Method for producing fungus-based food by providing a three-dimensional scaffold and fungus-based food obtainable by the method

Abstract

A method for preparing a fungus-based food by providing a three-dimensional edible substrate as a scaffold for fungal growth is described. The three-dimensional edible scaffold is formed by first providing a continuous edible substrate and then subsequently converting the substrate into a three-dimensional scaffold by introducing voids into the continuous edible substrate by foaming or perforation or a combination thereof. By this fungal growth in the provided three-dimensional edible scaffold, a fibrous fungus-based food is formed.

Description

[0001] The present invention relates to a method for preparing a fungus-based food by providing a three-dimensional edible matrix as a scaffold for fungal growth. By such fungal growth in the provided three-dimensional edible scaffold, a fibrous fungus-based food is formed.

[0002] Current meat consumption is depleting our natural resources while stimulating climate change (Foley 2011). Our current meat consumption is unsustainable, so meat substitutes must be developed to offset the growing meat consumption. Current approaches envision the use of meat analogues because such foods allow consumers to stick to existing cooking recipes and cultural habits. Therefore, alternative food protein sources such as plants, algae, and fungi are becoming increasingly popular as sustainable solutions to replace animal products. Fungus-based meat substitutes are interesting new meat substitutes because fungal mycelium (the vegetative part) naturally grows into long, branched fibers that resemble the structure of muscle tissue. In addition, given the naturally occurring flavor components, fungus-based products do not need to mask the unpleasant taste known from plant-based meat substitutes. Current methods for producing fungus-based meat substitutes typically rely on growing mycelium in tanks with nutrients, followed by harvesting and various post-processing, including cutting, heat treatment, extrusion, mixing with additives and forming into food products, as performed by Emergy Foods, Boulder, Colo., Prime Roots, disclosed in WO2007 / 139321 or known from Quorn production. In addition to requiring intensive post-processing, this method does not allow the full potential of the naturally grown fiber structure to be utilized. Another method disclosed in WO2020 / 023450 proposes growing a porous mat of fungal mycelium on a growth medium, which is infused with protein, fat, micronutrients and flavoring ingredients of plant origin in a second step by applying a vacuum. The porous mat is thus processed into a food product resembling animal meat. The growth of fungal mycelium in free space is limited in terms of height growth, which limits the production of a "whole cut". Moreover, the fiber structure, direction and thickness are given by the natural growth pattern of the fungal mycelium in free space, which does not allow customization of the product texture, which is critical for food products. As a result, they lack the organoleptic properties associated with common foods, lacking overall fiber cohesion, bite and juiciness.While fungus-based foods show potential as highly sustainable alternatives to animal meat, better solutions are needed to tailor product properties to meet consumer needs.

[0003] The need for better solutions is addressed by the embodiments as reflected in the claims.

[0004] Therefore, in a first aspect, the present invention relates to a method for preparing a fungus-based food product, the method comprising the following steps:

[0005] (a) providing a three-dimensional edible scaffold comprising an edible matrix, wherein the edible matrix comprises micronutrients and / or macronutrients required for the growth of fungal mycelium;

[0006] (b) inoculating the three-dimensional edible scaffold with at least one fungus to produce an inoculated scaffold;

[0007] (c) incubating the inoculated scaffold under growth conditions that allow mycelial growth of the at least one fungus such that the at least one fungus grows as mycelium through the scaffold to form a fungus-based food product.

[0008] Such a method allows the production of fungi-based foods whose texture and flavour can be modified. The method allows, for example, the production of fungi-based foods that can be used as mock meats, having:

[0009] i) Oriented and aligned fiber-like texture through mycelium growth throughout the food product;

[0010] ii) meat-like bite produced by the growth of fungal mycelium throughout the food product;

[0011] iii) juiciness, chewiness and elasticity produced by a three-dimensional scaffold that is tailored to achieve desired textural properties; and

[0012] iv) Forming into meat products by using a meat-resembling scaffold as a template for the product.

[0013] These properties can be achieved by providing a three-dimensional edible scaffold that is inoculated with and colonized by the fungus. As will be described in further detail below, the three-dimensional edible scaffold can be designed in any desired form to guide the growth of the fungal mycelium, thereby being able to provide a fungal mycelium with a desired structure, which structure can, for example, be similar to the structure of animal food. The fungal mycelium will grow along the gaps provided by the scaffold and will form a coherent product with the edible scaffold having the desired texture and form. Due to the fact that the three-dimensional scaffold provides gaps for the growth of the fungal mycelium, it also allows the absorption of liquids before or after the growth of the fungus, thereby allowing the juiciness of the resulting product to be improved. The method according to the present invention is Figure 1 It is schematically shown in FIG.

[0014] The term "fungus-based" means that the product obtained by the process always contains fungal components, in particular fungal mycelium, while the origin of other ingredients can be different. Preferably, the product obtained by the process contains at least 1 wt%, more preferably at least 5 wt%, at least 10 wt%, at least 20 wt%, at least 30 wt% or at least 40 wt%, particularly preferably at least 50 wt% of fungal components.

[0015] The method of the present invention is characterized in that a three-dimensional edible scaffold is provided in step (a). The term "provided" herein refers to the use of a preformed three-dimensional edible scaffold or the preparation of such a three-dimensional edible scaffold. The following further describes an apparatus and method for preparing a three-dimensional edible scaffold.

[0016] The term "scaffold" as used in the present invention refers to a structure that supports the formation of another structure (in this case, the formation of fungal mycelium), and the scaffold structure is made of a non-gaseous material, preferably a solid, non-liquid material, surrounding a gas-filled void space or multiple gas-filled void spaces. The non-gaseous material constituting the scaffold is denoted as a "matrix".

[0017] In the context of the present invention, the term "void" means an empty space surrounded by the matrix. Such void or multiple voids in the support structure can be formed in any possible manner known to those skilled in the art. For example, voids can be formed by stacking the matrix, by introducing a gas phase into the matrix, by removing liquid or solid material from the matrix and / or by perforating the matrix.

[0018] The void represents any form of aerated space within the support structure, which can show any possible form, as long as it allows fungal mycelium to grow therein and / or through it. The form of the void can depend on the way they are formed. For example, if a void is formed by a filament made of stacked edible substrates, i.e., a void is formed at the void of the filament, then they will each be at least partially limited by the raised outer surface of a single filament. An example of this method can be found in WO2020 / 164680 A1. On the other hand, if a void is formed by introducing a gas phase (e.g., bubbles) into an edible substrate (e.g., a non-solid substrate), then the void will each be at least partially limited by the concave inner surface of the substrate.

[0019] Irrespective of the form of the voids, it is preferred that the scaffold comprises a plurality of voids that are (at least) partially interconnected to allow mycelium to grow into and through the scaffold. Preferably at least 50% or at least 75%, preferably at least 80% of the voids formed by perforation and / or at least 50% or at least 75%, preferably at least 80% of the voids formed by foaming form part of one or more interconnected void spaces, wherein preferably at least one of said interconnected void spaces has a volume of at least 0.05 mm 3 , preferably at least 0.1 mm 3 , even more preferably at least 0.5 mm 3 , even more preferably at least 1 mm 3 or at least 5mm 3 .

[0020] In particular, the term "void" includes "channel". The term "channel" means a void space connected to the outer surface of the support and thus allowing penetration from the outside. In a preferred embodiment, the channel is understood to be an elongated tunnel-like void structure. It can have a straight form or a curved form. In one embodiment, the three-dimensional support comprises a plurality of channels. This channel can penetrate the entire support or only a part of the support. The channels can be organized as parallel channels, or their directions can be irregular. They can also cross each other and can be interconnected. This channel can be produced, for example, by punching with an object (e.g., a needle) leading to the channel, or by using a mold for preparing the support, wherein the mold shows a structure leading to the channel when the matrix material is applied to the mold. Alternatively, the pores can be interconnected by subjecting the foam to pressure fluctuations, or by drying, preferably under vacuum or by microwave heating.

[0021] The term "void" also includes holes. A hole represents a void space that is not necessarily connected to an outer surface. A hole can be closed and completely surrounded by a matrix, or open and therefore connected to other holes and / or open to an outer surface. Preferably, the three-dimensional edible scaffold comprises perforations such that at least 50%, preferably 75%, more preferably 80% of all holes are connected to at least one different hole. Preferably, the three-dimensional edible scaffold comprises perforations such that at least 5%, preferably at least 10%, more preferably at least 20% of all holes are directly or indirectly connected to an outer surface by being connected to one or more other perforations.

[0022] The three-dimensional scaffold may display channels or pores or channels and pores or channels and / or pores in combination with any other type of possible voids that may allow fungal mycelium to grow into and within the scaffold.

[0023] The pores may be closed, referred to as being completely surrounded by the edible matrix, or open, referred to as being connected to another pore and / or channel or to several other pore and / or channel spaces. Thus, the void space may be a partially or completely continuous network of pores and / or channels (possibly in combination with any other kind of possible voids that may allow the growth of fungal mycelium).

[0024] Preferably, the void space within the three-dimensional edible scaffold accounts for more than 10 vol%, more preferably more than 20 vol%, more preferably more than 30 vol%, more preferably more than 40 vol%, more preferably more than 50 vol%, more preferably more than 60 vol%, more preferably more than 70 vol%, and even more preferably more than 80 vol% of the three-dimensional edible scaffold. In this article, the term "void space" means the sum of all voids in the scaffold. When expressed as a "void fraction", this means the volume fraction of voids in the edible scaffold. The void space is the total volume (scaffold plus voids) measured, for example, by three-dimensional scanning minus the volume of the matrix constituting the scaffold, measured by weight divided by the density of the material or by pycnometry.

[0025] Thus, the void spaces present in the three-dimensional edible scaffold may be a partially or completely continuous void network, which may be, for example, pores and / or channels as described above. Such a void network allows fungal mycelium to grow into and through the scaffold, thereby filling the voids with the fungal mycelium, thus creating a structure in which the voids in the three-dimensional edible scaffold are filled with fungal mycelium.

[0026] Preferably, the average size of the voids is from 20 μm to 4 cm, preferably from 50 μm to 2 cm, even more preferably from 100 μm to 1 cm, particularly preferably from 100 μm to 0.5 cm. As used herein, the term "average size of the voids" is a parameter measured by taking an image (e.g., obtained by optical microscopy, electron microscopy or x-ray tomography), and the shortest diameter of the pores is measured by image analysis (in a layer or in a 3D reconstruction). Preferably, most of the voids (at least 50%, at least 70% or at least 90%) have a size greater than 20 μm, more preferably 50 μm. Preferably, most of the voids (at least 50%, at least 70% or at least 90%) have a size less than 4 cm, more preferably 2 cm.

[0027] Preferably, for a three-dimensional scaffold provided for inoculation with at least one fungus belonging to the class Basidiomycetes, the average size of the voids is from 100 μm to 4 cm, preferably from 500 μm to 2 cm, even more preferably from 1 mm to 1 cm.

[0028] Preferably, for a three-dimensional scaffold provided for inoculation with at least one fungus belonging to the class Ascomycetes, Deuteromycetes, Deuteromycetes or Zygomycetes and not belonging to the class Basidiomycetes, the average size of the voids is between 20 μm and 1 cm, preferably between 50 μm and 500 μm, even more preferably between 100 μm and 500 μm.

