laboratory Microscope) at 10x magnification. The protein fibers are stained black. The protein fibers are continuous throughout the image, with a length much greater than 1 mm. The protein fibers are mostly arranged parallel to each other. Cross-linking is low, with only a few connections between adjacent fibers.

[0209] Figure 6B is a microscope image of a sample taken from sample No. 2. The sample was stained with a dilute iodine solution, such as 1:5 diluted Sigma-Aldrich Lugol’s solution stabilized with polyvinylpyrrolidone, which is used for Gram staining. The sample was observed with an optical microscope at 10x magnification. The dark black material (masses) indicate starch-rich material that forms a dark blue iodine-starch complex with the iodine stain. Figure 6CAlso shown in grey is the protein fiber matrix, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material appears in circular or random shapes, not tightly embedded within the protein fiber matrix, and not evenly distributed throughout the structure. These findings suggest that the starch is in a clustered form, a separate phase from the protein phase, and not emulsified with the protein.

[0210] Figure 6A is a microscope image of a sample taken from Sample No. 2. The sample was stained with a protein dye as in Figure 6D and viewed at 20x magnification. The protein fibers are mostly aligned parallel to each other. Cross-linking is low, with only few connections between adjacent fibers.

[0211] Figure 6D is a microscope image of a sample taken from Sample No. 2. The sample was stained with a dilute iodine solution, such as Sigma-Aldrich Lugol’s solution stabilized with polyvinylpyrrolidone at 1 :5 dilution, used for Gram staining, and viewed at 20x magnification. The dark black material (masses) indicates starch-rich material, which forms a dark blue iodine-starch complex with the iodine stain. Figure 6E Also shown in grey is the protein fiber matrix, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material appears in circular or random shapes, not tightly embedded within the protein fiber matrix, and not evenly distributed throughout the structure. These findings suggest that the starch is in a clustered form, a separate phase from the protein phase, and not emulsified with the protein. There are starch clusters (shown as dark spots) with sizes (e.g., lengths) greater than 30 pm.

[0212] Figure 6E is a microscope image of a sample taken from Sample No. 6. The sample was stained with a protein dye such as Thermo Scientific Pierce Coomassie Brilliant Blue R-250 and viewed at 10x magnification. The protein fibers are stained black. The protein fibers are continuous throughout the image, with lengths much greater than 1 mm. The protein fibers are mostly aligned parallel to each other. Cross-linking is higher: Figure 6A than in Figure 6E . Figure 6A The interstitial spaces between adjacent fibers in Figure 6F are significantly narrower and smaller than in .

[0213] Figure 6Fis a microscope image of a sample taken from sample No. 6. The sample was stained with a dilute iodine solution, such as a 1 :5 dilution of Sigma-Aldrich Lugol's solution stabilized with polyvinylpyrrolidone, as used for Gram staining, and viewed at 10x magnification. The dark black material (blocks) indicates starch-rich material, which forms a deep blue iodine-starch complex with the iodine stain. Figure 6G The protein fiber matrix is also shown in grey, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material appears in narrow lines and is tightly embedded within the protein fiber matrix, and is apparently substantially uniformly distributed throughout the structure between and along the protein fibers, the shape and distribution of the starch-rich material is highly ordered. These indicate emulsification of the starch with the protein.

[0214] Figure 6G is a microscope image of a sample taken from sample No. 6. The sample was stained with a protein dye such as Thermo Scientific Pierce Coomassie Brilliant Blue R-250 and viewed at 20x magnification. The protein fibers are stained black. The protein fibers are mostly arranged parallel to each other. The cross-linking is high: Figure 6C The connections between adjacent fibers in Figure 6G are apparently more abundant than in Figure 6C . Figure 6H The interstitial spaces between adjacent fibers in are apparently narrower and smaller than in

[0215] . Figure 6H is a microscope image of a sample taken from sample No. 6. The sample was stained with a dilute iodine solution, such as a 1 :5 dilution of Sigma-Aldrich Lugol's solution stabilized with polyvinylpyrrolidone, as used for Gram staining, and viewed at 20x magnification. The dark black material (blocks) indicates starch-rich material, which forms a deep blue iodine-starch complex with the iodine stain. Figure 7A The protein fiber matrix is also shown in grey, which is lighter in color than the starch material, more transparent than the starch material, but not completely transparent. The starch-rich material appears in narrow lines and is tightly embedded within the protein fiber matrix, and is apparently substantially uniformly distributed throughout the structure between and along the protein fibers, the shape and distribution of the starch-rich material is highly ordered. These indicate emulsification of the starch with the protein.

[0216] Figure 7A is a microscope image of a sample taken from the washable starch washed out from sample No. 2 at 50°C with water. Figure 7BInsoluble washable starch (black material in image) is shown in the form of clusters, with sizes between 50 pm and 800 pm. Each cluster contains more than 5 individual starch granules (circular). Within each cluster, the individual starch granules are tightly bound to each other. The sample was observed with an optical microscope at 5x magnification.

[0217] Figure 7B is a microscope image taken from a sample of washable starch washed out from Sample 2 with water at 50°C. Figure 10 Insoluble washable starch (black material in image) is shown in the form of clusters, with sizes of about 100 pm. Each cluster contains more than 5 individual starch granules (circular). Within each cluster, the individual starch granules are tightly bound to each other. There is starch leaching out of the cluster of aggregated starch granules into the water. This kind of leached starch makes these clusters "washable" by water at 50°C. Those starches embedded in such clusters are insoluble in water at 50°C, but soluble in water at 110°C. The sample was observed with an optical microscope at 20x magnification.

[0218] Figure 14A Gelation of pea protein affected by heating temperature. To observe how heating temperature can affect pea protein gelation, pea protein was mixed with water in a 1 : 1 ratio, then packed into a vacuum bag, then heated at different temperatures (50°C to 110°C). Then the texture of the gel / blocks was measured. As can be seen from the results in the table, samples heated to 90°C and above have significantly higher hardness. These indicate that significantly stronger gels are formed when heated to 90°C or above.

[0219] Figure 14A Starch coating on the inner surface of the cavity of an extruded product observed by iodine staining and visual inspection. Left: slice of Sample 2. Right: slice of a sample produced under similar conditions as Sample 2, but using hulled but unpolished whole grain oat groats instead of steel-cut oats used in Sample 2. The right sample has an unacceptable texture: e.g. compression force exceeds 20 000 g.

[0220] Two samples were cut into slices of about 1 mm thick, about 10 mm wide, and 40 mm long. The direction of the length is mainly parallel to the direction of the fiber orientation. One slice of each sample was stained with diluted Lugol’s solution (iodine solution for staining) for 45 minutes, the amount of diluted Lugol’s solution was between 1 mL and 3 mL, and could cover the sample in all directions. Then the stained sample was gently moved and immersed in 50 ml of water for 5 minutes. We then placed the slices on white paper for visual observation.

[0221] Figure 14BThe grey matter in the photo refers to the overall structure (protein matrix structure and all other material embedded in the protein matrix structure). The dark colour (black) indicates starch content rich material.

[0222] The slice of sample number 2 (i.e. left side) has a distinct dark coloured coating material on the inner wall of the cavity of the extruded product as well as the outer wall (surface).

[0223] The slice of another sample (i.e. right side) has dark coloured large dots (e.g. 1 mm dots) within the structure. The dark dots should be unbroken oat seeds. The sample contains visible unbroken seeds as inclusion particles, but it has an unacceptable texture.

[0224] No distinct dark coloured coating material was found in number 1, 3, 5, 6 and 7.

[0225] Figure 14C The inner surface of the cavity of the extruded product as observed by iodine staining and microscopy (5x magnification using a stereomicroscope, e.g. Zeiss Stemi 305 stereomicroscope) is shown. The sample specimen was taken from sample number 2. The specimen was stained with diluted Lugol’s solution (iodine solution for staining) for 30 minutes before observation. The grey matter in the photo refers to the overall structure (protein matrix structure and all other material embedded in the protein matrix structure). The dark colour (black) indicates starch content rich material. When observed via the microscope, the colour view is blue or dark blue or black.

[0226] Figure 14D The inner surface of the cavity of the extruded product as observed by microscopy at 20x magnification with iodine staining is shown. The sample specimen was taken from sample number 2. The specimen was stained with diluted Lugol’s solution (iodine solution for staining) for 30 minutes before observation. The dark grey matter in the picture (from left to middle of the picture) with the specific fibrous (anisotropic) structure refers to the overall structure (protein matrix structure and all other material embedded in the protein matrix structure). There are clusters of black dots at the left side of the picture, indicating clusters of gelatinised starch. The light grey matter adjacent to very bright white and empty areas (at the right side of the picture) indicates starch content rich material. The starch at the cavity wall observed with this magnification and angle has a lighter colour than the protein matrix structure, as the cavity wall is more directly exposed to the microscope light. When observed via the microscope, the starch at the cavity wall observed with this magnification and angle is light blue.

[0227] Figure 14E and Figure 15The inner surface of the cavities of the extruded product as observed by iodine staining and examination with a microscope (40x magnification) is shown. The sample specimen was taken from sample no. 2. The sample was stained with diluted Lugol’s solution for 30 minutes before observation. The dark grey matter with specific fibrous (anisotropic) structure in the picture refers to the overall structure (protein matrix structure and all other materials embedded in the protein matrix structure). The light grey matter of the non-fibrous structure adjacent to the very bright white and blank area (middle of the picture) indicates starch content rich material. The starch at the cavity wall as observed with this magnification and angle has a lighter colour than the protein matrix structure. The starch at the cavity wall as observed with this magnification and angle is light blue when observed via the microscope.

[0228] Example 3 (Sample Nos. 10, 11, 12, 13) - Hardening and compressibility of the extruded product affected by extrusion temperature setting are pictures of sample no. 2 (reference 1) before (top picture) and after (bottom two pictures, reference 2) swelling by cooking in water in a pressure cooker for 10 minutes at 110 °C.

[0229] V: Further experiments (examples 3 and 4)

[0230] With examples 3 and 4 we further demonstrate exemplary parameters of the manufacturing process (impact heating) and their influence on the quality of the resulting meat substitute product (e.g. in terms of specific physical properties such as compressibility, firmness, swelling, cavity structure).

[0231] Example 4 (Sample Nos. 14, 15, 16, 17) - Structure and compressibility of the extruded product affected by extrusion temperature setting Extrusion impact heating

[0232] Sample no. 10, 11, 12, 13 comprise 70 wt% pea protein, 5 wt% steel cut oats, 24 wt% oat flour, 1 wt% salt. Sample no. 10, 11, 12, 13 were each processed with a different extrusion temperature setting in extruder 13.

[0233] Table IV shows that when using mechanically processed starch-containing cereal grains (e.g. steel cut oats) in the ingredients, the impact heating temperature setting of the extrusion conditions leads to a good compressibility (compression force 10234 g) and a moderate firmness (129%) of the produced product (sample no. 13).

[0234] However, when the liquid feed water temperature is low (e.g. 25°C, as is commonly used in the extruder 12 known), and / or when the temperature in the extruder is not distributed using impact heating (zone 2 temperature below 100°C, and / or zone 4 temperature below 160°C), the products thus produced (sample no. 10, sample no. 11 and sample no. 12) have more severe hardening problems (186% - 232%) and poor compressibility (compression force 17803g - 20844g). Although they have a lower hardness (lower than sample no. 13) when fresh (5 minutes after extrusion), they have a much higher hardness (higher than sample no. 13) after 5 hours of storage.

[0235] Table IV. Texture of sample no. 10, 11, 12, 13

[0236]

[0237] • The protein in example 3 is pea protein isolate. It can be replaced by other proteins in the manner explained in the context of example 1.

[0238] • Steel cut oats are used in example 3 as the mechanically processed starch-containing grain. Oat flour is used as the flour. Steel cut oats and oat flour can be replaced by other mechanically processed starch-containing grains and flours in the manner explained above and in the context of example 1.

[0239] • In particular, steel cut oats can be replaced by steel cut barley, rice kernels, broken rice, polished barley, polished rye, polished wheat, polished oat, etc. or mixtures thereof. The results are comparable. Oat flour can be replaced by barley flour, wheat flour, rice flour, pea flour, lentil flour, broad bean flour, quinoa, pigeon pea, sorghum, buckwheat, etc. and mixtures thereof. The results are comparable.

[0240] • In this example, the steel cut oats are not immersed in hot water before extrusion.

