Strengthened fungal material

Abstract

The present invention relates to strengthened fungal mats having enhanced mechanical properties compared to naturally occurring fungal mats produced by one or more fungi of the same species. The invention also relates to a material comprising said reinforced fungal mat, and to the use of said reinforced fungal mats and materials. The invention also relates to a method for enhancing the mechanical properties of a fungal mat.

Description

Field of the Invention

[0001] The present invention relates to a reinforced fungal mat. The reinforced fungal mat of the present invention has enhanced mechanical properties compared to a naturally occurring fungal mat produced by one or more fungi of the same species. The present invention also relates to a material comprising the reinforced fungal mat, and uses of the reinforced fungal mat and material. The present invention also relates to a method for enhancing the mechanical properties of a fungal mat. Background of the Invention

[0003] Fungal biomass can be used to produce a variety of products. For example, fungal cultures can be used to produce renewable and biodegradable materials that have the potential to be used in a variety of applications, such as clothing, furniture upholstery, and medical settings. Fungal cultures also have an important role in the food industry, such as in the production of meat substitutes.

[0004] The ability to produce materials or other products containing fungal biomass depends in part on the growth pattern of the fungal strain used, and the type of growth substrate (referred to herein as "medium") used to grow the fungal strain. For some applications, it is desirable to grow fungal cultures under conditions that allow the formation of fungal mats (also referred to as "biofilms"). This is typically achieved using solid growth media. However, industrial-scale growth of fungal cultures on solid media is challenging, for example, due to the cost and time associated with sterilizing solid media. In addition, fungal cultures grown on solid media may exhibit reduced homogeneity.

[0005] Methods for growing fungal mats in liquid culture media can be more easily scaled up for industrial applications. However, fungal mats grown in liquid culture media often exhibit inferior mechanical properties compared to fungal mats grown on solid culture media, which in turn limits the applicability of fungal mats grown in liquid culture media. For example, fungal mats grown in liquid culture media are often more fragile than fungal mats produced on solid culture media, and they may be too thin to provide sufficient structural integrity.

[0006] There is an urgent and unmet need for fungal mats that exhibit enhanced mechanical properties and industrially applicable methods for producing such reinforced fungal mats. SUMMARY OF THE INVENTION

[0008] The inventors have overcome the above challenges by developing methods for enhancing the mechanical properties of fungal mats. Advantageously, the fungal mats of the present invention (referred to herein as "enhanced fungal mats") exhibit enhanced mechanical properties compared to naturally occurring fungal mats produced by one or more fungi of the same species. In more detail, the enhanced fungal mats may exhibit increased thickness, increased tensile strength, increased ultimate load and / or reduced surface adhesion compared to naturally occurring fungal mats produced by one or more fungi of the same species. Importantly, the methods of the present invention are industrially applicable, particularly because their production does not rely on the use of solid growth media. The enhanced fungal mats of the present invention and materials comprising the enhanced fungal mats have a wide range of applications, ranging from the textile and clothing industries to the food industry.

[0009] The present invention provides a reinforced fungal mat, wherein: (a) the reinforced fungal mat has enhanced mechanical properties compared to a naturally occurring fungal mat; and (b) the reinforced fungal mat comprises one or more fungi of the same species as the naturally occurring fungal mat.

[0010] In one embodiment, the reinforced fungal mat has an increased tensile strength compared to a naturally occurring fungal mat. In one embodiment, the tensile strength of the reinforced fungal mat is at least 10% greater than the tensile strength of a naturally occurring fungal mat. In one embodiment, the reinforced fungal mat has a tensile strength of at least 1000 Pa.

[0011] In one embodiment, the reinforced fungal mat has an increased ultimate load compared to a naturally occurring fungal mat. In one embodiment, the ultimate load of the reinforced fungal mat is at least 10% greater than the ultimate load of a naturally occurring fungal mat. In one embodiment, the reinforced fungal mat has an ultimate load of at least 3 Newtons.

[0012] In one embodiment, the reinforced fungal mat has an increased thickness compared to a naturally occurring fungal mat. In one embodiment, the thickness of the reinforced fungal mat is at least 3 times greater than the thickness of a naturally occurring fungal mat. In one embodiment, the reinforced fungal mat has a thickness of at least 5 mm.

[0013] In one embodiment, the strengthened fungal mat has reduced surface adhesion compared to a naturally occurring fungal mat. In one embodiment, the surface adhesion of the strengthened fungal mat is at least 10% lower than the surface adhesion of a naturally occurring fungal mat.

[0014] In one embodiment, the reinforced fungal mat further comprises a support material, optionally wherein the support material comprises a fabric.

[0015] The present invention also provides a material comprising the reinforced fungal mat of the present invention.

[0016] In one embodiment, the material comprises two or more reinforced fungal mats.

[0017] The present invention also provides a method for enhancing the mechanical properties of a fungal mat, the method comprising: (a) exposing the fungal mat to an unused culture medium; and / or (b) physically manipulating the fungal mat; to provide a reinforced fungal mat having enhanced mechanical properties relative to the fungal mat. In one embodiment, the method comprises (a) exposing the fungal mat to an unused culture medium; and (b) physically manipulating the fungal mat.

[0018] In one embodiment, exposing the fungal mat to unspent culture medium comprises moving the fungal mat to a culture container containing unspent culture medium.

[0019] In one embodiment, exposing the fungal mat to unused culture medium comprises adding unused culture medium to the surface of the fungal mat.

[0020] In one embodiment, the method further comprises exposing the fungal mat to additional fungal material, optionally wherein the additional fungal material comprises fungal spores and / or fungal hyphae.

[0021] In one embodiment, physically manipulating the fungus mat comprises folding the fungus mat, optionally wherein folding the fungus mat comprises folding the fungus mat approximately in half.

[0022] In one embodiment, physically manipulating the fungus mat comprises turning the fungus mat over.

[0023] In one embodiment, physically manipulating the fungal mat comprises contacting the fungal mat with a support material, optionally wherein the support material comprises a fabric.

[0024] In one embodiment, physically manipulating the fungal mat comprises combining the fungal mat with one or more additional fungal mats to produce a combined fungal mat.

