METHOD OF PRODUCING A FUNGUS-BASED FOOD PRODUCT BY PROVIDING A THREE-DIMENSIONAL SCAFFOLD AND A FUNGUS-BASED FOOD PRODUCT OBTAINABLE BY SUCH A METHOD

Abstract

Described is a method of making a fungus-based food product by providing a three-dimensional edible matrix as a scaffold for fungal growth. The three-dimensional edible scaffold is formed by first providing a continuous edible matrix which is then subsequently converted into the three-dimensional scaffold by introducing voids into the matrix by foaming or puncturing or a combination thereof. Through this fungal growth in the provided three-dimensional edible scaffold, a fibrous fungus-based food product is formed.

Claims

1. A method of making a fungus-based food product, said method comprising the steps of: (a) providing a three-dimensional edible scaffold comprising an edible matrix, the three-dimensional edible scaffold being formed by first providing a continuous edible matrix which is then subsequently converted into the three-dimensional scaffold by introducing voids into the matrix by foaming or puncturing or a combination thereof, wherein the edible matrix comprises micro- or macronutrients required for fungal mycelium growth; (b) inoculating the three-dimensional edible scaffold with at least one fungus to generate an inoculated scaffold; and (c) incubating the inoculated scaffold at growth conditions that allow for mycelium growth of the at least one fungus so that the at least one fungus grows through the scaffold as a mycelium to form a fungus-based food product.

2. The method of claim 1, wherein the three-dimensional edible scaffold comprises a multitude of said voids which are partly interconnected so as to allow mycelium growth into the scaffold and through it.

3. The method of claim 1, said voids each being bound by at least a concave surface of the scaffold.

4. The method of claim 1, wherein the fungal mycelium at the end of the incubation step (c) is contained in at least 10% of said voids.

5. The method of claim 1, said voids being configured to allow to create a global fiber-like texture across the whole food product.

6. The method of claim 1, wherein an average size of the voids is between 20 μm to 4 cm, preferably 50 μm to 2 cm.

7. The method of claim 1, not comprising an extrusion process to create multiple macrostructures or filaments.

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

9. The method of claim 1, wherein the step (a) comprises: (a1) introducing voids into the edible matrix; (a2) introducing, the edible matrix into a mould; and (a3) solidifying the edible matrix in the mould, wherein the edible matrix is introduced into the mould before the edible matrix is formed into a macrostructure.

10. The method of claim 9, wherein the edible matrix has not been subjected to extrusion to create multiple macrostructures or filaments before being introduced into the mould.

11. The method of claim 9, wherein distribution of the voids in the scaffold is fixed upon solidification of the edible matrix in the mould.

12. (canceled)

13. The method of claim 1, wherein the voids are introduced into the matrix by foaming and the foam is solidified by gelation.

14. The method of claim 1, wherein in the step (a) the edible matrix is foamed and the foamed edible matrix is poured into a mould and cooled down to induce gelation.

15. The method of claim 1, wherein the steps (a) and (b) are carried out in one step by inoculating the edible matrix which is used to form the scaffold or by adding the at least one fungus during the formation of the scaffold.

16. The method of claim 1, wherein the edible scaffold comprises more than 0.1 wt % protein and/or saccharide.

17-18. (canceled)

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

20. The method of claim 1, wherein the edible scaffold contains at least a certain percentage of components which cannot be consumed by the fungus and which are, therefore, maintained as a scaffold while the fungus is growing into and through the scaffold.

21-22. (canceled)

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

24. The method of claim 1, further comprising the step of exposing the obtained fungus-based food product to a liquid so that the food-product absorbs the liquid.

25. The method of claim 24, wherein the liquid is further enriched with flavour compounds, colorants, viscosifiers, fibers, vitamins, enzymes, trace elements, salts, acids, bases, fat, polysaccharides, or proteins.

26. The method of claim 1, wherein the obtained fungus-based food product is cut or pulled into pieces or slices and/or compressed to mimic the shape of meat products.

27. A fungus-based food product obtainable by the method of claim 1.

28-30. (canceled)

31. The fungus-based food product of claim 27, the diameter of the voids in the scaffold is not larger than double the growth height of the fungus in free space.

32. The method of claim 9, wherein step (a2) comprises pouring, the edible matrix into a mould.

33. The method of claim 9, wherein step (a3) comprises solidifying the edible matrix in the mould by gelation.

Description

BRIEF DESCRIPTION OF FIGURES

[0133] The invention is further illustrated by the following examples and figures, from which further embodiments and advantages can be drawn. These examples are meant to illustrate the invention but not to limit its scope.

[0134] FIG. 1: Description of scaffolding process. The chosen ingredients are shaped into an edible matrix (which may optionally already be inoculated with fungus). This matrix is scaffolded into a porous scaffold that has voids, e.g., made up of pores and/or channels. The scaffold is inoculated with at least one fungus (in particular if the matrix had not already been inoculated with a fungus). Fungus growth is triggered by environmental growth conditions referred to as incubation. After incubation, the fungus-based food product is covered internally and externally by fungal mycelium.