[0241] • Extrusion parameters:

[0242] (1) The moisture content of the slurry (material being extruded) during extrusion is about 50%;

[0243] (2) Some of the extruded products are immediately immersed in water (e.g. 20°C) for 2 hours to cool and prevent drying. They are then removed from the water. After 24 hours of storage at 5°C, they are analyzed for compression force;

[0244] (3) Some of the extruded products are immediately packaged in closed plastic bags to prevent drying, stored at room temperature, and analyzed for hardness and hardening;

[0245] (4) Extrusion productivity: about 18 kg of product per hour. The cooling die temperature was 90°C.

[0246] • Compression force in Example 3 represents the resistance to compression with a cylinder analyzed by the texture analysis method described above.

[0247] • Texture observations in this example represent texture property observations recorded by the expert panel sensory evaluation analysis.

[0248] • Hardness in this example represents the hardness of the un-soaked extruded product analyzed by a texture analyzer using the cylinder compression method described below.

[0249] • Hardening refers to the hardening rate after 5 hours of storage, which is calculated as:

[0250] Hardening rate = 100% x Hardness (5 hours) / Hardness (5 minutes)

[0251] Hot water as liquid feed Example 5. Starch that can be washed out and dissolved by warm water from the extruded product affected by extrusion conditions.

[0252] The ingredients used for samples 14, 15, 16, 17 were: 90 wt% pea protein isolate, 5 wt% steel cut oats, 4 wt% pea fiber and 1 wt% salt.

[0253] Table V shows that when mechanically processed starch-containing grains (now: steel cut oats) were used in the ingredients, the functional combination (of sample 16) of (a) using Extrusion impact heating temperature settings and (b) using Hot water as liquid feed resulted in a good compressibility of the produced product (compression force 16290 g).

[0254] When the extrusion temperature was changed to a slower heating profile (temperature decrease in zone 4, 130°C; and temperature increase in zone 6, 160°C), the compressibility of the produced product (sample 15) was much worse (26484 g).

[0255] When the extrusion heating temperature was changed to an “excessive” heating profile as in the production of sample 17, where the temperatures of zone 5 and zone 6 were increased (160°C and 160°C), the produced product (sample 17) no longer had the desired continuous or intact structure. Therefore, its compression force was not measurable. And the product did not have the desirable chewiness similar to sample 16. These made it impossible for sample 17 to produce a meat substitute product similar to drumsticks or chicken nuggets.

[0256] When the extrusion temperature becomes a "very slow" heating profile as for sample 14, where zone 2 temperature is below 80°C, zone 4 temperature is below 160°C, and the liquid feed water is cold (25°C), the product produced (sample 14) no longer has the desired continuous or intact structure. So the compression force is not measurable. And the product does not have the desirable chewiness similar to sample 16. These make sample 14 impossible to produce meat substitute products like chicken drumsticks or chicken nuggets like.

[0257] Table V. Texture of sample 14, 15, 16, 17

[0258]

[0259] • The protein in example 4 is pea protein isolate. It can be replaced by other proteins in the manner explained in the context of example 1. The results will be comparable.

[0260] • The same considerations as in example 3 apply for the possibility to replace the steel-cut oats and oat flour.

[0261] • In example 4, the steel-cut oats are not immersed in hot water before extrusion.

[0262] • Extrusion parameters:

[0263] (1) The moisture content of the slurry (material being extruded) during extrusion is about 50%;

[0264] (2) The extruded products are immediately immersed in water (e.g. 20°C) for 2 hours to cool and prevent drying. They are then removed from the water. They are analyzed for compression force after 24 hours of storage at 5°C;

[0265] (3) Extrusion productivity: about 18 kg of product per hour are produced. The cooling die temperature is 90°C.

[0266] • Compression force in this example represents the resistance to compression with a cylinder as analyzed with the texture analysis method described above.

[0267] VI - Advanced experiments (examples 5 and 6)

[0268] With examples 5 and 6, we demonstrate the effect of extrusion conditions and ingredients on the formation of cavities coated with gelatinized starch, which are closer to the mechanism of how these processing methods can lead to quality improvements. Some of the samples used in examples 5 and 6 are the same as in example 1.

[0269] Figure 7A Figure 7B

[0270] Table VI shows that when steel-cut oats are used in the ingredients, Sample No. 13 combines (a) the use of Example 6. Starch that can be washed out and dissolved by warm water from the extruded product affected by ingredients temperature setting and (b) the use of Figure 3 have the effect of increasing the solubility of the starch.

[0271] The presence of soluble starch in the extruded product is caused by the combined effect of (a) mixing the grain with water, (b) heating the grain with water early enough before the starch of the grain is emulsified with the protein matrix.

[0272] During extrusion, soluble starch can cause phase separation between the protein gel and the protein fibers, preventing the formation of a strong, fully isotropic (three-dimensional) cross-linked network structure. Soluble starch also forms a coating material between the interstices of the protein matrix, which later become cavities inside the extruded product. The coating material reinforces the cavities and prevents them from being sealed by protein cross-linking.

[0273] Table VI.

[0274]

[0275] • The ingredients and extrusion parameters used in this example are the same as described in Example 3.

[0276] • Total starch in Example 5 represents the total amount of starch in the extruded product, which can be analyzed by any standard starch analysis method or by the hot water extraction method. The hot water analysis method is described below.

[0277] • Washable starch in Example 5 (g of washable starch per 100 g of product) represents the amount of starch that can be washed out of the extruded product's cut-out slices by 50°C water, which is analyzed by the water wash test. This analysis method is described separately in another paragraph. Figure 3 and Example 7 - Manufacture of meat substitute products in the form of (preferably vegan) thick pieces microscope images of washable starch in

[0278] • Soluble starch in Example 5 (g of soluble starch per 100 g of product) represents the amount of starch that can be dissolved into 50°C water from the extruded product's cut-out slices, which is analyzed by the water dissolution test. This analysis method is described separately in another paragraph.

[0279] • Starch solubility in this example represents the ratio between soluble starch and total starch.

[0280] Starch solubility = 100% x Soluble starch / Total starch

[0281] Figure 8

[0282] Table VII shows that the use of oat flour in the ingredients (sample 1) results in a very low starch solubility (3.4%) and little washable starch (0.08 g / 100 g) of the extruded product. However, when the oat flour is replaced by steel-cut oats having the same chemical composition but a larger size, the resulting product (sample 2) has a much higher starch solubility (8.4%) and more washable starch (0.41 g / 100 g).

[0283] As shown in Table V and as shown in Example 1, sample 2 has a more tender and compressible texture than sample 1. This is attributed to the higher content of soluble starch and washable starch. This is in line with the results of Example 5. Example 8 - Manufacture of meat substitute products in the form of (preferably vegan) small pieces

[0284] Table VII. Analysis of washable starch and soluble starch

[0285]

[0286] • The ingredients and extrusion parameters used in this example are the same as described in Example 1.

[0287] • Washable starch in Example 6 (grams of soluble starch in 100 g of product) represents the amount of starch that can be washed out from a cut slice of the extruded product by 50 °C water.

[0288] Example 6 starch solubility represents the "ratio between soluble starch and total starch".

[0289] Figure 9 A mathematical model is shown in which an exponential curve is fitted to the measurements. It shows that there is a relationship between the starch solubility and the compression force needed to compress a meat substitute product made with high-moisture protein texturized extrusion.

[0290] VII: Manufacturing examples (Examples 8 and 9)

[0291] Method for measuring the cooking expansion rate of thickness

[0292] A meat substitute product in the form of a thick cut (imitating a chicken thick cut) is produced with the following steps. Method for observing visible air cavities in the extruded product: The results are shown in Table VII, which is an example of a food made from a meat substitute product (sample 2) that is shredded into pieces having a size of more than 5 cm length, 1 cm width, 0.8 cm thickness, the pieces are marinated and pan-fried. The food imitates a chicken drumstick thick cut or a meat slice.

[0293] Step 1) Produce a meat substitute product, for example sample 2 or 13.

[0294] ​Step 2) The extruded product is torn into elongated strips (e.g. about 2-4 cm long, 1-3 cm wide, 0.8 cm thick) with the fiber direction aligned with the length direction. The tearing can be done manually, or by a shredder.

[0295] Step 3) The shredded / chopped extruded product is immersed in a marinade (e.g. comprising water, oil, lemon juice, balsamic vinegar, sugar, salt, and other spices) for a suitable time (e.g. 2 hours), preferably immediately after extrusion.

[0296] Step 4) The extruded product is removed from the marinade, preferably it is pan-fried for 2-3 minutes until it is hot and the surface is golden and crispy.

[0297] The extruded product can be frozen or cooled after step 3). Step 4) can be done just before consumption, e.g. at home or at work, or at a restaurant after the product is purchased.

[0298] Method for soluble starch concentration measurement

[0299] Method for analysis of soluble and washable starch from the extruded product An example food made from a meat substitute product (e.g. sample no. 2 or 13) is shown after the extruded product is shredded into pieces having dimensions of preferably more than 3 cm length, 2 cm width, 0.8 cm thickness, the pieces are marinated (left side), the extruded product is battered, the extruded product is breaded and deep-fried in oil (right side). The food mimics chicken nuggets.

[0300] A (preferably vegan) meat substitute product in the form of small pieces can be produced with the following steps:

[0301] Step 1) A meat substitute product is produced, e.g. sample no. 2 or 13. The extruded product is immersed in water or a marinade (e.g. comprising water, oil, lemon juice, balsamic vinegar, sugar, salt, and other spices) for a suitable time (e.g. 24 hours) after extrusion;

[0302] Step 2) The immersed extruded product is cut into a size and shape similar to conventional or typical commercial small pieces (e.g. at least 3 cm long, 2 cm wide, 0.8 cm thick),

[0303] Step 3) A batter is prepared by mixing ingredients, e.g. with a recipe of 40% w / w chickpea flour and 60 w / w water;

[0304] Step 4) The cut extruded product is covered with the batter liquid;

[0305] Step 5) The battered extruded product is covered with a breading ingredient (e.g. a commercial wheat-based deep-fried breading ingredient), breadcrumbs, or with a commercial gluten-free breadcrumb ingredient;

[0306] Step 6) Deep frying the coated extruded product in oil, for example at 170°C, preferably in oil, for a suitable time, for example 3 minutes.

[0307] VIII: Advanced analysis methods

[0308] The following describes analysis methods for analyzing different properties such as compression force, swelling rate, starch solubility.

[0309] Figure 13

[0310] The extruded product is cut into a thick piece by cutting along a direction perpendicular to the direction of the protein fibers (the direction in which the extruded product is removed from the extruder die). The length of the thick piece is equal to the original width of the extruded product. The thickness of the thick piece is equal to the original thickness of the extruded product. The width of the thick piece is 20 mm. The width measurement direction is parallel to the fiber direction.

[0311] The thick piece is placed in a beaker-shaped container. Water is then added to the container to submerge the thick piece. The water and the thick piece are then boiled in a high pressure cooker (pressure cooker) at 110°C for 10 minutes.

[0312] After boiling, the thick piece is removed from the water and placed on a kitchen sieve to drain. The thickness of the thick piece is measured and compared before and after boiling. The swelling rate is calculated as the thickness after boiling divided by the thickness before boiling. The thickness of the thick piece is measured at the center in the length direction of the thick piece. Unless otherwise explicitly stated, for example "extrusion swelling rate", the thickness of the boiling swelling rate is expressed as "swelling" or "swelling rate" throughout this application.

[0313] Swelling rate = 100% x thickness (after boiling) / thickness (before boiling)

[0314] Method for measuring total starch in the extruded product

[0315] The extruded product is cut into a thick piece (thick piece A) by cutting along a direction perpendicular to the direction of the protein fibers (the direction in which the extruded product is removed from the extruder die). The length of the thick piece is equal to the original width of the extruded product. The thickness of the thick piece is equal to the original thickness of the extruded product. The width of the thick piece is 20 mm. The width measurement direction is parallel to the fiber direction.

[0316] The extruded product is cut into a thick piece (thick piece B) by cutting the extruded product to take the middle part (in the middle of the width of the extruded product), so that the thickness of the thick piece is its original thickness, the length in the direction parallel to the fiber direction of the extruded product is 40 mm, and the width in the direction parallel to the width of the extruded product is 20 mm.

[0317] Thick pieces A and B are placed in a beaker-shaped container. Water is then added to the container to submerge the thick pieces. The water and the thick pieces are then heated at 60°C for 24 hours.

