Method for preparing food containing fibrous fungi and the product

Abstract

1. A method of providing a filamentous fungus-containing food product, comprising the steps of: providing one or more elongated pieces made from a moist textured protein product; providing nutrients for fungal growth in or around the one or more elongated pieces; inoculating the one or more elongated pieces with at least one fungus; forming a scaffold for mycelial growth using the one or more elongated pieces; and incubating the scaffold under growth conditions permitting mycelial growth of the at least one fungus such that the at least one fungus forms a mycelium, wherein the mycelium grows along and through the scaffold to form the filamentous fungus-containing food product.

Description

[Technical field]

[0001] The present invention relates to a method of providing a filamentous fungus-containing food product by providing elongated pieces of moist textured protein product that are attached to each other through surface growth of fungal mycelium, thus forming a fungus-containing food product. [Background technology]

[0002] Current meat consumption is depleting our natural resources while driving climate change. Our current meat consumption is unsustainable and therefore meat substitutes must be developed to combat the ever-increasing consumption of meat. Various techniques are applied to produce meat substitute products, resulting in wide differences in texture and nutritional value.

[0003] Some more modern products, such as tofu and plant-based sausages or meatloaf, are produced by gelling aqueous dispersions / solutions of proteins and / or polysaccharides. This technique results in a fairly low protein content that is not at all comparable to meat, and a soft, silky, elastic, rather juicy texture that does not have the fibrous qualities that are comparable to animal sausage meat.

[0004] In another approach, dry textured vegetable protein (TVP), fungal fragments, laboratory-grown animal cells, or mushrooms, grains, beans, nuts, or seeds are glued or sintered together, as known from traditional products such as tempeh, or more modern products such as plant-based burger patties, sausages, nuggets, and minced meats. In tempeh, protein-rich grains, beans, nuts, or seeds are grown by the growth of mycelium-forming fungi (e.g., Rhizopus spp.) between the particles. The tempeh is baked by growth of Oligosporus, resulting in a firm, granular texture that is less fibrous but has a protein content (up to 20g / 100g) closer to animal meat, as disclosed, for example, in https: / / www.thekitchn.com / how-to-make-tempeh-cooking-lessons-from-the-kitchn-202369 (accessed November 1, 2021), EP 2835058, and U.S. Pat. No. 3,885,048. In a similar approach disclosed in WO 2020 / 232347, grains are combined with vegetable protein concentrates or isolates before baking with fibrous fungi. Dry textured vegetable protein concentrates or isolates are often used in various plant-based meat-like products. The dry / porous extrudates, called TVPs, are baked together with a gelling agent (e.g., methylcellulose) (Kyriakopoulou et al., Foods, 10(3), 2021) to then form products such as burger patties or nuggets. In some other approaches, such TVPs, extruded pellets, extruded strands or extrudates are baked with the addition of mycelium-forming fungi, as disclosed in WO 2021 / 030412, WO 2020 / 164680, WO 2013 / 087558 and JP 2006-129703 A. Such porous / dry strands, extruded strands, extruded pellets or dry TVPs provide pores or channels for the fungi to grow and receive sufficient oxygen and nutrients.Alternatively, WO 2021 / 030412 suggests producing mycelium in liquid fermentation, thereby providing the resulting fungal biomass as a scaffold for baking based on fungal growth. The final baked product is elastic, spongy, and succulent, with a structure comparable to ground / minced / sausage meat based products, but lacking oriented fibrous and muscle-like structure. Fibrous meat substitutes, such as plant-based chicken pieces, are generally produced by high moisture extrusion cooking (HMEC) or shear cell (SC) processing, in which the protein is melted at high temperature, pressure and moisture content of 40-80% as described by Osen et al. ("High moisture extrusion cooking of pea protein isolates: Raw material characteristics, extruder responses, and texture properties", J. of Food Engineering, 127 (2014) 67-74), and then cooled under shear resulting in the formation of a solidified fibrous structure. WO 2021 / 195175 discloses further examples of using wet extrusion to produce fibrous meat analogues. Although the resulting product is closer to animal meat in terms of fibrousness, the fairly homogeneous, meaty structure formed in these processes is considered to lack juiciness and elasticity and to be overly sticky and meaty. When such products are processed into small pieces, such as pieces similar to chicken pieces or pulled pork, the fibrous texture appears to dominate the sensory perception, and the crust formed during cooking or the marinade or sauce added during cooking appears to cover the lack of stickiness and juiciness. However, when biting into larger pieces, the stickiness and lack of juiciness becomes more dominant, and therefore the texture is perceived as less pleasant. Furthermore, compared to animal meat, such fibrous structures based on vegetable proteins lack fat and collagen / connective tissue phases. In animal meat, fat and connective tissue are located between the muscle tissues, contributing to a more juicy and more marbling texture. Additionally, meat substitutes to larger meat pieces (whole cuts) and more succulent meat pieces such as chicken breasts are needed to meet consumer needs, transition to animal-free diets, and reduce the environmental footprint of food systems. However, HMEC and SC processing is limited in size to a few centimeters, typically 3 cm or less, due to thermodynamics and fluid dynamics. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] European Patent Application Publication No. 2835058 [Patent Document 2] U.S. Pat. No. 3,885,048 [Patent Document 3] International Publication No. 2020 / 232347 [Patent Document 4] International Publication No. 2021 / 030412 [Patent Document 5] International Publication No. 2020 / 164680 [Patent Document 6] International Publication No. 2013 / 087558 [Patent Document 7] Specification of JP2006-129703 [Patent Document 8] International Publication No. 2021 / 195175 [Non-patent literature]

[0006] [Non-Patent Document 1] https: / / www.thekitchn.com / how-to-make-tempeh-cooking-lessons-from-the-kitchn-202369 (Accessed November 1, 2021) [Non-Patent Document 2] Kyriakopoulou et al., Foods, 10(3), 2021) [Non-Patent Document 3] Osen et al. (“High moisture extrusion cooking of pea protein isolates: Raw material characteristics, extruder responses, and texture properties”, J. of Food Engineering, 127 (2014) 67-74) Summary of the Invention [Means for solving the problem]

[0007] In order to construct larger pieces of meat substitutes that combine the pleasing fibrous qualities of HMEC-processed or SC-processed wet textured protein products, but avoid the high density, stickiness, limited juiciness and limited structural complexity and size constraints, the present invention is proposed, which is a method for providing a fibrous fungal-containing food product, the method comprising: (i) providing one or more elongated pieces made from a wet textured protein product produced, for example, by high moisture extrusion cooking or shear cell processing; (ii) providing nutrients for fungal growth in or around the one or more elongate segments; (iii) inoculating said one or more elongated segments with at least one fungus; (iv) using one or more elongated pieces to form a scaffold for mycelial growth; (v) incubating the scaffold under growth conditions that permit mycelial growth of the at least one fungus such that the at least one fungus forms a mycelium, which mycelium grows along and through the scaffold to form a filamentous fungus-containing food product; The present invention relates to a method comprising the steps of:

[0008] The resulting fungus-containing food product exhibits a meat-like fibrous texture from fibrous elongated pieces or bundles, while the fungal mycelium at the interface of the pieces constitutes a connective tissue comparable to the complex animal meat structure. The method also results in a product having a lower density than the initial wet textured protein product, thus reducing unpleasant meaty ride and sticky texture.

[0009] Surprisingly, the texture and taste of the product can be adapted by modifying the distance between adjacent elongated pieces, the available nutrients and the growth conditions. For example, if the distance between two elongated pieces is greater, mycelium growth leads to a weaker attachment between the elongated pieces. Availability of sufficient nutrients, strain, composition of the elongated pieces and growth conditions can also lead to pleasant aroma formation. Furthermore, the amphiphilic nature and porous structure of the mycelium allows it to absorb oil and water into the interstitial spaces between the elongated pieces, resembling the fatty and / or connective tissue of animal meat. The liquid phase in the interstitial spaces is released during mastication, thus contributing to the succulence.

[0010] Even more surprisingly, coating the strips with carbohydrates, such as glucose-rich agar and / or starch, or adding carbohydrate-based strips, such as starch-based strips or glucose-rich agar strands between the strips, enhances fungal growth in the interstitial spaces, the strips attach better, and the remaining starch in the interstitial spaces contributes to a sliminess after cooking that is comparable to collagen or connective tissue found in animal meats. Such a structure interconnected with a collagen / fat-like matrix has not been achieved before in plant / fungal based meats. Furthermore, providing carbohydrates on or between the strips causes the fungus to metabolize the carbohydrates rather than the proteins, reducing ammonia formation and protecting the texture of the strips.

[0011] In other words, according to one aspect of the present invention, the lack of oriented fibrousness in known products can be addressed by providing elongated pieces of wet textured protein product, assembling such pieces into a scaffold having a fibrous structure such that the fungus grows through the scaffold, and binding the elongated pieces together, for example in a muscle-like fashion. Such muscle-like structures are highly desired by consumers as they are more reminiscent of high value cuts of animal meat compared to "minced products".

[0012] Since fungal growth requires oxygen and certain humidity and temperature conditions, it is difficult to provide sufficient conditions for fungi to grow through larger bodies. However, large pieces of meat-like fibrous food are desired by consumers to meet the needs of whole cuts, such as products similar to beef tenderloin or chicken breast. Surprisingly, in the present invention, this limitation can be overcome by placing a tube in the scaffold. Preferably, the tube communicates the inner part of the scaffold with the surrounding environment, allowing air transport from the surrounding environment to the inner part of the scaffold. Preferably, the tube is an oxygen-permeable and / or conduit tube, such as a plastic tube with small holes. Alternatively, rods may be placed in the scaffold, which are then preferably removed after an initial incubation period, thus creating channels for increasing oxygen permeability at a time when the scaffold is partially filled with mycelium and the oxygen concentration may already be reduced in the core due to fungal activity.

[0013] Even more surprising, the tubes or needles inserted into the scaffold can be connected to a source of pressurized gas, such as pressurized air or oxygen, or to a pump to force or pump oxygen-containing gas into the core. Thus, large pieces with a shortest dimension of at least 10 cm can be produced while growing mycelium throughout the entire scaffold. Interestingly, these rods, needles or tubes also help to avoid overheating of the core, which is necessary to ensure ideal growth conditions for the fungus.

[0014] The term "fungal-containing" means that the product obtained by the process always contains fungal components, in particular fungal mycelium, although other components may be of different origin.

[0015] The method of the invention is characterized in that in step (i) elongated fragments are provided. The term "provided" in this context means that elongated fragments are used or that such elongated fragments are prepared. Means and methods for preparing elongated fragments are further described below.

[0016] "Anisotropic" refers to the internal structure of the elongated pieces. Preferably, each elongated piece comprises an anisotropic internal structure, also called "fibrous structure", which is the result of a wet texturing process, such as high moisture extrusion cooking or shear cell processing, as described, for example, in R. Osen, Dissertation, 2017 (https: / / d-nb.info / 116838026X / 34) or K. Grabowska, Food Research International, 64, 2014. These processes result in a fibrous internal structure that is substantially aligned in the direction of the flow field. As in high moisture extrusion cooking and shear cell processing, the protein-containing matrix is ​​mixed with water, heated to above 100°C, and then solidified under shear, and the fibrous structure is formed as known to those skilled in the art. In the present invention, the pieces are provided with an elongated shape, which means that one dimension is substantially longer than the other two dimensions (e.g., "strand") or two dimensions are substantially longer than the third dimension (e.g., "sheet"). Importantly, the longest dimension or one of the longest dimensions is preferably substantially aligned with the fibrous internal structure. This feature of the elongated pieces in combination with the fibrous structure is advantageous in creating a highly fibrous texture and appearance in the final product.

