Composite material and method for manufacturing the same

Abstract

Provided herein are mycelial materials and methods of making the same. In some embodiments, the mycelial material comprises: a cultivated mycelial material comprising one or more clumps of branched hyphae, wherein the one or more clumps of branched hyphae can be disrupted or pressurized and / or an adhesive can be combined with the cultivated mycelial material. Also provided are methods of making mycelial materials.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 851,867, filed May 23, 2019; and U.S. Provisional Application No. 63 / 024,368, filed May 13, 2020; each of which is hereby incorporated by reference in its entirety. Technical Field

[0003] The present disclosure generally relates to various mycelial materials having a grown mycelial component and methods of making the same to provide advantageous mechanical and aesthetic qualities. Background Art

[0004] Mycelium is of increasing interest in next generation sustainable materials due to its bioefficiency, strength, and low environmental footprint. To this end, methods for growing networks of entangled mycelium (e.g., entangled with particles, fibers, fiber networks, solid matrix adhesives, or non-woven thin layers) have been discussed for a variety of applications, both alone and as composite materials. However, mycelium materials currently under development have poor mechanical qualities, including susceptibility to delamination and tearing under pressure, and uneven aesthetic qualities. Therefore, there is a need for improved mycelium materials having favorable mechanical properties, aesthetic properties, and other advantages, as well as materials and methods for preparing improved mycelium materials. Summary of the invention

[0005] According to some embodiments, various mycelium materials and methods of making the same are provided herein to provide mycelium materials and composite mycelium materials with advantageous mechanical and aesthetic qualities and related advantages.

[0006] In one aspect, a composite mycelium material is provided herein. In one aspect, a composite mycelium material is provided herein, comprising: a cultivated mycelium material comprising one or more clumps of branched hyphae, wherein the one or more clumps of branched hyphae are disrupted; and a binder.

[0007] In some embodiments, the cultivated mycelial material has been produced on a solid substrate.

[0008] In some embodiments, the cultivated mycelial material comprises one or more clumps of disrupted branched hyphae. In some embodiments, the one or more clumps of disrupted branched hyphae have a length of 0.1 mm to 5 mm. In some embodiments, the one or more clumps of disrupted branched hyphae have a length of 2 mm.

[0009] In some embodiments, the one or more masses of branched hyphae are entangled, wherein entangling the hyphae comprises needle punching, felting, or hydroentangling. In some embodiments, the entangled hyphae are hydroentangled.

[0010] In some embodiments, the adhesive comprises one or more reactive groups. In some embodiments, the one or more reactive groups react with active hydrogen-containing groups. In some embodiments, active hydrogen-containing groups include amines, hydroxyls and carboxyls. In some embodiments, the adhesive comprises an adhesive, a resin, a crosslinking agent and / or a matrix. In some embodiments, the adhesive is selected from the group consisting of vinyl acetate-ethylene (VAE) copolymers, vinyl acetate-acrylic copolymers, polyamide-epichlorohydrin resins (PAE), copolymers, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives and synthetic adhesives. In some embodiments, the adhesive is a copolymer having a characteristic selected from the group consisting of: a particle size less than or equal to 1 μm, a subzero glass transition temperature and a self-crosslinking function. In some embodiments, the adhesive is a vinyl acetate-ethylene (VAE) copolymer. In some embodiments, the adhesive is a vinyl acetate-acrylic copolymer. In some embodiments, the adhesive is PAE. In some embodiments, the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of the hyphae.

[0011] In some embodiments, the natural adhesive comprises a natural latex-based adhesive. In some embodiments, the natural latex-based adhesive is leather glue or weld.

[0012] In some embodiments, the composite mycelium material further comprises a support material. In some embodiments, the support material has a pore size of 1 / 16 inch.

[0013] In some embodiments, the support material comprises a reinforcing material. In some embodiments, the reinforcing material is entangled within the composite mycelium material. In some embodiments, the support material comprises a base material. In some embodiments, the base material is located on one or more surfaces of the composite mycelium material. In some embodiments, the support material is selected from the group consisting of a mesh fabric, a gauze, a fabric, a knitted textile, a woven textile, and a non-woven textile.

[0014] In some embodiments, the one or more masses of branched hyphae are disrupted by mechanical action. In some embodiments, the mechanical action comprises mixing the one or more masses of branched hyphae. In some embodiments, the mechanical action comprises breaking the one or more masses of branched hyphae. In some embodiments, the mechanical action comprises applying a physical force to the one or more masses of branched hyphae so that at least some of the masses of branched hyphae are aligned in a parallel form. In some embodiments, the physical force is a pulling force.