[0318] After heating, the slabs were removed from the water and placed on a kitchen sieve to drain. The cut sections (length x thickness) of slab A and slab B were then observed by visual inspection and by taking photographs.

[0319] The slabs were then air dried at room temperature for 7 days. The dried slabs were analyzed by X-ray micro-tomography (Micro-CT) scanning.

[0320] Method for measuring cutting force and compression force

[0321] The method used a modification of [Reference 10] and [Reference 11].

[0322] A solution containing soluble starch (1 mL) was mixed with diluted Lugol’s solution* (1 mL) and water (4 mL). The mixture was hand-shaken for about 10 seconds and then left to stand for 10 minutes. The absorbance of the mixture solution at a wavelength of 600 nm (the wavelength of the light beam used in the spectrophotometric measurement) was then measured**.

[0323] *The diluted Lugol’s solution was prepared by mixing 1 part of Lugol’s solution (synonyms: iodine / potassium iodide solution, aqueous solution of potassium iodide with iodine, iodine concentration between 3% and 10%) or stabilized Lugol’s solution (complex of iodine-polyvinylpyrrolidone (PVP) (homopolymer from 1 -vinyl-2-pyrrolidone) complexed with iodine) with 5 parts of water. One example of the final concentration after dilution: iodine concentration of 0.0100 mol / L and potassium iodide concentration of 0.0260 mol / L.

[0324] **The absorbance was measured by a UV / Vis spectrophotometer (one example of a UV / Vis spectrophotometer can be the UV-1600PC from supplier VWR Collection).

[0325] A standard curve of absorbance and soluble starch concentration was prepared as follows: potato starch (0.05 g, 0.1 g and 0.2 g) was dispersed in 200 mL of cold water by hand-shaking for 1 minute. The dispersion was then boiled twice in a pressure cooker (10 minutes at 110 °C each time, hand-shaking for 1 minute after each boiling when the mixture was still above 60 °C). In this way, the potato starch was completely dissolved in the water. The potato starch dispersion was centrifuged at room temperature at 644 g (g is the unit of RCF = relative centrifugal force). The supernatant was then taken as the starch solution for further analysis. The centrifugation can be performed by a centrifuge used in this study, i.e. a Heraeus Megafuge 8 small benchtop centrifuge, equipped with a rotor of 50 mL conical buckets (supplier’s product number 75005703). TM Megafuge TM 8 small benchtop centrifuge, equipped with a rotor of 50 mL conical buckets (supplier’s product number 75005703).

[0326] Based on the standard curve and the absorbance value at 600 nm wavelength, the concentration of soluble starch in the starch solution can be calculated.

[0327] Reference McGrance (1998) [Ref. 10], "The reaction between starch and iodine has been known for over a century. Over fifty years ago, Rundle and Baldwin proposed that the iodine component of the complex exists in a one-dimensional array within the amylose helix, with six glucose residues per turn. Two important aspects of the colorimetric method using the iodine reaction are its versatility and simplicity. It can be used for starches from a variety of plant sources, and no special equipment is required beyond a simple spectrophotometer capable of measuring absorbance near 600 nm. Samples with high and low amylose content can be analyzed, and only the volume of the aliquot selected needs to be changed for optimal results. The sensitivity of the iodine-starch reaction is quite high." Although the iodine colorimetric method is not used as an official method very often, it is reliable for starch quantification and is known by the person skilled in the art.

[0328] Method for measuring hardness

[0329] The method for extracting and defining soluble starch and washable starch employs a modification of [Ref. 12]. Soluble starch is starch that can be extracted (extracted = washed out) from a product by water at 50°C, passed through a sieve with a pore size of 1200 pm, and is soluble in water. Washable starch is starch and starch-containing material that can be extracted (extracted = washed out) from a product by water at 50°C and passed through a sieve with a pore size of 1200 pm. Soluble starch is part of washable starch, in other words, soluble starch is a synonym for "soluble washable starch". Washable starch comprises soluble washable starch and insoluble washable starch. Insoluble washable starch can be dissolved in water when it is cooked in water above its gelatinization temperature, preferably about 100°C. Soluble ingredients are ingredients in solution that are well dispersed in the liquid and do not precipitate during centrifugation at 644g (g is the unit for RCF = relative centrifugal force).

[0330] Mechanism study 1 The method for analyzing soluble starch and washable starch from the extruded product 61 is illustrated:

[0331] (Step 62) Cutting away the edges (5% of the width) to sample 63 from approximately the middle of the extruded product 62;

[0332] (Step 64) Cut sample 63 into thin slices 65, the thin slices 65 of the extruded product have dimensions of about 1 mm x 10 mm x 40 mm, wherein the length (40 mm) direction of the slices is parallel to the fiber orientation direction of the extruded product;

[0333] (Step 66) Immerse thin slices 65 in water at 50°C for 24 hours, hand shake for 2 minutes;

[0334] (Step 67) Sieve with 1.2 mm aperture;

[0335] Reference number 68 refers to the insoluble washable content in the wash extract;

[0336] (Step 69) Centrifuge at 644 g (RCF) for 30 minutes;

[0337] Reference number 70 refers to the supernatant resulting from the centrifugation, which comprises soluble starch;

[0338] (Step 71) Autoclave at 110°C for 10 minutes, hand shake;

[0339] (Step 72) Centrifuge at 644 g (RCF) for 30 minutes;

[0340] Reference number 73 refers to the supernatant resulting from the centrifugation, which comprises washable starch.

[0341] Measurements were performed on 20 g sliced extrudates, which were soaked (step 66) in 200 mL of water and kept at 50°C for 24 hours.

[0342] g is the unit for RCF = Relative Centrifugal Force.

[0343] Starch solubility of the extruded product = (soluble starch content / total starch content in the extruded product) x 100%

[0344] Starch washability of the extruded product = (washable starch content / total starch content in the extruded product) x 100%

[0345] Figure 4

[0346] The total amount of starch in the extruded product can be analyzed by standard starch analysis methods, such as AACCI method 76-13.01 “Total Starch Determination Procedure” (Megazyme Amyloglucosidase / alpha-amylase method). And it can also be measured by a hot water analysis method, which has the following steps: (1) cut the extruded product into thin slices of about 1 mm 3(1) a 10 mL aliquot of the slurry was added to a 50 mL centrifuge tube; (2) 200 mL of water was added to the centrifuge tube; (3) the centrifuge tube was placed in a boiling water bath at 110 °C for 10 minutes; (4) the centrifuge tube was removed from the boiling water bath and shaken by hand; (5) the centrifuge tube was placed in the boiling water bath for another 10 minutes; (6) the centrifuge tube was removed from the boiling water bath and shaken by hand; (7) the centrifuge tube was centrifuged at 644 g (RCF) for 30 minutes; and (8) the soluble starch concentration of the supernatant was measured. The total amount of starch in the supernatant was equal to the total starch content of the extrudate, which could be calculated from the volume of water and the soluble starch concentration value.

[0347] Figure 4

[0348] For the cutting force measurement, we measured the resistance of the sample in the compression test with a blade. The measurement was performed with a TA.XT Plus texture analyser (Supplier: Stable Micro Systems) equipped with a 294.2 N (30 kg) load cell (detector sensor) and a sharp blade. The knife was of the “double bevel (ground) Scandi” type. The blade of this knife has a total wedge angle of about 16 degrees at the sharpest part (edge), which means that the primary angle of the bevel of the knife is about 8 degrees. The knife has a flat part (ridge) of 0.6 mm thickness above the blade part. The height of the sample was between 7.0 mm and 12.0 mm. The width of the sample was 20 mm. The sample was placed stable and horizontal on the plate and the orientation of the sample was adjusted so that the blade compressed (i.e. cut) in cross direction to the elongated fibres (length direction of the fibres). The downward speed before the blade contacted the fibres was 4 mm / s (pre-test speed). The speed of the compression when the blade contacted the fibres was 20 mm / s (test speed), and the compression was performed until a cutting depth of 90% of the height of the sample was reached. For samples with a height above 9.0 mm, the compression was performed until a cutting depth of 8.0 mm. For this study, the peak force positive (peak force positive is a term used in the software of the equipment and it refers to the maximum force detected during the measurement) was taken as the cutting force.

[0349] For compression force measurements, we measured the resistance of the sample during compression testing with a cylindrical probe (model "P / 36R", 36 mm radius edge cylinder probe - aluminium - AACC standard probe (for bread hardness), supplier Stable Micro Systems). The TA.XT Plus texture analyser was equipped with a 294.2 N (30 kg) load cell (detector sensor) and the cylindrical probe when the measurements were performed. The height of the sample was between 7.0 mm and 12.0 mm. The width and length of the sample was 40 mm. The sample was placed stable and horizontal on the plate and the orientation of the sample was adjusted so that the cylinder was compressed towards the centre of the sample. The downward speed before the blade contacted the fibre was 2 mm / s (pre-test speed). The speed of the compression when the blade contacted the fibre was 0.5 mm / s (test speed) and the compression was performed until a cut depth of 40% of the height of the sample was reached. For this study, the peak force (peak force is a term used in the software of the equipment and it refers to the maximum force detected during the measurement) was taken as the compression force. There was a "trigger force" setting, which was set to 1000 g for this study. The trigger force was set to control the machine (texture analyser) so that when the detected resistance was lower than the trigger force, the probe was not in the position where the sample was contacted by the top surface and the probe was moved downwards with the pre-test speed of 2 mm / s. When the detected resistance was not lower than the trigger force, the probe reached the sample and the probe was moved downwards with the test speed of 0.5 mm / s.

[0350] Mechanism study 2. Comparison of oat flour, oat flakes, steel cut oats and whole oat seeds for their particle size, seed coat, seed structural integrity and starch extractability

[0351] For hardness measurements, we measured the resistance of the sample during compression testing with a cylindrical probe (model "P / 36R", 36 mm radius edge cylinder probe - aluminium - AACC standard probe (for bread hardness), supplier Stable Micro Systems). The TA.XT Plus texture analyser was equipped with a 294.2 N (30 kg) load cell (detector sensor) and the cylindrical probe when the measurements were performed. The height of the sample was between 7.0 mm and 12.0 mm. The width and length of the sample was 40 mm. The sample was placed stable and horizontal on the plate and the orientation of the sample was adjusted so that the cylinder was compressed towards the centre of the sample.

[0352] The test procedure uses the standard TPA measurement protocol (taken from the measurement equipment manual) "Texture Profile Analysis (TPA)" is an objective method of sensory analysis pioneered in 1963 by Szczesniak [Ref. 6] who defined the texture parameters first used in this analysis method. Later in 1978, Bourne [Ref. 7] adapted the Instron to perform TPA by compressing twice a standard sized food sample. TPA is based on the identification of texture as a multi-parameter attribute. For research purposes, it can be desirable to determine texture analysis on several parameters from a small homogeneous sample. The test consists of a reciprocating motion that mimics the action of the jaw compressing twice a bite-sized piece of food, and extracting from the resulting force-time curve a number of texture parameters that are very relevant to those parameters of sensory evaluation [Ref. 8]. The mechanical texture properties of a food that determine to a large extent the choice of the rheological procedure and equipment can be divided into primary parameters of hardness, cohesiveness, springiness and adhesiveness, and secondary (or derived) parameters of fracturability (brittleness), chewiness and gumminess [Ref. 9].

[0353] The downward speed before the blade contacts the fiber is 5 mm / s (pre-test speed). When the blade contacts the fiber, the speed of compression is 2 mm / s (test speed), and the compression proceeds until a cut depth of 30% of the sample height is reached. For this study, the peak force positive (peak force positive is a term used in the equipment software that refers to the maximum force detected during the measurement) is taken as the compression force. There is a "trigger force" setting, which in this study was set to 5000 g. The waiting time between the first compression and the second compression is 1 second. The hardness is calculated by the software of the measurement equipment. Hardness is equal to the peak force positive during the first compression.

[0354] IX: Advanced mechanism study

[0355] Mechanism study 1 shows the effect of the processing methods (ingredients, impact heating) on the properties of the test extrusion (extrusion without cooling die) material (particle size distribution), which reveals the mechanisms of how these processing methods affect the extruded product. This can also be used as an evaluation method to select processing parameters.

[0356] Further mechanism studies show the relevant knowledge of the differences between the properties of the grains and the flour, between the grains processed with cold water and the grains processed with warm water.