[0029] It is particularly preferred that the three-dimensional edible scaffold provided by the method according to the present invention consists of a continuous edible matrix, wherein the voids of the scaffold are introduced into the continuous edible matrix by foaming and / or perforation.

[0030] The three-dimensional support used in the method according to the present invention is characterized in that it is edible.This means that the non-gaseous material forming the " matrix " of the support around the gap is a material suitable for human intake, and is therefore palatable and nontoxic.About the source of the edible material of the edible matrix forming the support, there is no particular restriction.Every kind of edible material that can form support is suitable in this article.The matrix of support can be, for example, plant-derived, fungal-derived and / or cell-derived, particularly comprising protein, polysaccharide and / or lipid.

[0031] In one embodiment, the edible substrate is of plant origin. The term "plant-derived" refers to that at least part of the material forming the substrate is derived from a plant. In this article, the term "plant" includes higher plants and lower plants, such as algae. Therefore, in a preferred embodiment, the substrate comprises plant material.

[0032] In one embodiment, the edible substrate is of fungal origin. The term "of fungal origin" means that at least part of the material forming the substrate is derived from fungi / fungi. In this context, it is preferred that the substrate itself is not mycelium, but contains (at least partly) components derived from fungi / fungi.

[0033] In one embodiment, the edible matrix is ​​cell-derived. The term "cell-derived" refers to that at least part of the material forming the matrix is ​​derived from cultured cells, i.e., derived from cultured cells. This includes cells from higher organisms such as plants and animals, which are cultured in vitro, such as in fermenters or tissue cultures. It also includes the cultivation of unicellular organisms, such as bacteria, including cyanobacteria, unicellular fungi (e.g., yeast) and microalgae.

[0034] In one embodiment, the matrix comprises a combination of plant-derived and / or fungal-derived and / or cell-derived materials.

[0035] In a preferred embodiment, the edible substrate is of plant origin and may (additionally) contain edible materials from organisms other than plants (e.g., fungi or animals). However, in a preferred embodiment, the edible substrate does not contain any materials derived from animal slaughter or farming. In particular, the edible substrate may be vegan.

[0036] If edible substrate is plant-derived, i.e. at least part of the material forming substrate comes from plant, then vegetable material can be any material suitable for the intended purpose of forming edible support.For example, plant parts, such as seeds, grains or fruits, are preferably rich in protein, oil and / or sugar parts, and these parts are processed in a manner that allows vegetable material to form substrate.As described in the attached embodiment, for example, beans (of different sources) or peas can be used to smash them and use gained mashed material, slurry or paste to form edible support.But, certainly also can use purified or partially purified vegetable material, such as vegetable protein or plant polysaccharide extract, isolate or concentrate or from the crude extract of vegetable material, to prepare corresponding edible substrate.

[0037] Thus, the three-dimensional scaffold can be made, for example, from a mash, slurry, paste, solution, suspension or emulsion of edible material, preferably material of plant and / or fungal origin and / or cell origin. Such a mash, slurry, paste, solution, suspension or emulsion can be filled into a mold. In particular, such a mash, slurry, paste, solution, suspension or emulsion can be filled into a mold before providing it with a defined three-dimensional shape.

[0038] In some embodiments, the edible matrix of the present invention is a mash, a slurry, a paste, a solution, a suspension or an emulsion, i.e., an edible matrix, can be foamed before, after or simultaneously filled in a mold, and the foamed or not yet foamed edible matrix produces and / or flows when being filled in a mold, so that it fills the profile of the mold. Particularly preferably, the stress applied in the process of the foamed or unfoamed edible matrix being filled in the mold overcomes the yield stress of the edible matrix. Very particularly, the yield stress of the foamed or unfoamed edible matrix is ​​lower than 100Pa. Even more particularly, the temperature of the foamed or unfoamed edible matrix is ​​adjusted to avoid or slow down solidification, and is particularly adjusted to a solidification temperature higher than or lower than the gelling agent included in the edible matrix.

[0039] In one embodiment, the edible scaffold comprises protein. The protein can in principle be derived from any possible source, i.e. any possible organism or cell, such as animal, plant, bacterial or fungal or animal cell, plant cell or fungal cell. In a preferred embodiment, the protein is not derived from animal slaughter or farming. In another preferred embodiment, the protein is derived from plants.

[0040] The protein may be a naturally occurring protein or a recombinantly produced protein. Preferably, the protein is a naturally occurring protein. The protein may be in the form of an isolated protein which has previously been purified or partially purified from a source organism, a part thereof, a tissue or from a cell and / or it may be in the form of a directly processed plant material. The protein may also be in the form of a fermentation product based on a plant material containing the protein.

[0041] If the protein is derived from plants, it is preferred to use plant materials that contain a large amount of protein. Examples are plant parts known to be rich in protein, such as storage organs or seeds. Particularly suitable are proteins derived from leguminous plants such as peas, lentils, beans, etc. Examples of leguminous plants from which proteins present in edible matrices can be obtained are peas (e.g., yellow peas, chickpeas), beans (e.g., soybeans, kidney beans) and lentils. Other plants from which protein-containing materials can be obtained are cereals (e.g., rice, wheat, oats, rye), corn, oilseeds (e.g., rapeseed, sunflower seeds and pumpkin seeds) and all kinds of vegetables, such as pumpkin.

[0042] The protein may also be from seaweed or algae, such as Chlorella. Proteins from other unicellular organisms, such as yeast, may also be used, for example as yeast extract or as yeast protein isolate or as whole yeast cells.

[0043] In one embodiment, the edible support contains at least one sugar.Sugar can be derived from any possible source in principle, i.e. any possible organism or cell, such as animal, plant, bacterium or fungus or animal cell, plant cell, fungal cell or bacterial cell.Sugar can also be chemically synthesized and / or non-naturally occurring sugar.Sugar can be, for example, selected from monosaccharides, disaccharides, oligosaccharides and polysaccharides, such as complex polysaccharides, such as glucose, maltose, dextrose, mannose, alginate, alginate sulfate, gellan sulfate, starch, modified starch, gelatin, pectin, cellulose, bacterial cellulose, chitosan, agar, gellan gum, acylated and / or sulfated gellan gum, guar gum, cassia gum, konjac gum, gum arabic, ghatti gum, locust bean gum, xanthan gum, xanthan sulfate, carrageenan, carrageenan sulfate salt and any combination or mixture of the above substances.

[0044] In one embodiment, the edible scaffold comprises a protein and a sugar.

[0045] The amount of protein in the edible scaffold is not particularly limited. Preferably, the amount of protein in the edible scaffold is greater than 0.1 wt%, even more preferably at least 0.5%, even more preferably greater than 1%, greater than 2%, greater than 3%, greater than 5%, greater than 8%, greater than 10%, greater than 20% or greater than 30%.

[0046] Likewise, the amount of sugar (if present) in the edible scaffold is not particularly limited. Preferably, the amount of sugar in the edible scaffold is greater than 0.1 wt%, even more preferably at least 0.5%, even more preferably greater than 1%, greater than 2%, greater than 3%, greater than 5%, greater than 8% or greater than 10%.

[0047] In one embodiment, the protein and / or sugar are part of the edible matrix used to form the scaffold. They can be incorporated directly into the matrix during the process of forming the matrix, or they can be added to the matrix later, for example, by absorbing a liquid containing the protein and / or sugar into the matrix.

[0048] Alternatively, the protein and / or sugar may be incorporated into the three-dimensional edible scaffold by absorbing a liquid containing the protein and / or sugar into the voids of the scaffold.

[0049] In some embodiments, the three-dimensional edible scaffold (also) comprises macronutrients and / or micronutrients required for fungal growth. Such macronutrients and / or micronutrients may be part of the edible matrix. They may, for example, consist of proteins and / or sugars present in the scaffold / matrix. They may be incorporated directly into the matrix during the formation of the matrix, or they may be subsequently added to the matrix, for example by absorbing a liquid comprising the macronutrients and / or micronutrients into the matrix.

[0050] Alternatively, macronutrients and / or micronutrients, in particular proteins and / or sugars, can be added to the three-dimensional edible scaffold by absorbing a liquid containing the macronutrients and / or micronutrients into the interstices of the scaffold. Typically, macronutrients required for fungal growth include, for example, proteins and / or sugars. Micronutrients required for fungal growth include, for example, trace elements, such as organic and inorganic salts and vitamins required by the corresponding fungi or fungi.

[0051] The nutrients required may depend on the fungus / fungal species used to seed the scaffold. The selection of the required nutrients is within the skill of the artisan.

[0052] Advantageously, the three-dimensional edible scaffold contains water in an amount required for fungal growth. Preferably, the water activity of the scaffold is above 0.3, most preferably above 0.6. The term "water activity" means the vapor pressure (fraction) of water above the material divided by the standard partial vapor pressure of water.

[0053] In some embodiments, the three-dimensional edible scaffold further comprises fibers, viscosity enhancers, fats, vitamins, trace elements, flavoring compounds, colorants, spices, herbs, and / or salts.

[0054] These fibers, viscosity enhancers, fats, vitamins, trace elements, flavoring compounds, colorants and / or salts may be added to the matrix separately, or may be part of a material, such as a plant material, used to form the edible matrix.

[0055] In one embodiment, the edible scaffold contains only components that cannot be consumed by the fungi used to inoculate the scaffold, and the fungi grow into and through the scaffold. In this case, the edible scaffold serves as a scaffold to guide fungal growth.

[0056] In another embodiment, the edible scaffold comprises components that can be consumed by fungi when growing, such as components that fungi can digest and use for growth. Preferably, the scaffold is not only composed of components that can be consumed by fungi, but also contains at least a certain percentage of components that cannot be consumed by fungi, so that when fungi grow into and through the scaffold, the components are retained as a scaffold. Preferably, the percentage of scaffold components that cannot be consumed by fungi is at least 1 wt%, more preferably at least 10 wt%, at least 20 wt%, even more preferably at least 30 wt%, or at least 40 wt%, particularly preferably at least 50 wt%.

[0057] In a preferred embodiment, the edible matrix comprises water, agar, pea protein concentrate and / or pea protein isolate, sugar and oil or fat. The weight percentages based on the respective components of the edible matrix are preferably pea protein concentrate and / or isolate: 0.1-15 wt%; agar: 0.1-5 wt%; sugar: 0.01-15 wt%; and oil or fat: 0.1-10 wt%. The pH of the edible matrix is ​​preferably adjusted to a pH between 2 and 7, preferably 3.7 and 10, more preferably 4 and 7 by adding an acid or a base. Alternatively, the edible matrix may comprise water, vegetable protein, at least one gelling agent, at least one nutrient source selected from starch, sugar and mono / disaccharide / oligosaccharide and at least one ingredient that is not consumed by fungi. The weight percentages of the components based on the edible matrix are preferably vegetable protein: 0.1-30 wt%; at least one gelling agent: 0.1-5 wt%; at least one nutrient source: 0.01-52 wt%, preferably 5-20 wt%; and at least one ingredient not consumed by fungi: 0.1-5 wt%. Preferably, the water activity of the scaffold is adjusted to above 0.7, more preferably above 0.8, even more preferably above 0.9. Particularly advantageously, for a scaffold for inoculation of at least one fungus belonging to the class Ascomycetes, Deuteromycetes, Oomycetes or Zygomycetes, the water activity of the scaffold is adjusted to above 0.8.