[0025] In one embodiment, the fungal mat and the additional fungal mat each comprise opposing upper and lower surfaces, and wherein combining the fungal mat with the additional fungal mat comprises: (a) contacting the lower surface of the fungal mat with the lower surface of the additional fungal mat; (b) contacting the upper surface of the fungal mat with the upper surface of the additional fungal mat; (c) contacting the lower surface of the fungal mat with the upper surface of the additional fungal mat; or (d) contacting the upper surface of the fungal mat with the lower surface of the additional fungal mat.

[0026] In one embodiment, the method includes placing a support material between the fungal mat and the additional fungal mat.

[0027] In one embodiment, the method further comprises applying a force to an outer surface of the composite fungal mat.

[0028] In one embodiment, the fungal mat is grown in a culture container, and wherein physically manipulating the fungal mat comprises separating an edge of the fungal mat from a side of the culture container.

[0029] In one embodiment, the method further comprises producing a fungal mat by culturing at least one fungal strain in a liquid culture medium. In one embodiment, the method comprises culturing at least one fungal strain in the presence of a support material, and wherein the fungal mat is adhered to the support material. In one embodiment, the method comprises inoculating a liquid growth medium with fungal spores, fungal hyphae and / or fungal biomass. In one embodiment, culturing at least one fungal strain in a liquid growth medium comprises culturing at least one fungal strain under static conditions. In one embodiment, the liquid growth medium is selected from potato dextrose broth and Sabouraud dextrose broth.

[0030] The present invention also provides a reinforced fungal mat obtainable by the method of the present invention. The present invention also provides a material comprising a reinforced fungal mat obtainable by the method of the present invention.

[0031] The present invention also provides use of the reinforced fungus mat of the present invention or the material of the present invention as a clothing material or a furniture decoration material.

[0032] The present invention also provides the use of the reinforced fungal mat of the present invention or the material of the present invention as food.

[0033] The present invention also provides use of the reinforced fungal mat of the present invention or the material of the present invention as a medical device.

[0034] The present invention also provides the use of a reinforced fungal mat of the present invention or a material of the present invention as a dermal or transdermal delivery system for metabolites.

[0035] The present invention also provides the use of the reinforced fungal mat of the present invention or the material of the present invention as a thermal insulation material.

[0036] The invention also provides the use of the reinforced fungal mat of the invention or the material of the invention as packaging material.

[0037] The invention also provides the use of the reinforced fungal mat of the invention or the material of the invention as a plastic substitute.

[0038] The invention also provides the use of the reinforced fungal mat of the invention or the material of the invention as a leather substitute.

[0039] The present invention also provides the use of the reinforced fungal mat of the present invention or the material of the present invention as a rubber substitute.

[0040] The invention also provides a clothing material comprising the reinforced fungal mat of the invention or the material of the invention.

[0041] The present invention also provides a furniture upholstery material comprising the reinforced fungal mat of the present invention or the material of the present invention.

[0042] The invention also provides a food product comprising the reinforced fungal mat of the invention or the material of the invention.

[0043] The present invention also provides a medical device comprising the reinforced fungal mat of the present invention or the material of the present invention.

[0044] The present invention also provides a dermal or transdermal delivery system for a metabolite comprising the reinforced fungal mat of the present invention or the material of the present invention.

[0045] The present invention also provides a thermal insulation material comprising the reinforced fungal mat of the present invention or the material of the present invention.

[0046] The invention also provides a packaging material comprising the reinforced fungal mat of the invention or the material of the invention.

[0047] The invention also provides a plastic substitute comprising the reinforced fungal mat of the invention or the material of the invention.

[0048] The invention also provides a leather substitute comprising the reinforced fungal mat of the invention or the material of the invention.

[0049] The present invention also provides a rubber substitute comprising the reinforced fungal mat of the present invention or the material of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 Fungal mat thickness is expressed as fold change relative to control (A). BL represents fortified fungal mat according to the present invention (treatment details are provided in Table 1). Error bars show standard error of measurement (SEM).

[0052] Figure 2 (i) Key points and structure of a fungal mat, depicting the upper (air-exposed / upward facing) and lower (medium-exposed / downward facing) surfaces of the fungal mat. (ii) Experimental setup for producing an initial fungal mat (also referred to herein as a "naturally occurring fungal mat").

[0053] Figure 3.Exemplary fungal mat operations, (i) turning over the initial fungal mat, thereby exposing the upper surface to the culture medium. (ii) moving the initial fungal mat to unused culture medium. (iii) turning over the initial fungal mat and moving to unused culture medium. (iv) adding unused culture medium to the upper surface of the initial fungal mat. (v) adding support material and unused culture medium to the upper surface of the initial fungal mat. (vi) adding additional fungal material (e.g., fungal spores, fungal hyphae, or other fungal biomass) to the upper surface of the initial fungal mat. (vii) adding support material and additional fungal material (e.g., fungal spores, fungal hyphae, or other fungal biomass) to the upper surface of the initial fungal mat. "(xn)" indicates that the steps can be repeated at least once. After each operation, the fungal mat is typically allowed to grow for a predetermined period of time.

[0054] Figure 4 . An exemplary fungus mat operation in which the initial fungus mat is folded in half. Following the operation, the fungus mat is typically allowed to grow for a predetermined period of time.

[0055] Figure 5 .Exemplary fungal mat manipulations include forming a combined fungal mat. Two initial fungal mats are grown separately and then brought together to provide a combined fungal mat using: (i) the upper side of each fungal mat in contact; (ii) the lower side of each fungal mat in contact; or (iii) the upper side of a first fungal mat in contact with the lower side of a second fungal mat. Each of these combined fungal mats can be exposed to further manipulations, such as: (a) adding a support material between the fungal mats; (b) applying a force to the outer surface of the combined fungal mat; or (c) a combination of (a) and (b). After each manipulation, the combined fungal mat is typically grown for a predetermined period of time, such as in a culture container containing a liquid culture medium.

[0056] Figure 6 Exemplary fungal mat manipulation methods involve using a support material in the production of an initial fungal mat by (i) adding the support material to a growth medium prior to inoculating the growth medium with the fungal strain, or (ii) directly inoculating the support material with the fungal strain prior to adding the support material to the growth medium.