[0357] Mechanism study 3: Effect of soaking of steel cut oats on its mechanical properties - Effect of ingredients and extrusion temperature profile on the weight distribution of the particles

[0358] To study the influence of ingredients and extrusion temperature on the results, the inventors performed a large number of further experiments. Table VIII lists the ingredients and the tested extrusion parameters. A test extrusion means that during these test periods no dies were installed in the extruder, but only the ingredients were processed through the screw running in the heating chamber. A summary of the results and findings can be found in Table IX. Figure 5 The measured particle weight distribution of the extruded material influenced by the composition of the ingredients and the extrusion heating temperature profile is shown for experiments 1 to 6.

[0359] Table VIII. Sample preparation for test extrusion

[0360]

[0361] Oat flakes were used as the mechanically processed starch-containing cereal grains in experiments 2 and 3. In experiments 4, 5 and 6, steel-cut oats were used. The steel-cut oats were not soaked prior to the test extrusion.

[0362] The test extrusion did not form thick lumps with a long continuous fibrous matrix. Instead, the material produced was an agglomerate with different sizes (hence the weight per particle ranged from 0.1 g to 10 g). The agglomerates (i.e. the particles) were divided into different size (weight) groups (small, medium, large, etc.), then each size group was weighed and the percentage of it relative to the total weight of the produced agglomerate was calculated. Figure 5 The particle weight distribution curves are shown in the middle.

[0363] Table IX: Results and findings of the test extrusion

[0364]

[0365] The comparison should mainly be made between samples with the same chemical composition (protein content, starch content, etc.), for example between experiment 1, experiment 2 and experiment 3, or between experiment 4, experiment 5 and experiment 6, respectively.

[0366] Furthermore, there is a similarity between experiment 1 and experiment 4, which have parameters that can produce a product with good compressibility and flexibility. They both produce medium-sized particles (0.5 g - 4 g) with a percentage between 26% - 30%; large particles (> 4 g) with a percentage between 0% - 5%.

[0367] XI - Summary Figure 11

[0368] The results in Table X show that the starch extractability in water (9-26 g / 100 g) of oat flakes, steel cut oats, and whole grain oat seeds is much lower than that of oat flour (40 g / 100 g) due to the better integrity of the seed structure and seed coat. The starch extractability of whole grain oat seeds is very low (9 g / 100 g) due to the intact seed coat of the whole grain oat seeds.

[0369] Steel cut oats can absorb more water and faster when the water is hot (375%, 110°C, 10 minutes) than when the water temperature is lower (136%, 50°C, 12 hours). These explain why impact heating and soaking in hot water can change the behavior and effect of oat flakes, steel cut oats in high moisture extrusion. Hot water can allow the starch-containing kernels to absorb water faster and more completely, become gelatinized and more solubilized.

[0370] The functionality / substitutability of whole grain oat seeds is not as good as that of oat flakes and steel cut oats in the examples disclosed above. At the time of writing, the inventors are still testing other treatments to enable whole grain oat seeds to function. For example, boiling in excess water.

[0371] Table X: Oat-based starting materials, starch extractability in water

[0372]

[0373] To measure extractable starch, 10 g of starting material was boiled in 100 g of water in a pressure cooker for 10 minutes, and the boiled mixture was centrifuged at 644 g (RCF) for 30 minutes. The soluble starch concentration of the supernatant. Extractable starch was calculated as:

[0374] Extractable starch = 100% x soluble starch in supernatant / weight of starting material

[0375] To measure water uptake at 50°C, 20 g of starting material was immersed in 200 g of water, then held at 50°C for 24 hours of soaking, then sieved to remove water that was not absorbed by the material. The weight of the material before and after 24 hours of soaking was recorded.

[0376] Water uptake = 100% x (weight after soaking - weight before soaking) / weight before soaking

[0377] To measure water uptake at 50°C, 20 g of starting material was added to 200 g of water, then boiled in the water at 110°C in a pressure cooker for 10 minutes, then sieved to remove water that was not absorbed by the material. The weight of the material before and after boiling was recorded.

[0378] Water uptake = 100% x (weight after boiling - weight before boiling) / weight before soaking

[0379] Steel-cut oats having different sizes can be produced, ranging in size from 6 mm 3 to 15 mm 3 per particle. Those steel-cut oats having 8 mm 3 per particle were used in Mechanism Study 2.

[0380] Example

[0381] The inventors investigated the effect of soaking steel-cut oats. Example 9 (Sample Nos. 6, 18, GER 19) - Effect of germination treated oats on the textural properties of the extruded product. The results of the compression test of dry (unsoaked) steel-cut oats versus soaked steel-cut oats (soaked in hot water) are shown in Table;

[0382] As can be seen from Figure 12B the steel-cut oats without soaking water were significantly more brittle and less compressible than the steel-cut oats soaked in hot water. When the compression rate reached 27% (0.47 mm deep compression of 1.78 mm thick steel-cut oats), the steel-cut oats without soaking had already cracked and split. On the other hand, the steel-cut oats soaked in hot water (80°C, 2 hours) became soft, sticky and pasty. The soaked steel-cut oats did not crack or split throughout the compression (0%-90% compression during the test).

[0383] This reveals that starch-containing grains can be split into smaller pieces by a compression force, which is sufficient during extrusion.

[0384] Treating starch-containing grains with hot water can soften the grains and help prevent the grains from being split into smaller pieces by compression or extrusion.

[0385] Table XI: Effect of soaking of steel-cut oats on its mechanical properties

[0386]

[0387] As an overview of the comparison of the soluble starch content, the washable starch content, the starch solubility and the starch washability properties when the protein content is the same, the inventors summarized and classified the results and calculated the change in these values. In Table XII, S1, S3, S4, S5 and S6 have the same ingredients and extrusion conditions as in Sample No. 1, Sample No. 2, Sample No. 6, Sample No. 11 and Sample No. 13, respectively. S2 has the same ingredients as Sample No. 2, but it has different extrusion conditions. In S2, the steel-cut oats were not soaked in hot water before extrusion, and impact heating was achieved by using a hot water (60°C) liquid feed and an extruder temperature profile of 100-125-160-145-130 (°C) at zones 2-3-4-5-6.

[0388] Table XII shows that S2 has 52% higher starch solubility and 63% higher starch washability than SI. These differences are attributed to impact heating and ingredient differences (e.g. use of steel cut oats). S3, which uses steel cut oats, soaking and impact heating, has even higher starch solubility and starch washability. The impact of ingredients (e.g. use of steel cut oats) and impact heating is even greater when the pea protein content is reduced from 90% to 70%. S6 has 261% higher starch solubility and 58% higher starch washability than S4. Due to the differences in impact heating, S5 does not have as high starch solubility and starch washability as S6.

[0389] Table XII: Effect of extrusion conditions and ingredients on soluble starch content, washable starch content, starch solubility and starch washability properties

[0390]

[0391] X: Conclusions

[0392] The inventors have surprisingly found that starch added in the form of a starch-containing powder or flour can indeed cause individual parts of the protein matrix to stick together to form even larger pieces or more complete structures during extrusion with or without a long cooling die.

[0393] The extruded products produced with the addition of starch-containing powder also have much higher isotropic properties and much lower anisotropic properties (anisotropic fiber structure, anisotropic texture).

[0394] The inventors have also found that small particle size starch can emulsify into and / or between the protein fibers, become a filler material in the protein-based emulsion gel-like system, and be able to improve the uniformity and coverage (area, space, volume) of the protein material distribution. As a result, the proteins can form more isotropic interactions with each other throughout the extrusion process. Starch gelation can also bind different parts of the material to be connected to each other.

[0395] The inventors have also found that when a long cooling die is used in extrusion, the addition of such materials with higher content of starch-containing powder can form thicker, denser and more isotropic chunks with a specific fiber structure. When no cooling die is used in extrusion, the addition of such materials with higher content of starch-containing powder can form larger extruded product connection groups (pieces) without a fiber structure.

[0396] The inventors have also found that, as described in the appended method claims, the problem of protein matrix hardening can be prevented or at least further delayed when starch-containing grains are added to the protein material and extruded.

[0397] Without wishing to be bound by any theory and taking into account the very limited amount of knowledge in the field, the inventors found and there is a possible explanation that when the starch-containing kernels have a larger particle size than conventional starch-containing powders, the starch-containing kernels break up into smaller parts at a much slower rate. Furthermore, the broken kernel parts are not easily emulsified by the protein matrix. The broken kernel parts can still be gelatinized by sufficient heat, shear and water. Furthermore, the naturally occurring kernel cell wall structure and material can limit the full leaching, alignment and retrogradation of the starch molecules.

[0398] The naturally occurring kernel cell wall structure and the gel effect of gelatinized starch can also prevent the kernels from being fully powdered into small particles (e.g. particle size below 100 pm). As a result, a large number of gelatinized starch clusters are formed and remain residual throughout the extrusion process and in the final product.

[0399] The inventors surprisingly found that when the extruded product is cut into thin slices but not necessarily fully breaking the protein fibers, at least some of these clusters can be washed out of the extruded product by warm water (50°C) without the need for further gelatinization of the starch. These starch clusters have a much larger particle size than the starch in conventional processes, which are individualized in the protein matrix and emulsified in the conventional processes. These starch clusters are typically larger than 100 pm in at least one of their dimensions. Therefore, these starch clusters can behave like large particles, which separate the protein fibers far apart from each other, preventing the formation of hydrogen bond type protein-protein interactions and texture hardening.

[0400] The large starch clusters as large particles also often result in the formation of holes (cavities) or empty spaces next to them. This can be due to the flow behavior of the extruded material and the protein fiber strength during the extrusion process, allowing the protein fibers to flow far apart from each other after encountering the large particle barrier formed by the starch clusters. Then, after a period of continuous flow far apart from each other, the bundles of protein material (protein fibers) come close to each other and form interactions again. During this period of protein flow far apart from each other, empty spaces are formed behind the large particle of the starch clusters. The protein fibers separated by the empty spaces cannot form hydrogen bonds. The inventors believe that this can contribute to improved mouthfeel that lasts longer, even in cooled or frozen meat substitute products.

[0401] Furthermore, the inventors have found that the earlier the starch in the starch-containing grain is gelatinized before it is emulsified by the protein matrix, the higher the concentration of the gelatinized starch clusters, which can prevent the formation of a continuous protein matrix to a greater extent. Without wishing to be bound by any theory, the inventors have one explanation that the gelatinized starch clusters that are not emulsified by the protein matrix are immiscible with the protein phase, and thus can separate the phase from the protein phase, and thus can form a considerable connected phase, and can disrupt the protein-protein interactions to form, so they can prevent the formation of a continuous protein fiber matrix to some extent. This explanation is very consistent with the test results in the mechanism study experiments in selected examples, which will be described below. The observed differences between the number of samples tested by the inventors also seem to support this explanation.

[0402] After the formation of the gelatinized starch clusters, the melting, cross-linking and gelation of the protein material should be initiated within a certain short time window. If this happens too late, there will be two types of unacceptable results, namely (1) the gelatinized starch clusters are eventually homogenized, broken up and emulsified by the protein matrix, which is particularly likely when the starch-containing grain is added in small amounts, or the starch-containing grain is relatively easy to break up, and the starch powder content in the ingredient is high; (2) the gelatinized starch clusters completely prevent the formation of long continuous protein fiber structures by excessively separating and covering the protein material into individual clusters, and prevent protein-protein coagulation, aggregation and gelation, which is particularly likely when the starch-containing grain is added in large amounts, and the starch powder content in the ingredient is low.

[0403] In addition, the inventors have found that when the starch-containing grain is not soaked in hot water or mixed with hot water in the extruder at a very early stage (e.g. between 0 seconds and 15 seconds, preferably between 1 second and 15 seconds, after feeding into the extruder), the starch-containing grain is more easily ground into powder in the extruder. In this way, the starch-containing grain behaves similarly to its flour, which has the same chemical composition but smaller particle size and broken cell wall structure.

[0404] On the contrary, the starch-containing grain soaked in hot water before extrusion and the starch-containing grain mixed with hot water in the extruder at a very early stage (e.g. between 0 seconds and 15 seconds, preferably between 1 second and 15 seconds, after feeding into the extruder), will be less brittle, more extensible, and thus more resistant to emulsification by the protein matrix, and more easily remain as large particles throughout the extrusion. Therefore, this is part of the reason for the importance and necessity of using extrusion conditions with impact heating settings while using starch-containing grain in the ingredients for extrusion, in order to produce an extruded product of acceptable quality.