[0058] The three-dimensional edible scaffold may have any form suitable for providing a scaffold for the growth of fungal mycelium. Preferably, the matrix forming the scaffold is a solid material. In the context of the present invention, the term "solid" also includes materials that can be considered as semi-solid, such as gels or glassy or crystalline materials. In some embodiments, the matrix may include polysaccharides and / or proteins that form a gel, wherein the gel is formed when heated, cooled, enzymatically cross-linked and / or ions are added. Preferably, the edible matrix is ​​moldable before, simultaneously with or after the three-dimensional edible scaffold is formed. In particular, the edible matrix is ​​preferably molded before the edible matrix forms a macroscopic structure. The macroscopic structure can be a structure formed by a plurality of solid or semisolid elements having a width and / or length greater than 2 mm or greater than 4 mm.

[0059] Preferably, a gelling agent which is not consumed by fungi is used, in particular a plant-based gelling agent which is not consumed by fungi. These gelling agents may be agar or carrageenan.

[0060] Preferably, after solidification, the edible matrix as the continuous phase of the scaffold is viscoelastic with an elastic modulus of at least 0.01 Pa, preferably at least 0.1 Pa, more preferably at least 1 Pa. The elastic modulus is determined by oscillatory rheology in the linear viscoelastic regime at a frequency of 1 rad / s.

[0061] The external form of the support is not critical. The support can be provided, for example, in the form of a sphere, a cube, a cuboid, a cylinder, a cone, a plate, a pyramid, a sheet or any three-dimensional shape.

[0062] The total volume of the scaffold provided in step (a) (matrix plus voids) may be any volume suitable for obtaining the desired product. Preferably, the total volume is at least 1 mm 3 , even more preferably 1 cm 3 , or at least 5cm 3 , or at least 10cm 3 , or at least 20cm 3 , or particularly preferably at least 50 cm 3 .

[0063] In addition, the total volume is preferably not greater than 1 m 3 , even more preferably not more than 1000 cm 3 .

[0064] The three-dimensional edible scaffold provided in step (a) of the method according to the present invention may be prepared by any tools and methods available to those skilled in the art for preparing such scaffolds.

[0065] A possibility is, for example, by dispersing gas or by dissolving gas under pressure, followed by bubble nucleation, making the edible matrix foamed. Preferably, pressure can be released subsequently to further expand the bubble contained in the edible matrix, thereby increasing the void fraction. Term " foaming " herein refers to the method that bubble is introduced into the matrix by mechanical, chemical or physical means. This foaming step increases volume and therefore reduces the density of product by introducing air usually. This step can be used to produce three-dimensional support as described above by the empty array of channel and hole or other voids produced in the material interior. Preferably, the foam is solidified by gelation and / or drying. More preferably, the foam can be solidified by gelation and / or drying in a mold.

[0066] Preferably, the foaming is carried out at a temperature above the gelling temperature of the cold setting gelling agent or below or above the temperature at which the cross-linking enzyme causes gelling. More preferably, for an edible substrate containing agar as a gelling agent, the foaming is carried out at a temperature above 30° C., even more preferably above 45° C. and below 95° C. In particular, for an edible substrate containing transglutaminase or laccase as a cross-linking enzyme, the foaming is carried out at a temperature not higher than 4° C.

[0067] Preferably, step (a) of the method according to the present invention comprises the following steps: (a1) introducing voids into the edible matrix, preferably by foaming; (a2) introducing the edible matrix into the mold, preferably by pouring the edible matrix into the mold; and (a3) ​​curing the edible matrix in the mold, preferably by gelling. Preferably, step (a1) is performed before, after or simultaneously with step (a2). Preferably, steps (a1) and (a2) are performed before step (a3). In other words, the matrix can be introduced into the mold before curing (e.g., gelling). After the edible matrix has been cured to a certain extent, the matrix can be further perforated to provide more voids and / or more interconnected voids.

[0068] Preferably, the distribution of voids in the scaffold is fixed when the edible matrix solidifies in the mold. Preferably, when initially introduced into the mold, the edible matrix may contain a gas / solid interface, but not a macroscopic solid / solid interface. The term "gas / solid interface" refers to the interface between voids and the edible matrix material. The term "macroscopic solid / solid interface" refers to the interface between two separate solid structures (e.g., of an edible matrix) that are in contact with each other, wherein the minimum dimension of each solid structure is at least 0.1 mm, preferably at least 1 mm, at least 3 mm or at least 5 mm, more preferably at least 10 mm.

[0069] The three-dimensional scaffold can also be prepared by 3D printing, for example by printing filaments with gaps between printed lines to produce a porous network containing holes and / or channels. However, preferably, the three-dimensional edible scaffold provided by step (a) of the method according to the present invention is not formed by the method or other similar methods involving assembling filaments. In particular, step (a) of the method according to the present invention preferably does not include the step of assembling the filaments formed by extrusion. In an embodiment involving step (a1), the edible matrix is ​​preferably not subjected to extrusion before being introduced into the mold. It is possible that step (a) of the method according to the present invention does not include an extrusion process that produces multiple filaments or macro structures. It is also possible that the method for manufacturing fungus-based food according to the present invention does not include an extrusion process at all.

[0070] Three-dimensional scaffolds can also be prepared by using a sacrificial template in an edible matrix. Here, the sacrificial template is removed from the matrix to allow the porous network to be generated. An example is the use of a sacrificial template that is liquefied and / or removed before inoculation or before or during incubation. An example is the use of one or more threads in a mold and the removal of the threads after the edible matrix is ​​poured into the mold and dissolved.

[0071] Another possibility for preparing three-dimensional scaffolds is to use additive manufacturing, such as sintering-based techniques, or by freeze-drying of edible matrices.Another possibility is the drying of solutions, suspensions or foams, in particular microwave drying.

[0072] The three-dimensional scaffold can also be formed by perforating the matrix by methods such as punching, cutting or forming with a sharp object. Preferably, the puncture holes or channels point in one direction to produce an anisotropic structure. These holes or channels can each be at least partially defined by the concave inner surface of the matrix. The term "anisotropic" / "anisotropy" refers to a material property that allows it to change or exhibit different properties in different directions opposite to isotropy. In this particular case, it means that the growth of the mycelium is "guided" to a specific direction rather than another direction. This has a direct impact on the mechanical properties of the resulting food, showing a fiber-like or muscle-like texture in a defined direction. In other words, the direction of the mycelium fibers is mainly in one direction rather than another.

[0073] In an alternative approach, three-dimensional scaffolds can also be formed by foaming followed by perforation to create pores or holes and channels of varying sizes and shapes.

[0074] The combination of several of the above methods or techniques allows the creation of primary and secondary pores or voids with different sizes and shapes.

[0075] In a preferred embodiment, the three-dimensional edible support is composed of a continuous edible matrix, wherein the void in the support is introduced into the continuous edible matrix, for example, by foaming and / or perforation. The term "continuous (continuous)" means that the material including the void or void space forms an uninterrupted overall space within the scope of the present invention, wherein each partial volume of the material is in contact with at least two adjacent volumes, except at the surface layer. The continuous material interconnected by physical and / or chemical bonds and / or steric hindrance at the contact point is expressed as "coherent (coherent)". Therefore, in this embodiment, the three-dimensional edible support is formed by first providing a continuous edible matrix, and then subsequently by introducing the void into the continuous edible matrix and the continuous edible matrix is ​​converted into a three-dimensional support. Preferably, the void is introduced into the matrix by foaming or perforation or a combination thereof. When initially introduced into the mold, the edible matrix can be continuous.

[0076] In one embodiment, the three-dimensional edible scaffold is a dried scaffold that is re-wetted / rehydrated prior to inoculation with fungi according to step (b) of the method of the present invention. In particular, the scaffold may be a scaffold dried to a water activity below 0.6, preferably below 0.3, and re-wetted to a water activity required for growth of the respective fungi. The term "re-wetting / rehydration" refers to the addition of water to the material by capillary suction and / or swelling. Rehydration should increase the water activity to a level required for fungal growth.

[0077] Preferably, rewetting / rehydration is performed by soaking in water and / or spraying water onto the outer surface of the three-dimensional edible scaffold or by increasing the relative humidity of the surrounding gas phase. This water can be enriched with nutrients for fungal growth or human consumption, or enriched with flavoring compounds, coloring compounds, salts, acids and / or bases. In this way, a three-dimensional edible scaffold can be provided, which contains nutrients for fungal growth and optionally nutrients for human consumption.

[0078] Thus, in this embodiment, the step of providing a scaffold according to step (a) of the method comprises providing a dry three-dimensional edible scaffold and wetting the scaffold prior to the inoculation step (b) of the method.

[0079] According to step (b) of the method of the present invention, the three-dimensional edible scaffold is inoculated with at least one fungus to produce an inoculated scaffold.

[0080] Inoculation can be performed by tools and methods known to those skilled in the art. In particular, inoculation can be achieved by contacting the fungus or fungi with one surface, multiple surfaces or all surfaces of the three-dimensional edible scaffold. The term "surface" herein refers to an outer surface representing the surface of the voids in the scaffold as well as an "inner surface". Optionally, the term "outer surface" when used in conjunction with an entity refers to a surface that substantially encapsulates the entity, or when considered as a whole, substantially encapsulates the surface of the entity, while the term "inner surface" when used in conjunction with an entity refers to one or more surfaces that are substantially located within the encapsulating surface.

[0081] Typically, inoculation can be achieved by applying at least one fungus in any form to a support capable of growing mycelium. In one embodiment, inoculation is achieved by applying mycelium and / or spores of at least one fungus to at least one surface of the support. If mycelium is used, the mycelium is preferably applied in wet form, such as in water or in another suitable liquid. Spores can be applied, for example, as dry spores or as spores dispersed in a liquid phase such as water. When spores are used for inoculation, inoculation can also be achieved by mechanically introducing the spores into the support, such as by pressing them into the matrix of the support.

[0082] If more than one fungus is used for inoculation, the fungi may be added simultaneously or at different times.

[0083] Inoculation can be performed after the three-dimensional scaffold has been formed. However, inoculation of the scaffold can also be achieved while the scaffold is being produced. This can be achieved, for example, by already inoculating an edible substrate for forming the scaffold or by adding at least one fungus during the formation step of the scaffold, for example during the foaming or perforation process. In this context, spores and / or mycelium can be used to incorporate them into the substrate, so that the substrate already contains spores and / or mycelium and is thus inoculated with the fungus.

[0084] Therefore, steps (a) and (b) of the process according to the invention can also be carried out in one step.

[0085] The at least one fungus may also be added before or after the foaming and / or perforating process.Most preferably, the at least one fungus is added to the edible substrate before, simultaneously with or after foaming, but before perforating.

[0086] Inoculation can also be performed as directed inoculation. This term means that inoculation is performed only in a partial area of ​​the three-dimensional scaffold, in particular at a plane of the outer volume, preferably at the top plane or at the bottom plane. Thus, the fungal mycelium grows from the inoculation plane into the free void space of the scaffold, resulting in the formation of a directed mycelial structure.

[0087] The fungus used in the method for inoculating an edible scaffold according to the present invention is not particularly limited as long as it can grow fungal mycelium and as long as it is edible. The term "fungal mycelium" means the vegetative part of a fungus composed of a large number of branched filamentous hyphae.