[0015] In some embodiments, the mechanical action comprises applying the physical force in one or more directions such that at least some of the branched hyphal masses are aligned in parallel in the one or more directions, wherein the physical force is applied repeatedly.

[0016] In some embodiments, the one or more masses of branched hyphae are disrupted by chemical treatment. In some embodiments, the chemical treatment comprises contacting the one or more masses of branched hyphae with a base or other chemical agent in an amount sufficient to cause disruption. In some embodiments, the base comprises an alkaline peroxide.

[0017] In some embodiments, the composite mycelium material comprises one or more proteins from a species other than the fungal species that produced the cultivated mycelium material. In some embodiments, the one or more proteins are from a plant source. In some embodiments, the plant source is a pea plant. In some embodiments, the plant source is a soybean plant.

[0018] In some embodiments, the composite mycelium material further comprises a dye. In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes. In some embodiments, the dye is a reactive dye. In some embodiments, the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform on one or more surfaces of the composite mycelium material. In some embodiments, the dye is present throughout the interior of the composite mycelium material.

[0019] In some embodiments, the composite mycelium material further comprises a plasticizer. In some embodiments, the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglyceride and epoxidized soybean oil. In some embodiments, the plasticizer is a fatliquor.

[0020] In some embodiments, the composite mycelium material is flexible.

[0021] In some embodiments, an external element is applied to the cultivated mycelial material. In some embodiments, the external element is applied via heating and / or pressing. In some embodiments, the external element is a hot press.

[0022] In some embodiments, the composite mycelium material further comprises tannin.

[0023] In some embodiments, the composite mycelium material further comprises a finish. In some embodiments, the finish is selected from the group consisting of urethane, wax, nitrocellulose, and a plasticizer.

[0024] In some embodiments, the composite mycelium material comprises mechanical properties.

[0025] In some embodiments, the mechanical properties include wet tensile strength, initial modulus, elongation at break, thickness, and / or slit tear strength.

[0026] In some embodiments, the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa. In some embodiments, the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa. In some embodiments, the composite mycelium material has a wet tensile strength of 7 MPa. In some embodiments, the composite mycelium material has an initial modulus of 1 MPa to 100 MPa. In some embodiments, the composite mycelium material has an elongation at break of 1% to 25%. In some embodiments, the composite mycelium material has a thickness of 0.5 mm to 3.5 mm. In some embodiments, the composite mycelium material has a thickness of 2 mm. In some embodiments, the composite mycelium material has a crack tear strength of 5N to 100N. In some embodiments, the composite mycelium material has a crack tear strength of 50N.

[0027] In some embodiments, the composite mycelium material is made using conventional papermaking equipment.

[0028] In another aspect, provided herein is a composite mycelium material comprising: a cultivated mycelium material comprising one or more clumps of branched hyphae, wherein the one or more clumps of branched hyphae are disrupted; a binder comprising vinyl acetate-ethylene copolymer; and a reactive dye.

[0029] In another aspect, provided herein is a method for manufacturing a composite mycelium material, the method comprising: producing a cultivated mycelium material comprising one or more masses of branched hyphae; disrupting the cultivated mycelium material comprising the one or more masses of branched hyphae; and adding a binder to the cultivated mycelium material; thereby manufacturing the composite mycelium material.

[0030] In some embodiments, the binder is added before the branched hyphae mass is disrupted, during the branched hyphae mass is disrupted, or after the branched hyphae mass is disrupted.

[0031] In some embodiments, the producing comprises producing the cultivated mycelial material on a solid substrate.

[0032] In some embodiments, the cultivated mycelial material comprises one or more masses of disrupted branching hyphae.

[0033] In some embodiments, the one or more clumps of disrupted branched hyphae have a length of 0.1 mm to 5 mm.

[0034] In some embodiments, the one or more clumps of disrupted branched hyphae have a length of 2 mm.

[0035] In some embodiments, the method further comprises entangling the one or more masses of branched hyphae, wherein entangling the hyphae comprises needle punching, felting, or hydroentangling.

[0036] In some embodiments, entangling the hyphae comprises hydroentangling.

[0037] In some embodiments, hydroentangling the one or more masses of branched hyphae comprises hydroentangling using a liquid jet configured to eject liquid at a pressure of about 700 psi to about 1000 psi.

[0038] In some embodiments, hydroentangling comprises using liquid jets configured to eject liquid at a flow rate of about 100 mL / min to 300 mL / min.

[0039] In some embodiments, the disrupting comprises disrupting the one or more clumps of branched hyphae by mechanical action.