[0057] Figure 7 . Exemplary fungal mat manipulation methods involve combining two or more combined fungal mats. As with the combined fungal mats, a support material and / or force can be applied after combining the combined fungal mats. The combined fungal mats produced by these manipulation methods can be allowed to grow for a predetermined period of time, such as in a culture container containing a liquid culture medium, (xn) indicating that the combination can be repeated "n" times (n is typically selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, although a greater number of combinations can also be performed). DETAILED DESCRIPTION OF THE INVENTION

[0059] The present invention provides a reinforced fungal mat, wherein the reinforced fungal mat has enhanced mechanical properties compared to a naturally occurring fungal mat. The reinforced fungal mat of the present invention has enhanced mechanical properties compared to a naturally occurring fungal mat formed by one or more fungi of the same species. The present invention also provides a method of producing a reinforced fungal mat. The present invention also provides a material comprising the reinforced fungal mat of the present invention.

[0060] Fungal mats typically contain a large number of fungal cells enclosed in an extracellular matrix. This extracellular matrix typically includes a combination of macromolecules (such as polysaccharides, proteins, nucleic acids, and lipids) and contributes to various functions, including cell and surface adhesion.

[0061] The fungal mats described herein are grown on liquid culture medium. It should be understood that the liquid culture medium referred to herein includes any fluid culture medium, including liquid and slurry-based culture medium. In some embodiments, the fungal mats described herein are grown by submerged fermentation.

[0062] As described herein, a fungal mat formed on the surface of a liquid growth medium generally includes an "upper side" and a "lower side". In this article, the "upper side" refers to the surface of the fungal mat that is exposed to the air during growth and / or the surface of the fungal mat that faces upward. The "lower side" refers to the surface of the fungal mat that is exposed to the liquid medium during growth and / or the surface of the fungal mat that faces downward. It should be understood that "face up" and "face down" and the like refer to the top surface and bottom surface of the fungal mat during static growth on a liquid medium, respectively. The upper side and the lower side of the fungal mat generally have different properties, such as different extracellular matrix compositions. The methods described herein can change the properties of the upper side and the lower side of the fungal mat, thereby producing a reinforced fungal mat.

[0063] As used herein, a "reinforced fungal mat" is a fungal mat having enhanced mechanical properties compared to a naturally occurring fungal mat produced by one or more fungi of the same species. The enhanced mechanical properties include, but are not limited to, increased tensile strength, increased ultimate load, increased thickness, and reduced surface adhesion. Due to the enhanced mechanical properties, the reinforced fungal mat of the present invention is advantageously suitable for a wider range of applications than a naturally occurring fungal mat.

[0064] As used herein, "naturally occurring fungal mat" refers to a fungal mat produced by one or more fungi that has not been manipulated by the methods of the present invention. An exemplary naturally occurring fungal mat is a fungal mat grown in a liquid culture medium under static conditions without any physical manipulation. For comparison purposes, a naturally occurring fungal mat is typically grown for the same period of time on the same type of growth medium as the fortified fungal mat. In some embodiments, a naturally occurring fungal mat is a fungal mat formed on the surface of a liquid culture medium under static growth conditions.

[0065] In some embodiments, the reinforced fungus mat has an increased ultimate load compared to a naturally occurring fungus mat. The ultimate load of a material (e.g., a fungus mat) is the maximum force that the material can withstand without breaking when stretched or pulled. The ultimate load is usually measured in Newtons. The ultimate load of a fungus mat can be measured using any suitable method known in the art, such as a universal testing machine (UTM). For example, the ultimate load of a fungus mat can be tested by placing the fungus mat between the clamps of a UTM and applying tension to the fungus mat until it breaks. The ultimate load is the ultimate tension that the fungus mat can withstand before breaking. As used herein, a fungus mat having an increased ultimate load relative to a naturally occurring fungus mat is a fungus mat having a higher ultimate load than a naturally occurring fungus mat.

[0066] In some embodiments, the ultimate load of the reinforced fungus mat is at least 10% greater than the ultimate load of the naturally occurring fungus mat. In some embodiments, the ultimate load of the reinforced fungus mat is at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% greater than the ultimate load of the naturally occurring fungus mat.

[0067] In some embodiments, the ultimate load of the reinforced fungal mat is at least 2 times greater than the ultimate load of a naturally occurring fungal mat. In some embodiments, the ultimate load of the reinforced fungal mat is at least 2.5 times, at least 3 times, at least 3.5 times, at least 4 times, at least 4.5 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 20 times greater than the ultimate load of a naturally occurring fungal mat.

[0068] In some embodiments, the reinforced fungal mat has an ultimate load of at least 1, at least 1.5, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 150, or at least 200 Newtons. In some embodiments, such as where the reinforced fungal mat does not include a support material, the reinforced fungal mat has an ultimate load of 10-70, 20-70, 30-70, 40-70, 40-60, or 45-55 Newtons.

[0069] In some embodiments, the reinforced fungal mat has increased tensile strength compared to a naturally occurring fungal mat. Tensile strength can be measured by indexing the ultimate load of a material with its cross-sectional area. Tensile strength is typically measured in Pascals (Pa). As used herein, a fungal mat having increased tensile strength relative to a naturally occurring fungal mat is a fungal mat having a higher tensile strength than a naturally occurring fungal mat.

[0070] In some embodiments, the tensile strength of the reinforced fungal mat is at least 10% greater than the tensile strength of a naturally occurring fungal mat. In some embodiments, the tensile strength of the reinforced fungal mat is at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% greater than the tensile strength of a naturally occurring fungal mat.

[0071] In some embodiments, the tensile strength of the reinforced fungal mat is at least 2 times greater than the tensile strength of a naturally occurring fungal mat. In some embodiments, the tensile strength of the reinforced fungal mat is at least 2.5 times, at least 3 times, at least 3.5 times, at least 4 times, at least 4.5 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 20 times greater than the tensile strength of a naturally occurring fungal mat.

[0072] In some embodiments, the reinforced fungus mat has a tensile strength of at least 1000, at least 1500, at least 2000, at least 3000, at least 4000, at least 5000, at least 6000, at least 7000, at least 8000, at least 9000, at least 10000, at least 11000, at least 12000, at least 13000, at least 14000, at least 15000, at least 20000, at least 30000, at least 40000, at least 50000, at least 75000, at least 100000, at least 150000, or at least 200000 Pa.

[0073] In some embodiments, the reinforced fungal mat has an increased thickness compared to a naturally occurring fungal mat. The thickness of the fungal mat can be measured using any suitable method known in the art, such as using a ruler or a thickness gauge. As used herein, the thickness of a fungal mat refers to the distance between the upper surface and the lower surface of the fungal mat.