[0088] In particular, any mushroom that provides a filamentous structure may be used. Thus, any filamentous fungus from any division of the phylum Fungi may be used. In one embodiment, at least one fungus is a filamentous fungus selected from the class Ascomycetes, Basidiomycetes, Deuteromycetes, Oomycetes, and Zygomycetes. If more than one fungus is used, the same type or different types of fungi listed above may be used.

[0089] Preferably, the at least one fungus is selected from the group consisting of Zygomycota and Ascomycota

[0090] (Ascomycota), preferably selected from the genus Aspergillus, Rhizopus, Penicillum and Fusarium. Even more preferably, at least one fungus belongs to a species selected from the group consisting of Aspergillus oryzae, Rhizopus oligosporus, Penicillum camemberti and Fusarium venenatum.

[0091] In another preferred embodiment, the at least one fungus is selected from the divisions of the class Basidiomycetes, preferably from the genera Pleurotus, Laetiporus, Lentinula, Ganoderma

[0092] Ganoderma, Grifola, Agaricus, Flammulina

[0093] The genera include Flammulina, Morchella, Hypholoma, Macrolepiota and Cantharellus. Even more preferably, the at least one fungus belongs to a species selected from the group consisting of Pleurotus sapidus, Pleurotus ostreatus, Pleurotus djamor, Laetiporus sulphureus, Lentinula edodes, Ganoderma lucidum, Grifola frondosa, Agaricus bisporus, Flammulina velutipes, Morchella angusticeps, Hypholoma capnoides, Macrolepiota procera and Cantharelus cibarius.

[0094] In another preferred embodiment, the at least one fungus belongs to the class Agaricomycete.

[0095] After the inoculation step (b) of the method according to the invention, the inoculated scaffold is incubated under growth conditions allowing mycelial growth of the at least one fungus, such that the at least one fungus grows as mycelium through the scaffold to form a fungus-based food product.

[0096] This incubation step may be directly after the inoculation step. Alternatively, if immediate mycelial growth is not desired, the inoculated scaffold may also be treated or subjected to conditions that prevent mycelial growth. By this measure, the inoculated scaffold may be stored until the incubation step begins. For example, the inoculated scaffold may be dried to a water activity below that required for fungal growth and stored under the respective conditions prior to incubation for mycelial growth. It is also possible to store the inoculated scaffold at a temperature below that required for fungal growth prior to incubation for mycelial growth.

[0097] The incubation according to step (c) of the method of the invention is carried out under conditions that allow mycelial growth of the fungus / fungi used for inoculation in step (b). The growth conditions depend on the fungus / fungi and can be adjusted accordingly by a person skilled in the art. The term "allow for mycelium growth" means that the conditions allow mycelium growth but do not allow the formation and / or differentiation into fruiting bodies. Thus, under the conditions used for incubation, only mycelial growth occurs, but no formation and / or differentiation into fruiting bodies is formed.

[0098] Incubation allows the mycelium to grow along the surface of the scaffold, and in particular also into the interstices of the scaffold, thereby filling the interstices in the scaffold with the mycelium.

[0099] Preferably, the incubation is performed for at least 6 hours, more preferably at least 12 hours, even more preferably at least 24 hours.

[0100] In an alternative embodiment, after a certain incubation duration, preferably when the oxygen concentration has decreased by more than 10%, more preferably by more than 50%, the seeded scaffold is sheared or cut or flushed with air or oxygen-containing gas or expanded by applying a vacuum, in particular after at least 30 minutes of incubation, more preferably after 1 hour of incubation. When shearing or cutting, the seeded scaffold is broken into fragments or chips with a width and / or length of at least 1 mm, preferably fragments or chips with a width and / or length of at least 2 mm, more preferably fragments or chips with a width and / or length of at least 5 mm.

[0101] Preferably, more oxygen and / or nutrients, such as nitrogen-containing materials, preferably proteins and / or sugars, are added to the sheared or cut incubation scaffold.

[0102] The fragments or crumbs may be filled into another mold for incubation before or after the addition of additional oxygen and / or nutrients. Additional gelling agents may be added to the fragments or crumbs before they are filled into the mold, and the gelling agents may be gelling polysaccharides, gelling proteins or simply cross-linking agents, such as cross-linking enzymes or ions, preferably cations. In particular, the inoculated, incubated crumbs may be mixed with more inoculated scaffolds or edible substrates before solidification.

[0103] According to the edible substrate selected for forming the scaffold, mycelium can also grow into the edible substrate of the three-dimensional edible scaffold and penetrate into the substrate. Mycelium can also grow into the substrate and consume the substrate or part thereof. If the edible substrate contains a large amount and / or micronutrients required for fungal growth, this is especially true. In this embodiment, it is preferred to control the growth of fungal mycelium to ensure that the substrate is not completely consumed by the fungus during the growth of the mycelium, but to maintain at least a portion of the substrate that builds the edible scaffold. Preferably, at least 5wt% of the substrate, more preferably at least 10wt% or 20wt%, even more preferably at least 30wt% or at least 40wt%, particularly preferably at least 50wt% of the edible substrate that builds the scaffold is maintained and is not consumed by the fungus during the growth process.

[0104] As long as the edible matrix forming the scaffold contains components that are not required for fungal growth and these components are not or cannot be consumed by the fungi during growth, these components remain in the resulting fungus-based food and contribute to the texture and nutritional value of the food. Therefore, the product structure can be fine-tuned by adjusting the mechanical properties of the undigested scaffold.

[0105] Preferably, the incubation step (c) of the method according to the invention is carried out for a time and / or under conditions sufficient to achieve mycelial growth so that it completely fills the interstitial spaces in the three-dimensional edible scaffold, preferably each interstitial space contains mycelium. However, if desired, the incubation and / or fungal growth step can also be stopped at an earlier point in time when the fungal mycelium has not completely filled the interstitial spaces. However, preferably, at least 10% of the interstitial spaces, more preferably at least 20%, even more preferably at least 30%, at least 40%, at least 50%, at least 60%, at least 70% or at least 80%, particularly preferably at least 90% of the interstitial spaces contain fungal mycelium at the end of the incubation step (c).

[0106] The term "filled" with respect to void spaces filled with fungal mycelium preferably means that at least 2 vol% of the void spaces are occupied by fungal mycelium, more preferably at least 5 vol%, more preferably at least 10 vol%, most preferably at least 20 vol% of the void spaces are occupied by fungal mycelium.

[0107] During the incubation step, fungal mycelial growth can be controlled by varying environmental growth conditions, including time, humidity and temperature, that affect mycelial density, morphology and growth. In any case, the applied environmental growth conditions are selected in a manner that allows mycelial growth without forming / differentiating into fruiting bodies.

[0108] Since most fungi require oxygen, the fungal mycelium grows into the void along the surface of the support and fills the void. In a preferred embodiment, it is ensured that the void diameter in the support is not greater than twice the fungal growth height in the free space. In this article, the term "growth height" means the height measured from the surface, which is the height reached when the fungal mycelium grows into the free space on the surface containing nutrients. Under a larger diameter, the void may not be completely filled by the fungal mycelium, resulting in a void space left in the fungus-based food. As the fungal mycelium grows along the surface through the entire support, and also enters the matrix according to the composition of the fungal species and / or edible matrix selected for inoculation, the initial shape and size of the support serve as a template for the size and shape of the fungus-based food. The size, shape and direction of the fungal mycelium in the fungus-based food are sequentially templated with the void space in the support. Therefore, the support with long channels allows the production of long fungal mycelium fibers similar to the muscle fiber structure in meat. In addition, the fungal growth rate, morphology and hyphae thickness can be fine-tuned by changing the temperature and general growth conditions such as humidity and oxygen / carbon dioxide supply accordingly. Fungi ferment the products around them by using various enzymes, which contributes to the product's flavor. Using one or several fungi in combination, the flavor can be adjusted to suit consumer preferences without the need for added flavorings.

[0109] When the fungal mycelium grows into the voids of the support, a bonded food (with a meat-like texture if desired) is formed. In addition, the voids provided by the support allow the absorption of liquids before or after the fungal growth, thereby allowing the juiciness to be enhanced. The texture of the product can be further controlled by micro-modulators and therefore fine-tuning the mechanical properties of the support. If necessary, the voids allow the overall fiber-like texture to be produced on the whole food. The growth of mycelium in the whole food matrix can be oriented by directional inoculation, nutrient gradients or by introducing directional / anisotropic voids such as parallel channels, thereby producing directionality and meat-like fibers. Therefore, for example, the mycelium growth path can be guided by anisotropic voids in a three-dimensional edible support and / or by providing a mycelium growth path through a limited channel of the three-dimensional support, or by providing a mycelium growth path through nutrition and oxygen gradients, or by inoculating fungi only in a partial region of the three-dimensional edible support, thereby achieving the directional growth of mycelium.

[0110] Using this directionality, mycelial growth can be directed in a specific direction, similar to the directionality observed in meat products.

[0111] In a preferred embodiment, the edible scaffold of the product comprises channels that are filled with fungal mycelium during incubation, thereby forming a fiber-like fungal structure with an aspect ratio greater than 1. As used herein, "aspect ratio" refers to the ratio of the length to the diameter of the grown mycelial structure templated by the void spaces.

[0112] The method according to the invention may also comprise a step (d): interrupting the growth of at least one fungus. This can be achieved by any means and methods known to those skilled in the art that are suitable for stopping the growth of the corresponding fungus / fungi selected for inoculation. Preferably, the interruption of fungal growth is achieved by changing the temperature and / or water activity to a temperature and water activity condition lower than or higher than that required for the growth of at least one fungus. Alternatively, growth is interrupted as micronutrients and / or macronutrients are depleted. In this case, the end of fungal growth is predetermined by providing the nutrients provided. Fungal growth can also be interrupted by subjecting the product obtained in the incubation step (c) to a step of exposing it to a liquid absorbed by the product (as further described below).

[0113] After the incubation step and after the fungal growth has ceased, the resulting product consisting of the scaffold filled with mycelium may also be subjected to a washing step. This washing step may be used to remove, for example, residual nutrients or loose edible substrate parts. This washing step may be carried out with any suitable solution, for example with water, oil and / or an organic solvent.

[0114] The method according to the present invention may further comprise a step of exposing the obtained fungus-based food to a liquid so that the food absorbs the liquid, for example by capillary absorption into the remaining void space not filled by the fungal mycelium or by swelling into the edible matrix. This step may be performed after the incubation step (thereby preferably resulting in the interruption of fungal growth) or after the interruption step (d). As long as it is edible, any liquid suitable for absorption by the obtained mycelium-filled support may be used. In some embodiments, the absorbed liquid is water-based and / or oil-based. In a preferred embodiment, the liquid is further enriched with flavoring compounds, colorants, viscosity enhancers, fibers, vitamins, trace elements, salts, polysaccharides and / or proteins.

[0115] This absorption of liquid can be used to increase the juiciness of the food, which is often lacking in meat substitutes. In this article, the term "juiciness" refers to the sensory attribute felt when eating, for example, fresh fruits and vegetables (water release) or meat (water and oil release), and describes the amount of liquid released during chewing, the force with which the juice is expelled, the amount of juice released at the first bite and over time, the consistency of the juice, and the contrast between liquid and solid. Therefore, juiciness requires that the food can retain liquid and release liquid when compressed or broken (e.g., during chewing).