[0040] In some embodiments, the mechanical action comprises admixing the one or more masses of branched hyphae.

[0041] In some embodiments, the mechanical action comprises breaking the one or more clumps of branched hyphae.

[0042] In some embodiments, the mechanical action comprises applying a physical force to the one or more masses of branched hyphae such that at least some of the masses of branched hyphae are aligned in a parallel fashion.

[0043] In some embodiments, the physical force is a tensile force.

[0044] In some embodiments, the mechanical action comprises applying the physical force in one or more directions such that at least some of the branched hyphal masses are aligned in parallel in the one or more directions, wherein the physical force is applied repeatedly.

[0045] In some embodiments, the one or more clumps of branching hyphae are disrupted by chemical treatment.

[0046] In some embodiments, the chemical treatment comprises contacting the one or more masses of branching hyphae with an amount of alkali or other chemical agent sufficient to cause disruption.

[0047] In some embodiments, the base comprises a basic peroxide.

[0048] In some embodiments, the binder comprises one or more reactive groups.

[0049] In some embodiments, the one or more reactive groups react with active hydrogen-containing groups.

[0050] In some embodiments, active hydrogen-containing groups include amine, hydroxyl, and carboxyl groups.

[0051] In some embodiments, the adhesive comprises a sticker, a resin, a cross-linking agent, and / or a matrix.

[0052] In some embodiments, the adhesive is selected from the group consisting of vinyl acetate-ethylene copolymers, vinyl acetate-acrylic copolymers, polyamide-epichlorohydrin resins (PAE), copolymers, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives, and synthetic adhesives.

[0053] In some embodiments, the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

[0054] In some embodiments, the binder is vinyl acetate-ethylene copolymer.

[0055] In some embodiments, the binder is a vinyl acetate-acrylic copolymer.

[0056] In some embodiments, the binder is PAE.

[0057] In some embodiments, the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of the hyphae.

[0058] In some embodiments, the natural adhesive comprises a natural latex based adhesive.

[0059] In some embodiments, the natural latex-based adhesive is leather glue or weld.

[0060] In some embodiments, the method further comprises incorporating a support material into the composite mycelium material.

[0061] In some embodiments, the support material has a pore size of 1 / 16 inch.

[0062] In some embodiments, the support material comprises a reinforcement material.

[0063] In some embodiments, the reinforcement material is entangled within the composite mycelium material.

[0064] In some embodiments, the support material comprises a base material.

[0065] In some embodiments, the base material is located on one or more surfaces of the composite mycelium material.

[0066] In some embodiments, the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted textile, woven textile, and non-woven textile.

[0067] In some embodiments, the method further comprises perforating, sonicating, and / or vacuum processing the cultivated mycelial material.

[0068] In some embodiments, the method further comprises perforating and sonicating the cultivated mycelial material.

[0069] In some embodiments, the method further comprises adding one or more proteins from a species other than the fungal species that produced the cultivated mycelial material.

[0070] In some embodiments, the one or more proteins are from a plant source.

[0071] In some embodiments, the plant source is a pea plant.

[0072] In some embodiments, the plant source is a soybean plant.

[0073] In some embodiments, the method further comprises adding a dye to the cultivated mycelial material or the composite mycelial material.

[0074] In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

[0075] In some embodiments, the dye is a reactive dye.

[0076] In some embodiments, the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

[0077] In some embodiments, the dye is present throughout the interior of the composite mycelium material.

[0078] In some embodiments, the method further comprises adding a plasticizer to the cultivated mycelial material or the composite mycelial material.

[0079] In some embodiments, the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

[0080] In some embodiments, the plasticizer is a fatliquor.

[0081] In some embodiments, the composite mycelium material is flexible.

[0082] In some embodiments, the method further comprises applying an external element to the cultivated mycelial material.

[0083] In some embodiments, the external element is applied via heat and / or pressure.

[0084] In some embodiments, the external element is a heat press.

[0085] In some embodiments, the method further comprises adding tannin to the cultivated mycelial material or the composite mycelial material.

[0086] In some embodiments, the method further comprises adding a finish to the composite mycelium material.

[0087] In some embodiments, the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

[0088] In some embodiments, the method further comprises determining a mechanical property of the composite mycelium material.

[0089] In some embodiments, the mechanical properties include wet tensile strength, initial modulus, elongation at break, thickness, and / or slit tear strength.

[0090] In some embodiments, the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

[0091] In some embodiments, the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

[0092] In some embodiments, the composite mycelium material has a wet tensile strength of 7 MPa.