[0017] The term "fibrous structure" refers to a structure in which fiber bundles / fiber aggregates / aggregated fibers / fiber sheets, especially made of proteins, result in anisotropic properties regarding the structure and mechanical properties of the fibrous structure. Preferably, the fibrous structure has a high degree of alignment in one direction. The fibrous structure is formed in the wet texturing process when proteins and other components are stretched and / or aligned by the application of shear. The fibrous structure is composed of multiple fiber bundles / fiber aggregates / aggregated fibers / fiber sheets, sometimes referred to as "fibers". The fibrous structure resulting from wet texturing is known to those skilled in the art and results in a chewy animal meat-like texture and appearance.

[0018] Preferably, the elongated pieces are composed of wet textured protein products made by wet texturing, for example by high moisture extrusion cooking or shear cell processing. Wet texturing means mixing a protein-containing formulation with an aqueous phase and subjecting it to a temperature preferably above 100° C. under shear, followed by cooling under shear resulting in the formation of a fibrous structure. Wet texturing refers in particular to texturing at a moisture content of 40-80% by weight. In contrast, dry / porous extrudates, TVP, and extruded pellets, for example as utilized in WO 2020 / 164680 or WO 2021 / 030412, are processed to produce a porous structure. As noted in WO 2021 / 030412, TVP is produced, for example, by forcing the molten protein mixture through a die to cause a fairly sudden pressure release at the extruder outlet, resulting in a less fibrous and more spongy structure. The moisture content in extrusion to form such structures is typically less than 35-40% by weight to reach high enough pressure and provide sufficient cohesion of the mixture upon expansion. Such techniques do not result in the desired muscle-like oriented fibrous structure achieved in the present invention. Notably, in wet textured protein products, the fibers are substantially aligned due to controlled cooling and solidification under flow, which is particularly important to the present invention.

[0019] Preferably, said wet textured protein product is produced by high moisture extrusion cooking in an extruder, more preferably a twin screw extruder, at a moisture content of more than 40% and less than 80%, even more preferably 45% to 70%, and a protein content of more than 10%, preferably more than 15%, even more preferably more than 20% by weight. In particular, a mixture of protein, water and other ingredients is sheared in the extruder and heated to more than 100°C, preferably more than 120°C, and then cooled to less than 100°C in a cooling die before exiting the machine to avoid puff formation and form a fibrous structure. Elongated pieces are then prepared from the wet textured protein product.

[0020] Alternatively, elongated segments are produced by fiber spinning or 3D printing.

[0021] In particular, said wet textured protein products comprise at least 10% by weight of protein selected from the group consisting of pea, soy, wheat, sunflower, broad bean, pumpkin, rice, cereals, pulses, oilseeds, algae, single cell, fungi, and fermented ingredients or mixtures thereof, preferably at least 15% by weight, and most preferably at least 20% by weight of protein selected from the group consisting of pea, soy, wheat, sunflower, broad bean, pumpkin, rice, cereals, pulses, oilseeds, algae, single cell, fungi, and fermented ingredients or mixtures thereof.

[0022] "Protein" refers to a protein isolate, concentrate or flour or combination thereof that may also contain other macronutrients such as carbohydrates, fat, dietary fiber, salt, or residual water. The isolate, concentrate, flour or combination thereof preferably contains a pure protein content of at least 40% by weight, preferably at least 50% by weight, and even more preferably at least 60% by weight. The protein isolate, concentrate or flour ("protein") may also be referred to as a "protein composition" or "protein powder" in the context of the present invention. Preferably, the aforementioned protein also contains sufficient carbohydrate to act as a nutrient source for the fungus.

[0023] In one embodiment, said wet textured protein product comprises pea protein as the sole protein source.

[0024] In an alternative embodiment, said wet textured protein comprises not only pea protein but also at least one other protein source, preferably of plant origin.

[0025] In an alternative embodiment, said wet textured protein product comprises at least pea protein, sunflower protein and oat protein.

[0026] In an alternative embodiment, said moist textured protein product comprises at least pea protein and yeast protein.

[0027] In an alternative embodiment, said moist textured protein product comprises at least pea protein and soy protein.

[0028] In an alternative embodiment, said moist textured protein product comprises soy protein as the only protein.

[0029] In addition to water and protein / protein composition / protein powder, the wet textured protein product may contain any other edible ingredients added before or during the wet texturing process, such as oils or fats, dietary fiber, flavor ingredients, colorants, or carbohydrates, which other ingredients are preferably of non-animal origin, and even more preferably derived from plants, fungi, fermentation processes, laboratory grown animal cells, or single cell organisms.

[0030] Optionally, one of the elongated pieces is made from one formulation of the wet textured protein product and another elongated piece is made from a different formulation of the wet textured protein product, or there may be more than two different formulations of elongated pieces used in one scaffold.

[0031] Elongated pieces can be made from the wet textured protein product by, but not limited to, cutting, tearing, rolling, or by immersion in a liquid to the extent that the fibrous protein product disintegrates into anisotropic fibrous pieces. In particular, the fibrous structure is substantially aligned in the longest dimension of the elongated pieces. By "substantially aligned" is meant that thinner fiber bundles can be pulled away from the elongated pieces in the direction of the longest dimension.

[0032] For example, moist textured protein products produced by high moisture extrusion cooking can be peeled or cut in the direction of alignment of the fibrous structure into strips or sheets representing elongated segments.

[0033] Preferably, the elongated segments are prepared so that their longest dimension is at least two times longer than their other two dimensions, and most preferably at least five times longer (i.e., (i) perpendicular to the longest dimension, and (ii) the other two dimensions perpendicular to each other).

[0034] Preferably, said elongated pieces have a minimum dimension, e.g., diameter, of 2 cm or less, preferably 1 cm or less, more preferably 0.5 cm or less, even more preferably 0.3 cm or less. Preferably, said elongated pieces have a maximum dimension, e.g., length, of 1 cm or more, preferably 2 cm or more, more preferably 4 cm or more. The resulting filamentous fungus-containing food product closely resembles meat structure when the elongated pieces have dimensions falling within the above ranges.

[0035] Preferably, the elongated pieces are further processed prior to inoculation, including but not limited to soaking or cooking in water at 0-120°C, soaking in acidic or alkaline solutions, drying, pressing or smoking. In particular, the water activity or moisture content of the elongated pieces is adjusted to conditions desirable for fungal growth, preferably the water activity is 0.8-1.0, more preferably 0.9-1.0, even more preferably 0.96-1. Preferably, the pH of the elongated pieces is lowered to below 7, preferably below 6, even more preferably below 5 to support fungal growth. This may be achieved by spraying an acid, such as lactic acid, acetic acid, citric acid or other acid, onto the surface of the elongated pieces, or by soaking said elongated pieces in acid.

[0036] The elongated pieces may be dried to reach a moisture content of less than 40% by weight, preferably less than 30% by weight, prior to incubation.

[0037] Optionally, at least one of the elongated pieces is further processed to be different from at least one other elongated piece. In particular, some elongated pieces can be dried more than one or more other elongated pieces or immersed in liquid for a longer time than one or more other elongated pieces. Furthermore, some elongated pieces can be treated with an edible fungal growth inhibitor, such as an essential oil, while other elongated pieces used in the same scaffold are not treated with a fungal growth inhibitor.

[0038] If desired, some of the elongate strips are processed differently than other portions of the elongate strips.

[0039] Preferably, the elongate pieces are sterilized or pasteurized prior to inoculation to ensure inactivation of any coexisting surface microorganisms, for example, the elongate pieces can be sterilized in an autoclave at up to 135°C.

[0040] Preferably, the surface of the elongate pieces is treated with an acid, preferably a food grade acid, more preferably lactic acid, acetic acid, malic acid, citric acid or succinic acid, preferably to reach a pH at the surface of less than 6, even more preferably less than 5, most preferably not exceeding 4.6. Even more preferably, the pH of the surface is adjusted to ensure the growth of the utilized fungus and to reduce the growth of other microorganisms.

[0041] Nutrients such as carbohydrates, preferably starches or sugars, can be provided as nutrients within or on the elongated pieces, such as by adding starch or sugar-containing ingredients to a high moisture extrusion cooking or shear cell process, or by immersing the wet textured protein product or elongated pieces in a starch or sugar solution, or by coating the elongated pieces with a starch or sugar-containing liquid or powder. Preferably, at least 1% by weight of nutrients is included, more preferably at least 2% by weight of nutrients is included, and most preferably at least 5% by weight of nutrients is included, based on the total weight of the one or more elongated pieces. Such starch-containing ingredients added to the wet texturing process can be protein flours, protein concentrates or protein isolates containing starch or sugar residues.

[0042] Preferably, said nutrients are selected from the group of carbohydrates comprising monosaccharides, disaccharides, oligosaccharides or polysaccharides acting as nutrients for the respective fungi, preferably from the group of starch, glucose, sucrose or malt starch.

[0043] Alternatively or additionally, said nutrients may be selected from the group of proteins or fats or oils. Preferably, the nutrients are a mixture of carbohydrates, also called sugars, with proteins and / or fats or oils.

[0044] Preferably, the starch is swollen or pregelatinized prior to coating, or the starch-coated elongated pieces are heat treated to cause swelling and gelatinization on the surface of the elongated pieces.

[0045] Starch refers to native, unprocessed, malted, modified, pregelatinized or other types of starch or starch derivatives, preferably selected from rice, wheat, corn, root crops such as potato or cassava sources.

[0046] Sugar refers to any monosaccharide, disaccharide, or oligosaccharide that can be metabolized by fungal metabolism, and in particular sugar can refer to glucose, sucrose, and maltose.

[0047] Preferably, the surface of the elongate pieces is at least partially coated with a carbohydrate, preferably a starch, prior to incubation, to the extent that the carbohydrate or starch is not completely metabolized by the fungus, preferably with at least 2% by weight of carbohydrate or starch, more preferably at least 5% by weight of carbohydrate or starch, even more preferably at least 10% by weight of carbohydrate or starch. Said starch may be pregelatinized or partially or completely swollen, for example by heat treating the starch-coated elongate pieces.

[0048] The carbohydrates may be cellulose and / or hemicellulose and may be in the form of plant fibres, such as citrus fibre, banana fibre, bamboo fibre, oat fibre, carrot fibre, apple fibre, microcrystalline cellulose or microfibrillated cellulose, which may further comprise lignin or other plant components.

[0049] By "not completely metabolized" is meant that a portion of the carbohydrate or starch remains on the surface of the elongate fragments or in the fungal mycelium network between the elongate fragments once fungal growth has ceased or the final product has been consumed. Preferably, the remaining carbohydrate, preferably starch, contributes to a slimy, collagenous mouthfeel.

[0050] Alternatively or additionally, the elongate pieces may be coated with a hydrocolloid, preferably agar, carrageenan, pectin, gelatin, xanthan, gellan gum or alginate or a combination thereof, prior to inoculation and / or incubation. Preferably, a hydrocolloid is selected which is not metabolized at all or completely metabolized during incubation.

[0051] Such hydrocolloids may also be combined with starch, glucose, sucrose, malt starch or other carbohydrates, such as agar rich in glucose.

[0052] Preferably, any added ingredients, such as nutrients, are sterilized or pasteurized or treated by chemical or physical means to an extent that the microbial load is sufficiently reduced to avoid spoilage during incubation prior to inoculation.

[0053] The elongated pieces, and preferably all other pieces, e.g. starch-based pieces, can be inoculated with at least one fungus by mixing with a fungus-containing liquid or powder, said fungus being preferably selected from the group consisting of Ascomycetes, Basidiomycetes, Deuteromycetes, Oomycetes and / or Zygomycetes, in particular edible species belonging to the genera Rhizopus, Aspergillus, Penicillium, Ganoderma or Pleurotus, more particularly the species Rhizopus oligosporus, Rhizopus delemar, Rhizopus oryzae, Aspergillus oryzae, Aspergillus luchuensis, Aspergillus sojae, Penicillium nalgiovense, Penicillium camemberti, Penicillium roqueforti, Ganoderma lucidum, Pleurotus ostreatus, Pleurotus eryngii, or combinations thereof. The surface of the strips may be pre-wetted prior to inoculation with the fungus-containing powder.