[0116] The method according to the present invention may further comprise the step of cutting, pulling and / or pressing the obtained fungus-based food into sheets or slices to imitate the shape of meat products.

[0117] In another aspect, the present invention also relates to a fungus-based food obtainable or obtained by the above-described method of the present invention.

[0118] As described above, the method according to the present invention allows the preparation of a fungus-based food product comprising a three-dimensional edible scaffold comprising an edible solid matrix, wherein the voids in the three-dimensional edible scaffold are filled with fungal mycelium.

[0119] In a preferred embodiment, the fungus-based food contains at least 10 wt% edible scaffold, more preferably at least 20 wt% edible scaffold, even more preferably at least 30 wt% edible scaffold, or at least 40 wt% edible scaffold, particularly preferably at least 50 wt% edible scaffold.

[0120] The term "wherein the voids in the three-dimensional edible scaffold are filled with fungal mycelium" means that the mycelium fills at least part of the volume created by the voids in the scaffold. Preferably, the fungal mycelium fills at least 2 vol%, more preferably at least 5 vol%, even more preferably at least 10 vol%, at least 20 vol%, at least 30 vol% or at least 40 vol%, particularly preferably at least 50 vol% of the void space. In a particularly preferred embodiment, 100% of the voids contain mycelium.

[0121] In some embodiments, not the entire void space is filled with fungal mycelium. In these embodiments, it is preferred that at least part of the void space is filled with liquid. With regard to the liquid that may be introduced into the food, the same applies as described above in conjunction with the method of the invention.

[0122] In general, regarding the composition and structure of the three-dimensional scaffold and / or matrix and preferred embodiments, the same as has been described above in conjunction with the method of the invention also applies to the product.

[0123] Thus, in particular with regard to the composition of the matrix forming the scaffold framework, the same applies as described above in conjunction with the method according to the invention.

[0124] Furthermore, with regard to the fungus used for inoculation and the further treatment of the scaffold before, after or after inoculation, the preferred embodiments described above in conjunction with the method of the invention also apply to the product obtained.

[0125] As described above, in conjunction with the methods of the present invention, the structures formed by fungal mycelium and present in fungus-based foods are templated by the size and shape of the interstices of the three-dimensional edible scaffold. In a preferred embodiment, the edible scaffold of the product comprises channels filled with fungal mycelium, thereby forming a fiber-like fungal structure with an aspect ratio greater than 1.

[0126] In another preferred embodiment, the diameter of the voids in the scaffold is less than twice the growth height of the corresponding fungi contained in the product into the free space.

[0127] In some embodiments, the mycelial structure formed is anisotropic.

[0128] In some embodiments, the edible solid matrix present in the fungus-based food comprises more than 0.1 wt% protein or sugar.Regarding the definitions of protein and sugar, the same applies to the description of the scaffold and matrix in the context of the method of the present invention.

[0129] In some embodiments, the fungus-based food comprises fiber, viscosity enhancers, fats, vitamins, trace elements, flavoring compounds, colorants, acids, bases, spices, herbs, and / or salts.

[0130] The fungus-based food of the present invention comprises a three-dimensional edible scaffold made of an edible matrix with voids, wherein the voids in the three-dimensional edible scaffold are (completely or partially) filled with fungal mycelium. The edible matrix or food may contain residues of micronutrients and / or macronutrients required for fungal growth, water, and optionally other components that contribute to the texture, nutritional value, taste, color, and shelf life of the food. The method for producing fungus-based food (as described above) can be adjusted so that the mycelium grown through the void spaces of the scaffold and optionally through the edible matrix forming the scaffold has the same length scale as muscle fibers. Therefore, this mycelium is able to simulate the structure of meat. For example, the product has i) a directional or controlled fiber-like mycelium structure / network across the entire structure templated by the void spaces, ii) an overall meat-like texture produced by the combination of the mycelium structure / network and the edible scaffold adjusted, and iii) juiciness produced by the porous matrix. The texture of fungus-based foods can be adjusted by the size, shape and fraction of fungal mycelium, the selection of fungi and the growth conditions applied to the corresponding fungi or fungal combinations, as well as by the mechanical properties of the edible matrix of the preparation scaffold and by the scaffold mesostructure / microstructure itself. For the latter two, aspects such as the fraction of protein and sugar and other edible components, the network forming ability of the components used, the physical and chemical crosslinks formed between these components, and the shape of the voids and pillars can be used to change the product texture. Based on the method of the present invention, fungus-based foods similar to animal meat products can be provided and can be processed accordingly, such as by frying in a dish with slight changes in shape and water content. The juiciness of this product can be further improved by soaking the fungus-based food in a liquid based on water and / or oil before cooking / frying / heating and eating. In addition, the color and taste of fungus-based products can be adjusted not only by adding flavoring compounds and colorants, but also by the selection of fungi. Different fungi can produce products of different colors not only by mycelial growth but also by spore formation. Fungi can also form flavoring compounds by enzyme production, which in turn converts surrounding organic matter, preferably protein, into flavoring substances.

[0131] In a preferred embodiment, the fungus-based product is heated, pasteurized, sterilized and / or smoked prior to further processing or consumption.Preferably, the fungus-based product is heated or cooled or frozen to inactivate the fungus.

[0132] As used herein, the terms "comprising, comprises" and "comprised of" are synonymous with "including, includes"; or "containing, contains", and are inclusive or open-ended, and do not exclude additional, unrecited members, elements or steps. The terms "comprising, comprises" and "comprised of" also include the term "consisting of".

[0133] As used herein, the term "about" means approximately, within the range, roughly, or around. When the term "about" is used in conjunction with a numerical value or range, it modifies the numerical value or range by extending the boundaries above and below the numerical value or range. Typically, the term "about" is used herein to modify a numerical value that is 10% higher and lower than the stated value.

[0134] In this specification, many documents including patent applications are cited. Although the disclosures of these documents are considered irrelevant to the patentability of the present invention, their entire contents are incorporated herein by reference. More specifically, all cited documents are incorporated herein by reference, just as each individual document is specifically and individually indicated to be incorporated herein by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0135] The invention is further illustrated by the following examples and figures, from which further embodiments and advantages can be derived. These examples are intended to illustrate the invention but not to limit its scope.

[0136] Figure 1 : Description of the scaffolding process. The selected ingredients are formed into an edible matrix (which may optionally have been inoculated with fungi). The matrix is ​​scaffolded into a porous scaffold having interstices, such as composed of pores and / or channels. The scaffold is inoculated with at least one fungus (especially if the matrix has not been inoculated with fungi). Fungal growth is triggered by environmental growth conditions, which is called incubation. After incubation, the fungus-based food is covered with fungal mycelium on the inside and outside.

[0137] Figure 2: Comparison of mycelial growth on a substrate without voids and a scaffold made of a substrate with voids. (A) A substrate made of agar, malt extract and yeast extract without voids (and therefore not made into a scaffold) was inoculated with Aspergillus oryzae on the outer surface and incubated at 32°C and 80% relative humidity for 48 hours. Fungal mycelium grew mainly on the outer surface of the substrate and only minimally penetrated into the substrate. (B) A substrate made of pea protein, agar and malt extract was inoculated and incubated with Aspergillus oryzae under the same conditions as (A). Fungal mycelium filled the void spaces during incubation. The growth of (A) and (B) was terminated by removing the scaffold from the incubation conditions. (C) A scaffold composed of chickpea, yellow pea, agar and malt extract was inoculated with the same fungi as (A) and (B) and incubated under the same conditions as (A) and (B). As in (B), fungal mycelium grew into the void spaces of the scaffold. The schematic above illustrates the difference in mycelial growth with and without voids.

[0138] Figure 3 : Three-dimensional edible scaffolds made of pea proteins and agar were subjected to mechanical response in uniaxial compression in the absence of fungi by foaming and gelling, resulting in fungus-based food products, compared to the same scaffolds inoculated with Aspergillus oryzae and incubated for 48 h at 32 °C and 80% relative humidity. The growth of fungal mycelium into the interstices of the scaffolds resulted in increased stiffness and strength, manifested as a textural alteration.

[0139] Figure 4 : Chickpea-based scaffolds were inoculated with Aspergillus oryzae (A) and incubated at 30°C and 80% relative humidity for 48 hours for fungal growth (B). Fungal growth occurred throughout the structure, including on the surface and inside the scaffold (C). Close-ups of the pores show mycelium growing into the entire void space of the scaffold (D).

[0140] Figure 5 :Based on the proposed method, a variety of plant-based foams acting as scaffolds were inoculated and incubated to prepare fungus-based foods. Water-based foams made of (A) 2wt% agar and 10wt% maltose, (B) 2wt% agar containing Cremodan (a commercial surfactant based on fatty acids), (C) 10wt% soy protein isolate containing 2wt% agar and 10wt% maltose, (D) 10% kidney beans and 2wt% agar containing 10wt% maltose, (E) 10wt% pea protein isolate containing 2wt% agar and 1wt% maltose, (F) 10wt% oat protein containing 2wt% agar and 2wt% maltose. All foams were inoculated with Aspergillus oryzae and incubated at 32°C and 75% relative humidity for 48 hours.

[0141] Figure 6 : Alternative scaffolding methods. Scaffolding is achieved by mechanically introducing channels into an edible matrix. (A) The edible matrix can be perforated (or partially perforated) by mechanically perforating the channels or by molding to make the edible matrix in a perforated manner (B). As the fungal mycelium grows into the long, thin channels, meat-like fibers are formed. By removing the scaffold material (B, D), the individual fiber structures shown in (C) can be isolated and observed. The resulting mycelium-based fibers simulate the muscle fibers of meat.

[0142] Figure 7 : Frying and cooking stability of overgrown fungus-based foods. The stability of fungus-based foods grown with mycelium through the material when fried in rapeseed oil at high temperatures (>100°C) is shown. On the left side of the pan, an edible three-dimensional scaffold without any fungal growth is fried, while on the right side, a fungus-based food made from the same scaffold is fried. It can be seen that the fungus-based food remains stable when the mycelium forms a cohesive network throughout the product, while the scaffold without fungal mycelium loses its structure and collapses.

[0143] Figure 8 : Directionality of mycelium in fungus-based foods. (A) Demonstration of a three-dimensional edible scaffold with directionally inoculated from the top surface, so that fungal mycelium grows in one direction downward from the top surface. (B) Close-up of a single well, where mycelium grows in a controlled and defined direction.

[0144] Fig. 9 : Mycelialized scaffolds were molded in sausage-shaped molds and incubated to allow mycelium to grow through and on the three-dimensional scaffolds.

[0145] Fig.10 : Photographs of fungus-based foods mycelialized with Aspergillus oryzae and fried in oil.

[0146] Fig.11 : Comparison of two different foaming techniques according to Example 31, as shown in images in a) and uniaxial compression tests in b) and c).