[0093] In some embodiments, the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

[0094] In some embodiments, the composite mycelium material has an elongation at break of 1% to 25%.

[0095] In some embodiments, the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

[0096] In some embodiments, the composite mycelium material has a thickness of 2 mm.

[0097] In some embodiments, the composite mycelium material has a split tear strength of 5N to 100N.

[0098] In some embodiments, the composite mycelium material has a split tear strength of 50N.

[0099] In some embodiments, the composite mycelium material is made using conventional papermaking equipment.

[0100] In another aspect, provided herein are methods comprising: producing a cultivated mycelium material; pressurizing the cultivated mycelium material; and contacting the pressurized cultivated mycelium material with a solution comprising a binder.

[0101] In some embodiments, the binder is added before the pressurizing step, during the pressurizing step, or after the pressurizing step.

[0102] In some embodiments, the producing comprises producing the cultivated mycelial material on a solid substrate.

[0103] In some embodiments, the method further comprises incorporating a support material into the composite mycelium material.

[0104] In some embodiments, the support material has a pore size of 1 / 16 inch.

[0105] In some embodiments, the support material comprises a reinforcement material.

[0106] In some embodiments, the reinforcement material is entangled within the composite mycelium material.

[0107] In some embodiments, the support material comprises a base material.

[0108] In some embodiments, the base material is located on one or more surfaces of the composite mycelium material.

[0109] In some embodiments, the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted textile, woven textile, and non-woven textile.

[0110] In some embodiments, the binder comprises one or more reactive groups.

[0111] In some embodiments, the one or more reactive groups react with active hydrogen-containing groups.

[0112] In some embodiments, active hydrogen-containing groups include amine, hydroxyl, and carboxyl groups.

[0113] In some embodiments, the adhesive comprises a sticker, a resin, a cross-linking agent, and / or a matrix.

[0114] In some embodiments, the adhesive is selected from the group consisting of vinyl acetate-ethylene copolymers, vinyl acetate-acrylic copolymers, polyamide-epichlorohydrin resins (PAE), copolymers, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives, and synthetic adhesives.

[0115] In some embodiments, the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

[0116] In some embodiments, the binder is vinyl acetate-ethylene copolymer.

[0117] In some embodiments, the binder is a vinyl acetate-acrylic copolymer.

[0118] In some embodiments, the binder is PAE.

[0119] In some embodiments, the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of the hyphae.

[0120] In some embodiments, the natural adhesive comprises a natural latex based adhesive.

[0121] In some embodiments, the natural latex-based adhesive is leather glue or weld.

[0122] In some embodiments, the method further comprises perforating, sonicating, and / or vacuum processing the cultivated mycelial material.

[0123] In some embodiments, the method further comprises perforating and sonicating the cultivated mycelial material.

[0124] In some embodiments, the method further comprises adding one or more proteins from a species other than the fungal species that produced the cultivated mycelial material.

[0125] In some embodiments, the one or more proteins are from a plant source.

[0126] In some embodiments, the plant source is a pea plant.

[0127] In some embodiments, the plant source is a soybean plant.

[0128] In some embodiments, the method further comprises adding a dye to the cultivated mycelial material or the composite mycelial material.

[0129] In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

[0130] In some embodiments, the dye is a reactive dye.

[0131] In some embodiments, the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

[0132] In some embodiments, the dye is present throughout the interior of the composite mycelium material.

[0133] In some embodiments, the method further comprises adding a plasticizer to the cultivated mycelial material or the composite mycelial material.

[0134] In some embodiments, the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

[0135] In some embodiments, the plasticizer is a fatliquor.

[0136] In some embodiments, the composite mycelium material is flexible.

[0137] In some embodiments, the method further comprises applying an external element to the cultivated mycelial material.

[0138] In some embodiments, the external element is applied via heat and / or pressure.

[0139] In some embodiments, the external element is a heat press.

[0140] In some embodiments, the method further comprises adding tannin to the cultivated mycelial material or the composite mycelial material.

[0141] In some embodiments, the method further comprises adding a finish to the composite mycelium material.

[0142] In some embodiments, the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

[0143] In some embodiments, the method further comprises determining a mechanical property of the composite mycelium material.

[0144] In some embodiments, the mechanical properties include wet tensile strength, initial modulus, elongation at break, thickness, and / or slit tear strength.

[0145] In some embodiments, the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

[0146] In some embodiments, the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

[0147] In some embodiments, the composite mycelium material has a wet tensile strength of 7 MPa.