[0054] At least three elongated pieces can be assembled with or without a mold to form a scaffold. The distance between adjacent pieces can be adjusted by vibration or by applying pressure or vacuum so that each piece is at least 2 mm or less away from at least one adjacent piece, more preferably 1 mm or less, and even more preferably 0.5 mm or less. Said adjacent pieces can be elongated pieces or other elements, such as starch-based pieces. Each elongated piece can be in contact with at least one other elongated piece or other elements, such as starch-based pieces.

[0055] At least 5, preferably at least 10, elongate fragments may be substantially aligned in a fibrous body comprising bundles of elongate fragments. The scaffold may also be composed of some bundles that are not aligned with each other.

[0056] The elongated pieces may be strands or sheets. Preferably, the scaffold is formed by assembling multiple strands into a bundle, rolling up one or more sheets into a tube or multi-layer roll, or folding or stacking one or more sheets. Fibers and sheets may be combined within the same scaffold.

[0057] Carbohydrate-based fragments, preferably starch-based fragments, may be provided and added to the scaffold, preferably said carbohydrate-based fragments being arranged between two or several elongated fragments. Preferably said carbohydrate-based fragments comprise a carbohydrate selected from the group of starch, starch derivatives, agar, alginate, carrageenan, pectin, konjac gum or sugar or combinations thereof. Preferably, each of said starch-based fragments comprises at least 5% by weight starch, preferably at least 10% by weight, more preferably at least 20% by weight starch, based on the weight of said starch-based fragment. Even more preferably, one of the carbohydrates contained is gelled and / or gelatinized, thus allowing to form the fragment.

[0058] Said carbohydrate-based, preferably starch-based, pieces may for example be in the form of cubes, rods, cylinders, sheets or stripes.

[0059] Said carbohydrate-based fragments may be plant fibres containing cellulose and / or hemicellulose, such as citrus fibre, banana fibre, bamboo fibre, oat fibre, carrot fibre, apple fibre, microcrystalline cellulose or microfibrillated cellulose.

[0060] The starch-based fragments may be made from at least water and starch that has been gelatinized by heating above the gelatinization temperature. For example, the starch may be mixed with water in a ratio of 1:5 to 5:1. The carbohydrate-based or starch-based fragments may be cut as needed.

[0061] The carbohydrate-based or starch-based pieces may have a thickness of 2 cm or less, preferably 1 cm or less, more preferably 5 mm or less, even more preferably 2 mm or less. The carbohydrate-based or starch-based pieces may be provided by forming the carbohydrate- or starch-containing composition into a layer having a thickness of 2 cm or less, preferably 1 cm or less. The layer may then be cut into pieces.

[0062] For example, starch-based pieces can be produced by mixing starch with water in a ratio of 1:5 to 5:1, preferably 1:1 to 2:1, forming the mixture into a layer less than 1 cm thick, and heating the mixture to a temperature above the gelatin temperature of the starch, preferably above 60°C, more preferably above 70°C, even more preferably above 80°C. The aforementioned temperatures cause gelation of the starch and a reduction in the microbial count. The layer can be cut into cubes or rods or stripes or sheets that are preferably no more than three times the size of the elongated pieces.

[0063] For example, the starch-based pieces may be starch-containing noodles or pasta.

[0064] The carbohydrate-based fraction may further comprise water, lipids, proteins and flavor or color components.

[0065] Differently processed or treated elongated pieces can be assembled into a scaffold to create a pH or nutrient gradient across the scaffold. For example, elongated pieces with higher nutrient concentrations can be placed in the center of the scaffold, and pieces with lower nutrient concentrations can be placed on the outside of the scaffold. Fragments with lower nutrient concentrations can be placed between areas of fragments with higher nutrient concentrations to create intended cutoff points and distinguish compartments of the body. Differently colored pieces can be placed in groups in different areas of the body.

[0066] Alternatively, the elongated pieces may be combined with another material, which is a fragment or a continuous phase, to fill one or several spaces between the elongated pieces. Preferably, such material has texture properties different from the elongated pieces. Such a material may be, for example, a solution, a paste, a suspension, an emulsion or a foam, the aforementioned material comprising a carbohydrate. Preferably, the solution, suspension, emulsion, gel or foam comprises a thickening or gelling agent to form a layer between the elongated pieces resembling connective tissue. The carbohydrate included may be selected from the group of agar, carrageenan, konjac, starch, alginate, pectin or other carbohydrates.

[0067] All the pieces may be substantially aligned in the scaffold, and thus the internal fiber structure of the elongated pieces is substantially aligned. Alternatively, three or more pieces may be substantially aligned and assembled to form a fiber bundle, which is then assembled with one or more other fiber bundles. Several fiber bundles may be joined together to form a scaffold. Two or more fiber bundles may be substantially aligned or not aligned to mimic the muscle structure of animal meat. Two or more fiber bundles may be in direct contact with or separated by other pieces, such as layers of starch-based pieces, resembling the layers of connective tissue between muscle bundles of animal meat.

[0068] The two or more fiber bundles may differ in the properties of the three or more constituent elongated fragments with respect to orientation, hardness, elasticity, texturing, color, moisture content, growth nutrient composition, flavor, size, surface coating, and source of the elongated fragments.

[0069] Preferably, pressure or vacuum is applied when assembling the elongated segments and other segments or fiber bundles to form the scaffold. Pressure may be applied manually or by machine. Preferably, pressure is applied to an extent that keeps the density of the scaffold lower than the average density of the elongated segments.

[0070] Preferably the scaffold, prior to incubation, is at least 1cm in at least one dimension, preferably at least 2cm, more preferably at least 4cm, even more preferably between 4cm and 15cm, and / or preferably has a shortest dimension of 30cm or less, even more preferably 20cm or less.

[0071] Preferably, the scaffold is incubated at a temperature between 4 and 70°C, in particular between 10 and 50°C, even more preferably between 14°C and 20°C or between 22°C and 38°C, for a minimum of 2 hours, in particular for a minimum of 12 hours, especially for the minimum period required until the mycelium becomes visible to the human eye on the surface of the elongated pieces. Preferably, the incubation time is short enough to avoid fungal sporulation and / or the formation of fruiting bodies. Thus, the incubation can be terminated before fungal sporulation or before the formation of fruiting bodies.

[0072] Preferably, scaffolds inoculated with filamentous fungi such as Aspergillus oryzae, Rhizopus oligosporus, Rhizopus delemar, or Rhizopus oryzae are incubated at 10° C. to 45° C., while scaffolds inoculated with other fungi are incubated at temperatures between 10° C. to 45° C., and even more preferably between 22° C. to 38° C. for Ascomycetes, Imperfect Fungi, Oomycetes, and / or Zygomycetes. Scaffolds inoculated with Basidiomycetes can be incubated at temperatures between 14° C. to 20° C.

[0073] In another embodiment, Basidiomycete species such as Ganoderma species or Pleurotus species can be used as inoculants.

[0074] Incubation temperature refers to the temperature set in the incubator.

[0075] Even more preferably, the incubation temperature is adjusted as a function of metabolic heat production by the fungal growth such that the temperature of the scaffold, referred to as the substrate temperature, does not rise above 45° C. during incubation. For scaffolds inoculated with Aspergillus oryzae, the substrate temperature is preferably maintained between 25° C. and 45° C. by adjusting the incubation temperature.

[0076] Regardless of other incubation conditions, the water activity of the scaffold is preferably in the range of 0.8 to 1.0, preferably 0.9 to 1.0, even more preferably 0.96 to 1.0. Preferably, such water activity is maintained throughout the incubation.

[0077] The preferred oxygen concentration during incubation may depend on the fungus.

[0078] The scaffold may be wrapped in an outer layer, said layer may have holes or may be semi-permeable or permeable to oxygen, preferably said outer layer is a plastic foil with holes, such as a polyethylene-based cling film, even more preferably the outer layer is permeable to oxygen but not to water.

[0079] The scaffold may be incubated by covering the scaffold with a material, preferably having an oxygen permeability coefficient less than 1 but greater than 0. In particular, the scaffold may be wrapped in a cling film.

[0080] Alternatively or additionally, microaerobic conditions can be created by incubating the scaffold in a space having an oxygen concentration less than that in the ambient atmosphere. For example, the oxygen concentration can range from 4000 to 10000 ppm for scaffolds, e.g., scaffolds inoculated with R. oligosporus.

[0081] Preferably, holes and / or channels are created in the scaffold before or during incubation to provide sufficient oxygen for fungal growth.

[0082] Preferably, to provide such one or more holes or channels, one or more placeholders may be placed in the scaffold during assembly of the scaffold or before or during incubation.

[0083] Such placeholders may provide holes and / or channels in or through the scaffold.

[0084] For example, one or more tubes or one or more rods, as examples of such placeholders, can be placed in the scaffold before or during incubation. The tubes or rods can extend in any direction, such as axially or radially. Preferably, the tubes or rods extend along the shortest dimension of the scaffold.

[0085] The tubes or rods may be straight or curved.

[0086] The one or more rods or tubes can have any cross-sectional shape, such as circular or polygonal (e.g., rectangular). The one or more rods can be solid and / or lack a lumen.

[0087] The placeholders (eg tubes or rods) preferably have a diameter or width (ie perpendicular to their longitudinal extension) of at least 0.1 mm, preferably at least 1 mm, even more preferably at least 2 mm.

[0088] The placeholder (eg, tube or rod) preferably has a shortest dimension or width of 15 mm or less, more preferably 10 mm or less, and most preferably 5 mm or less.

[0089] One or more placeholders (e.g., one or more rods) may be withdrawn from the scaffold before the start of incubation or before the end of incubation. In this way, one or more channels may be formed in the scaffold along which oxygen may diffuse during incubation.

[0090] One or more of the tubes may have a lumen extending therethrough, for example, to deliver oxygen to the scaffold. One or more of the tubes may be at least partially oxygen permeable and / or may include one or more openings along the portion thereof that is inserted into the scaffold. For example, one or more of the tubes may be hollow needles. One or more of the tubes may remain in the scaffold during incubation.

[0091] One or more tubes can connect the inner part of the scaffold with the surroundings, especially during incubation, thus providing a higher oxygen concentration in the inner part of the scaffold, especially when compared to a scaffold incubated without tubes or channels.

[0092] One or more of the tubes can be removed during or after incubation. Preferably, one or more of the tubes are edible. In this case, one or more of the tubes do not need to be removed prior to consumption.

[0093] One or more of the tubes may be covered with a protective culture or anti-fungal material that prevents fungi from growing within the tube.

[0094] Alternatively, the tubes may be fully or partially metabolised by the fungus during incubation and do not need to be removed prior to consumption.

[0095] Preferably, one or more tubes are connected to an oxygen source, preferably pressurized air or oxygen, or a pump. The oxygen source preferably provides an oxygen-containing gas. Preferably, the oxygen-containing gas is humidified before being delivered (e.g., pumped) into the scaffold.

[0096] Preferably, one or more rods or tubes are inserted before or at the beginning of incubation.Preferably, one or more rods or tubes are removed after at least 2 hours of incubation, preferably after at least 6 hours of incubation, even more preferably after at least 10 hours of incubation, or even more preferably when mycelium growth in the scaffold becomes visible.Preferably, one or more rods or tubes are removed at a time when the holes or cavities left behind by the rods or tubes do not completely disappear after the rods or tubes are removed.

[0097] One or more tubes may be interconnected and may be used to supply nutrients to the fungus during incubation by pumping nutrients to the scaffold and removing waste products.