[0135] FIG. 2: Comparison of mycelium growth on a matrix without voids and scaffold made of a matrix with voids. (A) A matrix made of agar, malt extract, and yeast extract without voids, hence not made into a scaffold, was inoculated with Aspergillus oryzae at the outer surface and incubated at 32° C. and 80% relative humidity for 48 hours. The fungal mycelia grew mainly on the outer surface of the matrix and had only little penetration into the matrix. (B) A matrix made of pea protein, agar, and malt extract was inoculated and incubated with Aspergillus oryzae at the same conditions as in (A). Fungal mycelium filled up the void space during incubation. Growth of (A) and (B) was terminated by removing the scaffold from incubation conditions. (C) Scaffold consisting of chickpeas, yellow peas, agar and malt extract was inoculated with the same fungus and incubated at same conditions as in (A) and (B). As in (B) the fungal mycelium grew into the void space of the scaffold. Schematics above illustrate the difference in mycelium growth with and without voids.

[0136] FIG. 3: Mechanical response in uniaxial compression of a three-dimensional edible scaffold made from pea protein and agar by foaming and gelation without fungi compared to the same scaffold inoculated with Aspergillus oryzae and incubated at 32° C. and 80% relative humidity for 48 hours resulting in a fungus-based food product. The growth of fungal mycelium into the voids of the scaffold led to an increase in stiffness and strength, perceived as an alteration in texture.

[0137] FIG. 4: A chickpea-based scaffold (A) is inoculated with Aspergillus oryzae and incubated for fungal growth at 30° C. and 80% relative humidity for 48 hours (B). Fungal growth occurred through the entire structure, including surface and interior of the scaffold (C). A close-up of the pores reveals that the mycelia grew into the entire void space of the scaffold (D).

[0138] FIG. 5: Based on the the presented approach, a variety of plant-based foams acting as scaffolds were inoculated and incubated to make fungus-based food products. Water-based foam made of (A) 2 wt % agar and 10 wt % maltose sugar, (B) 2 wt % agar with Cremodan (commercial surfactant based on fatty acids), (C) 10 wt % soy protein isolate with 2 wt % agar and 10 wt % maltose (D) 10% kidney beans and 2 wt % agar with 10 wt % maltose, (E) 10 wt % pea protein isolate with 2 wt % agar and 1 wt % maltose, (F) 10 wt % oat protein with 2 wt % agar and 2 wt % maltose. All foams were inoculated with Aspergillus oryzae and incubated at 32° C. and 75% relative humidity over 48 hours.

[0139] FIG. 6: Alternative scaffolding approach. Scaffolding was achieved by mechanically introducing channels into an edible matrix. (A) The edible matrix can be perforated (or partially perforated) by punching through channels mechanically or by making an edible matrix in a perforated fashion by moulding (B). As the fungal mycelium grows into the long, thin channels, meat-like fibers are formed. By removing the scaffold material (B, D), it is possible to isolate and visualize the single fiber structures shown in (C). The resulting mycelium-based fibers mimic the muscle fibers of meat.

[0140] FIG. 7: Frying and cooking stability of over-grown fungus-based food products. The stability of a fungus-based food product grown with mycelium through the material while frying in rapeseed oil at high temperatures (>100° C.) is shown. On the left side of the pan, the edible three-dimensional scaffold without any fungal growth is fried, while on the right side, the fungus-based food product made from the same scaffold is fried. As visible, the fungus-based food product stays stable as the mycelium forms a coherent network throughout the product, while the scaffold without fungal mycelium loses its structure and collapses.

[0141] FIG. 8: Directionality of mycelia in fungus-based food product. (A) demonstrates the three-dimensional edible scaffold with directed inoculation from the top plane, hence fungal mycelium grows from the top plane downward in one direction. (B) Close-up of a single pore with mycelia growing in one controlled and defined direction.

[0142] FIG. 9: A myceliated scaffold moulded in a sausage-shaped mould and incubated to allow mycelial growth through and on top of the three-dimensional scaffold.

[0143] FIG. 10: Photograph of a fungus-based food product myceliated by A. oryzae and pan fried in oil.

[0144] FIG. 11: Comparison of two different foaming techniques according to Example 31, as images in a) and uniaxial compression tests in b) and c).

[0145] FIG. 12: Comparison of three different formulation according to Example 32 with different nitrogen sources, being rice protein in a), pea protein concentrate in b), and yeast extract in c) and with and without sugar.