[0148] In some embodiments, the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

[0149] In some embodiments, the composite mycelium material has an elongation at break of 1% to 25%.

[0150] In some embodiments, the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

[0151] In some embodiments, the composite mycelium material has a thickness of 2 mm.

[0152] In some embodiments, the composite mycelium material has a split tear strength of 5N to 100N.

[0153] In some embodiments, the composite mycelium material has a split tear strength of 50N.

[0154] In some embodiments, the composite mycelium material is made using conventional papermaking equipment.

[0155] In another aspect, provided herein is a composite mycelium material comprising: a pressurized cultivated mycelium material; and a binder.

[0156] In some embodiments, the cultivated mycelial material has been produced on a solid substrate.

[0157] In some embodiments, the binder comprises one or more reactive groups.

[0158] In some embodiments, the one or more reactive groups react with active hydrogen-containing groups.

[0159] In some embodiments, active hydrogen-containing groups include amine, hydroxyl, and carboxyl groups.

[0160] In some embodiments, the adhesive comprises a sticker, a resin, a cross-linking agent, and / or a matrix.

[0161] In some embodiments, the adhesive is selected from the group consisting of vinyl acetate-ethylene (VAE) copolymers, vinyl acetate-acrylic copolymers, polyamide-epichlorohydrin resins (PAE), copolymers, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives, and synthetic adhesives.

[0162] In some embodiments, the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

[0163] In some embodiments, the binder is a vinyl acetate-ethylene (VAE) copolymer.

[0164] In some embodiments, the binder is a vinyl acetate-acrylic copolymer.

[0165] In some embodiments, the binder is PAE.

[0166] In some embodiments, the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of the hyphae.

[0167] In some embodiments, the natural adhesive comprises a natural latex based adhesive.

[0168] In some embodiments, the natural latex-based adhesive is leather glue or weld.

[0169] In some embodiments, the composite mycelium material further comprises a support material.

[0170] In some embodiments, the support material has a pore size of 1 / 16 inch.

[0171] In some embodiments, the support material comprises a reinforcement material.

[0172] In some embodiments, the reinforcement material is entangled within the composite mycelium material.

[0173] In some embodiments, the support material comprises a base material.

[0174] In some embodiments, the base material is located on one or more surfaces of the composite mycelium material.

[0175] In some embodiments, the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted textile, woven textile, and non-woven textile.

[0176] In some embodiments, the composite mycelial material comprises one or more proteins from a species other than the fungal species that produced the cultivated mycelial material.

[0177] In some embodiments, the one or more proteins are from a plant source.

[0178] In some embodiments, the plant source is a pea plant.

[0179] In some embodiments, the plant source is a soybean plant.

[0180] In some embodiments, the composite mycelium material further comprises a dye.

[0181] In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

[0182] In some embodiments, the dye is a reactive dye.

[0183] In some embodiments, the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

[0184] In some embodiments, the dye is present throughout the interior of the composite mycelium material.

[0185] In some embodiments, the composite mycelium material further comprises a plasticizer.

[0186] In some embodiments, the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

[0187] In some embodiments, the plasticizer is a fatliquor.

[0188] In some embodiments, the composite mycelium material is flexible.

[0189] In some embodiments, an external element is applied to the cultivated mycelial material.

[0190] In some embodiments, the external element is applied via heat and / or pressure.

[0191] In some embodiments, the external element is a heat press.

[0192] In some embodiments, the composite mycelium material further comprises tannin.

[0193] In some embodiments, the composite mycelium material further comprises a finish.

[0194] In some embodiments, the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

[0195] In some embodiments, the composite mycelium material comprises mechanical properties.

[0196] In some embodiments, the mechanical properties include wet tensile strength, initial modulus, elongation at break, thickness, and / or slit tear strength.

[0197] In some embodiments, the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

[0198] In some embodiments, the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

[0199] In some embodiments, the composite mycelium material has a wet tensile strength of 7 MPa.

[0200] In some embodiments, the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

[0201] In some embodiments, the composite mycelium material has an elongation at break of 1% to 25%.

[0202] In some embodiments, the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

[0203] In some embodiments, the composite mycelium material has a thickness of 2 mm.

[0204] In some embodiments, the composite mycelium material has a split tear strength of 5N to 100N.

[0205] In some embodiments, the composite mycelium material has a split tear strength of 50N.

[0206] In some embodiments, the composite mycelium material is made using conventional papermaking equipment.

[0207] Brief description of the attached figure

[0208] Figure 1 Schematic diagram depicting a method of making a composite mycelium material according to some embodiments described herein. Boxes with solid lines indicate required steps and boxes with dashed lines indicate optional steps.