[0405] The inventors have also surprisingly found that meat substitute products manufactured using high moisture protein texturized extrusion can have significantly higher levels of extrusion expansion shortly after the extruded product exits the extruder long cooling die when produced using the method described in the appended method claim.

[0406] At one second after the extruded product exits the extruder long cooling die during extrusion, the high extrusion expansion is clearly visible, the extruded product has air bubbles significantly within the interior of the expanded structure, and its thickness is much greater (e.g., 200-600% greater) than its original thickness (which is approximately the same as the height of the opening of the extruder long cooling die) immediately prior to exiting the extruder long cooling die. After the extruded product cools, the expanded structure can collapse substantially. However, there are still a significant number of cavity (in other words, air pocket) structural units remaining in the cooled extruded product. This difference can be attributed to the formation of clusters of gelatinized starch that are not emulsified by the protein matrix, which results from the method described in the appended method claim.

[0407] Gelatinized starch can cause greater expansion during high moisture extrusion. The increase in expansion can be attributed to a decrease in structural firmness and a decrease in viscosity of the extruded material.

[0408] In contrast, such extrusion expansion phenomena are essentially non-existent, or in other words, undetectable, in processing methods of such tests that do not use starch-containing grains or do not have an impact heat set in the extrusion conditions. It is found that extruded products produced by processing methods that are unable to produce products with textures close to cooked chicken drumstick meat tend to have a more dense and compact structure (a thickness at one second after exiting the extruder long cooling die that is 0% to 199% greater than the thickness immediately prior to exiting the extruder long cooling die), and retain significantly fewer cavity structural units (in other words, air pockets) after cooking. During high moisture extrusion, starch-containing flours can cause a higher amount of leached starch, a higher water uptake, and a higher increase in viscosity compared to starch-containing grains. This is found to be consistent with observations during the extrusion tests, and with mechanism studies experiments of cooking starch-containing materials in water in a pressure cooker.

[0409] The inventors have surprisingly found that for extruded products produced using the method described in the appended method claim, when the extruded product is cut into thin slices but not necessarily completely breaking the protein fibers, there are more starch molecules that can be dissolved out of the extruded product by warm water (50°C). The 50°C temperature is below the gelatinization temperature of starch. Generally, native (non-gelatinized) starch is not soluble in 50°C water. Pre-gelatinized starch and some modified starches can be soluble in 50°C water before they are extruded in high moisture protein texturized extrusion for meat substitute production, but they lose solubility after the extrusion process because they are quickly emulsified with the protein matrix after being extruded with the protein material.

[0410] The starch dissolved in the extruded products described herein and below is soluble washable starch, which is a portion of the washable starch. Soluble starch (soluble washable starch) is more completely gelatinized, more leached from the starch granule shell and the grain cell wall structure (without being limited thereto), has a higher affinity for water, and their molecules have a more swollen structure (e.g., volume and surface area) than insoluble washable starch. Soluble starch has even less affinity for the protein matrix and is even less tightly embedded in or captured by the long continuous protein fiber structure. Soluble starch is more immiscible with the protein and so it is more completely separated from the protein phase by phase separation. Soluble starch compounds are the primary component of the coating of the inner walls of the cavities (pockets) of the acceptable extruded products. Soluble starch compounds are the primary component and the primary site of the extrusion expansion and the creation of the cavities. The coating material of the inner walls of the cavities in the acceptable quality extruded products can be seen by visual and microscopic observation after staining with a dilute iodine solution. After staining, the coating material turns a deep blue or black, which indicates a high concentration of starch. The cavities coated with the gelatinized starch clusters also act as novel disruptive compounds that prevent the formation of protein-protein interactions (e.g., hydrogen bonds) between the protein fibers after extrusion. The cavities coated with the gelatinized starch clusters are different from and perform better than other known disruptive particles, such as starch, flour, insoluble salt, dietary fiber, apparently because the starch clusters push the protein fibers away from each other with a volume that is larger than the size of the individual particles.

[0411] There is no background art teaching about the role and influence of soluble starch, washable starch, insoluble washable starch, starch solubility, starch washability in the production of meat substitute products with long continuous protein fiber structure by high moisture protein texturized extrusion or in low moisture protein texturized extrusion. There can be some research about starch solubility in starch extrusion processes which mainly process the starch component of starchy foods and have a very different configuration than protein texturized extrusion. However, starch solubility is highly correlated with crumb staling and texture quality. For example, Boyacioglu and D'Appolonia [Reference 5] reported that crumb staling (storage, aging) over 4 days can result in a continuous, progressive and significant decrease in starch solubility while the firmness value continues to increase significantly; recommended the use of soluble starch content to measure the rate and extent of staling since a decrease in soluble starch content indicates an increase in crumb staling and firming; the higher the amount of soluble starch in the crumb sample, the lower the rate of increase in firmness value. In crumb, a decrease in starch solubility indicates an increase in the rate of starch molecule retrogradation. Starch retrogradation is a well-known factor that usually results in a leathery mouthfeel and hard texture of starch-containing foods such as bread. Starch retrogradation occurs fastest at temperatures slightly above freezing (e.g. between 0°C and 6°C). Starch retrogradation is caused in part by recrystallization of starch amylose and amylopectin molecules and is a result of an increase in starch-starch hydrogen bonding and a decrease in starch-water affinity. It is possible but not obvious to make a correlation between the knowledge about starch solubility behavior in meat substitute products produced by high moisture protein texturized extrusion and the knowledge about crumb. Meat substitute products produced by high moisture protein texturized extrusion have a completely different ingredient formulation, structure and microstructure than crumb. The process and structure formation mechanism of protein texturized extrusion and bread baking are also completely different.

[0412] The inventors surprisingly found that meat substitute products manufactured with high moisture protein texturized extrusion and having low starch solubility and low starch washability, their starch is mostly homogenized and emulsified uniformly with the protein matrix. With microscopic observation, it was found that the emulsified starch in the products is linearly aligned so that the starch particles are parallel to each other. The protein fibers tightly cover and trap the starch compounds. The starch compounds are completely leached out. The original starch granule structure has essentially disappeared. Therefore, the starch can be subjected to severe retrogradation. These findings are consistent with the results that those samples have low starch solubility, more severe hardening during 5 hours storage, worse compressibility after overnight storage and worse ability to swell by cooking in water in a pressure cooker. In contrast, meat substitute products having a relatively high starch solubility and starch washability were found to have better texture properties (good compressibility, good swelling properties, mouthfeel close to chicken drumstick).

[0413] Starch solubility and starch washability are even more important than soluble starch content and washable starch content. Starch solubility and starch washability are calculated as the proportion of soluble starch content and washable starch content relative to the total amount of starch in the extruded product. Soluble starch and washable starch positively contribute to the quality (e.g. mouthfeel) of the extruded product. In contrast, a higher percentage and higher amount of insoluble starch and non-washable starch can lead to a poorer quality (e.g. mouthfeel) of the extruded product, because insoluble starch and non-washable starch are relatively more completely emulsified, trapped, embedded in the protein matrix, and have more retrogradation.

[0414] In view of this background art and the inventors' new findings, there is reason to believe in the importance of monitoring and controlling the levels of soluble starch content, washable starch content, starch solubility and starch washability in meat substitute products manufactured with high-moisture protein texturization extrusion.

[0415] A method of controlling and improving starch solubility and starch washability in meat substitute products produced by high-moisture protein texturization extrusion cannot be found in the background art, but is disclosed in the following description.

[0416] The inventors found that when a meat substitute product manufactured in an extruder configured to perform high-moisture protein texturization extrusion comprises a continuous protein fiber matrix structure, which is substantially linearly oriented and has disruptions forming cavities, wherein the walls of the cavities are at least partially coated by a cluster of gelatinized starch, then the mouthfeel tends to remain acceptable for a long time.

[0417] It is known that a decrease in starch solubility (e.g. in water at 50°C) and an increase in starch retrogradation are important factors in causing the texture of food (e.g. bread crumbs comprising starch gel structures) to become firm. See references (a) SOHO CH, T. J.; FRENCH, D. 1947. Studies on bread staling. 1. The role of starch. Cereal Chemistry, 24: 231-249; (b) T. Inagaki and P. A. 1992. Firming of Bread Crumb with Cross-Linked Waxy Barley Starch Substituted for Wheat Starch. Cereal Chem 69: 321-325; (c) K. Ghiasi, R. C. Hoseney, and D. R. Lineback. 1979. Characterization of Soluble Starch from Bread Crumb. Cereal Chem 56: 485-490.

[0418] Alternatively or additionally, the pasted starch clusters include starch that does not emulsify with the protein fiber matrix structure (unemulsified starch). The resulting advantages are: (1) an increase in the percentage of unemulsified starch results in a decrease in the percentage of emulsified starch. Unemulsified starch does not behave like a filler that fills the gaps between protein fibers and strengthens the overall extrudate structure, but emulsified starch does; (2) unemulsified starch is less ordered (more disordered or less molecular) than emulsified starch, and thus has less and / or delayed starch retrogradation, and improved softness at temperatures above freezing (e.g., between 0°C and 6°C) over long storage times; (3) unemulsified starch disrupts the ordering of the protein fiber matrix structure, and thus improves its softness at temperatures above freezing (e.g., between 0°C and 6°C) over long storage times by reducing and / or delaying the formation of intermolecular (e.g., protein-protein, starch-starch) hydrogen bonds in the extrudate.

[0419] Alternatively or additionally, a high-moisture protein texturized extrusion process can be used to make a meat substitute product in which starch-containing grain is pasted, and protein that forms the protein matrix is melted:

[0420] (a) before an emulsion is formed between the pasted starch-containing grain and the protein that forms the protein matrix; and

[0421] (b) before the pasted starch forms an impenetrable barrier that prevents the formation of a continuous protein fiber cross-linked matrix. The resulting advantages are that in this way the extrusion material is controlled in a good balance between (a) sufficient protein-protein cross-linking to form continuous protein fibers; and (b) prevention of cross-linking formation by the pasted starch. As a result, the extrudate can have a chewiness (cutting force above 300 g) in a particular threshold range, and at the same time have a compressibility (compression force below 17500 g) in a particular threshold range. If protein melting is not achieved before an emulsion is formed between the pasted starch-containing grain and the protein material, emulsification can still be achieved by continuous shearing, tearing, and homogenization of the protein-starch mixture, then the starch becomes emulsified, and cannot prevent an increase in undesirable interaction forces (e.g., hydrogen bonds) and hardening of the extrudate (e.g., compression force becomes above 17500 g). On the other hand, if protein melting is not achieved before the pasted starch forms an impenetrable barrier that prevents the formation of a continuous protein fiber cross-linked matrix, there will be a lack of protein-protein cross-linking. As a result, the chewiness will be too low and not in the threshold range (cutting force above 300 g).

[0422] The extrusion step can be performed with an extrusion die having a length of more than 300 mm, preferably more than 1000 mm. An advantage resulting therefrom is that such dies are typical set-ups for performing high moisture protein texturizing extrusion. This die allows the extruder to process extrusion cooking of material having a moisture content of more than 40% to form a texturized (cross-linked) structure before the material leaves the extruder. This die also allows the molten protein material to arrange into a long continuous fiber structure.

[0423] Preferably, the heating step d) is performed preferably between 140°C and 200°C. An advantage resulting therefrom is that this temperature allows the protein to melt, denature, form a gel and form the protein-protein cross-links needed to form a long continuous fiber structure.

[0424] Preferably, the mechanically processed starch-containing cereal grains comprise or consist of one or more of the following: oat, barley, rye, wheat, rice, corn, lentil, chickpea, mung bean, haricot bean, pea, quinoa, pigeon pea, sorghum, buckwheat. An advantage resulting therefrom is that these cereal grains are commercially available, contain a high amount of starch, are known to be palatable and nutritious, and are widely used in different other food applications.

[0425] Alternatively or in addition, the heating step d) is performed such that protein melting occurs between 1 s and 40 s, preferably between 10 s and 30 s after step b). An advantage resulting therefrom is that in this way the protein forming the protein matrix is melted:

[0426] (a) before the emulsion of the gelatinized starch-containing cereal grains and the protein of the protein matrix is formed; and

[0427] (b) before the gelatinized starch forms an intact barrier preventing the cross-linking of the continuous protein fiber matrix.

[0428] The time needed for the extruder to break the cereal grains (e.g. rolled oats, steel cut oats, rice) into a powder was observed in the test.