EXAMPLES

Example 1

[0146] FIG. 2 shows the comparison of fungal mycelium growth on a matrix without voids (A) compared to a scaffolded matrix with voids (B, C). For the preparation of the matrix without voids, 1.5 wt % agar, 10 wt % malt extract and 3 wt % yeast extract were dispersed in water followed by cooking and moulding. Upon cooling, the agar gelled resulting in a solid matrix without voids. This gelled matrix was inoculated at the outer surface with Aspergillus oryzae spores dispersed in water. After incubation at 32° C. and 80% relative humidity over 48 h, the fungal mycelium had only covered the surface of the matrix and only minorly penetrated into the matrix. The scaffolded matrices were made by dispersing (B) 10 wt % pea protein, 1.5 wt % agar and 10 wt % malt extract, and (C) 10 wt % mashed yellow peas and chickpeas with 1.5 wt % agar and 10 wt % malt extract in water followed by cooking, foaming in a cream whipping device, moulding, and gel-setting. Upon gelation a stable scaffold with pores and channels was formed, which was inoculated with spores of A. oryzae in water followed by incubation at same conditions as (A). In contrast to (A), the mycelium grew in the voids of the scaffolds and filled the whole scaffold. The fibrosity of this product was given directly by the porosity and the mycelium growth.

Example 2

[0147] An 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 dry spores of Aspergillus oryzae from every side and incubated for 48 hours at 32° C. and 75% relative humidity. To demonstrate how growth through the matrix affects the mechanical properties of the resulting fungi-based food product, compression tests were performed using a mechanical testing device (FIG. 3). The mechanical response of a cylindrical sample (2.5 cm in diameter, 2 cm in heights) in uniaxial compression at 2 mm/s showed a higher stiffness, higher energy absorption capacity and higher hardness for the obtained fungus-based food product compared to the corresponding scaffold which had not been inoculated with the fungus due to an interlinkage of the three-dimensional scaffold by mycelium. Through mycelia growth, the edible matrix is interconnected, thus resulting in enhanced mechanical stability, which in turn correlates to the perceived texture.

Example 3

[0148] An edible matrix with 30% mashed whole chickpeas and water was mixed with 2 wt % agar and with citric acid to reach a pH value of 4.5 and heated to 95° C. This hot slurry was then introduced into a cream whipping device and pressurized with N.sub.2O to reach a pressure of 15 bar. The device was agitated over 10 min to partly disperse and partly dissolve the gas in the slurry. The slurry was released from the device leading to an expansion of the gas and hence foaming of the slurry. Upon cooling, the foamed slurry gelled due to the contained agar. With this technique, a void fraction of about 50% was reached. The resulting porosity was further increased by mechanically introducing pores with a sharp object. The resulting porous scaffold was then inoculated with spores of Aspergillus oryzae. The foam was inoculated by slight compression of the scaffold in a water solution with fungal spores. After compression release, the water uptake into the scaffold led to an evenly distributed inoculation with fungi. The inoculated foam was then incubated at 32° C. under high ambient humidity of 80%. The resulting foam structure after 40 h was then fried (>100° C.) in oil to prevent further fungal growth. The resulting fungus-based food product has mycelia growth throughout the entire material thanks to the open porous structure across the entire fungus-based food product (FIG. 4).

Example 4

[0149] Three-dimensional scaffolds were created from various plant-based matrices. The compositions were as follows: (A) 2 wt % agar and 10 wt % maltose sugar, (B) 2 wt % agar with Cremodan® (commercial surfactant based on fatty acids), (C) 10 wt % soya protein isolate with 2 wt % agar and 10 wt % maltose (D) 10% kidney beans and 2 wt % agar with 10 wt % maltose, (E) 10 wt % pea protein isolate with 2 wt % agar and 1 wt % maltose, (F) 10 wt % oat protein with 2 wt % agar and 2 wt % maltose. The mixtures were heated to 95° C., introduced into a vessel and pressurized with N.sub.2O to reach a pressure of 15 bar. The slurries were mixed with the gas for around 10 minutes. The slurries were released from the vessel leading to an expansion of the gas, hence foaming of the slurries to reach void fractions of about 30 vol %-70 vol %. The resulting porosity was further increased by mechanically introducing pores with a sharp object. Inoculation with Aspergillus oryzae of these foams was done by a variety of techniques ranging from inoculation with alive mycelia in water, spores in water, and dry spores. All inoculated scaffolds were incubated at 30-35° C. at relative humidities of 65-80% over 40-48 hours. As shown in FIG. 5, the final products obtained after incubation are entirely overgrown with fungal mycelium to form a fungus-based food product.