[0147] Fig.12 : Comparison of three different formulations according to Example 32 with different nitrogen sources, namely rice protein in a), pea protein concentrate in b) and yeast extract in c) and with and without sugar. Example

[0148] Example 1

[0149] Figure 2Shown is a comparison of fungal mycelium (A) grown on a substrate without voids and a scaffolding substrate with voids (B, C). In order to prepare a substrate without voids, 1.5wt% agar, 10wt% malt extract and 3wt% yeast extract were dispersed in water, then cooked and molded. Agar gelled when cooled, resulting in a solid substrate without voids. The gelled substrate was inoculated with Aspergillus oryzae spores dispersed in water on the outer surface. After incubation at 32°C and 80% relative humidity for 48h, the fungal mycelium only covered the surface of the substrate and only penetrated into the substrate in small amounts. The scaffolding substrate was prepared by dispersing (B) 10wt% pea protein, 1.5wt% agar and 10wt% malt extract and (C) 10wt% mashed yellow peas and chickpeas with 1.5wt% agar and 10wt% malt extract in water, followed by cooking, foaming in a cream whipping apparatus, molding and gel solidification. After gelation, a stable scaffold with pores and channels was formed, which was inoculated with Aspergillus oryzae spores in water and then incubated under the same conditions as (A). In contrast to (A), mycelium grew in the interstices of the scaffold and filled the entire scaffold. The fibrous nature of this product is directly given by the pores and mycelium growth.

[0150] Example 2

[0151] The edible three-dimensional scaffold was formed by foaming a solution of 10 wt% mashed beans, 10% maltose, 2 wt% agar. The three-dimensional edible scaffold was inoculated with dried Aspergillus oryzae spores on each side and incubated at 32°C and 75% relative humidity for 48 hours. In order to show how the growth of the matrix affects the mechanical properties of the resulting fungus-based food, compression tests ( Figure 3 Due to the interconnection of the three-dimensional scaffolds through the mycelium, the mechanical response of the cylindrical samples (2.5 cm in diameter, 2 cm in height) in uniaxial compression at 2 mm / s showed higher stiffness, higher energy absorption capacity and higher hardness of the obtained fungus-based food products compared to the corresponding scaffolds without fungus inoculation. Through the mycelium growth, the edible matrix is ​​interconnected, resulting in enhanced mechanical stability, which in turn is related to the perceived texture.

[0152] Example 3

[0153] An edible matrix with 30% mashed whole chickpeas and water was mixed with 2wt% agar and citric acid to reach a pH of 4.5 and heated to 95°C. The hot slurry was then introduced into a cream whipping apparatus and pressurized with N2O to a pressure of 15 bar. The apparatus was stirred for 10 min to partially disperse and partially dissolve the gas in the slurry. The slurry was released from the apparatus, causing the gas to expand, thereby foaming the slurry. Upon cooling, the foamed slurry gelled due to the agar contained. With this technique, the void fraction reached about 50%. The resulting pores were further increased by mechanically introducing the pores with a sharp object. The resulting porous scaffold was then inoculated with spores of Aspergillus oryzae. The foam was inoculated by slightly compressing the scaffold in an aqueous solution with fungal spores. After the compression was released, the water absorbed into the scaffold resulted in a uniformly distributed inoculation of the fungus. The inoculated foam was then incubated at 32°C and a high ambient humidity of 80%. The foam structure obtained after 40h was then fried in (>100°C) oil to prevent further fungal growth. Due to the open porous structure of the entire fungus-based food, the resulting fungus-based food has mycelial growth throughout the material ( Figure 4 ).

[0154] Example 4

[0155] Three-dimensional scaffolds were generated from various plant-based matrices. The compositions were as follows: (A) 2 wt% agar and 10 wt% maltose, (B) (commercial surfactant based on fatty acids), (C) 10 wt% soy protein isolate containing 2 wt% agar and 10 wt% maltose, (D) 10% kidney bean and 2 wt% agar containing 10 wt% maltose, (E) 10 wt% pea protein isolate containing 2 wt% agar and 1 wt% maltose, (F) 10 wt% oat protein containing 2 wt% agar and 2 wt% maltose. The mixture was heated to 95°C, introduced into a container and pressurized with N2O to reach a pressure of 15 bar. The slurry was mixed with gas for about 10 minutes. The slurry was released from the container, causing the gas to expand, so that the slurry foamed to a void fraction of about 30 vol%-70 vol%. The resulting porosity was further increased by mechanically introducing holes with a sharp object. Inoculation of these foams with Aspergillus oryzae was carried out by a variety of techniques, including inoculation with live mycelium in water, spores in water, and dry spores. All seeded scaffolds were incubated at 30-35°C and 65-80% relative humidity for 40-48 hours. Figure 5 As shown, the final product obtained after incubation is completely overgrown with fungal mycelium to form a fungus-based food product.

[0156] Example 5

[0157] The scaffold was created by first creating an edible matrix composed of proteins and polysaccharides and then subsequently adding voids / pores to the matrix by mechanically perforating the matrix. The edible matrix with 2 wt% agar and 40% mashed chickpeas was heated to 90°C and cooled to room temperature. The resulting hardened agar was made porous by perforating the introduction channels with a sharp needle of 2 mm diameter. This was done selectively and randomly ( Figure 6 A). The scaffolds were inoculated with spores from Aspergillus oryzae at 30°C, 80% relative humidity for 40 hours. After growth, oriented mycelial growth was observed in the direction of the channels. These channels were able to form long fibers corresponding to the exact length of the perforated channels. The thickness of these channels directly corresponded to the thickness of the fibers formed. By dissolving the three-dimensional scaffold, a fleshy fiber structure was revealed showing fungal mycelial fibers aligned in a specific direction ( Figure 6 B, C, D). The same results were obtained by using a mold with spikes that allowed the immediate formation of a porous scaffold.

[0158] Example 6

[0159] Comparison of three-dimensional scaffolds with and without mycelium in a frying test ( Figure 7 ). While the fungus-based food (right) showed browning upon frying, remained stable and did not lose more than 20% of its volume, the same 3D scaffold without the fungus immediately lost its shape and melted upon frying. This suggests that the final fungus-based food is thermally stable and can be used for cooking purposes, showing frying behavior similar to that of meat chunks.

[0160] Example 7

[0161] A three-dimensional porous scaffold was produced by 3wt% agar, 10wt% pea protein and 10wt% malt extract in water. The scaffold was inoculated with Aspergillus oryzae by placing spores only on the outer plane of the scaffold. The inoculation conditions were 31°C and 80% relative humidity for 45 hours. By using this inoculation technique, mycelium grew from the surface into the scaffold. Through this guided growth, the mycelium formed an anisotropic mycelial matrix, so a meat-like texture was produced with this method. A close-up of the three-dimensional scaffold with mycelium shows that the growth direction of the mycelium leads to an anisotropic structure.

[0162] Example 8

[0163] By mixing 2wt% agar, 4% 1% maltose and 3% yeast extract are mixed with water and heated to 95°C to form a three-dimensional scaffold. After heating, the edible matrix (also referred to as "food matrix" in the context of this application) is foamed by using kitchen aids to achieve a high void fraction of 80%. The foamed edible matrix is ​​poured into a mold and slowly cooled to induce agar gelation. During this cooling step, the fungal mycelium of Aspergillus oryzae is stirred into the foamed edible matrix, which is called inoculation. After cooling, the gel is further perforated with a sharp needle to produce a porous network with a large open area, which is called an inoculated three-dimensional scaffold. After this cooling step, the inoculated scaffold is incubated at 31°C and 85% relative humidity. After incubation for 40h, a mycelial network is formed around and inside the entire food matrix, resulting in a fungus-based food.

[0164] Example 9

[0165] An edible matrix with 2wt% agar, 10% pea protein and 10% maltose was mixed in water and heated to 95°C. After heating, the food matrix was foamed by using a kitchen aid while cooling to room temperature. The foam was then poured into a mold with thorns and slowly cooled. A porous three-dimensional matrix was formed by the thorns in the mold. During this cooling step, the fungal mycelium of Aspergillus oryzae was added while cooling, which is called inoculation, to obtain an inoculated three-dimensional edible scaffold. After this cooling step, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and inside the entire food matrix to obtain vegetarian meat.

[0166] Example 10

[0167] Prepare a 4% Food matrix of 1% maltose, 3% yeast extract. Mycelial spores of Rhizopus oligosporus were added to the foam structure at 40°C. The foam was inoculated at 32°C. The fermented foam formed a closed cellular wet foam. The fungus grew only on the surface and did not penetrate into the matrix itself.

[0168] Embodiment 11

[0169] As in Example 9, a three-dimensional scaffold was formed and inoculated with fungi. However, here the fungi were inoculated only from the top surface, allowing for directed fungal growth through the interstices, thereby forming a directed fungal network, such as Figure 8 shown.

[0170] Embodiment 12:

[0171] The edible matrix consisting of water, 2wt% agar, 12wt% pea protein isolate and 10wt% sugar is mixed and heated to boiling point. After heating, the food matrix is ​​cooled to 55°C and spores of Aspergillus oryzae are added, and 400g of edible matrix is ​​filled in a 1L pressurized container, gaseous N2O or CO2 is added to reach a pressure of 6-10 bar, and then the edible matrix is ​​foamed by shaking for 10 minutes. The slurry is released from the container, causing the gas to expand when released from the gas pressurized container, reaching a gas volume fraction of 50-60vol%. The slurry is transferred to a mold and cooled to room temperature (20-25°C). After cooling, the matrix forms an inoculated three-dimensional scaffold. Additional gaps are provided in the resulting scaffold by perforating with a needle of about 1mm in diameter. After the perforation step, the inoculated scaffold is incubated at 31°C and 85% relative humidity with or without a mold. After growing for 40h, a mycelial network is formed around and inside the entire food matrix to obtain a fungus-based food.

[0172] Embodiment 13:

[0173] An edible matrix consisting of water, 2 wt% agar, 12 wt% pea protein isolate and 10 wt% sugar was mixed and heated to boiling point. After heating, the food matrix was cooled to 55°C, spores of Aspergillus oryzae were added, and the food matrix was foamed using the pressurized container of Example 12. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the gel formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforation with a needle of approximately 0.5 mm in diameter. After this perforation step, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40 h of growth, a mycelial network was formed around and inside the entire food matrix to obtain a fungus-based food.

[0174] Example 14.

[0175] An edible matrix consisting of water with 2wt% agar, 3wt% pea protein isolate, 8wt% pea protein concentrate and 10wt% sugar was mixed and heated to boiling point. The pH of the final mixture was reduced to 5 with 80% lactic acid solution. After heating, the food matrix was foamed by using a pressurized container according to Example 12 while cooling to room temperature. In this cooling step, the fungal mycelium of Aspergillus oryzae was added, referred to as inoculation. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the gel formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforation with a needle of approximately 0.5mm in diameter. After these cooling and perforation steps, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and inside the entire food matrix, resulting in a fungus-based food.

[0176] Example 15.

[0177] The edible matrix consisting of water, 2wt% agar, 8wt% pea protein isolate and 10wt% sugar was mixed and heated to 95°C. After heating, the food matrix was foamed by using a rotor-stator system (Polytron, 10'000rpm, 10 minutes) while cooling to room temperature to reach a gas volume fraction of 25-35vol%. After foaming for 9 minutes, the fungal mycelium of Aspergillus oryzae was added, which is called inoculation. The foam was then poured into a mold and slowly continued to cool to room temperature. After cooling, the gel formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforating with a needle having a diameter of about 0.1mm. After the perforation step, the inoculated scaffold was incubated at 27°C and 95% relative humidity. After growing for 27h, a mycelium network was formed around and in the entire food matrix to obtain a fungus-based food.