[0098] The placeholder may be branched, for example the placeholder has a branched shape resembling a shrub or tree in morphology.

[0099] Rods or tubes or other placeholders may be added during the assembly of the scaffold so that the scaffold is assembled around the placeholder. Preferably, the placeholder has holes or is porous, e.g., a porous solid. The porous solid may be an open-pore structure similar to bone. The open-pore placeholder can communicate the inner part to the outside during incubation to allow gas exchange and oxygen supply during incubation, and later act as the bone-like material of the final food. The open-pore placeholder may be connected to an oxygen source, so that oxygen-containing gas is pumped through the "bone-like material" and into the scaffold. This allows meat-like structures to grow with the bone-like structure in place. Such a placeholder can remain in the food until it is consumed.

[0100] In alternative embodiments, the atmosphere within and surrounding the scaffold during incubation is adjusted to support fungal growth of a particular fungus or fungi, for example, the composition is altered and / or gas pressure is increased and / or varied to ensure sufficiently high oxygen concentrations throughout the scaffold.

[0101] Preferably, the scaffold is incubated at least until mycelium is visible to the human eye. Preferably, incubation is terminated before sporulation. Incubation conditions and times can be adjusted to suit the fungal strain, available nutrients, composition of the elongated pieces and the desired outcome. Preferably, incubation conditions are adjusted to avoid the formation of unpleasant off-flavors, such as those resulting from the formation of ammonia.

[0102] If necessary, fungal growth is interrupted by changing the temperature and / or water activity conditions, preferably below or above the temperature and water activity conditions required for the growth of at least one fungus. Alternatively, fungal growth can be interrupted by lowering the oxygen concentration below the critical level required for growth by the respective fungus. Preferably, fungal growth is interrupted by heating the fungus-containing product to above 60°C, preferably above 80°C, more preferably above 90°C, with the temperature measured in the center of the product and maintained for at least 1 minute.

[0103] Alternatively, fungal growth is not interrupted. Instead, the fungus-containing product is stored in a refrigerator or freezer until it is to be consumed, preferably prior to sporulation or spoilage.

[0104] According to the described method a fungus-containing food product is obtained.

[0105] Preferably, the fungus-containing food product as obtained by any of the above-mentioned variants is exposed to a liquid such that the food product absorbs the liquid, said liquid being water-based or lipid-based or a combination thereof.

[0106] Preferably, at least 2% by weight of water, more preferably at least 5% by weight of water, even more preferably at least 10% by weight of water, based on the weight after water addition, is added to the fungus-containing food product after or before interrupting fungal growth, with or without heating the water to 99° C. Surprisingly, the water is absorbed by the fiber fragments, causing them to swell and thus shortening the distance between the fragments.

[0107] Preferably, at least 1% by weight of lipid phase, more preferably at least 2% by weight, even more preferably at least 5% by weight, most preferably at least 10% by weight, based on the weight after lipid addition, is absorbed into the fungus-containing food product. Preferably, the lipid phase is absorbed into the voids filled with mycelium, e.g., mostly absorbed into said void spaces. For example, lipid absorption can occur during or prior to frying in a pan. Lipid absorption can be provided at room temperature or at elevated temperatures in the case of fats with higher melting temperatures, preferably at temperatures higher than the melting temperature of the lipid. The lipid can be a fat or oil. Preferably, said lipid is of non-slaughter origin, including but not limited to plants, algae, fermentation, or laboratory-grown fat cells. Said lipid can solidify after absorption into the mycelium network to form a solid phase between the elongated pieces. Solidification can be caused by lowering the temperature or by enzymatic treatment.

[0108] The liquid may be enriched with flavor compounds, colorants, thickeners, fiber, vitamins, enzymes, trace elements, salts, acids, bases, fats, carbohydrates, polysaccharides, and / or proteins.

[0109] Additionally or alternatively, the absorbed liquid changes the pH of the fungus-containing food product, preferably lowers the pH, more preferably lowers the pH to an extent that the shelf life of the fungus-containing food product is longer than if the pH was not altered, and / or more preferably the altered pH growth conditions are selective for the inoculated strain or strains and / or prevent undesirable microbial growth.

[0110] Liquid absorption can be further achieved by applying vacuum or pressure or pressure fluctuation. Absorption of aqueous and lipid-based liquids can be performed in parallel or in any order at the same or different temperatures. The absorbed liquid can be an emulsion containing both aqueous and lipid-based liquids, or alternatively, when aqueous and lipid-based liquids are absorbed separately, they can form an emulsion in the interstitial phase.

[0111] Preferably, the liquid absorbed into the fungus-containing food product remains at least partially in the interstitial spaces between the elongated pieces until consumption, referred to as free liquid. Free liquid, in contrast to liquid that diffuses into the elongated pieces, can be pushed out of the fungus-containing food product and leave the fungus-containing food product during chewing, thus contributing to the perception of juiciness. Even more preferably, at least 10% by weight of the absorbed liquid is free liquid, even more preferably at least 20% by weight, and most preferably at least 50% by weight.

[0112] In an alternative embodiment, the absorbed liquid hydrates solubilize or liquefy some or all of the non-metabolized carbohydrates and / or hydrocolloids surrounding the elongated pieces. The absorbed liquid thus becomes more viscous or even gels in the interstitial spaces. In particular, after absorbing the liquid into the interstitial spaces and contacting the liquid with the non-metabolized carbohydrates and / or hydrocolloids such as agar or starch, the fungus-containing food product can be heated and cooled to gel the liquid with the carbohydrates or hydrocolloids in the interstitial spaces, which is referred to as "gelling in place". This may provide an additional phase similar to the collagen phase of animal meat products. Heating the fungus-containing food product prior to consumption melts or liquefies the gel in the interstitial spaces, again giving the perception of meat juices released during cooking or consumption.

[0113] The resulting fungus-containing food product may be cut and / or torn into pieces or slices and / or compressed to mimic the shape of a meat product.

[0114] Alternatively or additionally, the resulting fungus-containing food product may be washed with liquid water, steam or oil.

[0115] Alternatively or additionally, the resulting fungus-containing food product may be dried prior to further use.

[0116] Alternatively or additionally, the resulting fungus-containing food product may be marinated, spiced, smoked, salted, dehydrated, or post-processed in any other manner, as is typically done with animal meat products.

[0117] The resulting fungus-containing food may be cut into pieces, preferably mixed with nutrients and / or smaller pieces, reshaped or molded as required, and then re-incubated to allow further mycelium growth.

[0118] The fungus-containing food product may comprise a scaffold and at least one fungal mycelium growing along and through the scaffold, the scaffold being formed by one or more elongated pieces of wet textured protein product. The wet textured protein product preferably has a protein content of at least 10% by weight, more preferably at least 15% by weight.

[0119] Preferably, the first force required to pull apart the fibrous fungus-containing food product is lower than the second force required to pull apart the wet textured protein product. Preferably, the first force in this context is a force applied transverse to the longitudinal direction of the elongated pieces, or the first force is a force that pulls apart the fibrous fungus-containing food product along a direction parallel to the longitudinal direction of the elongated pieces. Preferably, the wet textured protein product comprises a fibrous structure in which the fibers are substantially aligned with one another, and the second force in this context is a force applied transverse to the direction of elongation of the fibers, or the second force is a force that pulls apart the wet textured protein product along the direction of elongation of the fibers.

[0120] Preferably the density of the fungus containing food product is lower than the density of the wet textured protein product, preferably the density of the fungus containing food product is less than 1 g / cm 3 or less, particularly immediately after incubation and prior to any subsequent steps of absorbing liquid into the food product or drying the food product.

[0121] Preferably, the oil absorption capacity of the fibrous fungus-containing food product is higher than the oil absorption capacity of the wet textured protein product, preferably the oil absorption capacity of the fibrous fungus-containing food product is at least 5% of its own weight, most preferably at least 10%.

[0122] Preferably, said fungus-containing food product is capable of absorbing at least 10% of its own weight of oil or water or a mixture of oil and water.

[0123] The fungus-containing food product preferably comprises elongated pieces of moist textured protein product interconnected with fungal mycelium having a protein content of at least 10% by weight, preferably at least 15% by weight, and at least 0.1% by weight starch between the elongated pieces.

[0124] Preferably, the fungus-containing food product is larger in at least one dimension than the wet textured protein product. Preferably, when prepared from a wet textured protein product produced by HMEC, the fungus-containing food product is larger in all three dimensions than the thickness of the HMEC wet textured protein product, said thickness of the HMEC wet textured protein product being given by the height of the cooling channels.

[0125] Preferably, the fungus-containing food product comprises at least 3, preferably at least 5, more preferably at least 10 elongate fragments.

[0126] Preferably, the fungus-containing food product comprises at least two distinct phases visible to the human eye, one phase consisting essentially of elongated fragments and one phase not consisting essentially of elongated fragments, preferably the latter phase comprises fungal mycelium, more preferably the latter phase comprises fungal mycelium and starch or lipids.

[0127] Preferably, the fungus-containing food product further comprises flavouring ingredients, colouring agents, spices, herbs, salts, acids or other food ingredients to adjust taste and appearance.

[0128] Preferably, the fungus-containing food product exhibits substantial shape stability and surface browning comparable to animal meat pieces when subjected to dry heat and oil, for example when baked or fried.

[0129] Preferably, said fungus-containing food product has at least two distinct attractive phases visible to the human eye, one phase consisting essentially of elongated fibrous fragments and one phase not consisting essentially of fibrous elongated fragments, more preferably the latter phase comprises fungal mycelium, and most particularly the latter phase comprises fungal mycelium and carbohydrates, e.g. starch, or lipids.

[0130] Preferably, said fungus-containing food product resembles an animal meat product, and more preferably, the colonized scaffold forming said fungus-containing food product resembles an animal muscle-like structure.

[0131] As used herein, the terms "comprising," "comprises," and "comprised of" are synonymous with "including" or "includes"; or "containing" or "contains," and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or steps. The terms "comprising," "comprises," and "comprised of," like the term "consisting of," should be understood to encompass certain embodiments where these terms are interpreted as closed-ended.

[0132] As used herein, the term "about" means approximately, within a region, roughly, or around. When the term "about" is used with a numerical value or range, it modifies the value or range by extending the boundaries above and below the stated numerical values. In general, the term "about" is used herein to modify a numerical value above and below the stated value by 10%. Whenever the term "about" is used, it should be understood that the specific value (i.e., without the use of the term "about") is also disclosed.

[0133] In this application, numerous documents, including patent applications, are cited. The disclosures of these documents are not considered relevant to the patentability of the present invention, but are incorporated herein by reference in their entirety. More specifically, all referenced documents are incorporated by reference to the same extent as if each individual document was specifically and individually indicated to be incorporated by reference. [Brief description of the drawings]

[0134] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] FIG. 1 shows a rectangular piece before incubation (left) and after incubation according to Example 1, bound into a coherent body by mycelium (right).

[0135] [Diagram 2] FIG. 2 shows the stress response of a cylindrical food product in uniaxial compression according to Example 2, with six different scaffolds differing in the fungus and nutrient supply used for binding.

[0136] [Diagram 3] Figure 3 shows a schematic of the preparation of the substrate before incubation in Example 3. The two pieces were sandwiched between two Petri dishes. a = Petri dish, b = rectangular piece, c = defined gap between the rectangular pieces.

[0137] [Figure 4] Figure 4 shows the stress-strain diagrams of uniaxial tensile tests of two pieces as shown in Figure 3 after being fused by mycelium at distances of 0, 1, and 2 mm. The results show higher bond strength with a lower initial distance between the pieces.

[0138] [Diagram 5] Figure 5 shows the oil uptake rates for samples of unbound loose strips and strips bound via mycelium from R. oligosporus, indicating an increased oil absorption capacity by the mycelium compared to the wet textured protein product alone.