Example 5

[0150] A scaffold was created by first producing an edible matrix composed of proteins and polysaccharides and then subsequently adding voids/pores to the matrix by mechanically punching through the matrix. An edible matrix with 2 wt % agar and 40% mashed chickpea beans was heated to 90° C. and cooled down to room temperature. The resulting hardened agar was made porous by introducing channels through the puncturing with a sharp needle with a diameter of 2 mm. This was done selectively and randomized (FIG. 6A). The scaffold was inoculated with spores from Aspergillus oryzae at 30° C., 80% relative humidity for 40 hours. After growth, the directed mycelia growth was observed in the direction of the channels. These channels enabled the formation of long fibers corresponding to the exact length of the punctured channels. The thickness of these channels directly corresponds to the thickness of the formed fibers. By solubilizing the three-dimensional scaffold, a meat-like fiber structure appears that reveals fibers of fungal mycelia that are aligned in a specific direction (FIG. 6 B, C, D). The same result was obtained by using a mould with spikes that allowed to form a porous scaffold immediately.

Example 6

[0151] A three-dimensional scaffold with mycelia and without mycelia were compared in a frying test (FIG. 7). While the fungus-based food product (right) showed browning upon frying, remained stable and did not lose more than 20% of its volume, the same three-dimensional scaffold without fungus lost its shape and melted immediately upon frying. This demonstrated that the final fungus-based food product is heat stable and can be used for cooking purposes showing similar frying behavior as a piece of meat.

Example 7

[0152] A three-dimensional porous scaffold was created by 3 wt % agar, 10 wt % pea protein, and 10 wt % malt extract in water. This scaffold was inoculated with Aspergillus oryzae by only placing spores on the exterior planes of the scaffold. Inoculation conditions were at 31° C., 80% relative humidity for 45 hours. Through the use of this inoculation technique, the mycelium grew from the surface into the scaffold. Through this guided growth, the mycelium formed an anisotropic mycelium matrix, therefore creating a meat-like texture with this approach. A close-up of the three-dimensional scaffold with mycelium is presented to demonstrate that mycelia growth direction led to an anisotropic structure.

Example 8

[0153] A three-dimensional scaffold was formed by mixing 2 wt % agar, 4% Cremodan®, 1% malt sugar, and 3% yeast extract with water and heating to 95° C. After heating, the edible matrix (which may also be referred to as a “food matrix” in the context of the present application) was foamed by using a kitchen aid to reach a high void fraction of 80%. The foamed edible matrix was poured into a mould and slowly cooled down to induce agar gelation. During this cooling step, the fungal mycelia of Aspergillus oryzae was stirred into the foamed edible matrix, referred to as inoculation. After cooling, the gel was additionally perforated with sharp needles, rendering a porous network with a large open area, referred to as inoculated three-dimensional scaffold. After this cooling step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After incubation over 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 9

[0154] An edible matrix with 2 wt % agar, 10% pea protein and 10% malt sugar, was mixed in water and heated to 95° C. After heating, the food matrix was foamed by using a kitchen aid while cooling down to room temperature. This foam was then poured into a mould with spikes and slowly cooled down. Through the spikes in the mould a porous three-dimensional matrix was formed. During this cooling step, the fungal mycelia of Aspergillus oryzae was added while cooling down, referred to as inoculation, resulting an inoculated three-dimensional edible scaffold. After this cooling step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth over 40 h, a mycelium network was formed around and in the entire food matrix, rendering a meat analogue.

Example 10

[0155] A food matrix with 4% Cremodan®, 1% malt sugar, 3% yeast extract was produced by heating to 95° C. and cooling down to 40° C. At 40° C. the mycelia spores of Rhizopus oligosporus were added into the foam structure. The foam was inoculated at 32° C. The fermented foam formed a closed porous wet foam. The fungi only grew on the surface and did not penetrate into the matrix itself.

Example 11

[0156] Just as in Example 9, a three-dimensional scaffold was formed and inoculated with fungi. However, here the fungi were only inoculated from the top surface, allowing to form directed fungal growth across the voids and thus a directed fungal network, as shown in FIG. 8.

Example 12

[0157] An edible matrix composed of water with 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. and spores of A. oryzae were added and the edible matrix was foamed by filling 400 g of the edible matrix in a 1 L pressurized vessel, adding gaseous N.sub.2O or CO.sub.2 to reach a pressure of 6-10 bar, followed by shaking for 10 min. The slurry was released from the vessel leading to an expansion of the gas upon release from the gas pressurized vessel to reach a gas volume fraction of 50-60 vol %. This slurry was transferred into a mould and cooled down to room temperature (20-25° C.). After cooling, the matrix formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforation with needles with a diameter of around 1 mm. After this perforation step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity, either with or without the mould. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 13

[0158] An edible matrix composed of water with 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 A. oryzae were added and the food matrix was foamed by using a pressurized vessel according to Example 12. This foam was then poured into a mould and slowly cooled down to room temperature. After cooling, the gel formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforation with needles with a diameter of around 0.5 mm. After this perforation step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 14