[0178] Embodiment 16:

[0179] An edible matrix consisting of water containing 2wt% agar, 6wt% pea protein isolate, spices, colorants and 10wt% sugar was mixed and heated to boiling temperature. After heating and initial cooling to 55°C, spores of Aspergillus oryzae were added. The inoculated edible matrix was foamed using a pressurized container as described in Example 12 while continuing to cool to 45°C. The foam was then poured into a sausage-shaped mold and slowly cooled to a final temperature of 20°C to solidify the foam. After cooling, additional voids were provided in the resulting scaffold by roughening the surface. After this roughening step, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and in the entire food matrix, resulting in a fungus-based food similar to a sausage (see Fig. 9 ).

[0180] Embodiment 17:

[0181] An edible matrix consisting of water and 2 wt% agar, 5 wt% pea protein concentrate, 5 wt% pea protein isolate, 5 wt% rapeseed oil and 10 wt% maltose was mixed and heated to boiling temperature. After heating, the food matrix was cooled to 55 ° C, spores of Aspergillus oryzae were added, and the food matrix was foamed by using a pressurized container as described in Example 12 while cooling to room temperature. The foam was poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold was provided with additional voids by perforating with needles of several different sizes ranging in diameter from 0.1 mm to 1 cm. After this perforation step, the inoculated scaffold was incubated at 31 ° C and 85% relative humidity. After 40 h of growth, a mycelial network was formed around and inside the entire food matrix, resulting in a fungus-based food.

[0182] Embodiment 18:

[0183] An edible matrix consisting of water with 2wt% agar, 10wt% pea protein, 10wt% coconut fat, tomato paste and 10wt% maltose is mixed and heated to boiling temperature. After heating, the food matrix is ​​foamed by using a pressurized container (see Example 12) while cooling to 45°C. The foam is then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold is provided with additional voids by perforating with a needle having a diameter of 0.1 mm. After this perforation step, the food matrix is ​​inoculated with spores of Aspergillus oryzae. The inoculated scaffold is incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network is formed around and inside the entire food matrix, resulting in a fungus-based food. The fungus-based food is fried in oil, producing a juicy sensation when eaten, such as Fig.10 shown.

[0184] Embodiment 19:

[0185] An edible matrix consisting of water with 2wt% agar, 7wt% pea protein concentrate, 2.5wt% pea protein isolate, yeast extract and 10wt% maltose is mixed and heated to boiling temperature. According to Example 12, after heating, the food matrix is ​​foamed by using a pressurized container while cooling to 45°C. The foam is then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold is provided with additional voids by perforating with a needle having a diameter of 0.1mm. After this perforation step, the scaffold is inoculated with spores of Aspergillus oryzae. The inoculated scaffold is incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network is formed around and inside the entire food matrix, resulting in a fungus-based food.

[0186] Embodiment 20:

[0187] An edible matrix consisting of water with 2wt% agar, 7wt% pea protein concentrate, 2.5wt% pea protein isolate, 10wt% texturized protein and 10wt% maltose was mixed and heated to boiling temperature. After heating and cooling to 55°C, spores of Aspergillus oryzae were added. According to Example 12, the food matrix was foamed by using a pressurized container while cooling to 45°C. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold was removed from the mold and provided with additional voids by perforating with a needle having a diameter of 0.1mm. After this perforation step, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and inside the entire food matrix, resulting in a fungus-based food.

[0188] Reality Example 21:

[0189] The edible matrix consisting of water, 2wt% agar, 7wt% pea protein concentrate, 2.5wt% pea protein isolate and 10wt% native starch was mixed and heated to boiling temperature. After heating and cooling, spores of Aspergillus oryzae were added and the food matrix was foamed by using a pressurized container while cooling to room temperature. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold was made porous by perforating with a needle. After this perforation step, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and inside the entire food matrix to obtain a fungus-based food.

[0190] Embodiment 22:

[0191] The edible matrix consisting of water, 2wt% agar, 7wt% pea protein concentrate, 2.5wt% pea isolate and 10wt% pregelatinized starch was mixed and heated to boiling temperature. After heating and cooling, spores of Aspergillus oryzae were added and the food matrix was foamed by using a pressurized container while cooling to room temperature. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold was removed from the mold and provided with additional voids by perforating with a needle having a diameter of 0.1mm. After the perforation step, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and inside the entire food matrix to obtain a fungus-based food.

[0192] Embodiment 23:

[0193] An edible matrix consisting of water with 2wt% agar, 7wt% pea protein concentrate, 2.5wt% pea protein isolate and 10wt% pregelatinized starch was mixed and heated to boiling temperature. After heating and cooling to 55°C, spores of Aspergillus oryzae were added and the food matrix was foamed by using a pressurized container according to Example 12 while cooling to 45°C. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold was removed from the mold and made porous by perforating with a needle having a diameter of 0.1mm. After these cooling and perforation steps, the inoculated scaffold was incubated at 31°C and 85% relative humidity. After 40h of growth, a mycelial network was formed around and inside the entire food matrix, resulting in a fungus-based food.

[0194] Embodiment 24:

[0195] An edible matrix consisting of water with 2wt% agar, 5wt% pea protein concentrate, 5wt% pea protein isolate and 10wt% sugar was mixed and heated to boiling temperature. After heating, the food matrix was foamed by using a rotating membrane device while cooling to 45°C. With pressurized air, a membrane with holes of 3 microns in diameter was foamed to reach a gas volume fraction of 30vol%. During this process, spores of Aspergillus oryzae were added to the process. The foam was then poured into a mold and slowly cooled to room temperature. After cooling, the resulting scaffold had additional holes by perforation with a needle of 0.1mm in diameter. After these cooling and perforation steps, the inoculated scaffold was incubated at 27°C and 95% relative humidity. After 30h of growth, a mycelial network was formed around and in the entire food matrix, resulting in a fungus-based food.

[0196] Embodiment 25:

[0197] An edible matrix consisting of water with 27 wt% pea protein isolate, 0.5 wt% baking powder and 10 wt% starch was mixed and heated in a microwave. The resulting foam was inoculated with spores of Aspergillus oryzae. Incubation was carried out at 27°C and 95% relative humidity. After 30 h of growth, a mycelial network was formed around and in the entire food matrix, resulting in a fungus-based food.

[0198] Embodiment 26:

[0199] An edible matrix consisting of water, 20 wt% pea protein isolate, 10 wt% sugar, Aspergillus oryzae spores and transglutaminase was mixed and foamed at room temperature using a pressurized container according to Example 12. The foam was then poured into a mold and cooled to 5°C for 12 h. The resulting porous scaffold had additional voids by perforating with a needle with a diameter of 0.5 mm. The inoculated scaffold was incubated at 31°C and 85% relative humidity for 30 h.

[0200] Embodiment 27:

[0201] The edible matrix consisting of water, 2wt% agar, 10wt% pea protein isolate and 10wt% sugar was mixed and heated to 95°C. After heating, the food matrix was foamed and released into a beaker using a pressurized container according to Example 12 and cooled to below 55°C. At below 55°C, the fungal mycelium of Aspergillus oryzae was added to the foam and beaker and mixed manually with a spatula, which is called inoculation. At a temperature above 45°C, the foam was then poured into a mold and slowly continued to cool. After cooling, the gel formed an inoculated three-dimensional scaffold. The resulting porous scaffold was removed from the mold, the outer layer (thickness 1mm) was cut off to produce a more open surface, and additional voids were provided for the scaffold by perforating with a needle of about 0.3mm in diameter. After this perforation step, the inoculated scaffold was incubated for 20h at 31°C and 85% relative humidity. After growth, the matrix was mixed and blended, and incubated for another 20h. By this step, the pieces formed by mixing are reconnected by the fungal growth, so they are able to grow through the entire food matrix, resulting in a fungus-based food.

[0202] Embodiment 28:

[0203] In another example, the properties of the fungus-based food of Example 14 are demonstrated. The resulting fungus-based food has high water and oil uptake capabilities. In this example, the 3D porous edible scaffold had a water content of 75 wt% before fermentation, 68 wt% after fermentation, and was able to exceed the initial water content by up to 87 wt% after soaking.

[0204] Embodiment 29:

[0205] In another example, the characteristics of the fungus-based food of Example 14 are demonstrated by its juiciness. In contrast to water that can be expelled from the matrix by mechanical forces, juiciness is defined as the total amount of liquid chemically bound by the matrix. Unbound liquid is trapped in the channels of the porous network of the edible matrix. For this experiment, a disc of fungus-based food with a height of 3 cm and a diameter of 8 cm was soaked in water at room temperature for 1 hour to fully saturate the fungus-based food with water. The fungus-based food was able to increase its water content by min.90wt% of the initial weight. After soaking, squeezing the final product by applying a weight of 3 kg to the cross-section of the disc resulted in a 13wt% reduction in absorbed water, so the final product was able to bind 77wt% of additional water. Squeezing resulted in a reduction in water because the unbound water representing juiciness was expelled due to mechanical forces.

[0206] Embodiment 30:

[0207] In another example, the properties of the fungus-based food of Example 14 are demonstrated by its juiciness. For this experiment, a disc of the fungus-based food having a height of 3 cm and a diameter of 8 cm was soaked in oil for 1 hour at room temperature to completely saturate the fungus-based food with oil. The fungus-based food was able to increase its oil content by 42 wt % of the initial weight. Extrusion of the final product after soaking by applying a weight of 3 kg to the cross section of the disc resulted in a 3 wt % reduction in absorbed oil, so the final product was able to bind 39 wt % of additional oil. Extrusion results in a reduction in oil because free oil representing unbound is expelled due to mechanical forces.

[0208] Embodiment 31:

[0209] In another example, two different foaming techniques and thus pore sizes were compared. The three-dimensional scaffolds prepared according to Example 18 were foamed by pressure foaming or by stirring / mechanical foaming and then inoculated with Aspergillus oryzae and incubated at 30°C and 85% relative humidity for 40 h. Fig.11 a), c1) and d1) as shown in Fig.11 Compared to the mechanically foamed substrates of b), c2) and d2), the pressure foamed substrates have larger and more open cells and thus lead to a higher stability during frying and cooking and an overall higher mechanical stability during mycelium growth.

[0210] Embodiment 32:

[0211] In another example, fungus-based foods having three different formulations were compared. Fig.12 As shown, a mixture containing 2 wt% agar and 5 wt% rice protein in water ( Fig.12 , a), a mixture containing 2 wt% agar, 5 wt% pea protein concentrate and 5 wt% glucose in water ( Fig.12 , b), and one containing 2wt% agar, 15wt% glucose and 0.2wt% yeast extract in water. All edible matrices were pressure foamed, inoculated with Aspergillus oryzae (A. oryzae), incubated at 30°C and 85% relative humidity for 40 hours and molded in cylindrical molds. The fungal mycelium after incubation was the densest in the product containing pea protein concentrate and sugar (b), because it provided both a carbon source and a nitrogen source, while in the absence of almost no nitrogen (in c), it grew fast but not dense. As a result, compared with a) and c), the volume was significantly reduced, Fig.12 The fungus based food product in b) showed the best shape retention after frying and boiling in water.

[0212] The present invention may be defined by the following aspects:

[0213] 1. A method for preparing a fungus-based food, the method comprising the following steps:

[0214] (a) providing a three-dimensional edible scaffold comprising an edible matrix, wherein the edible matrix comprises micronutrients and / or macronutrients required for the growth of fungal mycelium;

[0215] (b) inoculating the three-dimensional edible scaffold with at least one fungus to produce an inoculated scaffold; and

[0216] (c) incubating the inoculated scaffold under growth conditions that allow mycelial growth of the at least one fungus, such that the at least one fungus grows as mycelium through the scaffold to form a fungus-based food product.