[0139] [Figure 6] Figure 6 shows the water uptake rates of samples of unbound loose strips and strips bound via mycelium from R. oligosporus and A. oryzae, indicating a higher water uptake rate by mycelium compared to the wet textured protein product alone.

[0140] [Figure 7] Figure 7 shows the fibrous food product after incubation, in which the starch fragments are incorporated into the structure and attached to the elongated pieces by mycelium. The food product comprises elongated pieces and gelatinized starch strips. A cross section perpendicular to the longitudinal direction of the elongated pieces is shown.

[0141] [Figure 8] FIG. 8 shows a schematic diagram of the assembly of elongated and starch-based pieces into anisotropic bodies.

[0142] [Figure 9] FIG. 9 shows the food product of Example 6 containing elongated pieces and starch-based pieces after frying in a pan with oil, showing that the internal structure is fibrous, with the starch pieces providing a stable collagen-like layer between the fibrous pieces.

[0143] [Figure 10] FIG. 10 shows strips of various thicknesses and aspect ratios bonded to a food product to create different textures as described in Example 7.

[0144] [Figure 11] FIG. 11 shows stress-strain diagrams of strips with various thicknesses attached to food products measured under uniaxial compression as described in Example 7.

[0145] [Figure 12]FIG. 12 is a side schematic diagram showing, by way of example, how rods or tubes can be positioned within a generally cylindrical scaffold, with a=radial insertion and b=axial insertion.

[0146] [Figure 13] FIG. 13 shows pre-incubation scaffolds radially positioned within a 4 mm diameter rod scaffold as described for sample (iii) in Example 8, where a=metal rod, b=approximately cylinder.

[0147] [Figure 14] Figure 14 shows a close-up of the core of the food product after fermentation, where oxygen was delivered to the core during fermentation. The close-up shows good mycelium growth in the cavities between the strips (white / light cavities) indicating sufficient oxygen supply, as described in Example 9, a = mycelium growth in cavities, b = strips.

[0148] [Figure 15] Figure 15 shows a close-up of the cores of the food products after fermentation, where no oxygen was delivered to the cores during fermentation. The close-up shows no or only weak mycelial growth in the cavities between the strips (black / dark holes) indicating insufficient oxygen supply, as described in Example 9, a = no or only weak growth in the cavities, b = strips. EXAMPLES

[0149] Working Example Example 1 A wet textured protein product produced by high moisture extrusion cooking (HMEC) was provided, comprising 10% by weight glucose as fungal nutrient and 25% by weight pea protein isolate, 15% by weight pea fibre and 50% by weight water, cut into strips with a rectangular cross section of 2-4 mm x 2-4 mm and a length of at least 90 mm. A pea protein isolate content of 25% by weight corresponds to a pure protein content of 20% by weight. The wet textured protein products in all examples were produced by high moisture extrusion cooking in a twin screw extruder at a moisture content of more than 40% by weight and at a temperature of more than 120°C, followed by cooling in a cooling die to less than 100°C before exiting the die. The strips contained 4.8 x 10 granules per gram of substrate. 4 The plants were inoculated by covering their surfaces evenly with a powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram. The starter culture contained Rhizopus oligosporus spores and organic rice flour. 6 The inoculated elongated pieces were then arranged in parallel in a cylindrical shape and held together by a PVC pipe acting as a mold, thereby forming a scaffold. The scaffold was incubated in a fermenter (Hera Cell240i) at a temperature of 30° C. and a relative humidity of 95% for 38 hours. Although the pieces had to be held together by the mold before incubation, the pieces attached to each other after incubation through mycelium growth supplied by nutrients available in the pieces.

[0150] Figure 1 shows the fragments before (left) and after (right) incubation. After incubation, the product can be cut into fragments without losing adhesion. The elongated fragments and the mycelium-containing phase can be clearly distinguished by eye.

[0151] Example 2 Four different elongated pieces were prepared, all measuring 2-4 mm x 2-4 mm x 90 mm: (i) starch-containing elongated shreds made from a wet textured protein product produced by HMEC, comprising 12% by weight corn starch and 25% by weight pea protein isolate, 15% by weight pea fiber, and 48% by weight water. (ii) glucose-containing elongated pieces made from a wet textured protein product produced by HMEC, comprising 10% glucose by weight and 25% pea protein isolate, 15% pea fiber, and 50% water by weight; (iii) elongated pieces made from a wet textured protein product produced by HMEC, comprising 25% by weight pea protein isolate, 15% by weight pea fiber and 60% by weight water. (iv) Elongated fragments like (iii) that have been surface coated with glucose to achieve the same glucose concentration as (ii) but with potentially higher availability.

[0152] The strips were inoculated with either Aspergillus oryzae or Rhizopus oligosporus. The inoculum was 4.8 × 10 per gram of substrate for Rhizopus oligosporus. 4 viable spores and 3 × 10 for A. oryzae 5 This was done by covering all surfaces evenly with powdered starter cultures by mechanical mixing at a concentration of 8 × 10 viable spores per gram. The starter cultures consisted of Rhizopus oligosporus (oligosprus) spores and 8 × 10 viable spores per gram. 6 Organic rice flour with a spore concentration of 10 viable spores or Aspergillus oryzae and 7–9 × 10 per gram 8 The inoculated pieces were assembled into cylindrical scaffolds of 20 mm diameter according to Example 1 and incubated in a fermenter (Hera Cell240i) at a temperature of 30° C. and a relative humidity of 95% RH for 38 hours.

[0153] The resulting products with a diameter of approximately 20 mm were cut into samples with a height of 20 mm and compressed in a texture analyzer (TA.XT Plus, Stable Micro Systems) with a flat surface in the longitudinal direction of the elongated pieces at a compression speed of 0.5 mm / s. Uniaxial compression in the aforementioned direction of the pieces caused buckling of the elongated pieces. Thus, the stress response during buckling depended on the strength of the mycelial bonds between the pieces. As shown in Figure 2, the samples prepared with Rhizopus oligosporus showed a higher stress response overall, indicating stronger bonds. The sample prepared according to (i) showed the lowest adhesion, instead crumbling into pieces upon compression, indicating that the mycelium did not bond to the pieces, as in the case with glucose or without additional nutrients. Thus, the texture of the resulting food product can be adapted by adjusting the choice of nutrients and fungi according to the needs of the customer. The density of the resulting fibrous bodies was 1000 kg / m2 as summarized in Table 1. 3 The elongated pieces were less than 1000 kg / m 3 It has a higher solid density. [Table 1]

[0154] Example 3 A wet textured protein product produced by HMEC, containing 30% by weight soy protein concentrate, 10% by weight pea fiber, and 60% by weight water, was cut into pieces measuring 5×7×13 mm to give a pure protein content of 20% by weight. The pieces were coated with a glucose solution to give a glucose concentration of 10% by weight, and then 4.8×10 per gram of substrate was added. 4 The surfaces were inoculated by covering evenly with powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram of Rhizopus oligosporus (oligosprus) spores and organic rice flour. 6The two fragments contained a spore concentration of 1000 viable spores. The two fragments were placed with their radial cross-sections facing the Petri dish, as shown in Figure 3. The distance between the two fragments was adjusted to 0 mm, 1 mm, or 2 mm. A second Petri dish was placed on top of the fragments to prevent warping of the fragments. The Petri dish containing the assembled fragments was incubated in a fermenter (Hera Cell 240i) at 95% RH and 30 °C for 39 h.

[0155] After incubation, the two opposing pieces were fused at the opposing surfaces through mycelium growth. Tensile tests were performed on an Anton Paar Dynamic Mechanical Analyzer MCR 702 MultiDrive at 1% s -1 The adhesion between the two fragments was evaluated by stretching at an extension rate of 100 μm to 100 μm. As shown in Figure 4, the stiffness decreased with increasing distance between the fragments, while the strain at break increased with increasing distance, both of which indicated that a shorter distance results in a denser mycelial network and better adhesion.

[0156] Example 4 The oil absorption of the wet textured protein products produced by HMEC was compared between the mycelium bound fraction, i.e. the food product according to the invention, and the fraction without mycelium binding, i.e. the pure wet textured protein product.

[0157] Two different samples were prepared according to samples (ii) and (iii) in Example 2, all with sizes of 2-4 mm × 2-4 mm × 90 mm.

[0158] The aforementioned samples were inoculated with Rhizopus oligosporus. The inoculum was 4.8 × 10 per gram of substrate. 4 This was done by covering all surfaces evenly with a powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram of Rhizopus oligosporus (oligosprus) spores and organic rice flour. 6The inoculated pieces were assembled into cylindrical scaffolds with a diameter of 20 mm according to Example 1 and incubated in a fermenter (Hera Cell240i) at a temperature of 30° C. and a relative humidity of 95% RH for 38 hours. The resulting product was bound via mycelium, had a diameter of about 20 mm, and was cut into samples with a height of 20 mm.

[0159] Additionally, a control group of unbound loose fragments without mycelium, measuring 2-4 mm x 2-4 mm x 20 mm, was prepared from the same wet textured protein product as samples (ii) and (iii) of Example 2. The weight of the control group was adjusted to match the weight of the fungus-bound cylinders produced from fragments (ii) and (iii).

[0160] The oil absorption of the samples was tested by soaking the prepared pieces in sunflower oil for 120 minutes and measuring their weight before and after soaking. The control group without fungal attachment disintegrated during the test, whereas the products bound via mycelium remained whole. As shown in Figure 5, the oil absorption was increased in the fungal-bound products. Thus, the interstitial space in the product improves oil uptake. Without wishing to be bound by theory, it is believed that this effect can be used to enhance the juiciness, texture and flavor of consumer products.

[0161] Example 5 The water absorption of the wet textured protein product produced by HMEC was compared between the mycelium bound pieces, i.e. the food product according to the invention, and the pieces without mycelium binding, i.e. the pure wet textured protein product.

[0162] Elongated pieces, all measuring 2–4 mm × 2–4 mm × 90 mm, were prepared from a wet textured protein product produced by HMEC, containing 25% by weight pea protein isolate, 15% by weight pea fiber, and 60% by weight water. The aforementioned fibrous pieces were inoculated with either Aspergillus oryzae or Rhizopus oligosporus. The inoculum was 4.8 × 10 per gram of substrate for Rhizopus oligosporus. 4 viable spores and 3 × 10 for A. oryzae 5 This was done by covering all surfaces evenly with powdered starter cultures by mechanical mixing at a concentration of 8 × 10 viable spores per gram. The starter cultures consisted of Rhizopus oligosporus (oligosprus) spores and 8 × 10 viable spores per gram. 6 Organic rice flour with a spore concentration of 10 viable spores or Aspergillus oryzae and 7–9 × 10 per gram 8 The inoculated pieces were assembled into cylindrical scaffolds of 20 mm diameter according to Example 1 and incubated in a fermenter (Hera Cell240i) at a temperature of 30° C. and a relative humidity of 95% RH for 38 hours. The resulting product was bound via mycelium, had a diameter of about 20 mm, and was cut into samples of 20 mm height.

[0163] In addition, a control group of mycelium-free unbound loose strips measuring 2-4 mm x 2-4 mm x 20 mm was prepared from the same wet textured protein product as the mycelium-bound samples, produced by HMEC containing 25% by weight pea protein isolate, 15% by weight pea fiber, and 60% by weight water. The weight of the control group was adjusted to match the weight of the mycelium-bound cylinders produced from the strips.

[0164] The water absorption of the samples was tested by immersing the prepared products in distilled water for 14 minutes and measuring their weight before and after immersion.

[0165] The control group without fungal binding fell apart during the test, while the product bound via mycelium remained whole. As shown in Figure 6, water absorption was increased through mycelium binding of the fragments. Thus, the interstitial space in the product improves water absorption. Without wishing to be bound by theory, it is believed that this effect can be used to enhance the juiciness, texture and flavor of consumer products.