[0159] An edible matrix composed of water with 2 wt % agar, 3 wt % pea protein isolate, 8 wt % pea protein concentrate, and 10 wt % sugar, was mixed and heated to boiling point. The final mixture was lowered to a pH of 5 with a 80% lactic acid solution. After heating, the food matrix was foamed by using a pressurized vessel according to Example 12 while cooling down to room temperature. During this cooling step, the fungal mycelia of Aspergillus oryzae was added, referred to as inoculation. This foam was then poured into a mould and slowly cooled down to room temperature. After cooling, the gel formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforation with needles with a diameter of around 0.5 mm. After these cooling and perforation steps, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 15

[0160] An edible matrix composed of water with 2 wt % agar, 8 wt % pea protein isolate, and 10 wt % sugar, was mixed and heated to 95° C. After heating, the food matrix was foamed by using a rotor-stator system (Polytron, 10,000 rpm, 10 minutes) while cooling down to room temperature to reach a gas volume fraction of 25-35 vol %., After 9 minutes of foaming, the fungal mycelia of Aspergillus oryzae was added, referred to as inoculation. This foam was then poured into a mould and slowly continued to cool down to room temperature. After cooling, the gel formed an inoculated three-dimensional scaffold. Additional voids were provided in the resulting scaffold by perforation with needles with a diameter of around 0.1 mm. After this perforation step, the inoculated scaffold was incubated at 27° C. and 95% relative humidity. After growth of 27 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 16

[0161] An edible matrix composed of water with 2 wt % agar, 6 wt % pea protein isolate, spices, coloring, and 10 wt % sugar, is mixed and heated to boiling temperature. After heating and initial cooling down to 55° C., spores of A. oryzae were added. The inoculated edible matrix is foamed by using a pressurized vessel as described in Example 12 while continuing cooling down to 45° C. This foam is then poured into a mould in the shape of a sausage and slowly cooled down to final temperature of 20° C. to solidify the foam. After cooling, additional voids were provided in the resulting scaffold by roughening up the surface. After this roughening step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product resembling a sausage (see FIG. 9).

Example 17

[0162] An edible matrix composed of water with 2 wt % agar, 5 wt % pea protein concentrate, 5 wt % pea protein isolate, 5 wt % rapeseed oil, and 10 wt % malt sugar is mixed and heated to boiling temperature. After heating, the food matrix is cooled down to 55° C., spores of A. oryzae were added, and the food matrix was foamed by using a pressurized vessel while cooling down towards room temperature as described in Example 12. This foam is poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was provided with additional voids by perforation with several needles of different sizes ranging from 0.1 mm to 1 cm in diameter. After this perforation step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 18

[0163] An edible matrix composed of water with 2 wt % agar, 10 wt % pea protein, 10 wt % coconut fat, tomato paste and 10 wt % malt sugar was mixed and heated to boiling temperature. After heating, the food matrix is foamed by using a pressurized vessel (see Example 12) while cooled down to 45° C. This foam is then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was provided with additional voids by perforation with a needle of 0.1 mm in diameter. After this perforation step, the food matrix was inoculated with spores of A. oryzae. The inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product. The fungus-based food product was pan-fried in oil resulting in a juicy sensation upon consumption, as depicted in FIG. 10.

Example 19

[0164] An edible matrix composed of water with 2 wt % agar, 7 wt % pea protein concentrate, 2,5 wt % pea protein isolate, yeast extract, and 10 wt % malt sugar was mixed and heated to boiling temperature. After heating, the food matrix was foamed by using a pressurized vessel while cooling down to 45° C., according to Example 12. This foam was then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was provided with additional voids by perforation with a needle of 0.1 mm in diameter. After this perforation step, the scaffold was inoculated with spores of A. oryzae. The inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 20

[0165] An edible matrix composed of water with 2 wt % agar, 7 wt % pea protein concentrate, 2,5 wt % pea protein isolate, 10 wt % texturized protein, and 10 wt % malt sugar was mixed and heated to boiling temperature. After heating and cooling down to 55° C. spores of A. oryzae were added. The food matrix was foamed by using a pressurized vessel, according to Example 12, while cooling down towards to 45° C. This foam was then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was removed from the mould and was provided with additional voids by perforation with a needle of 0.1 mm in diameter. After this perforation step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network is formed around and in the entire food matrix, rendering a fungus-based food product.

Example 21

[0166] An edible matrix composed of water with 2 wt % agar, 7 wt % pea protein concentrate, 2,5 wt % pea protein isolate, and 10 wt % native starch is mixed and heated to boiling temperature. After heating and cooling down spores of A. oryzae are added, and the food matrix is foamed by using a pressurized vessel while cooling down towards room temperature. This foam is then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold is made porous by perforation with a needle. After this perforation step, the inoculated scaffold is incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network is formed around and in the entire food matrix, rendering a fungus-based food product.