[0209] Figure 2 Depicted are stress-strain curves for a pressed sample with polyamide-epichlorohydrin resin (PAE) and Scaffold 3 (dashed line) and a pressed sample with PAE and Scaffold 4 (solid line). Standard stress (MPa) is plotted against elongation (%).

[0210] Figure 3 Different scaffold materials are depicted. From left to right: Scaffold 1, a gauze scaffold with pores slightly smaller than 1 / 16 inch; Scaffold 2, a cotton textile scaffold with pores smaller than 1 / 32 inch; Scaffold 3, a non-textile scaffold with pores sized 1 / 16 inch; and Scaffold 4, a cotton textile scaffold with large pores sized 1 / 8 inch.

[0211] Figure 4Depicted are samples containing 5 g of cultivated mycelial material (indicated by arrows), 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 1 g of pea protein, Scaffold 4, and weldwood adhesive after wet tensile testing.

[0212] Figure 5 Plot of Crack Tear (N) versus Thickness (mm) for stressed samples (HM1-4-3 based HM1-1-11_120p) and unstressed samples (HM1-1-1, HM1-1-7, and HM1-1-11).

[0213] Figure 6 Stress-strain curves were drawn, which plot engineering stress (MPa) versus nominal strain (%). Prior to stretching, the strain was cycled from 10% to 80% in 10% increments to a maximum elongation at break.

[0214] Fig. 7A Depicted is a SEM micrograph of mycelial hyphae prior to drawing. Figure 7B SEM micrograph depicting mycelial hyphae after drawing. Fig. 7A Scale bar = 50 μm; Figure 7B Scale bar = 200 μm.

[0215] Figure 8 Fourier transform images of SEM images of mycelium before (black squares) and after (grey circles) stretching are shown.

[0216] Fig. 9 The normalized absorbance versus wavenumber 1 / cm of aligned mycelial hyphae is shown along with polarized (0 degrees) and polarized Fourier transform infrared spectroscopy (FTIR) spectra perpendicular to the polarization (90 degrees). The spectrum of pure chitin is shown for comparison.

[0217] Fig.10 Plotting the normalized absorbance versus wave number for non-aligned and aligned mycelial hyphae versus the second order Legendre parameter ( <p2>) at a wave number of 1 / cm.

[0218] Fig.11A Depicted is a Scanning Electron Microscopy (SEM) micrograph of two aligned mycelium sheets bonded with a polyurethane hot melt adhesive at 150X. Fig. 11B Depicted is a scanning electron microscopy (SEM) micrograph of two aligned mycelium sheets bonded with a polyurethane hot melt adhesive at 500X magnification.

[0219] Fig. 12A Plotted are stress-strain curves of aligned mycelia bonded with a polyurethane hot melt adhesive tested in the dry state after conditioning at 65% relative humidity (RH). Fig. 12B Plotted are stress-strain curves of aligned mycelia bonded with a polyurethane hot melt adhesive tested in a wet state after conditioning at 65% relative humidity (RH).

[0220] Fig.13 Flowchart showing a method of making a material comprising mycelium

[0221] Fig.14 A flow chart showing a method for converting raw mycelial material into processed material.

[0222] Fig.15 The mycelial hyphae are shown to form a 3D network containing open pores between the hyphal structures.

[0223] Fig.16 It is shown that most pore diameters fall within the range between 0.36 and 0.84 microns, with a sharp peak of very high pore size distribution between 0.72 and 0.78 microns.

[0224] Fig.17 Particle size of 5% aqueous solution of Mimosa tannin is shown

[0225] Fig.18 The particle size of a 5% wt / wt fatliquor (DXV / LEX=3:1) emulsion is shown.

[0226] Fig.19 The particle size distribution of a 10 wt% gum arabic solution is shown.

[0227] Fig. 20 Showing the particle size of a crusting solution containing gum arabic, mimosa tannins and a fatliquor

[0228] Fig.21 10wt% is shown Particle size distribution of 2833 emulsion (Celanese).

[0229] Fig. 22 The particle size distribution of VAE S-10 emulsion (US Adhesive) is shown.

[0230] Fig.23 Tensile strength results for PAE cross-linked samples are shown.

[0231] Fig.24 Shown are cross-sectional SEM images of processed mycelium panels treated with gum arabic and latex.

[0232] Fig.25 Shown are the normalized flexural moduli of mycelium panels treated with gum arabic and APS or latex.

[0233] Fig.26 The normalized split tear maximum strength of mycelium panels treated with gum arabic and APS or latex is shown.