[0166] Example 6 A wet textured protein product containing >20% by weight pea protein isolate and 10% by weight sunflower protein concentrate, 10% by weight pea fiber, and 60% by weight water was torn into pieces ranging from 2–7 mm × 2–7 mm × 40–80 mm and mixed with pregelatinized potato starch powder to reach a starch concentration of 10% by weight. The starch-coated pieces were placed in vacuum bags and pasteurized in a 95°C water bath for 25 minutes. The pieces were partially glued together due to partial swelling of the starch granules caused by the heat treatment. The pieces were loosened and 4.8 × 10 per gram of substrate were mixed. 4 The plants were inoculated by covering their surfaces evenly with a powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram. The starter culture contained Rhizopus oligosporus spores and organic rice flour. 6 The pieces were assembled into a sausage-like shape with the orientation of each piece parallel to the length of the sausage-like shape, resulting in an elongated structure.

[0167] As a further test sample, the same wet textured protein product containing 20% ​​by weight of pea protein isolate and 10% by weight of sunflower protein concentrate, 10% by weight of pea fibre and 60% by weight of water was torn into pieces in the range of 2-7 mm x 2-7 mm x 40-80 mm. The fibre pieces were placed in vacuum bags and pasteurised in a water bath at 95°C for 25 minutes. A crumbly dough was formed by mixing 100% native potato starch (Agrana Starke GMBH, Starkina, 20001) with 50-65% tap water, where the percentages (%) refer to the total weight of native potato starch (100%). The starch-water dough was spread into a layer 1-3 mm thick, vacuum packed and pasteurised in a water bath at 95°C for 25 minutes. Starch gelatinization was achieved during pasteurization, resulting in elastic, pliable starch plates that were cut into long strips measuring 60–70 mm × 4–10 mm. The long strips and starch strips were mixed at 4.8 × 10 per gram of substrate. 4 The plants were inoculated by covering their surfaces evenly with a powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram of starter culture. The starter culture contained Rhizopus oligosporus spores and organic rice flour, with a concentration of 8 × 10 viable spores per gram of starter culture. 6 The pieces and starch strips were mixed and assembled into a sausage-like shape with the longitudinal orientation of the elongated pieces and starch strips parallel to the length of the sausage-like shape, resulting in an elongated scaffold structure as shown diagrammatically in FIG.

[0168] The scaffolds were wrapped in polyethylene film and compressed manually to decrease the distance between the fragments and increase the packing density. The polyethylene film was perforated with 1 mm wide holes every 7-10 mm in a square mesh pattern. After 48 h of incubation at 30 °C and 95% relative humidity, the surface and voids of the intermediate product were filled with fungal mycelium, and the body was very tightly packed. Some of the starch granules were not fermented by the fungus, thus filling some of the voids between the fungal mycelium. To terminate the fermentation, the products were either stored at 4 °C, frozen, pasteurized in a vacuum bag immersed in a 95 °C water bath for 25 min, or a combination of these. As can be seen in Figure 7, the gelatinized starch phase was bound to the product by the mycelium. Before and after preparation by pan-frying, the starch phase had the visual appearance and mechanical similarity of fat or connective tissue. As can be seen in Figure 9, the internal structure was highly fibrous. During consumption of the final product after pan frying, the mycelium contributed to the cohesive, firm texture, the fibrous, elongated pieces contributed to the meat-like fibrous texture, and the gelatinized starch granules melted in the mouth, providing the sliminess known from the connective tissue of meat.

[0169] Example 7 Elongated pieces of various thicknesses were all prepared from a wet textured protein product containing greater than 25% by weight soy protein isolate, 15% by weight sunflower protein, and 5% by weight soy protein concentrate, and 55% by weight water. The pieces were surface coated with glucose to achieve a glucose concentration of 10% by weight: (i) Size: 1-2 mm x 1-2 mm x 90 mm (ii) Size: 4-6 mm x 4-6 mm x 90 mm

[0170] The aforementioned fragments were inoculated with either Aspergillus oryzae or Rhizopus oligosporus, assembled into a 20 mm diameter cylinder according to Example 1, and incubated for 38 hours in a fermenter (Hera Cell240i) at a temperature of 30° C. and a relative humidity of 95% RH.

[0171] The resulting products with a diameter of about 20 mm were cut into samples with a height of 20 mm as shown in FIG. 10. The samples were compressed in the length direction of the elongated pieces in a texture analyzer (TA.XT Plus, Stable Micro Systems) at a compression speed of 0.5 mm / s. Uniaxial compression in the aforementioned direction of the pieces caused the buckling of the fibrous pieces. Thus, the stress response during buckling depended on the mycelial bond strength between the pieces as well as the mechanical and intrinsic mechanical properties of the pieces along their length. As shown in FIG. 11, varying the thickness (diameter) of the pieces results in different stress responses of the products. This leads to the sensory perception of the products in terms of texture. Thus, it is believed that the thickness (diameter) of the pieces allows adapting the hardness, elasticity, and bond of the products.

[0172] The main phase of chewing is dominated by the sensation of breaking the body into pieces, whereas the subsequent phases of chewing are dominated by the overall texture sensation of the fibrous structure, so that when eaten, the texture of products made from thicker pieces can be described as heterogeneous. The texture of products made from thinner pieces is perceived as more homogeneous, with less differentiation between the phases of breaking the body and chewing the fibers.

[0173] Example 8 To increase the oxygen concentration in the center of the body, continuous tubular pores (channels) were created within the body to facilitate the flow of fresh air into the interior of the body.

[0174] A wet textured protein product containing 20% ​​by weight pea protein isolate and 10% by weight sunflower protein concentrate, 10% by weight pea fiber, and 60% by weight water was torn into pieces ranging from 2-7 mm x 2-7 mm x 40-80 mm. The pieces were placed in vacuum bags and pasteurized in a 95°C water bath for 25 minutes.

[0175] For elongated fragments, 4.8 × 10 per gram of substrate 4The plants were inoculated by covering their surfaces evenly with powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram of starter culture. The starter culture contained Rhizopus oligosporus spores and organic rice flour, with a concentration of 8 × 10 viable spores per gram of starter culture. 6 The spore concentration of 1000 viable spores was 100%. The aforementioned fragments were mixed and assembled into a sausage-like shape with the longitudinal direction of the elongated fragments parallel to the length of the sausage-like body, resulting in an elongated scaffold structure. The scaffold was wrapped in polyethylene film and compressed manually to decrease the distance between the fragments and increase the packing density. The polyethylene film was drilled with 1 mm wide holes every 5-10 mm in a square mesh pattern.

[0176] The following bodies were produced, some of which had placeholders placed within the body to provide continuous tubular pores within the body: (i) A 1 cm 2 A hollow plastic tube of dimensions 4 x 15 mm perforated with holes of size 1-2 mm at a density of 5 holes per tube (see "b" in Figure 12). (ii) A metal rod of dimensions 4 x 15 mm arranged axially within the cylinder (see "b" in Figure 12). (iii) A metal rod with dimensions: 4×15 mm arranged radially within the cylinder as shown in FIG. 13 (see “a” in FIG. 12). (iv) A metal rod of dimensions 2 x 15 mm arranged radially within the cylinder (see "a" in Figure 12). (v) control sample without placeholder

[0177] During layering of the elongated pieces, the placeholders (i) and (ii) were placed axially within the body. In this way, the elongated pieces were arranged around the tubular placeholder. The placeholders (iii), (iv) were pierced radially into the sausage-like body after layering of the elongated pieces, so that the fibrous pieces were penetrated by the rods.

[0178] After incubation at 40°C for 6 hours, then at 30°C for 5 hours, then at 24°C for 3 hours, then at 20°C for 10 hours, and at 23°C for 14 hours, while maintaining the relative humidity at 95%, the surface and voids of the scaffold showed mycelium growth during a total fermentation time of 38 hours. After 12 hours of incubation time, the primary mycelium growth had given the product structural integrity, and the tubular placeholders (ii), (iii) and (iv) were removed from the body, leaving continuous tubular holes in the body to aid in internal aeration and increase oxygen diffusion to the growing mycelium. At this time, the polyethylene film was removed from the longitudinal end faces of the cylinder to further aid in aeration. The polyethylene film on the circumferential side of the cylinder was not removed. After 38 hours of incubation, the surface and voids of the scaffold were filled with fungal mycelium and the body was very tightly attached. To terminate the fermentation, the product was stored at 4°C, frozen, or a combination thereof. [Table 2]

[0179] The control without the placeholder did not show any discernible mycelium formation in the center of the body. As shown in Table 2, strong mycelium growth was detected at a depth of 20-25 mm from the lateral surface. Areas below this depth showed sparse mycelium growth or no visible mycelium.

[0180] All bodies (i)-(iv) showed mycelium growth up to the center of the sample. Depending on the characteristics of the pores brought by the placeholder, the mycelium binding in the center of the sample was stronger or weaker. The body with perforated tubes (i) and the body with radial 4 mm rods (iii) showed the most uniform growth and good internal mycelium binding of the substrate fragments throughout the sample. The body with axial 4 mm rods (ii) and the body with radial 2 mm rods (iv) showed mycelium growth in the center of the body, but the binding capacity was not as strong, which demonstrated the possibility of adapting the binding capacity of the mycelium by pore design. The results show that adding continuous tubular pores to the body, which connects the pores to the outside, increases the internal mycelium growth compared to the body without pores.

[0181] It was observed that in all samples, strong mycelial growth occurred towards the longitudinal ends of the cylinder, at a depth of 40-45 mm measured from the end face of the cylinder. This could be attributed to increased axial oxygen diffusion into the samples due to the removal of the polyethylene film after 12 hours of incubation. At the same time, the removal of the polyethylene film caused drying from the surface of the face of the cylinder, which weakened the mycelial growth on the surface of the aforementioned area. Therefore, additional supply of oxygen and adjustment of the relative humidity are desirable to produce larger pieces of meat analogue.

[0182] Example 9 A wet textured protein product prepared by high moisture extrusion cooking containing 20% ​​by weight soy protein isolate and 10% by weight sunflower protein concentrate, 10% by weight citrus fiber, and 60% by weight water was torn into elongated pieces in the range of 2-7 mm x 2-7 mm x 40-80 mm, the pieces were vacuum bagged and pasteurized in a 95°C water bath for 25 minutes.

[0183] For elongated fragments, 4.8 × 10 per gram of substrate 4The plants were inoculated by covering their surfaces evenly with powdered starter culture by mechanical mixing at a concentration of 8 × 10 viable spores per gram of starter culture. The starter culture contained Rhizopus oligosporus spores and organic rice flour, with a concentration of 8 × 10 viable spores per gram of starter culture. 6 The cylindrical body had a spore concentration of 10 ...

[0184] A plastic tube with a diameter of 4 mm, closed at its ends and measuring 1 cm for a length of 50 mm from its ends. 2 A plastic tube with 0.5 mm wide holes drilled at a density of approximately 30 holes per tube was affixed axially to the center of one of the bodies (see "a" in Figure 12). The other body was left unchanged and served as a control. The tube was then placed in a 200 lh -1 The scaffolds were connected to an aquarium pump (EHEIM air400) that continuously pumped air through the samples at a flow rate of 100 s. The pumped air was thermostated to the incubator temperature and humidified to 80–100% RH. The scaffolds were incubated at 30 °C and 95% relative humidity for the first 10 h, then at 20 °C for an additional 5 h, and then at 18 °C for 14 h.