Example 22

[0167] An edible matrix composed of water with 2 wt % agar, 7 wt % pea protein concentrate, 2,5 wt % pea isolate, and 10 wt % pre-gelatinized starch was mixed and heated to boiling temperature. After heating and cooling down spores of A. oryzae were added and the food matrix was foamed by using a pressurized vessel while cooling down to room temperature. This foam was then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was removed from the mould and was provided with additional voids by perforation with a needle of 0.1 mm in diameter. After this perforation step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 23

[0168] An edible matrix composed of water with 2 wt % agar, 7 wt % pea protein concentrate, 2,5 wt % pea protein isolate, and 10 wt % pre-gelatinized starch was mixed and heated to boiling temperature. After heating and cooling down to 55° C. spores of A. oryzae were added and the food matrix was foamed by using a pressurized vessel, according to Example 12, while cooling down to 45° C. This foam is then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was removed from the mould and made porous by perforation with a needle of 0.1 mm in diameter. After these cooling and perforation steps, the inoculated scaffold was incubated at 31° C. and 85% relative humidity. After growth of 40 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 24

[0169] An edible matrix composed of water with 2 wt % agar, 5 wt % pea protein concentrate, 5 wt % pea protein isolate, and 10 wt % sugar was mixed and heated to boiling temperature. After heating, the food matrix was foamed by using a rotating membrane device while cooled down to 45° C. Foaming was performed with pressurized air, a membrane with pores of 3 micrometer in diameter to reach a gas volume fraction of 30 vol %. During this process, spores of A. oryzae were added into the process. This foam was then poured into a mould and slowly cooled down to room temperature. After cooling, the resulting scaffold was provided with additional pores by needle perforation with needle of 0.1 mm in diameter. After these cooling and perforation steps, the inoculated scaffold was incubated at 27° C. and 95% relative humidity. After growth of 30 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 25

[0170] An edible matrix composed 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 A. oryzae and incubated at 27° C. and 95% relative humidity. After growth of 30 h, a mycelium network was formed around and in the entire food matrix, rendering a fungus-based food product.

Example 26

[0171] An edible matrix composed of water with 20 wt % pea protein isolate, 10 wt % sugar, Aspergillus oryzae spores and transglutaminase was mixed and foamed by using a pressurized vessel, according to Example 12 but at room temperature. The foam was then poured into a mould and cooled down to 5° C. for 12 h. The resulting porous scaffold was provided with additional voids by perforation with needles with diameter of around 0.5 mm. The inoculated scaffold is incubated at 31° C. and 85% relative humidity for 30 h.

Example 27

[0172] An edible matrix composed of water with 2 wt % agar, 10 wt % pea protein isolate and 10 wt % sugar, was mixed and heated to 95° C. After heating, the food matrix was foamed by using a pressurized vessel according to Example 12 and released into a beaker and cooled down to below 55° C. At below 55° C., the fungal mycelia of Aspergillus oryzae was added to the foamed and the beaker and mixed by hand with a spatula, referred to as inoculation. At a temperature of above 45° C., this foam was then poured into a mould and slowly continued to cool down. After cooling, the gel formed an inoculated three-dimensional scaffold. The resulting porous scaffold was removed from the mould, the outer layer (1 mm in thickness) was cut off to create a more open surface and the scaffold was provided with additional voids by perforation with needles with a diameter of around 0.3 mm. After this perforation step, the inoculated scaffold was incubated at 31° C. and 85% relative humidity for 20 h. After growth, the matrix was mixed and blended and re-incubated for another 20 h. Through this step, the pieces formed through mixing were re-connected by fungal growth and thus they were able to grow through the entire food matrix, rendering a fungus-based food product.

Example 28

[0173] In another example the properties of the fungus-based food product of Example 14 were demonstrated. The resulting fungus-based food products have a high water and oil uptake capability. In this example the 3D porous edible scaffold had a water content of 75 wt % before fermentation, 68 wt % after fermentation and is able to exceed the original water content up to 87 wt % after soaking.

Example 29

[0174] In another example, the properties of the fungus-based food product of Example 14 were demonstrated by its juiciness. The juiciness was defined as the total amount of liquid that was chemically bound by the matrix, as opposed to water that can be expelled from the matrix through mechanical force. The unbound liquid is entrapped in the channels of the porous network of the edible matrix. For this experiment, a disk of 3 cm in height and 8 cm in diameter of the fungus-based food product was soaked in water for 1 hour at room temperature to fully saturate the fungus-based food products with water. The fungus-based food product was able to increase its water content by min. 90 wt % of initial weight. Squeezing of the final product by applying a weight of 3 kg onto the cross-section of the disk after soaking led to a decrease of 13 20 wt % of uptaken water, so the final product was able to bind 77 wt % additional water. Squeezing led to a decrease of water because unbound water, representing the juiciness, is expelled due to the mechanical force.