[0234] Fig. 27 Shown are the mycelium content of the samples as a function of the binder concentration in the solutions of the samples impregnated with natural rubber latex and S-10 vinyl acetate-ethylene.

[0235] Fig.28 Shown is the maximum T-peel force of mycelium samples impregnated with Tear mender natural rubber latex. Points represent individual T-peel tests.

[0236] Fig.29 Shown are the maximum T-peel forces for mycelium samples impregnated with a range of S-10 vinyl acetate-ethylene concentrations. Points represent individual T-peel tests.

[0237] Fig.30 Representative scanning electron micrographs of untreated mycelium samples and mycelium samples impregnated with natural rubber latex or S-10 vinyl acetate-ethylene are shown.

[0238] Fig.31 The normalized crack tear maximum force for samples impregnated with various binders at various concentrations is shown. The dots represent individual crack tear tests. The diamonds represent the upper and lower 95% confidence intervals for the mean normalized crack tear force in each group.

[0239] Fig.32 Shows the impregnation with the same binder concentration Maximum T-peel force for mycelium samples of either 2833 vinyl acetate-acrylic or Dur-O-Set Elite 22 vinyl acetate-ethylene.

[0240] Fig.33 The percent weight change of the binder-infiltrated mycelium panels after sonication and soaking is shown.

[0241] Fig.34 Shown is the percent change in weight of the binder-infiltrated mycelium panels in the samples perforated by the needle perforation method and sonicated as compared to sonication only.

[0242] Fig.35 Shown is the percent change in weight of the binder-infiltrated mycelium panels in samples perforated by the water perforation method and sonicated as compared to sonication only.

[0243] Fig.36 The percent weight change of the binder infiltrated mycelium panels in Samples 1, 2 and 3 after vacuum assisted infiltration is shown.

[0244] Fig.37 The maximum force to tear at rips is shown for various mycelium samples and composites: a single layer mycelium sample; two layers of mycelium tested simultaneously; two layers of mycelium laminated with natural rubber latex; two layers of mycelium laminated with natural rubber latex with a woven scrim in between; the same woven scrim (without mycelium) coated with natural rubber latex.

[0245] Fig.38 Representative crack tear force-travel curves are shown for the indicated mycelium composite samples.

[0246] Fig.39 The maximum tear force of the mycelium samples laminated and impregnated in parallel ("double layer + latex") is shown, compared with the unlaminated ("single layer + latex") and unimpregnated ("single layer") samples. The dots represent individual tear tests. The diamonds represent the upper and lower 95% confidence intervals for the average maximum tear force.

[0247] Fig.40 The flexural initial modulus is shown for the indicated samples: two-ply sample laminated with latex (thickness t); two-ply sample laminated with scrim and latex (thickness t + scrim thickness); one-ply sample treated with gum arabic and APS (thickness 2t); and one-ply sample with scrim adhered to one side (thickness 0.5t + scrim thickness). The cowhide handbag sample is shown for reference. The dots represent individual measurements. The diamonds represent the upper and lower 95% confidence intervals for the mean.

[0248] Fig.41 The T-peel maximum force is shown for untreated, hot pressed, and hot pressed and bonded mycelial materials. Hot pressing was performed at 70°C for 2 min. For the hot pressed and bonded samples, 0.5 wt% S-10 vinyl acetate-ethylene binder was added to the crusting solution. The cross marks the mean and the bars represent the standard deviation.

[0249] Fig.42 The maximum force for tearing the cracks for untreated, hot pressed, and hot pressed and bonded mycelial materials is shown. Hot pressing was performed at 70°C for 2 min. For the hot pressed and bonded samples, 0.5 wt% S-10 vinyl acetate-ethylene binder was added to the crusting solution. The crosses mark the mean and the bars represent the standard deviation.

[0250] Fig.43 Shown are sample thicknesses after the indicated plasticizer treatments.

[0251] Fig.44 Shown are sample densities after the indicated plasticizer treatments.

[0252] Fig.45 The highest flexural modulus for each sample after the indicated plasticizer treatment is shown.

[0253] Fig.46 The maximum crack tear force for each sample after the indicated plasticizer treatment is shown.

[0254] Fig.47 Shown are the normalized crack tear forces for each sample after the indicated plasticizer treatment.

[0255] Fig.48 Shown are sample thicknesses after the indicated plasticizer treatments.

[0256] Fig.49 Shown are sample densities after the indicated plasticizer treatments.

[0257] Fig.50 The highest flexural modulus for each sample after the indicated plasticizer treatment is shown.