[0074] In some embodiments, the thickness of the reinforced fungal mat is at least 10% greater than the thickness of a naturally occurring fungal mat. In some embodiments, the thickness of the reinforced fungal mat is at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% greater than the thickness of a naturally occurring fungal mat.

[0075] In some embodiments, the thickness of the reinforced fungal mat is at least 2 times greater than the thickness of a naturally occurring fungal mat. In some embodiments, the thickness of the reinforced fungal mat is at least 2.5 times, at least 3 times, at least 3.5 times, at least 4 times, at least 4.5 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 20 times greater than the thickness of a naturally occurring fungal mat.

[0076] In some embodiments, the reinforced fungus mat has a thickness of at least 3 mm. In some embodiments, the reinforced fungus mat has a thickness of at least 4 mm, at least 5 mm, at least 6 mm, at least 7 mm, at least 8 mm, at least 9 mm, at least 10 mm, at least 11 mm, at least 12 mm, at least 13 mm, at least 14 mm, at least 15 mm, at least 16 mm, at least 17 mm, at least 18 mm, at least 19 mm, at least 20 mm, at least 25 mm, at least 30 mm, at least 35 mm, at least 40 mm, at least 45 mm, at least 50 mm, at least 75 mm, or at least 100 mm.

[0077] In some embodiments, the reinforced fungus mat has reduced surface adhesion compared to the naturally occurring fungus mat. The surface adhesion of the fungus mat can be measured using any suitable method known in the art. For example, the surface adhesion of the fungus mat can be measured using a tape test, in which a sticky tape (e.g., a transparent tape) is placed on the surface of the fungus mat and then pulled off. A scale of 0 to 5 (low surface adhesion) is used to characterize the surface adhesion level, wherein the surface of the fungus mat remains adhered to the tape when the tape is removed, and when the surface of the fungus mat continues to tear after the tape is removed, the surface adhesion level of the fungus mat is "0"; when the surface of the fungus mat remains adhered to the tape when removed, but the surface of the fungus mat does not continue to tear after the tape is removed, the surface adhesion level is specified as "1"; surface adhesion levels "2", "3" and "4" are intermediate levels specified based on how much fungus mat surface remains adhered to the tape when the tape is removed from the fungus mat surface; and when the surface of the fungus mat is not removed with the tape and the surface of the fungus mat remains completely undamaged, the surface adhesion level is specified as "5". The reinforced fungus mat can have a greater surface adhesion level than the naturally occurring fungus mat. In some embodiments, the reinforced fungal mat has a surface adhesion rating of 4 or 5. In some embodiments, the reinforced fungal mat has a surface adhesion rating of 5.

[0078] In some embodiments, the surface adhesion of the reinforced fungal mat is at least 10% lower than the surface adhesion of a naturally occurring fungal mat. In some embodiments, the surface adhesion of the reinforced fungal mat is at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% lower than the surface adhesion of a naturally occurring fungal mat.

[0079] In some embodiments, the surface adhesion of the reinforced fungal mat is at least 2 times lower than the surface adhesion of a naturally occurring fungal mat. In some embodiments, the surface adhesion of the reinforced fungal mat is at least 2.5 times, at least 3 times, at least 3.5 times, at least 4 times, at least 4.5 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, or at least 20 times lower than the surface adhesion of a naturally occurring fungal mat.

[0080] The present invention provides a material comprising a reinforced fungal mat. In some embodiments, the material comprises a single reinforced fungal mat. In some embodiments, the material comprises more than one reinforced fungal mat, such as at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 75 or at least 100 reinforced fungal mats. The reinforced fungal mats can be the same or different. In some embodiments, the material comprises at least 2 reinforced fungal mats produced by different strains of one or more fungi. In some embodiments, the material comprises at least 2 reinforced fungal mats produced by the same strain of one or more fungi.

[0081] The material or reinforced fungal mat of the present invention can be used for a wide range of applications. In some embodiments, the material or reinforced fungal mat of the present invention is used to produce biofibers, for example for clothing and / or furniture decoration. Therefore, the present invention provides biofibers comprising the material and / or reinforced fungal mat of the present invention. The present invention also provides the use of the reinforced fungal mat of the present invention or the material of the present invention as clothing material or furniture decoration material. Clothing material is a flexible material suitable for producing clothing. Furniture decoration material is a flexible material suitable for covering furniture such as sofas and chairs.

[0082] In some embodiments, the materials or reinforced fungal mats of the present invention are used to produce biomaterials for use in medicine. Thus, the present invention provides biomaterials comprising the materials and / or reinforced fungal mats of the present invention. In some embodiments, the present invention provides medical devices comprising the materials or reinforced fungal mats of the present invention.

[0083] In some embodiments, the material or the fortified fungi comprises at least one fungal strain modified to express a transgene. In some embodiments, the transgene encodes a medically relevant protein. In some embodiments, the protein is a therapeutic protein, such as an antimicrobial protein. Therefore, in some embodiments, the material or the fortified fungal mat of the present invention is used as a protein delivery system, such as a dermal or transdermal delivery system or protein. In some embodiments, the material or the fortified fungal mat comprises at least one fungal strain modified to increase metabolite production relative to the corresponding wild-type fungal strain. In some embodiments, the metabolite is a medically relevant metabolite. In some embodiments, the metabolite is a therapeutic metabolite, such as an antimicrobial metabolite. Therefore, in some embodiments, the material or the fortified fungal mat of the present invention is used as a metabolite delivery system, such as a dermal or transdermal delivery system for metabolites. In some embodiments, the protein is an industrially relevant protein. In some embodiments, the metabolite is an industrially relevant metabolite. Any suitable method can be used to modify the fungal strain, such as by genetic engineering, site-directed mutagenesis or directed evolution.

[0084] In some embodiments, the material or reinforced fungal mat of the invention is used to produce thermal insulation material, for example for use in buildings. Thus, the invention provides thermal insulation material comprising the material and / or reinforced fungal mat of the invention.

[0085] In some embodiments, the material or reinforced fungal mat of the present invention is used to produce a food product. The food product may be a meat substitute. Thus, the present invention provides a food product comprising the material and / or reinforced fungal mat of the present invention.