Example 30

[0175] In another example, the properties of the fungus-based food product of Example 14 were demonstrated by its juiciness. For this experiment, a disk of 3 cm in height and 8 cm in diameter of the fungus-based food product was soaked in oil for 1 hour at room temperature to fully saturate the fungus-based food product with oil. The fungus-based food product was able to increase its oil content by 42 wt % of initial weight. Squeezing of the final product by applying a weight of 3 kg onto the cross-section of the disk after soaking led to a decrease of 3 wt % of uptaken oil, so the final product was able to bind 39 wt % additional oil. Squeezing led to a decrease of oil because free oil, representing the unbound, is expelled due to the mechanical force.

Example 31

[0176] In another example, two different foaming techniques and thus pore sizes were compared. The three-dimensional scaffold made according to Example 18, was either foamed by pressure foaming or by whisking/mechanical foaming and then inoculated with A. oryzae and incubated for 40 h at 30° C. and 85% relative humidity. As shown in FIG. 11 a), c1) and d1), the pressure-foamed substrate had larger and more open pores and thus led to a higher stability upon frying and cooking and to an overall higher mechanical stability upon mycelial growth compared to the mechanically-foamed substrate in FIGS. 11 b), c2) and d2) with smaller and less interconnected or open pores.

Example 32

[0177] In another example, fungus-based food products with 3 different formulations were compared. As shown in FIG. 12, one contained 2 wt % agar and 5 wt % rice protein in water (FIG. 12, a), one comprised 2 wt % agar, 5 wt % pea protein concentrate and 5 wt % glucose in water (FIG. 12, b), and one comprised 2 wt % agar, 15 wt % glucose and 0.2 wt % yeast extract in water. All edible matrices were pressure-foamed, inoculated with Aspergillus oryzae (A. oryzae) and incubated for 40 hours at 30° C. and 85% relative humidity and moulded in cylindrical moulds. The fungal mycelium after incubation was the densest in the product containing both pea protein concentrate and sugar (b), as it provided both a carbon source and a nitrogen source, while instead with little nitrogen (in c), the growth was fast but not dense. As a result, the fungus-based food product in FIG. 12 b) showed best shape retention after frying and boiling in water, compared to a) and c), which decreased majorly in volume.