[0429] Alternatively or in addition, the heating step c) is performed such that starch gelatinization occurs between 0 s and 18 s, preferably between 1 s and 15 s. An advantage resulting therefrom is that in this way the starch-containing cereal grains can preferably be milled by the extruder screw to a particle volume of less than 5000 pm 3 before and preferably before the starch-containing cereal grains are milled by the extruder screw to a particle volume of less than 0.001 mm 3 before, the heating step c) is performed. The particle volume is more than 5000 pm 3The gelatinized starch clusters of the present application are non-emulsified starches, larger than those emulsified starches, and can provide a much greater disruptive force to prevent the formation of too much protein-protein interaction forces, and thus can prevent hardening of the extrudate during storage.

[0430] Preferably, after heating step d), the extrusion mixture is continued for more than 5 s, preferably more than 10 s, at a temperature not higher than the temperature of heating step c), preferably between 90 °C and the temperature of heating step d). The resulting advantage is that a heating level like this can lead to a good balance between (a) sufficient formation of protein-protein cross-link structures (forces) to provide an acceptable chewiness (cutting force higher than 300 g); and (b) having an acceptable compressibility (compression force lower than 17500 g). Higher temperatures can lead to too much cross-link formation, and thus poor compressibility. Temperatures lower than 90 °C can lead to too weak structures lacking well-aligned long-fiber structures and poor chewiness.

[0431] Table XIII

[0432] To improve the mouthfeel of meat substitute products, improvements have been invented to the meat substitute products and the high-moisture protein texturized extrusion. The inventors found that by proper selection of the extrusion parameters and the starting material comprising mechanically processed starch-containing cereal grains, the formation of an emulsion between starch and the protein matrix forming the protein melt can be prevented or reduced to such an extent that there is a substantial presence of unbound starch in the protein matrix. The presence of unbound starch in the protein matrix was observed to improve the mouthfeel and maintain an acceptable mouthfeel for a long time. This patent application contains a number of independent claims for meat substitute products and methods.

[0433] Summary of previous work

[0434] The specific examples disclosed in the section “Previous work” above are disclosed in co-pending international application PCT / EP2019 / 068926, which is not published at the time of writing, planned to be published on January 21, 2021, under publication number WO 2021 / 008680 Al, the content of which is incorporated herein by reference. Summary:

[0435] The mouthfeel of meat substitute products manufactured with high-moisture protein texturized extrusion can be improved such that the improved mouthfeel is comparable to boiled chicken drumstick meat, and the improved mouthfeel is further maintained for a longer time, for example overnight or 24 hours, without the need to freeze the meat substitute product.

[0436] The mouthfeel can be said to be comparable to boiled chicken drumstick when the linear compressibility of the sample is relatively high and the cylindrical compressibility is relatively low. The linear compressibility is preferably between 300 g and 1500 g when measured with a texture analyser model TA.XT Plus of Stable Micro Systems, Salisbury, UK, equipped with a 294.2 N (30 kg) load cell (detector sensor) and sharp blade. The cylindrical compressibility is preferably between 7000 g and 17500 g when measured with a texture analyser TA.XT Plus of Stable Micro Systems, equipped with a 294.2 N (30 kg) load cell (detector sensor) and a cylindrical probe (model "P / 36R", 36 mm radius edge cylinder probe - aluminium - AACC standard probe for bread firmness). For the measurements, samples with a height between 7.0 mm and 12.0 mm should be used. The width and length of the sample are preferably 40 mm. Conclusions: A cutting force and compression force analysis method is shown which preferably should be used.

[0437] Alternatively, the mouthfeel of the meat substitute product can be said to be comparable to boiled chicken drumstick when a panel of test persons identifies the experienced compressibility and chew characteristics as being similar to boiled chicken drumstick.

[0438] Furthermore, the starch solubility in a meat substitute product manufactured with high moisture protein texturization extrusion can be increased.

[0439] The starch solubility in a meat substitute product manufactured with high moisture protein texturization extrusion can be controlled.

[0440] A meat substitute product manufactured with high moisture protein texturization extrusion and comprising an extrudate having a continuous protein fiber matrix structure with a substantially linear orientation, the extrudate comprising starch, at least 5.1 %, preferably at least 5.2 % of the starch being soluble starch, said meat substitute product showing a long lasting improved mouthfeel.

[0441] Respectively, a meat substitute product showing a long lasting improved mouthfeel can be manufactured by using a manufacturing method with an extruder configured to perform high moisture protein texturization extrusion, wherein starch containing cereal grains are gelatinized and the protein of the protein matrix is melted, such meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch, at least 5.1 %, preferably at least 5.2 % of the starch being soluble starch.

[0442] The soluble starch is preferably located at the breakage of the matrix structure and is not emulsified therewith. Most preferably, some of the breakage in the matrix structure is in the form of cavities, the walls of said cavities being at least partially coated with gelatinized starch clusters, the gelatinized starch clusters being formed from starch, preferably from soluble starch.

[0443] According to a second aspect, which is alternative to or in addition to the previous aspect, a meat substitute product manufactured with high moisture protein texturization extrusion and comprising an extrudate having a continuous protein fiber matrix structure with a substantially linear orientation, the extrudate comprising starch, such that in the extrudate,

[0444] i) when the protein content of the extrudate is greater than 55 wt% but less than 70 wt%, at least 10.5% of the starch is washable starch,

[0445] ii) when the protein content of the extrudate is at least 70 wt% but less than 90 wt%, at least 15% of the starch is washable starch,

[0446] iii) when the protein content of the extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of the starch is washable starch,

[0447] wherein the wt% indicated is based on dry basis,

[0448] showing a long lasting improved mouthfeel.

[0449] Respectively, a meat substitute product showing a long lasting improved mouthfeel can be manufactured by a manufacturing method using an extruder configured to perform high moisture protein texturization extrusion, wherein starch-containing cereal grains are gelatinized and protein melt forming a protein matrix, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch, such that in the extrudate,

[0450] i) when the protein content of the extrudate is greater than 55 wt% but less than 70 wt%, at least 10.5% of the starch is washable starch,

[0451] ii) when the protein content of the extrudate is at least 70 wt% but less than 90 wt%, at least 15% of the starch is washable starch,

[0452] iii) when the protein content of the extrudate is at least 90 wt% but equal to or less than 99 wt%, at least 16% of the starch is washable starch,

[0453] wherein the wt% indicated is based on dry basis.

[0454] Preferably, the washable starch is located in disruptions of the matrix structure and is not emulsified therewith. Most preferably, some of the disruptions in the matrix structure are in the form of cavities, the walls of which are at least partially coated by gelatinized starch clusters formed by the washable starch. The washable starch is washable in water at a temperature of 50 °C, which is lower than the gelatinization temperature of starch.

[0455] According to a third aspect, which is alternative to or in addition to the first and second aspects, a meat substitute product manufactured with high moisture protein texturization extrusion and comprising an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch,

[0456] and wherein the extrudate is manufactured using a high moisture protein texturization extrusion method in which starch-containing cereal grains are gelatinized and protein forming the protein matrix is melted such that:

[0457] the starch-containing cereal grains are gelatinized before they are substantially pulverized by the extruder screw.

[0458] Respectively, a meat substitute product showing long-lasting improved mouthfeel can be manufactured by using a manufacturing method of an extruder configured to perform high moisture protein texturization extrusion, in which a meat substitute product is produced that is an extrudate having a continuous protein fiber matrix structure, starch-containing cereal grains are gelatinized and protein forming the protein matrix is melted, the extrudate comprising starch, wherein: the step of heating the slurry in the extruder is performed with heating in such a way that the starch-containing cereal grains are gelatinized before they are substantially pulverized by the extruder screw.

[0459] The manufacturing method of the meat substitute product increases the starch solubility, and accordingly, the meat substitute product has increased starch solubility.

[0460] According to a fourth aspect, which is alternative to or in addition to the first, second and third aspects, a meat substitute product manufactured with high moisture protein texturization extrusion and comprising an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the extrudate comprising starch, and wherein: the extrudate is manufactured using a high moisture protein texturization extrusion method in which starch-containing cereal grains are gelatinized and protein forming the protein matrix is melted such that:

[0461] the protein is melted:

[0462] (a) before the starch-containing cereal grains are gelatinized and the protein forming the protein matrix is melted to form an emulsion,

[0463] and

[0464] (b) before the starch is gelatinized to form a complete barrier preventing the formation of a continuous protein fiber cross-linked matrix,

[0465] showing long-lasting improved mouthfeel.

[0466] Respectively, a meat substitute product exhibiting long-lasting improved mouthfeel manufactured by a manufacturing method using an extruder configured to perform high-moisture protein texturization in which starch-containing cereal is gelatinized and protein forming a protein matrix is fused, the meat substitute product being an extrudate having a continuous protein fiber matrix structure, the extrudate comprising starch, such that the protein forming the protein matrix is fused:

[0467] (a) before the gelatinized starch-containing cereal is emulsified with the protein forming the protein matrix,

[0468] and

[0469] (b) before the gelatinized starch forms a complete barrier preventing the formation of a continuous protein fiber cross-linked matrix.

[0470] The manufacturing method of the meat substitute product enables control of starch solubility, and accordingly, the meat substitute product can have a controllable starch solubility.

[0471] According to a fifth aspect, alternatively to or in addition to the first, second, third, and fourth aspects, a meat substitute product manufactured with high-moisture protein texturization, and comprising:

[0472] an extrudate having a substantially linearly oriented continuous protein fiber matrix structure, the

[0473] extrudate comprising starch, the starch being located at disruptions of the matrix structure without being emulsified therewith; exhibiting long-lasting improved mouthfeel.

[0474] Respectively, a meat substitute product exhibiting long-lasting improved mouthfeel manufactured by a manufacturing method using an extruder configured to perform high-moisture protein texturization in which starch-containing cereal is gelatinized and protein forming a protein matrix is fused, the meat substitute product being an extrudate having a continuous protein fiber matrix structure,

[0475] the extrudate comprising starch, the starch being located at disruptions of the matrix structure without being emulsified therewith.

[0476] The manufacturing method of the meat substitute product increases starch solubility, and accordingly, the meat substitute product has an increased starch solubility.

[0477] Particularly advantageously, some of the disruptions in the matrix structure can be in the form of cavities, the walls of the cavities being at least partially coated by gelatinized starch clusters formed from starch, preferably from soluble starch or leachable starch.

[0478] In particular, the advantages resulting from the fifth aspect are that the disruption sites and in particular the cavities at least partially, preferably completely, coated with starch clusters (and phase-separated starch clusters) prevent hardening of the extrusion (resulting from increased gel firmness). The disruption sites formed by the cavities at least partially coated with starch clusters (and phase-separated starch clusters) act as novel disruptive compounds preventing further protein-protein interactions between the protein fibers after extrusion. They differ from and are superior to other disruptive microparticles known to the inventors, such as starch, flour, insoluble salt, dietary fiber, pregelatinized starch, gas, which (a) disappear after extrusion (e.g. gas), or (b) are emulsified by the protein matrix during extrusion (e.g. insoluble salt, dietary fiber, flour, starch), or (c) become a factor accelerating or worsening the deterioration (hardening) of the extrudate (e.g. the retrogradation of starch, which is the rearrangement of amylose and amylopectin molecules and the resulting recrystallization, which often leads to a leathery mouthfeel and a hard texture of starch-containing foods (e.g. bread). These phenomena occur fastest at air temperatures slightly above freezing).

[0479] According to a sixth aspect, which is alternative to or in addition to the first, second, third, fourth and fifth aspects, a meat substitute product showing a long-lasting improved mouthfeel can be manufactured by the following manufacturing method:

[0480] a) feeding a mixture into an extruder configured to perform a high-moisture protein texturization extrusion, the mixture comprising:

[0481] a1) at least one protein matrix-forming ingredient, such as an isolate protein or a concentrate protein, and

[0482] a2) a mechanically processed starch-containing cereal grain having a microparticle volume of at least 0.125 mm 3 , preferably at least 1 mm 3 , most preferably at least 6 mm 3 ;

[0483] b) feeding water into the extruder;

[0484] c) heating the mixture in the extruder to gelatinize the starch-containing cereal grain;

[0485] d) further heating the mixture in the extruder to melt the at least one protein matrix-forming ingredient after starch gelatinization has been achieved; and

[0486] e) extruding the mixture through an extrusion die at a temperature of 70°C to 100°C

[0487] wherein:

[0488] i) heating step c) is performed with impact heating such that the starch-containing grains are gelatinized before they are substantially pulverized by the extruder screw;

[0489] and

[0490] ii) heating step d) is performed with impact heating such that the protein melting temperature of the protein matrix-forming ingredient is reached:

[0491] (a) before the gelatinized starch and protein matrix-forming ingredient form an emulsion, and

[0492] (b) before the gelatinized starch forms a complete barrier to the formation of a continuous protein fiber cross-linked matrix.