[0185] After 29 hours of incubation, the incubation was terminated by cooling the samples in a refrigerator. In the sample with continuous aeration by the inserted tube connected to the pump, the surface and voids of the product were uniformly filled with fungal mycelium and the body was very tightly packed and rigid, as shown enlarged in Figure 14. The control sample without continuous aeration showed mycelium growth only at the edges of the sample and had no mycelium attachment in the center, as shown enlarged in Figure 15. This experiment demonstrates the possibility of increasing internal mycelium growth and uniform growth of mycelium within large bodies via pressurized air injection.

[0186] Preferred embodiments of the present invention are defined by the following aspects: 1. A method for providing a food product containing a filamentous fungus, comprising: (i) providing one or more elongated pieces made from a wet textured protein product; (ii) providing nutrients for fungal growth in or around the one or more elongate segments; (iii) inoculating said one or more elongated segments with at least one fungus; (iv) using one or more elongated pieces to form a scaffold for mycelial growth; (v) incubating the scaffold under growth conditions that permit mycelial growth of the at least one fungus such that the at least one fungus forms a mycelium, which mycelium grows along and through the scaffold to form a filamentous fungus-containing food product; A method comprising:

[0187] 2. The method of embodiment 1, wherein the wet textured protein product comprises at least 10% by weight protein, preferably at least 15% by weight protein.

[0188] 3. The method of any of the preceding embodiments, wherein the wet textured protein product is produced by subjecting a material having at least 10% by weight protein and at least 35% by weight water to shear and a temperature greater than 100°C, preferably greater than 120°C, preferably by high moisture extrusion cooking (HMEC) or shear cell processing (SC).

[0189] 4. The method of any of the previous aspects, wherein the wet textured protein product comprises a moisture content of more than 35% by weight, preferably at least 40% by weight.

[0190] 5. The density of the wet textured protein product is less than 0.8 g / cm 3 Equal to or greater than 0.9 g / cm 3 more preferably 1 g / cm 3 20. The method of any one of the preceding aspects.

[0191] 6. The method of any of the previous aspects, wherein the wet textured protein product comprises a fibrous structure in which the fibers are substantially aligned with one another.

[0192] 7. one or more elongate segments each have a longest dimension, e.g., a length, equal to or greater than 1 cm, preferably equal to or greater than 2 cm, more preferably equal to or greater than 4 cm; and / or The method of any of the preceding embodiments, wherein the one or more elongate segments each have a shortest dimension, e.g., diameter, equal to or less than 2 cm, preferably equal to or less than 1 cm, more preferably equal to or less than 0.5 cm.

[0193] 8. The method of embodiment 6 or 7, wherein the fibers of the elongate segment are substantially aligned with the longitudinal direction of said elongate segment. 9. The fibers of the plurality of elongated segments are aligned along substantially the same direction; or A method according to any of aspects 6 to 8, wherein the fibers of the first plurality of elongate segments are substantially aligned along a first direction and the fibers of the second plurality of elongate segments are substantially aligned along a second direction that is different from the first direction, preferably at an angle of at least 30°.

[0194] 10. The method of any of the preceding embodiments, wherein said wet textured protein product comprises at least 10% by weight of protein selected from the group consisting of pea, soybean, wheat, sunflower, pumpkin, rice, cereals, bulus, oilseed, algae, single cell, fungus, and fermented ingredients.

[0195] 11. The method of any of the previous aspects, wherein the scaffold is formed by at least 2 elongate fragments, preferably at least 10 elongate fragments.

[0196] 12. The method of any of the preceding embodiments, wherein the wet textured protein product is treated into one or more elongated pieces by cutting, tearing, rolling, or by immersion in a liquid to the extent that the wet textured protein product disintegrates into one or more elongated pieces.

[0197] 13. The method of any of the preceding embodiments, wherein the one or more elongate fragments are in the form of a strand or sheet, respectively.

[0198] 14. The method of embodiment 13, wherein forming the scaffold comprises assembling a plurality of strands into a bundle, rolling up one or more sheets into a tube or multi-layer roll, or folding or stacking one or more sheets.

[0199] 15. The method of any of the previous aspects, wherein forming the scaffold comprises assembling differently engineered or processed elongated pieces to create a gradient of a substance, such as a nutrient gradient, throughout the scaffold.

[0200] 16. The method of any of the preceding aspects, wherein one dimension of each elongate segment is longer than the other dimension, preferably at least 2 times longer, and most preferably at least 5 times longer.

[0201] 17. The method of any of the preceding aspects, wherein nutrients are provided inside or on the one or more elongated pieces, for example by adding nutrient-containing ingredients to a high moisture extrusion cooking or shear cell process whereby the wet textured protein product is produced, or by immersing the wet textured protein product or the one or more elongated pieces in a nutrient solution, or by coating the one or more elongated pieces with a nutrient-containing liquid or powder.

[0202] 18. The method according to any of the preceding aspects, wherein the nutrients are selected from the group of monosaccharides, disaccharides, or oligosaccharides acting as nutrients for the respective fungi, preferably the nutrients are selected from the group of starch, glucose, sucrose or malt starch.

[0203] 19. At least partially coating one or more surfaces, preferably one or more outer surfaces, of the one or more elongated pieces with a carbohydrate and / or hydrocolloid, the carbohydrate and / or hydrocolloid preferably being selected from the group of starch, carrageenan, konjac, agar, alginate, xanthan, gellan gum, pectin, gelatin or combinations thereof, preferably the one or more elongated pieces being coated with starch, more preferably coated with gelatinized starch, prior to incubation; Preferably, one or more surfaces are coated with a carbohydrate and / or hydrocolloid, preferably starch, to the extent that the carbohydrate and / or hydrocolloid is not completely metabolized by the fungus during incubation, and / or A method according to any of the preceding aspects, wherein the one or more elongate pieces are preferably coated with at least 2 wt.-% carbohydrate and / or hydrocolloid, preferably starch, more preferably at least 5 wt.-% carbohydrate and / or hydrocolloid, preferably starch, prior to incubation, based on the total weight of the one or more elongate pieces.

[0204] 20. The method of any of the preceding embodiments, wherein the one or more elongated pieces are dried or hydrated prior to inoculation to adjust the water activity to the conditions required for fungal growth.

[0205] 21. The method of any of the preceding embodiments, wherein the one or more elongated pieces are sterilized or pasteurized prior to inoculation.

[0206] 21.a. The method of any of the preceding aspects, wherein the surface of the elongate pieces is treated with an acid, preferably a food grade acid, more preferably lactic acid, acetic acid, malic acid, citric acid or succinic acid, preferably to reach a pH at the surface of less than 6, even more preferably less than 5, and most preferably not exceeding 4.6.

[0207] 22. The method according to any of the preceding aspects, wherein the one or more elongated pieces are inoculated with at least one fungus by mixing them with a fungus-containing liquid or powder, preferably said fungus being selected from the group consisting of Ascomycetes, Basidiomycetes, Deuteromycetes, Oomycetes and / or Zygomycetes, in particular Rhizopus oligosporus, Rhizopus delemar, Rhizopus oryzae, Penicillium nalgiovense, Penicillium camemberti, Penicillium roqueforti, Aspergillus oryzae, Aspergillus luchuensis, Aspergillus sojae, Ganoderma lucidum, Pleurotus ostreatus, Pleurotus eryngii, or a combination thereof.

[0208] 23. The method of any of the preceding aspects, wherein one or more surfaces, preferably one or more outer surfaces, of the one or more elongate pieces are pre-wetted prior to inoculation with the fungus-containing powder.

[0209] 24. The method of any of the preceding embodiments, wherein at least three elongate pieces are assembled with or without a mold to form a scaffold, and preferably the distance between adjacent pieces is adjusted by vibration or application of pressure or vacuum such that each piece is adjacent to an adjacent piece by a distance of 2 mm or less, preferably 1 mm or less, and even more preferably 0.5 mm or less.

[0210] 25. The method of any of the preceding aspects, wherein at least 5, preferably at least 10, elongate fragments are assembled to form bundles in which the elongate fragments are substantially aligned with one another, preferably the scaffold comprises some bundles that are not aligned with one another.

[0211] 26. The method according to any of the preceding aspects, wherein one or more carbohydrate-based fragments, preferably starch-based fragments, are provided to the scaffold, preferably before incubation, preferably said one or more carbohydrate-based fragments, preferably starch-based fragments, are arranged between two or several elongate fragments, and even more preferably said one or more starch-based fragments comprise at least 5 wt.-% starch, preferably at least 10 wt.-%, more preferably at least 20 wt.-% starch, based on the total weight of the one or more starch-based fragments.

[0212] 27. The method of the preceding embodiment, wherein the one or more carbohydrate-based pieces, preferably starch-based pieces, are each in the form of a cube, rod, cylinder or stripe and are made from at least water and a carbohydrate selected from the group of starch, carrageenan, konjac, agar, alginate, pectin or combinations thereof, preferably starch being gelatinized, e.g. by heating to a temperature above the gelatinization temperature, and optionally cut.

[0213] 28. Before incubation, the scaffold at least 1 cm, preferably at least 2 cm, more preferably at least 4 cm, even more preferably between 4 cm and 15 cm in at least one dimension; and / or 13. The method of any preceding embodiment, wherein the shortest dimension is no more than 30 cm, even more preferably no more than 20 cm.

[0214] 29. The method of any of the preceding aspects, wherein the scaffold is incubated at a temperature between 4°C and 70°C, preferably between 10°C and 50°C, even more preferably between 14°C and 20°C or between 22°C and 38°C for a minimum of 2 hours, preferably for a minimum of 12 hours, more preferably for a minimum period of time such that mycelium becomes visible to the human eye on the surface of one or more elongated fragments, preferably wherein incubation is terminated before fungal sporulation or fruiting body formation.

[0215] 30. The method of any of the preceding aspects, further comprising the step of wrapping the scaffold in an outer layer prior to incubation, said layer may have holes or may be oxygen semi-permeable or oxygen permeable, preferably said outer layer is a plastic foil having holes, and / or said outer layer is oxygen permeable but not water permeable.

[0216] 31. The method according to any of the preceding aspects, wherein the scaffold is incubated in hypoaerobic conditions, where the oxygen concentration is lower than under atmospheric conditions, in particular by covering the scaffold with a material such that the oxygen permeability coefficient is less than 1 but greater than 0.

[0217] 32. Holes and / or channels are created in the scaffold before or during incubation; The method of any of the preceding aspects, wherein the holes and / or channels are closed after incubation, more preferably by subjecting the filamentous fungus-containing food product to a heat treatment or by adding a liquid to the filamentous fungus-containing food product.

[0218] 33. The method of any of the preceding aspects, wherein prior to or during the incubation, one or more placeholders, such as one or more tubes or one or more rods or one or more structures having a branched shape, are inserted into the scaffold, wherein the one or more tubes or rods extend axially, radially or in any direction within the scaffold, preferably along the shortest dimension of the scaffold.

[0219] 34. The method of embodiment 33, wherein the one or more tubes or rods have a diameter of at least 0.1 mm, preferably at least 1 mm, even more preferably at least 2 mm, preferably no greater than 15 mm, more preferably no greater than 10 mm, and most preferably no greater than 5 mm.

[0220] 35. The method of the preceding embodiment, wherein one or more tubes are oxygen permeable and / or one or more tubes comprise a vent tube, preferably one or more tubes comprise one or more hollow needles.

[0221] 36. The method of any of aspects 33-35, wherein one or more tubes communicate the interior portion of the scaffold with the surrounding environment.

[0222] 37. The method of any of aspects 33-36, wherein one or more tubes are removed during or after incubation, or one or more tubes are edible and are not removed prior to consumption.

[0223] 38. The method of any of aspects 33-37, wherein one or more tubes are covered with a protective culture to prevent fungi from growing within the respective tube.

[0224] 39. The method according to any of aspects 33-38, wherein at least one tube is connected to a source of oxygen, preferably pressurized air or oxygen, or a pump.