[0086] The present invention provides a method for enhancing the mechanical properties of a fungal mat, the method comprising: (a) exposing the fungal mat to virgin culture medium; and / or (b) physically manipulating the fungal mat; to provide a reinforced fungal mat with enhanced mechanical properties. After exposure to virgin culture medium and / or physical manipulation, the fungal mat is typically allowed to grow for a predetermined period of time, such as until the fungal mat reaches a desired thickness. The inventors have discovered that the fungal mat structure can be improved by the methods of the present invention, and that such structural improvements achieve improvements in various mechanical properties of the fungal mat (e.g., thickness, tensile strength, ultimate load and / or surface adhesion). Without wishing to be bound by theory, the inventors believe that these improved mechanical properties are achieved by changes in gene expression (e.g., at the transcriptional and / or translational level) of a fungus / multiple fungi induced by the methods described herein.

[0087] The present invention also provides a reinforced fungal mat obtained by the method of the present invention.

[0088] An "initial fungal mat" is typically a fungal mat grown in a liquid medium under static conditions (e.g., Figure 2(ii)). The "initial fungal mat" without further manipulation generally corresponds to a naturally occurring fungal mat.

[0089] As used herein, growing "in" a liquid medium generally refers to growing on or substantially at the surface of a liquid medium.

[0090] In some embodiments, the fungal mat grows for a "predetermined period of time". In some embodiments, the predetermined period of time is the period of time required for the fungal strain to establish a visible fungal mat. In some embodiments, the predetermined period of time is the period of time required for the fungal mat to reach a desired thickness. One skilled in the art will appreciate that the predetermined period of time can be routinely optimized based on the growth rate of a particular fungal strain.

[0091] As used herein, "virgin culture medium" generally refers to a growth medium that has not been previously used to support microbial growth (e.g., fungal growth). In some embodiments, exposing the fungal mat to virgin culture medium comprises moving the fungal mat to a culture container (e.g., such as a culture vessel) containing virgin culture medium. Figure 3 (ii)). In some embodiments, exposing the fungal mat to an unused culture medium comprises replacing the growth medium in which the fungal mat is grown with the unused culture medium. In some embodiments, exposing the fungal mat to an unused culture medium comprises adding the unused growth medium to the culture container in which the fungal mat is already grown. In some embodiments, the unused culture medium has been previously used to support microbial growth, but retains the ability to support further microbial growth. In some embodiments, where the unused culture medium has been previously used to support microbial growth, the unused culture medium has been sterilized (e.g., autoclaved) prior to use in the methods of the present invention. In some embodiments, where the unused culture medium has been previously used to support microbial growth, the unused culture medium has not been sterilized after supporting microbial growth and prior to use in the methods of the present invention.

[0092] In some embodiments, exposing the fungal mat to unused culture medium comprises adding unused culture medium to the upper surface of the fungal mat (e.g., Figure 3(iv)). In some embodiments, the fungal mat can be submerged so that both the upper and lower surfaces of the fungal mat are exposed to the culture medium. In some embodiments, a sufficient volume of unused culture medium is added to the surface of the fungal mat so that the edge of the fungal mat is separated from the side of the culture container. In some embodiments, exposing the fungal mat to the unused culture medium comprises: (i) replacing the growth medium in which the fungal mat has been grown in the culture container with the unused culture medium; and (ii) adding the unused culture medium to the upper surface of the fungal mat. In some embodiments, exposing the fungal mat to the unused culture medium comprises: (i) moving the fungal mat to a culture container containing unused culture medium; and (ii) adding the unused culture medium to the upper surface of the fungal mat.

[0093] In some embodiments, the method further comprises exposing the fungal mat to additional fungal material (e.g., Figure 3 (vi)). In some embodiments, the additional fungal material is selected from fungal spores, fungal hyphae, or other fungal biomass. The fungal biomass can be a fungal colony isolated from a solid growth medium (e.g., an agar plate). In some embodiments, the fungal mat is exposed to unused culture medium containing additional fungal material. As used herein, additional fungal material includes any reproductive form of fungal biomass, including but not limited to spores and hyphae.

[0094] As used herein, physically manipulating a fungus mat means that the fungus mat is moved in some manner, e.g., it is moved, displaced, and / or rotated. In some embodiments, physically manipulating a fungus mat comprises folding the fungus mat. When folding the fungus mat, portions of the same surface of the fungus mat are in contact with each other. In some embodiments, folding the fungus mat comprises folding the fungus mat approximately in half, e.g., along an approximate line of symmetry (e.g., such as Figure 4 (i)). In some embodiments, folding the fungus mat comprises folding an edge of the fungus mat toward a center of the fungus mat.

[0095] In some embodiments, physically manipulating the fungus mat comprises turning the fungus mat over. When turning the fungus mat over, the surfaces are generally inverted so that the upper surface becomes the lower surface (e.g., Figure 3 In some embodiments, the fungal mat is turned over and then exposed to fresh culture medium (e.g., Figure 3 (iii)).

[0096] In some embodiments, physically manipulating the fungal mat includes combining the fungal mat with one or more additional fungal mats to produce a combined fungal mat. The combined fungi are then typically grown for a predetermined period of time, such as until the combined fungi reaches a desired thickness. As used herein, a "combined fungal mat" is a composite fungal mat formed by combining at least two fungal mats that have been grown separately.

[0097] In some embodiments, the fungal mat and the additional fungal mat are combined in opposite orientations, for example, the method comprises contacting: (a) contacting the lower surface of the fungal mat with the lower surface of the additional fungal mat (e.g., Figure 5 (ii); or (b) contacting the upper surface of the fungal mat with the upper surface of another fungal mat (e.g., Figure 5 (as shown in (i)).

[0098] In some embodiments, the fungal mat and the additional fungal mat are combined in the same orientation such that opposing surfaces are in contact, e.g., (a) the lower surface of the fungal mat is in contact with the upper surface of the additional fungal mat; and / or (b) the upper surface of the fungal mat is in contact with the lower surface of the additional fungal mat (e.g., Figure 5 (iii)).

[0099] In some embodiments, the method comprises combining the fungal mat with at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, or at least 20 additional fungal mats. In some embodiments, one or more additional fungal mats are initial fungal mats as described herein. In some embodiments, one or more additional fungal mats are reinforced fungal mats as described herein. In some embodiments, the additional fungal mats comprise one or more initial fungal mats and one or more reinforced fungal mats.

[0100] In some embodiments, the method further comprises combining the combined fungal mat with at least a second combined fungal mat (e.g., Figure 7 shown).