[0493] "Particle volume" and "per particle volume" are terms that describe the size of a particle. They can be calculated based on the outer dimensions of the particle, for example:

[0494] - when the particles are mostly close to cuboid in shape, their particle volume can be calculated as length times width times thickness;

[0495] - when the particles are close to spherical, the particle volume can be calculated using the diameter of the particle. For example, the Dv0.5 value in a conventional particle size distribution analysis method can be used to calculate the average value of the particle size (diameter).

[0496] The particle volume is at least 0.125 mm 3 The average volume of the particles is 0.125 mm 3 . A typical commercial oat flour has a particle size diameter of less than 0.300 mm as measured by sieving, from which it can be calculated that the average particle volume is no more than 0.014 mm 3 .

[0497] Conventionally, in high-moisture protein texturizing extrusion, a heating temperature profile is used that gradually increases in temperature from the material feed side of the extruder to the other end of the screw chamber, as protein melting is expected to occur at the end of the extruder, where the ingredients gradually absorb heat and increase in temperature. Under the current impact heating concept, the material in the extruder to be heated to the target temperature is heated significantly faster, preferably within a few seconds after they are fed into the extruder, that is, before they are transported to the last part of the screw chamber of the extruder.

[0498] Preferably, the elevated temperature water is fed to the starch-containing grains. The specific heat capacity of water is about 220% higher than that of protein powders and flours. Therefore, feeding elevated temperature water can heat the material in the extruder to reach the target temperature in a significantly shorter time.

[0499] Preferably, the starch-containing grain is treated prior to feeding into the extruder such that the starch is gelatinized prior to feeding into the extruder, in this way the size of the grain (volume of the particle) is kept at least the same or even increased.

[0500] The inventors observed a permanent consistency of the five first aspects in the samples studied, having an improved mouthfeel. Furthermore, the object of the application can be solved according to the method of the sixth aspect.

[0501] Common to the meat substitute product and method according to any of the aspects is that the extrudate is an extrudate manufactured using a high moisture protein texturizing extrusion method, preferably a twin-screw extruder with a long cooling die (the length of the cooling die is preferably more than 300 mm, most preferably more than 1000 mm). In the extrusion, mechanically processed starch-containing grain is processed together with at least one of the isolated protein / concentrated protein / combination thereof, oil, and spices to manufacture a slurry, which is subsequently extruded.

[0502] The term "mechanically processed" refers to flakes - e.g. compressed, rolled or flaked steel cut grains, dehulled polished grains, crushed grains, or dehulled but not polished grains, but excludes: dehulled but not polished oat grains, dehulled but not polished rye grains, dehulled but not polished barley grains, dehulled but not polished corn grains.

[0503] The mechanically processed starch-containing grain preferably comprises or consists of one or more of the following: oat, barley, rye, wheat, rice, corn, lentil, chickpea, mung bean, fava bean, pea, quinoa, pigeon pea, sorghum, buckwheat, but excludes: dehulled but not polished oat grains, dehulled but not polished rye grains, dehulled but not polished barley grains, dehulled but not polished corn grains.

[0504] Preferably, the meat substitute product is further processed such that it can be sold in the form of a thick slab, a mince, a chicken nugget, a meat slice, a meat steak, or a turkey roast-like slice, or in the form of a layered stratified layer of a yogurt or a vegan yogurt and spices in the form of a turkey roast skewer.

[0505] The use of insoluble washable starch in the form of clusters in food products can open interesting possibilities for the food industry.

[0506] The inventors observed using a microscope equipped with polarized light that the starch in the extruded product does not have the "Maltese cross" characteristic that the starch has before being extruded or soaked in hot water. This indicates that the starch in the extruded product is gelatinized.

[0507] The protein fiber matrix structure of the minced extruded product remains insoluble and unbroken after testing with the starch washability test. The protein fiber matrix structure of the meat substitute product also remains insoluble and unbroken after cooking in a pressure cooker in water at 110°C for 10 min. The cutting force of the pressure cooked meat substitute product remains between 40% to 50% of the cutting force before pressure cooking. These are important differences in performance of products produced by other extrusion methods than high moisture protein texturized extrusion. Products produced by other extrusion methods generally substantially dissolve, soften or collapse after cooking in water or soaking in warm water overnight.

[0508] According to yet another aspect, the method of manufacturing a meat substitute product by high moisture protein texturized extrusion can be improved by selecting the extrusion parameters and the starting material containing at least i) a protein ingredient, preferably an isolated protein or a concentrated protein or a mixture thereof; ii) a mechanically processed starch-containing cereal grain and iii) a flour, such that the formation of emulsions between starch and the protein matrix forming protein melt is substantially prevented or reduced to an extent that the amount of starch not adhering to the protein matrix is present in the meat substitute product after extrusion.

[0509] The controlled extrusion parameters preferably include the water feed temperature and / or the heat distribution, for example along the extrusion screw and in the cooling die, such that an impact heating of the starting material in the extruder is obtained.

[0510] Advantageously, the hardness or compressibility of the meat substitute product is controlled by controlling the starch solubility in the meat substitute product. Most advantageously, the starch solubility is controlled such that the linear compressibility is between 300 g and 1500 g and the cylindrical compressibility is between 7000 g and 17500 g. Preferably, the linear compressibility and the cylindrical compressibility are measured at least 24 hours after extrusion.

[0511] Advantageously, the amount of starch not adhering to the protein matrix is determined as soluble starch. Preferably, the compressibility is controlled by changing the extrusion parameters such that the proportion of the amount of soluble starch in relation to the total amount of starch (starch solubility) in the meat substitute product after extrusion is between 3 wt% and 10 wt%. In this case, the soluble starch content in the meat substitute product after extrusion is between 0.03 wt% and 1.10 wt%.

[0512] Current work - detailed description

[0513] The inventors of the present patent application continue their previous work. An important finding is that malted grain, germinated grain, malt or any combination of two or three of them can be used as one of the ingredients in the extrusion to manufacture a meat substitute food product.

[0514] Furthermore, the inventors have found that the methods and meat substitute products published in previous work can be made using starch-containing grains selected in this way. The inventors suspect that the theoretical conclusions drawn from previous work that explain the effects also apply to malted grains, germinated grains, malt or any combination of two or three, respectively.

[0515] Germinated grains are commercially available and are mainly used in breweries. They are also used in the production of bakeries and cookies, cereal bars and confectionery. Compared to dry whole grains, germinated grains have the advantage of a softer kernel structure, provide new flavors, increase the nutritional profile, and reduce the content of anti-nutritional compounds.

[0516] Germination is a natural process that starts with the uptake of water from a viable dry seed and ends with the elongation of the embryo axis. After imbibition, the seed rapidly resumes metabolic activity. The biochemical composition of the grain changes dramatically. Enzymes degrade storage macromolecules, such as starch, proteins to some extent. During germination, the kernel structure is softer and new compounds are produced [Ref. 13]. Some of the new compounds are flavor precursors that contribute to the formation of the delicious malt flavor. Almost all nutrients (e.g. phenolic compounds, phytosterols, folates, and GABA) become fully available, while anti-nutritional compounds (e.g. phytate, trypsin inhibitors, tannins) are substantially reduced [Ref. 14].

[0517] The traditional malting process is a controlled germination process for brewing purposes and food applications. It comprises three steps: steeping, germination, and kilning. During steeping, the kernel’s moisture content is increased to start germination. During germination, conditions are strictly controlled to allow enzyme synthesis and kernel modification to occur. The kilning step is to dry the kernel so that biochemical reactions stop or slow down; aroma and flavor compounds are produced; and it is microbiologically stable. After the kilning step, the germinated oat produces a roasted aroma and flavor as well as a sweet taste.

[0518] Although some starch will be degraded by the degrading enzymes synthesized during the germination process, even after the malting process, germinated grains contain functional and / or unaltered starch. Compared to other common germinated grains (e.g. barley, rye, wheat, rice, corn, lentil, chickpea, mung bean, fava bean, pea, quinoa, pigeon pea, sorghum, buckwheat), the germination of oats is much more limited due to the strongest resistance of oat starch granules to degradation by a-amylolytic enzymes. Therefore, germinated oats are the best choice.

[0519] In addition to germinated grains, malted grains and malt can also be used individually or in any combination.

[0520] The inventors tested this invention and found that germinated oats have a similar effect to steel-cut oats. The germinated oats are preferably hulled, meaning the outer shell has been removed. We found that during the extrusion process, the hard and difficult-to-break outer shell contributes to an unpleasant texture in the final product.

[0521] In addition to germinated grains, malt or sprouted grains can be used. Alternatively, soaked grains can be used. The grains can be whole grains or mechanically processed germinated starchy grains.

[0522] Starchy grains used for soaking / germination / malting / sprouting preferably contain one or more of the following or are composed of one or more of the following: oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, and buckwheat.

[0523] In addition, flour and / or bran and / or starch and / or fiber may be used. Preferably, the flour is derived from oats, barley, rye, wheat, rice, corn, lentils, chickpeas, mung beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat, potatoes, sweet potatoes, lupins, or any mixture thereof. Preferably, the bran is oat bran, barley bran, wheat bran, rice bran, rye bran, corn bran, millet bran, or any mixture thereof. Preferably, the starch is derived from oat starch, barley starch, rye starch, wheat starch, rice starch, corn starch, lentil starch, chickpea starch, mung bean starch, broad bean starch, pea starch, quinoa starch, pigeon pea starch, sorghum starch, buckwheat starch, potato starch, sweet potato starch, lotus root starch, or any mixture thereof. Preferably, the fiber is derived from oat fiber, barley fiber, rye fiber, wheat fiber, rice fiber, corn fiber, lentil fiber, chickpea fiber, mung bean fiber, broad bean fiber, pea fiber, quinoa fiber, pigeon pea fiber, sorghum fiber, buckwheat fiber, potato fiber, sweet potato fiber, lupin fiber, apple fiber, or any mixture thereof.

[0524] ​

[0525] ​ ​

[0526] The inventor prepared to use ​ Three samples (6, 18, and GER19) were processed by extruder 13 for high-moisture protein tissue extrusion.

[0527] Sample 6 was made with the same recipe as Sample 6, containing 70% by weight pea protein and 30% by weight oat flour.

[0528] Sample 18 comprises 70 wt% of pea protein, 20 wt% of oat flour, 10 wt% of steel cut oats.

[0529] Sample GER19 comprises 70 wt% of pea protein, 20 wt% of oat flour, 10 wt% of sprouted oats.

[0530] The mechanical properties of sample 6, 18, GER19 were measured a) within 5 min after extrusion; b) after cooling and storage in a sealed bag for 5h.

[0531] The results are shown in Table XIII.

[0532] ​ . Texture of sample 6, 18, 19 GER.

[0533]

[0534] The results of Table XIII show that sample 6, produced with ingredients comprising a starch-containing flour (oat flour), has a hard and rubbery texture and a high resistance to cylindrical compression.

[0535] The results of Table XIII further show that sample 18 and sample GER19 are more flexible and compressible than sample 6, for which the starch-containing flour (oat flour) part is replaced by a starch-containing grain (by steel cut oats in sample 18, by sprouted oats in sample GER19).

[0536] Sample GER19 has a better flavor than sample 6 and sample 18, for example, with good improvements in sweet, nutty and slightly toasted flavors.

[0537] • The protein in Example 9 is pea protein isolate. It can be replaced by other proteins in the manner explained in the context of Example 1.

[0538] • Steel cut oats are used in Example 9 as mechanically processed starch- containing grains. Steel cut oats can be replaced by other mechanically processed starch-containing grains in the manner explained above and in the context of Example 1. In particular, barley flakes, oat flakes, steel cut barley, rice kernels, broken rice, polished barley, polished rye, polished wheat, etc. and mixtures thereof can be used. The results are comparable.

[0539] • Sprouted oats are used in Example 9 as sprouted grains. Sprouted oats can be replaced in the manner explained above. In particular, barley, rye, wheat, rice, corn, lentils, chickpeas, green beans, broad beans, peas, quinoa, pigeon peas, sorghum, buckwheat. The results are comparable.

[0540] • In Example 9, the mechanically processed starch-containing grain and the malted starch-containing grain were not soaked in hot water prior to extrusion.