[0225] 40. The pressurized air or oxygen is preferably humidified to a relative humidity of at least 80%, at least 90%, or at least 95% before entering the scaffold; and / or 2. The method according to the preceding aspect, wherein said pressurized air or oxygen is conditioned to a temperature of preferably at least 4°C, at least 10°C, or at least 14°C and / or below 50°C, preferably below 45°C, more preferably below 40°C, prior to entering the scaffold.

[0226] 41. A method according to any of aspects 33-40, wherein the one or more rods are inserted before or at the start of incubation and removed after at least 2 hours of incubation, preferably after at least 6 hours of incubation, even more preferably after at least 10 hours of incubation, or even more preferably when mycelium growth within the scaffold becomes visible to the human eye.

[0227] 42. The method of any of the preceding aspects, further comprising the step of (vi) interrupting the growth of the at least one fungus, preferably by changing the temperature and / or water activity to lower or higher temperature and water activity conditions than are necessary for the growth of the at least one fungus, said interruption preferably occurring before sporulation or fruiting body formation of the fungus.

[0228] 43. The method of any of the preceding aspects, comprising exposing the resulting fungus-containing food product to a liquid such that the food product absorbs the liquid, said liquid being water-based or lipid-based, and the liquid completely or partially filling the interstitial spaces between the elongated pieces and the mycelium-filled spaces and allowing it to diffuse into the elongated pieces.

[0229] 44. The method of the preceding aspect, wherein the liquid is further enriched with flavor compounds, colorants, thickeners, fiber, vitamins, enzymes, trace elements, salts, acids, bases, fats, carbohydrates, polysaccharides, or proteins.

[0230] 45. The method according to the two preceding aspects, when according to aspect 19, the absorbed liquid completely or partially solubilizes or liquefies not completely metabolized carbohydrates and / or hydrocolloids, preferably said carbohydrates and / or hydrocolloids increase the viscosity of said absorbed liquid in the interstitial spaces or gel said absorbed liquid.

[0231] 46. ​​The method of any of the preceding aspects, wherein the resulting fungus-containing food product is cut and / or torn into pieces or slices and / or compressed to mimic the shape of a meat product.

[0232] 47. The method of any of the preceding aspects, wherein the resulting fungus-containing food product is dried before further use.

[0233] 48. A filamentous fungus-containing food product obtainable by the method according to any of the preceding aspects.

[0234] 49. A filamentous fungus-containing food product comprising a scaffold and at least one fungal mycelium growing along and through the scaffold, the scaffold being formed by one or more elongated pieces made from a wet textured protein product.

[0235] 50. A filamentous fungus-containing food according to aspect 47 or 48 or a method according to any one of aspects 1 to 46, wherein a first force required to separate the filamentous fungus-containing food is lower than a second force required to separate the elongated pieces.

[0236] 51. The filamentous fungus-containing food product of aspect 49, wherein the first force and the second force are applied transversely to the longitudinal direction of the elongated piece.

[0237] 52. The wet textured protein product comprises a fibrous structure in which the fibers are substantially aligned with one another; A fibrous fungus-containing food product according to aspect 49 or 50, wherein the bonds between the substantially aligned fibers are stronger than the bonds between the elongated fragments in the fibrous fungus-containing food product.

[0238] 53. The density of the fungus-containing food product is less than the density of the wet textured protein product, preferably the density of the fungus-containing food product is less than 1 g / cm 3 52. The method according to any of aspects 47 to 51 or the method according to any of aspects 1 to 46, wherein the filamentous fungus-containing food product is further treated by subjecting the food product to a subsequent step of absorbing liquid into the food product or drying the food product, in particular immediately after incubation and before a subsequent step of absorbing liquid into the food product or drying the food product.

[0239] 54. A filamentous fungus-containing food according to any of aspects 47 to 52 or a method according to any of aspects 1 to 46, wherein the oil absorption capacity of the filamentous fungus-containing food is higher than the oil absorption capacity of the wet textured protein product, preferably the oil absorption capacity of the filamentous fungus-containing food is at least 5% of its own weight, most preferably at least 10%.

[0240] 55. A filamentous fungus-containing food product according to any of aspects 47 to 53, further comprising at least 0.1% by weight of carbohydrate, preferably starch, and / or carbohydrate-based fragments, preferably starch-based fragments, in the interstitial spaces of the scaffold.

[0241] 56. The filamentous fungus-containing food product of any of aspects 47 to 54, further comprising one or more channels extending from the surface of the food product to its interior, the channels having a diameter of at least 0.1 mm, preferably at least 1 mm, even more preferably at least 2 mm, and / or preferably no greater than 15 mm, more preferably no greater than 10 mm, and most preferably no greater than 5 mm.

[0242] 57. The filamentous fungus-containing food product of the preceding embodiment, wherein the one or more channels are filled with mycelium of at least one fungus.

[0243] 58. A fibrous fungus-containing food product comprising elongated pieces of moist textured protein product interconnected with fungal mycelium, the elongated pieces having a protein content of at least 10% by weight, preferably at least 15% by weight, and at least 0.1% by weight starch.

[0244] 59. A fungus-containing food product according to any of aspects 47 to 57, wherein the product is capable of absorbing at least 5%, preferably at least 10%, of its own weight in oil or water or a mixture thereof.

[0245] 60. A fungus-containing food product according to any of aspects 47 to 58, comprising at least two distinct attractive phases visible to the human eye, a first phase consisting essentially of one or more elongated fragments and a second phase not consisting essentially of one or more elongated fragments, preferably the second phase comprising fungal mycelium, more preferably the second phase comprising fungal mycelium and a carbohydrate, preferably starch or lipid.

[0246] 61. The food product is at least 2 cm in at least one dimension, more preferably at least 4 cm, and even more preferably between 4 cm and 15 cm; and / or 60. The fungus-containing food according to any one of aspects 47 to 59, having a shortest dimension of 30 cm or less, even more preferably 20 cm or less.

[0247] 62. A fungus-containing food product according to any one of aspects 47 to 60, wherein said food product contains fungal mycelium throughout the product.

[0248] 63. A fungus-containing food product obtainable by the method according to any of aspects 1 to 46, comprising at least 10% by weight of vegetable protein, preferably at least pea protein, at least one unfermented starch, and fungal mycelium at the interface between at least two elongated pieces.

Claims

1. 1. A method of providing a food product containing a filamentous fungus, comprising: (i) providing one or more elongated pieces made from a wet textured protein product; (ii) providing nutrients for fungal growth in or around the one or more elongate segments; (iii) inoculating the one or more elongate segments with at least one fungus; (iv) using the one or more elongated pieces to form a scaffold for mycelial growth; (v) incubating the scaffold under growth conditions that permit mycelial growth of the at least one fungus such that the at least one fungus forms mycelium, the mycelium growing along and through the scaffold to form a filamentous fungus-containing food product; A method comprising:

2. the wet textured protein product is produced by high moisture extrusion cooking (HMEC) or shear cell processing (SC); Preferably, the wet textured protein product comprises a moisture content of more than 35% by weight, more preferably at least 40% by weight; More preferably, the wet textured protein product comprises a fibrous structure in which the fibers are substantially aligned with one another.

3. the one or more elongate segments each have a longest dimension, e.g., length, of at least 1 cm, preferably at least 2 cm, more preferably at least 4 cm; or 2. The method of claim 1, wherein the one or more elongated segments each have a shortest dimension, e.g., diameter, of less than 2 cm, preferably less than 1 cm, more preferably less than 0.5 cm.

4. i) the fibers of the elongate segment are substantially aligned with the longitudinal direction of said elongate segment; ii) the fibers of the plurality of elongate segments are aligned along substantially the same direction; or iii) the fibers of the first plurality of elongate segments are substantially aligned along a first direction, and the fibers of the second plurality of elongate segments are substantially aligned along a second direction that is different from the first direction, preferably by an angle of at least 30°.

5. the one or more elongate segments are each in the form of a strand or a sheet; 10. The method of claim 1, wherein forming the scaffold preferably comprises assembling a plurality of strands into a bundle, winding one or more sheets into a tube or multi-layer roll, or folding or stacking one or more sheets into a stack.

6. 10. The method of claim 1, wherein forming the scaffold comprises assembling differently processed or treated elongated segments to create a gradient of a substance, such as a gradient of a nutrient, throughout the scaffold.

7. one or more surfaces, preferably one or more outer surfaces, of said one or more elongate pieces are at least partially coated with a carbohydrate and / or a hydrocolloid, preferably with starch, preferably with gelatinized starch, prior to incubation, Preferably, said one or more surfaces are coated with a carbohydrate and / or hydrocolloid, preferably starch, to the extent that said carbohydrate, hydrocolloid or starch is not completely metabolised by said fungus during incubation, and / or 2. The method of claim 1, wherein the one or more elongated pieces are preferably coated with at least 2% by weight of carbohydrate, hydrocolloid or starch, more preferably at least 5% by weight of carbohydrate, hydrocolloid or starch, based on the total weight of the one or more elongated pieces, prior to incubation.

8. one or more carbohydrate-based fragments, preferably starch-based fragments, are provided in the scaffold, preferably prior to said incubation; 2. The method of claim 1, wherein the one or more carbohydrate-based fragments, preferably starch-based fragments, are preferably arranged between two or several elongated fragments, and even more preferably the one or more starch-based fragments comprise at least 5% by weight of starch, preferably at least 10% by weight, more preferably at least 20% by weight of starch, based on the total weight of the one or more starch-based fragments, and most preferably the one or more starch-based fragments are made from at least water and gelatinized starch.

9. holes and / or channels are created in the scaffold before or during incubation; The method of claim 1, wherein the holes and / or channels are preferably closed after incubation, more preferably by subjecting the filamentous fungus-containing food to a heat treatment or by adding a liquid to the filamentous fungus-containing food.

10. one or more tubes or rods are inserted into the scaffold before or during incubation, the one or more tubes or rods extending axially, radially or in any direction within the scaffold, preferably along the shortest dimension of the scaffold; 2. The method of claim 1, wherein the one or more tubes or rods preferably have a diameter of at least 0.1 mm, preferably at least 1 mm, even more preferably at least 2 mm, and preferably no greater than 15 mm, more preferably no greater than 10 mm, and most preferably no greater than 5 mm.

11. 11. The method of claim 10, wherein at least one tube is connected to a source of oxygen, preferably pressurized air or pressurized oxygen, or to a pump.

12. 10. The method of claim 1, further comprising exposing the resulting fungus-containing food product to a liquid such that the food product absorbs the liquid, the liquid being water-based or lipid-based.

13. A filamentous fungus-containing food product comprising a scaffold and mycelium of at least one fungus grown along and through said scaffold, said scaffold being formed by one or more elongated pieces made from a wet-textured protein product.

14. the fungus-containing food product is composed of at least two distinct phases, preferably the two distinct phases are distinguishable by the human eye, more preferably a first phase consisting essentially of the one or more elongated fragments and a second phase not consisting essentially of the one or more elongated fragments, 14. The fungus-containing food product of claim 13 or the method of any of claims 1 to 12, wherein the second phase preferably comprises fungal mycelium, more preferably the second phase comprises fungal mycelium and a carbohydrate, preferably a starch or a lipid.

15. i) a first force required to separate the filamentous fungus-containing food product is less than a second force required to separate the moist textured protein product; ii) the density of the fungus-containing food product is lower than the density of the wet textured protein product, preferably the density of the fungus-containing food product is less than 1 g / cm, especially immediately after incubation and before a subsequent step of absorbing liquid into the food product or drying the food product. ３ and / or iii) The oil absorption capacity of the filamentous fungus-containing food product is higher than the oil absorption capacity of the wet textured protein product, preferably the oil absorption capacity of the filamentous fungus-containing food product is at least 5%, most preferably at least 10% of its own weight.

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