[0101] In some embodiments, physically manipulating the fungus mat includes folding the fungus mat and then combining the folded fungus mat with an additional fungus mat to produce a combined fungus mat. In some embodiments, the additional fungus mat is positioned to cover an edge of the folded fungus mat. In some embodiments, the additional fungus mat is positioned to cover an edge of the folded fungus mat, and excess portions of the additional fungus mat are removed (e.g., Figure 4 (ii)).

[0102] In some embodiments, the method further comprises applying a force to the combined fungal mat (e.g., Figure 5(b) and (c)). The force is typically used to press the fungus mat together, and is typically applied prior to the growth of the combined fungus mat, and is typically continued for a predetermined period of time, such as until the combined fungus mat reaches a desired thickness. In some embodiments, the force is applied by placing the combined fungus mat in a fixture. In some embodiments, the force is applied by placing a weight on the surface of the combined fungus mat.

[0103] The force can be measured relative to the surface area of ​​the fungus mat, i.e., the pressure applied to the fungus mat. The pressure applied to the fungus mat is typically at least 100, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, at least 900, or at least 1000 Pascals.

[0104] In some embodiments, physically manipulating the fungal mat comprises contacting the fungal mat with a support material. In some embodiments, the support material comprises a fabric. In some embodiments, the method comprises placing the support material in contact with the upper surface of the fungal mat. In some embodiments, the method comprises: (i) placing the support material in contact with the upper surface of the fungal mat; and (ii) adding unused culture medium to the upper surface of the fungal mat (e.g., Figure 3 (v)).

[0105] In some embodiments, the method includes: (i) placing a support material in contact with the upper surface of the fungal mat; and (ii) exposing the fungal mat to additional fungal material (e.g., Figure 3 (vii)).

[0106] In some embodiments, when forming a combined fungal mat as described above, the method includes placing a support material between the fungal mat and another fungal mat (e.g., Figure 5 (as shown in (a)).

[0107] In some embodiments, physically manipulating the fungal mat includes separating the fungal mat from a culture container in which it has been grown, such as separating the edge of the fungal mat from the side of the culture container.

[0108] In some embodiments, the method includes: (i) turning over the fungus mat; and (ii) placing a support material on top of the fungus mat. In some embodiments, the method includes: (i) turning over the fungus mat; and (ii) placing a second fungus mat on top of the first fungus mat to provide a combined fungus mat. In some embodiments, the method includes: (i) turning over the fungus mat; and (ii) placing the second turned over fungus mat on top of the first fungus mat to provide a combined fungus mat. In some embodiments, the method includes: (i) turning over the fungus mat; (ii) placing a support material on top of the fungus mat; and (iii) placing the second turned over fungus mat on top of the support material to provide a combined fungus mat. In some embodiments, the method includes: (i) turning over the fungus mat; (ii) placing a support material on top of the fungus mat; and (iii) placing the second turned over fungus mat on top of the support material to provide a combined fungus mat. In some embodiments, the method includes turning over the fungus mat. In some embodiments, the method includes transferring the fungus mat to a container containing unused culture medium. In some embodiments, the method includes: (i) sliding the fungal mat; and (ii) transferring the fungal mat to a container containing unused culture medium. In each of these embodiments, the method also includes growing the fungal mat or combined fungal mats, typically for a predetermined period of time, such as until the fungal mat or combined fungal mat reaches a desired thickness.

[0109] In some embodiments, the method also includes producing a fungal mat by cultivating a fungal strain in a liquid culture medium. The method can include inoculating the liquid culture medium with other fungal materials (such as fungal hyphae and / or spores), and incubating the culture until a visible fungal mat is formed. In some embodiments, the culture is incubated until a fungal mat with a desired thickness is formed. The desired thickness can be at least 1 mm, at least 2 mm, at least 3 mm, at least 4 mm, or at least 5 mm. In some embodiments, the culture is incubated for a predetermined time period, such as about 4 days, about 7 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 8 weeks, about 10 weeks, or about 12 weeks.

[0110] In some embodiments, the method includes culturing the fungal strain in the presence of a support material. In some embodiments, as the fungal mat grows, it adheres to the support material. In some embodiments, the method includes directly inoculating the support material with the fungal strain and then adding the support material to a growth medium.

[0111] In some embodiments, the method further comprises drying the strengthened fungus mat. In some embodiments, the strengthened fungus mat is dried by heating the strengthened fungus mat to at least 30° C., at least 35° C., at least 40° C., at least 45° C., at least 50° C., at least 55° C., at least 60° C., at least 65° C., at least 70° C., at least 75° C., at least 80° C., at least 85° C., at least 90° C., at least 95° C., or at least 100° C. In some embodiments, the strengthened fungus mat is dried by heating the strengthened fungus mat for at least 30 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 6 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 24 hours, or at least 48 hours.

[0112] The fungus mat manipulation steps described above can be combined in any manner. Each fungus mat manipulation step described herein can be repeated at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 11 times, at least 12 times, at least 13 times, at least 14 times, at least 15 times, or at least 20 times.

[0113] A support material as described herein is any suitable material to which a fungal mat can adhere during growth. Exemplary support materials include, but are not limited to, fabrics made of cotton, hemp, polyester, nylon, denim, wool, and / or silk. Exemplary support materials also include food products, such as meat substitutes and / or tofu.

[0114] The fungal strain can be any fungal strain or a combination of fungal strains capable of forming a fungal mat. In some embodiments, the fungus is an Ascomycota or a Basidiomycota. In some embodiments, the fungus is a member of the Clavicipitaceae or Trichocomaceae families. In some embodiments, the fungus is a member of the genus selected from: Cordyceps, Metacordyceps, Pleurotus, Hericium, Grifola, Fistulina, Fomes, Fomitopsis, Inonotus, Trametes Ganoderma, Aspergillus, Penicillium, Chrysosporium, Myceliophthora, Trichoderma, Acremonium, and Fusarium. In some embodiments, the fungus is selected from Cordyceps sensu lato, Pleurotus ostreatus, Pleurotus eryngii, Hericium erinaceus, Grifola frondosa, Fistulina hepatica, Fomes fomentarius, Fomitopsis pinicola, Inonotus obliquus, Trametes versicolor, Ganoderma applanatum, and Ganoderma lucidum.