[0541] • The flour in Example 9 was oat flour. Barley flour, wheat flour, rice flour, pea flour, chickpea flour, lentil flour, quinoa, pigeon pea, sorghum, buckwheat, potato, sweet potato, lupin, etc. or mixtures thereof can be used instead. The results are comparable.

[0542] • Extrusion parameters:

[0543] (1) The moisture content of the slurry (material being extruded) during extrusion was about 50%;

[0544] (2) The compressibility of the extruded product a) within 5 min after extrusion; b) after 5 h storage in a sealed bag was measured;

[0545] (3) The production rate: about 18 kg product per hour.

[0546] • The cooling die temperature was 99°C.

[0547] ​

[0548] The inventors found that the use of malted starch-containing grain in extrusion has similar effects as the use of mechanically processed starch-containing grain, resulting in the prevention or delay of hardening of the protein matrix. No benefit was found when using dry whole grains without a malting process. The inventors have a possible explanation that malted grains have a softer core and can be broken into smaller parts more easily than dry whole grains without malting. The average particle size is larger than conventional starch-containing powders. The broken grain parts are less prone to be emulsified by the protein matrix. Under sufficient heat, shear and water action, the broken grain parts can still gelatinize. In addition, the naturally occurring grain cell wall structure and material can limit the complete leaching, alignment and resynthesis of starch molecules. Furthermore, the starch-containing malted grains, when cooked together with other ingredients in a high-moisture extrusion process, outperform mechanically processed starch-containing grains and whole grains in terms of flavor modification, such as sweet, nutty and toasty flavors.

[0549] CONCLUSION

[0550] It is apparent to a person skilled in the art that, as technology advances, the basic idea of the application can be implemented in various ways. The application and its embodiments are thus not limited to the examples described above; instead they can vary within the content of the claims and their legal equivalents.

[0551] In the claims of the present application and in the preceding description, the word "comprise" and variations thereof such as "comprise" or "comprising", will be understood to imply the inclusion of a stated feature or step or group of features or steps but not the exclusion of any other feature or step or group of features or steps that can be insubstantially non-essential to the operation of the application.

[0552] Reference list of publications:

[0553] [Reference 1] Tolstoguzov, V. B. (1993), Thermoplastic extrusion - the mechanism of the formation of extrudate structure and properties. J Am Oil Chem Soc, 70: 417-424. doi: 10.1007 / BF02552717

[0554] [Reference 2] Akdogan, H. (1999), High moisture food extrusion. International Journal of Food Science & Technology, 34: 195-207. doi: 10.1046 / j.1365-2621.1999.00256.x

[0555] [Reference 3] Lin, S., Huff, H. and Hsieh, F. (2000), Texture and Chemical Characteristics of Soy Protein Meat Analog Extruded at High Moisture. Journal of Food Science, 65: 264-269. doi: 10.1111 / j.1365-2621.2000.tb15991.x

[0556] [Reference 4] Xiang Dong Sun, Susan D. Arntfield. (2010) Gelation properties of salt-extracted pea protein induced by heat treatment. Food Research International. Volume 43, Issue 2, 2010, Pages 509-515.

[0557] [Reference 5]MHBoyacioglu and BLD'Appolonia.(1994)Characterization and utilization of durum wheat for breadmaking III.Staling properties ofbread baked from bread wheat flours and durum wheat flours.CerealChemistry.71:34-41

[0558] [Reference 6] SZCZESNIAK, AS (1963). Classification of textural characteristics. J. Food Sci, 28, 385-389.

[0559] [Reference 7] BOURNE, MC (1978).Texture Profile Analysis.Food Technol.,32(7),62-66,72.

[0560] [Reference 8] BOURNE, MC (1988).Basic Principles of Food TextureMeasurement.Lecture text of Dough Rheology and Baked Products TextureWorkshop-Chicago.

[0561] [Reference 9] SZCZESNIAK, AS(1966).Texture Measurements.Food Technol.,20,50,55-58.)

[0562] [Reference 10] McGrance, SJ, Cornell, HJ and Rix, CJ (1998), A Simple and Rapid Colorimetric Method for the Determination of Amylose in Starch Products. Starch / 50:158-163.doi:10.1002 / (SICI)1521-379X(199804)50:4<158::AID-STAR158>3.0.CO;2-7.

[0563] [Reference 11] Adedeji, O.E., Oyinloye, O.D., & Ocheme, O.B. (2014). Effects of germination time on the functional properties of maize flour and the degree of gelatinization of its cookies. African Journal of Food Science, 8(1), 42-47.

[0564] [Reference 12] Azarfar, A., Williams, B.A., Boer, H. and Tamminga, S. (2007) In vitro gas production profile and the formation of end products from non-washable, insoluble washable and soluble washable fractions in some concentrate ingredients. Journal of the Science of Food and Agriculture. 87: 1345-1355

[0565] [Reference 13] A Kaukovirta-Norja, A Wilhelmson, K Poutanen. (2004) Germination: a means to improve the functionality of oat. Journal of the Agricultural and Food Science. 12: 100-112.

[0566] [Reference 14] Paolo Benincasa et al., Sprouted Grains: A Comprehensive Review, Nutrients 2019, 11, 421; https: / / doi.org / 10.3390 / nu11020421.

Claims

1. A method of manufacturing a meat substitute food product, wherein: i. at least one protein matrix forming ingredient, ii. starch-containing grain selected from the group consisting of: ii.a) malted grain, ii.b) germinated grain, ii.c) malt, ii.d) sprouted grain, or ii.e) any combination of two, three or four of these starch-containing grains as one of the ingredients, and iii. water or aqueous liquid a. is fed to an extruder suitable for high moisture protein texturizing extrusion; and b. is extruded in the extruder under conditions resulting in a continuous protein fiber matrix structure comprising disruptions, the extrudate comprising starch located at the disruptions and not emulsified with the protein fiber matrix structure.

2. The method of claim 1, wherein: The extrusion is performed in a high moisture protein texturizing extrusion method in which starch-containing grain is gelatinized and the protein forming the protein matrix is melted.

3. The method of claim 1 or 2, wherein: In the extrusion, the starch-containing grain is gelatinized before they are substantially pulverized by the extruder screw to produce a substantially linearly oriented continuous protein fiber matrix structure with some starch not emulsified with the protein fiber matrix structure.

4. The method of claim 1 or 2, wherein: In the extrusion, the starch-containing grain is gelatinized and the protein forming the protein matrix is melted: i. before an emulsion is formed of the gelatinized starch-containing grain and the protein of the protein matrix, and / or ii. before the gelatinized starch forms a complete barrier preventing the formation of a continuous protein fiber cross-linked matrix.

5. The method of claim 1 or 2, wherein: In the extrudate after the extrusion i. at least 10.5% of the starch is washable starch when the protein content of the extrudate is greater than 55% but less than 70% by weight, ii. at least 15% of the starch is washable starch when the protein content of the extrudate is at least 70% but less than 90% by weight, iii. at least 16% of the starch is washable starch when the protein content of the extrudate is at least 90% but equal to or less than 99% by weight, wherein the % by weight is based on a dry basis.

6. The method of claim 1 or 2, wherein: The protein matrix structure comprises disruptions, wherein some of the disruptions are in the form of cavities, the walls of the cavities being at least partially coated by clusters of gelatinized starch, the clusters of gelatinized starch being formed from starch.

7. The method of claim 1 or 2, wherein: The at least one protein matrix forming ingredient comprises at least one isolate protein and / or at least one concentrate protein.

8. The method of claim 1 or 2, wherein: The starch-containing grain has an average cell volume or median cell volume of at least 0.125 mm 3 .

9. The method according to claim 1 or 2, wherein in the method the extrusion is performed: a. water or aqueous liquid is fed to the extruder; b. the mixture is heated in the extruder to gelatinize the starch-containing grain; c. after starch gelatinization is achieved, the mixture is further heated in the extruder to melt the at least one protein matrix forming ingredient; d. the mixture is extruded through an extrusion die at a temperature of 70°C to 100°C.

10. The method according to claim 9, wherein: e. heating step b. is performed with impact heating such that the starch-containing grain is gelatinized before they are substantially pulverized by the extruder screw; and / or f. the heating step c. is performed with impact heating such that the melting temperature of the protein of the protein matrix forming ingredient is reached; and / or g. after the heating step c. the extrusion of the mixture is continued at a temperature not higher than in the heating step c.

11. The method of claim 1 or 2, wherein: The starch-containing grain is treated before being fed into the extruder such that the starch is at least partially gelatinized before being fed into the extruder.

12. The method of claim 1 or 2, wherein: Water or an aqueous liquid is fed into the extruder at elevated temperature.

13. The method of claim 1 or 2, wherein: The starch-containing grain is selected from, comprises or consists of one or more of the following: oat, barley, rye, wheat, rice, corn, lentil, chickpea, green gram, horse bean, pea, quinoa, pigeon pea, sorghum, buckwheat.

14. The method of claim 1 or 2, wherein: The starch-containing grain consists of whole grains or comprises whole grains.

15. The method of claim 1 or 2, wherein: The starch-containing grain consists of mechanically processed starch-containing grains or comprises mechanically processed starch-containing grains.

16. The method according to claim 1 or 2, wherein: In addition to i. at least one protein matrix forming ingredient; ii. a starch-containing grain selected from the group consisting of: ii.a) steeped grain, ii.b) germinated grain, ii.c) malt, ii.d) sprouted grain, or ii.e) any combination of two, three or four of these starch-containing grains as one of the ingredients, and iii. water or an aqueous liquid Further iv. flour and / or bran and / or starch and / or fiber are used.

17. The method according to claim 16, wherein: - the flour comprises, consists of or is selected from at least one of the following: oat, barley, rye, wheat, rice, corn, lentil, chickpea, green gram, horse bean, pea, quinoa, pigeon pea, sorghum, buckwheat, potato, sweet potato, lupin, any mixture thereof; and / or - the bran comprises, consists of or is selected from at least one of the following: oat bran, barley bran, wheat bran, rice bran, rye bran, corn bran, millet bran, any mixture thereof; and / or - the starch comprises, consists of or is selected from at least one of the following: oat starch, barley starch, rye starch, wheat starch, rice starch, corn starch, lentil starch, chickpea starch, green gram starch, horse bean starch, pea starch, quinoa starch, pigeon pea starch, sorghum starch, buckwheat starch, potato starch, sweet potato starch, lotus root starch, any mixture thereof; and / or - the fiber comprises, consists of or is selected from at least one of the following: oat fiber, barley fiber, rye fiber, wheat fiber, rice fiber, corn fiber, lentil fiber, chickpea fiber, green gram fiber, horse bean fiber, pea fiber, quinoa fiber, pigeon pea fiber, sorghum fiber, buckwheat fiber, potato fiber, sweet potato fiber, lupin fiber, apple fiber, any mixture thereof.

18. The method of claim 1 or 2, wherein: The starch-containing cereal grains are selected such that the malted treated grains (ii.a) are used only in combination with the germinated treated grains (ii.b) and / or malt (ii.c) and / or germinated grains (ii.d), i.e. the option ii.a. of claim 1 is excluded alone.

19. The method of claim 1 or 2, wherein: The extrusion step is performed with an extrusion die having a length of more than 300 mm.

20. The method of claim 3 or 4, wherein: Some of the unemulsified starch is soluble starch.

21. A meat substitute food product, characterized in that: The meat substitute product is an extrudate manufactured according to the method of any one of claims 1-20 or the meat substitute product comprises an extrudate manufactured according to the method of any one of claims 1-20.

22. The meat substitute food product of claim 21, characterized in that: The extrudate comprises starch, at least 5.1% of which is soluble starch.

23. The meat substitute food product of claim 21 or 22, wherein: The meat substitute product is in the form of a loaf, a dice, a chicken nugget, a steak, a fillet, a kebab or a layered layering layer in a yogurt or a vegan yogurt and spice and a kebab string. The extrusion step is performed with an extrusion die having a length of more than 300 mm. Some of the unemulsified starch is soluble starch. The meat substitute product is an extrudate manufactured according to the method of any one of claims 1-20 or the meat substitute product comprises an extrudate manufactured according to the method of any one of claims 1-20. The extrudate comprises starch, at least 5.1% of which is soluble starch. The meat substitute product is in the form of a loaf, a dice, a chicken nugget, a steak, a fillet, a kebab or a layered layering layer in a yogurt or a vegan yogurt and spice and a kebab string.

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