[0178] The invention may be defined by the following aspects: [0179] 1. A method of making a fungus-based food product, said method comprising the steps of: [0180] (a) providing a three-dimensional edible scaffold comprising an edible matrix wherein the edible matrix comprises micro- and/or macronutrients required for fungal mycelium growth; [0181] (b) inoculating the three-dimensional edible scaffold with at least one fungus to generate an inoculated scaffold; and [0182] (c) incubating the inoculated scaffold at growth conditions that allow for mycelium growth of the at least one fungus so that the at least one fungus grows through the scaffold as a mycelium to form a fungus-based food product. [0183] 2. The method of aspect 1, the three-dimensional edible scaffold being formed by first providing a continuous edible matrix which is then subsequently converted into the three-dimensional scaffold by introducing voids into the matrix by foaming or puncturing or a combination thereof. [0184] 3. The method of aspect 2, wherein the three-dimensional edible scaffold comprises a multitude of said voids which are partly interconnected so as to allow mycelium growth into the scaffold and through it. [0185] 4. The method of aspect 1 or 2, wherein the three-dimensional edible scaffold comprises a multitude of voids which are partly interconnected so as to allow mycelium growth into the scaffold and through it, said voids each being bound by at least a concave surface of the scaffold, preferably a concave inner surface of the scaffold. [0186] 5. The method of any of aspects 2 to 4, wherein the fungal mycelium at the end of the incubation step (c) is contained in at least 10% of said voids, more preferably at least 50% and particularly preferred at least 90% of the voids. [0187] 6. The method of any of aspects 2 to 5, said voids being configured to allow to create a global fiber-like texture across the whole food product. [0188] 7. The method of any of aspects 2 to 6, wherein an average size of the voids is between 20 μm to 4 cm, preferably 50 μm to 2 cm, even more preferably 100 μm to 1 cm. [0189] 8. The method of any of the preceding aspects, not comprising an extrusion process to create multiple macrostructures or filaments. [0190] 9. The method of any of the preceding aspects, wherein the three-dimensional edible scaffold is not formed by assembling filaments, particularly extruded filaments, preferably the three-dimensional edible scaffold not being formed by printing filaments with voids in between printing lines thereby creating a porous network containing pores and/or channels. [0191] 10. The method of any of the preceding aspects, wherein the step (a) comprises: [0192] (a1) introducing voids into the edible matrix, preferably by foaming; [0193] (a2) introducing, preferably pouring, the edible matrix into a mould; and [0194] (a3) solidifying the edible matrix in the mould, preferably by gelation, [0195] preferably wherein the step (a1) is carried out before, after or at the same time with the step (a2), and/or [0196] preferably wherein the edible matrix is introduced into the mould before solidification thereof and/or before the edible matrix is formed into a macrostructure. [0197] 11. The method of aspect 10, wherein the edible matrix has not been subjected to extrusion to create multiple macrostructures or filaments before being introduced into the mould. [0198] 12. The method of aspect 10 or 11, wherein distribution of the voids in the scaffold is fixed upon solidification of the edible matrix in the mould. [0199] 13. The method of any of aspects 10 to 12, wherein the edible matrix, after being introduced into the mould and before being solidified, does not comprise macroscopic solid/solid interfaces. [0200] 14. The method of any of the preceding aspects, wherein the voids are introduced into the matrix by foaming and the foam is solidified by gelation, preferably voids are further introduced after gelation by puncturing. [0201] 15. The method of any of the preceding aspects, wherein in the step (a) the edible matrix is foamed and the foamed edible matrix is poured into a mould and cooled down to induce gelation, e.g. agar gelation. [0202] 16. The method of any of the preceding aspects, wherein the steps (a) and (b) are carried out in one step by inoculating the edible matrix which is used to form the scaffold or by adding the at least one fungus during the formation of the scaffold. [0203] 17. The method of any of the preceding aspects, wherein the edible scaffold comprises more than 0.1 wt % protein and/or saccharide. [0204] 18. The method of any of the preceding aspects, wherein the edible matrix comprises at least one protein and at least one saccharide. [0205] 19. The method of any of the preceding aspects, wherein the edible matrix comprises plant material. [0206] 20. The method of any 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. [0207] 21. The method of any of the preceding aspects, wherein the edible matrix or the scaffold after solidification is viscoelastic and exhibits an elastic modulus of at least 0.01 Pa, preferably of at least 0.1 Pa, more preferably at least 1 Pa. [0208] 22. The method of any of the preceding aspects, wherein the edible scaffold contains at least a certain percentage of components which cannot be consumed by the fungus and which are, therefore, maintained as a scaffold while the fungus is growing into and through the scaffold, preferably the components which cannot be consumed by the fungus comprising agar, carrageenan and/or alginate. [0209] 23. The method of any of the preceding aspects, wherein the three-dimensional edible scaffold comprises fibers, viscosifiers, oils, fats, vitamins, trace elements, enzymes, flavour compounds, colorants, acids, bases and/or salts. [0210] 24. The method of any of the preceding aspects, wherein said at least one fungus is selected from the group consisting of ascomycetes, basidiomycetes, deuteromycetes, oomycetes, and/or zygomycetes. [0211] 25. The method of any of the preceding aspects, further comprising the step of: [0212] (d) interrupting the growth of the 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 growth of the at least one fungus. [0213] 26. The method of any 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. [0214] 27. The method of aspect 26, wherein the liquid is further enriched with flavour compounds, colorants, viscosifiers, fibers, vitamins, enzymes, trace elements, salts, acids, bases, fat, polysaccharides, or proteins. [0215] 28. The method of any of the preceding aspects, wherein the obtained fungus-based food product is cut and/or pulled into pieces or slices and/or compressed to mimic the shape of meat products. [0216] 29. A fungus-based food product obtainable by the method of any one of aspects 1 to 28. [0217] 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. [0218] 31. The fungus-based food product of aspect 30, wherein the three-dimensional edible scaffold comprises a multitude of said voids which are partly interconnected so as to allow mycelium growth into the scaffold and through it, said voids each being bound by at least a concave surface of the scaffold, preferably a concave inner surface of the scaffold. [0219] 32. The fungus-based food product of aspect 30 or 31, the three-dimensional edible scaffold being formed by first providing a continuous edible matrix which is then subsequently converted into the three-dimensional scaffold by introducing the voids into the matrix by foaming or puncturing or a combination thereof. [0220] 33. The fungus-based food product of any of aspects 30 to 32, wherein the three-dimensional edible scaffold is not an assembly of filaments, particularly extruded filaments. [0221] 34. The fungus-based food product of any of aspects 30 to 33, wherein the voids in the three-dimensional edible scaffold are pores or channels or a combination thereof. [0222] 35. The fungus-based food product of any of aspects 30 to 34, wherein the edible scaffold comprises more than 0.1 wt % protein and/or saccharide. [0223] 36. The fungus-based food product of any one of aspects 30 to 35, wherein the edible matrix comprises plant material. [0224] 37. The fungus-based food product of any one of aspects 30 to 36, wherein the edible scaffold comprises fibers, viscosifiers, fats, vitamins, enzymes, trace elements, flavour compounds, colorants, acids, bases, and/or salts. [0225] 38. The fungus-based food product of any of aspects 30 to 37, wherein the food product is a meat analogue. [0226] 39. The fungus-based food product of any of aspects 30 to 38, the diameter of the voids in the scaffold is not larger than double the growth height of the fungus in free space.