[0115] In some embodiments, the reinforced fungus mat is a reinforced Cordyceps mat. In some embodiments, the reinforced fungus mat is a reinforced Aspergillus mat. In some embodiments, the reinforced fungus mat is a reinforced Pleurotus ostreatus mat. In some embodiments, the reinforced fungus mat is a reinforced Pleurotus eryngii mat. In some embodiments, the reinforced fungus mat is a reinforced Hericium erinaceus mat. In some embodiments, the reinforced fungus mat is a reinforced Grifola frondosa mat. In some embodiments, the reinforced fungus mat is a reinforced Liver-colored Beefsteak Mat. In some embodiments, the reinforced fungus mat is a reinforced Wood Hoof Foliar Mat. In some embodiments, the reinforced fungus mat is a reinforced Pinus pineoides mat. In some embodiments, the reinforced fungus mat is a reinforced Inonotus birchii mat. In some embodiments, the reinforced fungus mat is a reinforced Trametes versicolor mat. In some embodiments, the reinforced fungus mat is a reinforced Ganoderma lucidum mat. In some embodiments, the reinforced fungus mat is a reinforced Ganoderma lucidum mat.

[0116] The growth medium can be any suitable growth medium, and is typically a liquid growth medium. In some embodiments, the growth medium is selected from potato dextrose broth, Sabouraud dextrose broth, Czapek-Dox broth, and malt extract broth.

[0117] The fungal mat is typically grown for a predetermined period of time. In some embodiments, the predetermined period of time is, for example, about 4 days, about 7 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 8 weeks, about 10 weeks, or about 12 weeks. In some embodiments, the predetermined period of time is the period of time required for the fungal mat to grow to a desired thickness, such as at least 1 mm, at least 2 mm, at least 3 mm, at least 4 mm, at least 5 mm, at least 6 mm, at least 7 mm, at least 8 mm, at least 9 mm, or at least 10 mm.

[0118] Fungal mats are usually grown under static conditions.

[0119] The fungal mats typically grow between 15°C and 60°C, 15°C and 50°C, 15°C and 45°C, 15°C and 40°C, 15°C and 35°C, 15°C and 30°C, 20°C and 60°C, 20°C and 50°C, 20°C and 45°C, 20°C and 40°C, 20°C and 35°C, 20°C and 30°C, 30°C and 60°C, 30°C and 50°C, 30°C and 45°C, 30°C and 40°C, or 30°C and 35°C. Example

[0120] The present invention will be further illustrated by the following examples, which are intended to be purely illustrative of the present invention and are in no way limiting.

[0121] Example 1

[0122] Effect of operation method on fungal mat thickness

[0123] Fungal mats were produced by inoculating 80 ml of potato dextrose broth in an Erlenmeyer flask with a Cordyceps spore suspension containing approximately 2 million spores. The culture was incubated at 24° C. under dark, static conditions for 2 weeks to form a naturally occurring fungal mat, also referred to herein as an initial fungal mat. For control, hereinafter designated as "A", the initial fungal mat was grown for another 2 weeks without any additional manipulation.

[0124] The initial fungal mats were then exposed to various manipulations of the invention as described in Table 1. The experiments were performed in triplicate.

[0125] Following the procedures described in Table 1, the initial fungal mats and reinforced fungal mats (designated "B" - "L") were dried at 45°C for several hours and the thickness was measured.

[0126] Table 1: Fungus Mat Operation Methods

[0127]

[0128]

[0129] Table 2: Thickness of fungal mats provided as fold change relative to control mats

[0130] A B C D E F G H I J K L average 1 5.3 16 5.3 8 5.3 5 5 5.3 3.7 7.7 8 Rep 1 1 6 20 6 6 4 5 5 5 3 7 8 Rep 2 1 5 14 6 10 6 5 5 5 4 8 7 Rep 3 1 5 14 4 8 6 5 5 6 4 8 9

[0131] Each of the manipulations described in Table 1 produced a reinforced fungal mat with significantly improved thickness compared to the initial fungal mat (A). The average increase in thickness ranged from 3.7 to 16 times relative to the initial fungal mat (see Tables 2 and Figure 1 ).

[0132] These data clearly demonstrate that the process of the present invention significantly improves the mechanical properties (eg thickness) of the initial fungal mat, thereby providing a reinforced fungal mat that can be used in a range of applications.

[0133] Example 2

[0134] Production of fungus-based meat substitutes

[0135] The edible fungal strain or combination of edible fungal strains is selected from Pleurotus ostreatus, Pleurotus eryngii, Hericium erinaceus, Grifola frondosa, Hepatica serrata, Faecalibacterium arvense, Pseudomonas syringae, Inonotus birchii, Trametes versicolor, Ganoderma lucidum and / or Ganoderma lucidum. The fungal strain is grown in a liquid medium under static conditions until a visible fungal mat is formed.

[0136] The fungal mat was then exposed to one of the procedures described in Table 1 to provide a reinforced fungal mat. The wet thickness of the reinforced fungal mat was then measured.

[0137] The operating method of the present invention significantly improves the thickness of the fungus mat, thereby providing an edible fungus mat that can be used as a meat substitute.

Claims

1. A method for enhancing the mechanical properties of a fungal mat grown in a liquid medium, the method comprising: (a) exposing the fungal mat to virgin culture medium; and / or (b) physical manipulation of the fungal mat; To provide a reinforced fungus mat having enhanced mechanical properties relative to a fungus mat.

2. The method of claim 1, wherein the method comprises: (a) exposing the fungal mat to virgin culture medium; and (b) Physical manipulation of fungal mats.

3. The method of claim 1 or claim 2, wherein the method comprises exposing the fungal mat to unused culture medium and physically manipulating the fungal mat by turning the fungal mat over, wherein: (i) turning the fungal mat before exposure to unused culture medium; and / or (ii) turning the fungal mat after exposure to unused culture medium.

4. A method according to any preceding claim, wherein the method comprises exposing the fungal mat to unused culture medium and physically manipulating the fungal mat by placing a further fungal mat on the upper side of the fungal mat.

5. The method of any preceding claim, wherein exposing the fungal mat to unspent culture medium comprises moving the fungal mat to a culture container containing unspent culture medium.

6. The method of any preceding claim, wherein exposing the fungal mat to virgin culture medium comprises adding virgin culture medium to the surface of the fungal mat.

7. The method of any preceding claim, wherein the method further comprises exposing the fungal mat to further fungal material, optionally wherein the further fungal material comprises fungal spores and / or fungal hyphae.