[0217] 2. According to the method of aspect 1, the three-dimensional edible scaffold is formed by first providing a continuous edible matrix and then converting the continuous edible matrix into the three-dimensional scaffold by introducing voids into the continuous edible matrix through foaming or perforation or a combination thereof.

[0218] 3. The method according to aspect 2, wherein the three-dimensional edible scaffold comprises a plurality of said voids, said voids being partially interconnected to allow mycelium to grow into and through the scaffold.

[0219] 4. A method according to aspect 1 or 2, wherein the three-dimensional edible scaffold comprises a plurality of voids that are partially interconnected to allow mycelium to grow into and through the scaffold, each of the voids being defined at least by a concave surface of the scaffold, preferably a concave inner surface of the scaffold.

[0220] 5. The method according to any one of aspects 2 to 4, wherein at the end of incubation step (c) at least 10% of the interstices, more preferably at least 50%, particularly preferably at least 90% of the interstices contain fungal mycelium.

[0221] 6. The method according to any one of aspects 2 to 5, wherein the voids are configured to allow for an overall fiber-like texture to be created throughout the food product.

[0222] 7. The method according to any one of aspects 2 to 6, wherein the average size of the voids is from 20 μm to 4 cm, preferably from 50 μm to 2 cm, even more preferably from 100 μm to 1 cm.

[0223] 8. The method according to any of the preceding aspects, which does not comprise an extrusion process that produces a plurality of macrostructures or filaments.

[0224] 9. A method according to any one of the preceding aspects, wherein the three-dimensional edible scaffold is not formed by assembling filaments, in particular extruded filaments, preferably the three-dimensional edible scaffold is not formed by printing filaments with gaps between the printed lines to produce a porous network containing pores and / or channels.

[0225] 10. The method according to any one of the preceding aspects, wherein step (a) comprises:

[0226] (a1) introducing voids into the edible matrix, preferably by foaming;

[0227] (a2) introducing the edible substrate into the mould, preferably by pouring; and

[0228] (a3) solidifying the edible matrix in the mould, preferably by gelling,

[0229] Preferably, step (a1) is performed before, after or simultaneously with step (a2), and / or

[0230] Preferably, the edible substrate is introduced into the mould before it solidifies and / or before the edible substrate is formed into the macrostructure.

[0231] 11. The method according to aspect 10, wherein the edible substrate is not extruded to produce the plurality of macrostructures or filaments prior to being introduced into the mould.

[0232] 12. A method according to aspect 10 or 11, wherein the void distribution in the scaffold is fixed when the edible substrate is solidified in the mould.

[0233] 13. The method according to any one of aspects 10 to 12, wherein the edible substrate comprises no macroscopic solid / solid interfaces after being introduced into the mould and before being solidified.

[0234] 14. A method according to any one of the preceding aspects, wherein voids are introduced into the matrix by foaming and the foam is cured by gelling, preferably voids are further introduced by perforation after gelling.

[0235] 15. A method according to any one of the preceding aspects, wherein in step (a) the edible base is foamed and the foamed edible base is poured into a mould and cooled to induce gelling, such as agar gelling.

[0236] 16. The method according to any of the preceding aspects, wherein steps (a) and (b) are performed in one step by inoculating the edible substrate used to form the scaffold or by adding at least one fungus during the formation of the scaffold.

[0237] 17. The method according to any one of the preceding aspects, wherein the edible scaffold comprises greater than 0.1 wt% protein and / or sugar.

[0238] 18. The method according to any one of the preceding aspects, wherein the edible matrix comprises at least one protein and at least one sugar.

[0239] 19. The method according to any one of the preceding aspects, wherein the edible substrate comprises plant material.

[0240] 20. The method according to any one of the preceding aspects, wherein the edible matrix comprises a gel-forming polysaccharide and / or protein, wherein the gel is formed upon heating, cooling, addition of ions and / or enzymatic cross-linking.

[0241] 21. The method according to any one of the preceding aspects, wherein the edible matrix or scaffold is viscoelastic after curing and exhibits an elastic modulus of at least 0.01 Pa, preferably at least 0.1 Pa, more preferably at least 1 Pa.

[0242] 22. A method according to any one of the preceding aspects, wherein the edible scaffold contains at least a certain percentage of components that cannot be consumed by the fungus and therefore remain as a scaffold when the fungus grows into and through the scaffold, preferably, the components that cannot be consumed by the fungus include agar, carrageenan and / or alginate.

[0243] 23. The method according to any one of the preceding aspects, wherein the three-dimensional edible scaffold comprises fibers, viscosity enhancers, oils, fats, vitamins, trace elements, enzymes, flavoring compounds, colorants, acids, bases and / or salts.

[0244] 24. The method according to any one of the preceding aspects, wherein the at least one fungus is selected from the class Ascomycetes, Basidiomycetes, Deuteromycetes, Oomycetes and / or Zygomycetes.

[0245] 25. The method according to any one of the preceding aspects, further comprising the following steps:

[0246] (d) interrupting the growth of at least one fungus, preferably by changing the temperature and / or water activity to below or above the temperature and water activity conditions required for the growth of the at least one fungus.

[0247] 26. The method according to any one of the preceding aspects, further comprising the step of exposing the obtained fungus-based food product to a liquid so that the food product absorbs the liquid.

[0248] 27. The method according to aspect 26, wherein the liquid is also enriched with flavoring compounds, colorants, viscosity enhancers, fibers, vitamins, enzymes, trace elements, salts, acids, bases, fats, polysaccharides or proteins.

[0249] 28. The method according to any one of the preceding aspects, wherein the obtained fungus-based food product is cut and / or pulled into blocks or slices and / or compressed to simulate the shape of a meat product.

[0250] 29. A fungus-based food obtainable by the method of any one of aspects 1 to 28.

[0251] 30. A fungus-based food product comprising a three-dimensional edible scaffold comprising an edible matrix, wherein voids in the three-dimensional edible scaffold are filled with fungal mycelium.

[0252] 31. A fungus-based food according to aspect 30, wherein the three-dimensional edible scaffold comprises a plurality of said voids, said voids being partially interconnected to allow mycelium to grow into and through the scaffold, said voids each being defined at least by a concave surface of the scaffold, preferably a concave inner surface of the scaffold.

[0253] 32. The fungus-based food according to aspect 30 or 31, the three-dimensional edible scaffold being formed by first providing a continuous edible matrix and then converting the matrix into the three-dimensional scaffold by introducing voids into the matrix by foaming or perforation or a combination thereof.

[0254] 33. A fungus-based food product according to any one of aspects 30 to 32, wherein the three-dimensional edible scaffold is not an assembly of filaments, in particular extruded filaments.

[0255] 34. The fungus-based food according to any one of aspects 30 to 33, wherein the voids in the three-dimensional edible scaffold are pores or channels or a combination thereof.

[0256] 35. A fungus-based food according to any one of aspects 30 to 34, wherein the edible scaffold comprises greater than 0.1 wt% protein and / or sugar.

[0257] 36. A fungus-based food according to any one of aspects 30 to 35, wherein the edible substrate comprises plant material.

[0258] 37. A fungus-based food according to any one of aspects 30 to 36, wherein the edible scaffold comprises fibres, viscosity enhancing agents, fats, vitamins, enzymes, trace elements, flavouring compounds, colouring agents, acids, bases and / or salts.

[0259] 38. A fungus-based food product according to any one of aspects 30 to 37, wherein the food product is a mock meat.

[0260] 39. A fungus-based food according to any one of aspects 30 to 38, wherein the diameter of the voids in the scaffold is no greater than twice the height of fungal growth in free space.

Claims

1. A method for preparing a fungus-based food, the method comprising the following steps: (a) providing a three-dimensional edible scaffold comprising an edible matrix by first providing a continuous edible matrix, and then subsequently converting the matrix into a three-dimensional scaffold by introducing voids into the matrix by foaming or perforation or a combination thereof to form the three-dimensional edible scaffold, wherein the edible matrix comprises micronutrients and / or macronutrients required for the growth of fungal mycelium, wherein step (a) does not include an extrusion process that produces a plurality of filaments or macrostructures; (b) inoculating the three-dimensional edible scaffold with at least one fungus to produce an inoculated scaffold; and (c) incubating said inoculated scaffold under growth conditions that allow the growth of mycelium of said at least one fungus, The at least one fungus is allowed to grow as mycelium through the support to form a fungus-based food product.

2. The method of claim 1, wherein the three-dimensional edible scaffold comprises a plurality of the voids, the voids being partially interconnected to allow mycelium to grow into and through the scaffold.

3. A method according to claim 1 or 2, wherein each void is defined at least by a concave surface of the support.

4. The method of claim 1, wherein at least 90% of the interstices contain the fungal mycelium at the end of step (c).

5. The method of claim 1, wherein the voids are configured to allow for an overall fiber-like texture to be created on the food product. The method according to claim 1 , wherein the average size of the voids is from 100 μm to 1 cm.

7. The method of claim 1, wherein the three-dimensional edible scaffold is not formed by assembling filaments.

8. The method according to claim 1, wherein step (a) comprises: (a1) introducing voids into the edible matrix by foaming; (a2) introducing the edible substrate into a mould; as well as (a3) solidifying the edible substrate in the mold, wherein the edible substrate is introduced into the mould prior to forming the edible substrate into a macrostructure.

9. The method of claim 8, wherein the void distribution in the scaffold is fixed when the edible substrate solidifies in the mold.

10. The method of claim 1, wherein the voids are introduced into the matrix by foaming and the foam is cured by gelling.

11. The method according to claim 10, wherein the voids are further introduced by perforation after gelation.

12. The method of claim 1, wherein in the step (a), the edible base is foamed, and the foamed edible base is poured into a mold and cooled to induce gelation.

13. The method of claim 1, wherein the steps (a) and (b) are performed in one step by inoculating an edible substrate used to form the scaffold or by adding the at least one fungus during the formation of the scaffold.

14. The method of claim 1, wherein the edible scaffold comprises greater than 0.1 wt% of plant-derived protein and / or sugar.

15. The method of claim 1, wherein the edible matrix comprises a gel-forming polysaccharide and / or protein, wherein the gel is formed upon heating, cooling, addition of ions, and / or enzymatic cross-linking.

16. The method of claim 1, wherein the edible scaffold contains at least a percentage of components that are not consumed by the fungus and thus remain as a scaffold as the fungus grows into and through the scaffold.

17. The method of claim 16, wherein the components that cannot be consumed by the fungus comprise agar, carrageenan and / or alginate.

18. The method of claim 1, wherein the three-dimensional edible scaffold comprises fibers, viscosity enhancers, oils, fats, vitamins, trace elements, enzymes, flavoring compounds, colorants, acids, bases, and / or salts.

19. The method of claim 1, wherein the at least one fungus is selected from the group consisting of ascomycetes, basidiomycetes, deuteromycetes, oomycetes and / or zygomycetes.

20. The method according to claim 1, further comprising the steps of: (d) interrupting the growth of the at least one fungus by changing the temperature and / or water activity to below or above the temperature and water activity conditions required for the growth of the at least one fungus.

21. A fungus-based food obtainable by the method of any one of claims 1 to 20.

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