8. The method of any preceding claim, wherein physically manipulating the fungus mat comprises folding the fungus mat, optionally wherein folding the fungus mat comprises folding the fungus mat substantially in half.

9. The method of any preceding claim, wherein physically manipulating the fungal mat comprises turning the fungal mat.

10. A method according to any preceding claim, wherein physically manipulating the fungal mat comprises contacting the fungal mat with a carrier material, optionally wherein the carrier material comprises a fabric.

11. The method of any preceding claim, wherein physically manipulating the fungal mat comprises combining the fungal mat with one or more additional fungal mats to produce a combined fungal mat.

12. The method of claim 11, wherein the fungal mat and the additional fungal mat each comprise opposing upper and lower surfaces, and wherein combining the fungal mat with the additional fungal mat comprises: (a) contacting the lower surface of the fungal mat with the lower surface of another fungal mat; (b) contacting the upper surface of the fungal mat with the upper surface of another fungal mat; (c) contacting the lower surface of the fungal mat with the upper surface of another fungal mat; or (d) contacting the upper surface of the fungal mat with the lower surface of another fungal mat.

13. A method according to claim 11 or claim 12, wherein the method comprises placing a support material between the fungal mat and the further fungal mat.

14. The method of any one of claims 11-13, wherein the method further comprises applying a force to an outer surface of the composite fungal mat.

15. The method of any preceding claim, wherein the fungal mat is grown in a culture container, and wherein physically manipulating the fungal mat comprises separating edges of the fungal mat from sides of the culture container.

16. The method of the preceding claim, wherein the method further comprises producing the fungal mat by culturing at least one fungal strain in a liquid culture medium.

17. The method of claim 16, wherein the method comprises culturing at least one fungal strain in the presence of a support material, and wherein the fungal mat adheres to the support material.

18. The method of claim 16 or claim 17, wherein the method comprises inoculating a liquid growth medium with fungal spores, fungal hyphae and / or fungal biomass.

19. The method of any one of claims 16-18, wherein culturing the at least one fungal strain in a liquid growth medium comprises culturing the at least one fungal strain under static conditions.

20. The method of any one of claims 16-19, wherein the liquid growth medium is selected from potato dextrose broth and Sabouraud dextrose broth.

21. A reinforced fungal mat obtainable by the method of any one of the preceding claims.

22. A reinforced fungal mat, wherein: (a) the reinforced fungal mat has enhanced mechanical properties compared to naturally occurring fungal mats; and (b) The enhanced fungal mat comprises one or more fungi of the same species as the naturally occurring fungal mat.

23. The reinforced fungal mat of claim 22, wherein the reinforced fungal mat has increased tensile strength compared to a naturally occurring fungal mat.

24. The reinforced fungal mat of claim 23, wherein the tensile strength of the reinforced fungal mat is at least 10% greater than the tensile strength of a naturally occurring fungal mat.

25. The reinforced fungal mat of any one of claims 22-24, wherein the reinforced fungal mat has a tensile strength of at least 1000 Pa.

26. The reinforced fungal mat of any one of claims 22-25, wherein the reinforced fungal mat has an ultimate load of at least 3 Newtons.

27. The reinforced fungal mat of any one of claims 22-26, wherein the reinforced fungal mat has an increased thickness compared to a naturally occurring fungal mat.

28. The reinforced fungal mat of claim 27, wherein the thickness of the reinforced fungal mat is at least 3 times greater than the thickness of a naturally occurring fungal mat.

29. The reinforced fungal mat of any one of claims 22-28, wherein the reinforced fungal mat has a thickness of at least 5 mm.

30. The reinforced fungal mat of any one of claims 22-29, wherein the reinforced fungal mat has reduced surface adhesion compared to a naturally occurring fungal mat.

31. The reinforced fungal mat of claim 30, wherein the surface adhesion of the reinforced fungal mat is at least 10% less than the surface adhesion of a naturally occurring fungal mat.

32. The reinforced fungal mat according to any one of claims 22-31, wherein the reinforced fungal mat further comprises a carrier material, optionally wherein the carrier material comprises a fabric.

33. A material comprising the reinforced fungal mat of any one of claims 21-32.

34. The material of claim 33, wherein the material comprises two or more reinforced fungal mats.

35. The reinforced fungal mat of any one of claims 22-32 or the material of claim 33 or 34, wherein the reinforced fungal mat is obtained by the method of any one of claims 1-20.

36. Use of the reinforced fungal mat of any one of claims 21 to 32 or 35 or the material of any one of claims 33 to 35 as a clothing material or furniture upholstery material.

37. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a food.

38. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a medical device.

39. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a dermal or transdermal delivery system for a metabolite.

40. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a thermal insulation material.

41. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as packaging material.

42. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a plastic substitute.

43. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a leather substitute.

44. Use of a reinforced fungal mat according to any one of claims 21 to 32 or 35 or a material according to any one of claims 33 to 35 as a rubber substitute.

45. A clothing material comprising the reinforced fungal mat of any one of claims 21 to 32 or 35, or the material of any one of claims 33 to 35.

46. ​​A furniture upholstery material comprising the reinforced fungal mat of any one of claims 21-32 or 35, or the material of any one of claims 33-35.

47. A food product comprising the reinforced fungal mat of any one of claims 21 to 32 or 35, or the material of any one of claims 33 to 35.

48. A medical device comprising the reinforced fungal mat of any one of claims 21-32 or 35, or the material of any one of claims 33-35.

49. A dermal or transdermal delivery system for a metabolite comprising a reinforced fungal mat according to any one of claims 21 to 32 or 35, or a material according to any one of claims 33 to 35.

50. A thermal insulation material comprising the reinforced fungal mat of any one of claims 21 to 32 or 35, or the material of any one of claims 33 to 35.

51. A packaging material comprising the reinforced fungal mat of any one of claims 21 to 32 or 35, or the material of any one of claims 33 to 35.

52. A plastic substitute comprising the reinforced fungal mat of any one of claims 21-32 or 35, or the material of any one of claims 33-35.

53. A leather substitute comprising the reinforced fungal mat of any one of claims 21 to 32 or 35, or the material of any one of claims 33 to 35.

54. A rubber substitute comprising the reinforced fungal mat of any one of claims 21-32 or 35, or the material of any one of claims 33-35.

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