[0258] Fig.51 Shown are the average flexural moduli for each sample after treatment with the indicated plasticizer.

[0259] Fig.52 The maximum crack tear force for each sample after the indicated plasticizer treatment is shown.

[0260] Fig.53 Shown are the normalized crack tear forces for each sample after the indicated plasticizer treatment.

[0261] Fig.54 Shown are the average flexural moduli for each sample after treatment with the indicated plasticizer.

[0262] Fig.55A Shown are the absorbance of stained mycelial plates after 1 round of immersion. Fig.55B Shown are the absorbance of stained mycelial plates after 2 rounds of immersion. Fig.55C Shown are the absorbance of stained mycelial plates after 3 rounds of immersion. Fig.55D Shown are the absorbance of stained mycelial plates after 4 rounds of immersion. Fig.55E Shown are the absorbances after 5 rounds of soaking.

[0263] Fig.56 The maximum force of the crack tear is shown as compared to the starting thickness (cm) of the panel.

[0264] Fig.57 The maximum force at crack tear is shown as compared to the final thickness (cm) of the panel.

[0265] Fig.58 Shown are the coloration averages as compared to soak pH.

[0266] Fig.59 Shown is dye penetration as compared to soaking pH.

[0267] Fig.60 Shown is colorfastness as compared to soaking pH.

[0268] Fig.61 A bivariate fit of the average peel force (N / cm) versus the binder estimate (%) is shown.

[0269] Fig.62 Standard force values ​​for mycelium samples with various concentrations of abaca fiber are illustrated.

[0270] Fig.63 Shown are the tensile strength results for disrupted mycelium samples cross-linked with sodium periodate, ammonium persulfate or EDC.

[0271] Fig.64 Shown are the tensile strength results for disrupted mycelium samples cross-linked with a combination of EDC, APS / lignin, and heat pressing.

[0272] Fig.65 The tensile strength results for the ionic liquid treated samples relative to the control are shown.

[0273] Fig.66 Shown are the tensile strength results for disrupted mycelium cross-linked with PAE resin.

[0274] Fig.67 A linear model is shown that describes the effect of pressing time, temperature and pressure on the final thickness of the material. The solid line represents the mean of the reaction. The shaded area represents the 95% confidence interval of the mean. R 2 =0.96.

[0275] Fig.68 Representative scanning electron micrographs of fabricated webs at various points in the volume-concentration-vacuum parameter space are shown, along with their corresponding area densities. Scale bar is 1.5 mm.

[0276] Fig.69 Shown are scanning electron micrographs of webs prepared with and without incorporating 0.05 wt% sodium dodecyl sulfate (SDS) into the slurry.

[0277] Fig.70 A model is shown that relates the mass % of mycelium in a dry-bonded web to the volume of binder solution added to the web and the concentration of the binder solution.

[0278] Fig.71 A model describing the average binder:mycelium peak area ratio over the concentration-volume-Z position parameter space is shown.

[0279] Fig.72 The binder:mycelium ratio is shown as a function of Z position in the web for low and high binder addition volumes.

[0280] Fig.73 The effect of hydroentanglement on the wet tensile curve of a mycelial web is shown.

[0281] Fig.74 Shown are the flexural moduli of mycelium material samples with different curing conditions.

[0282] Fig.75 The crack tear strength of mycelium material samples with different curing conditions is shown.

[0283] Fig.76 Shown are the effects of curing and dyeing of a sample of mycelial material.

[0284] Fig.77 Shown are the effects of curing and dyeing of a sample of mycelial material.

[0285] Fig.78 The effect of dyeing a sample of mycelial material without curing is shown.

[0286] Fig.79 Shown is the leaching of binder solution onto a mycelial web over time using sonication and warm soak methods.

[0287] Fig.80 Shown are images of samples stained with 0 g / L, 25 g / L and 50 g / L sodium sulfate and 2 g / L Procion MX Medium Blue without fixation or rinsing. The bottom two rows show the penetration of the dye into the interior of the sample.

[0288] Fig.81 Images showing dye solution leached from stained mycelial samples after progressive immersion in water for more than one hour and fixation with 0.5 g / L sodium carbonate.

[0289] Fig.82 Images showing dye solution leached from stained mycelial samples after progressive immersion in water for more than one hour and fixation with 1 g / L sodium carbonate.

[0290] Fig.83 Images of spunlace samples dyed at 22° C., 40° C. and 60° C. for total dyeing times of 2 and 6 h are shown. The bottom section of each sample was scraped off to show dye penetration.

[0291] Fig.84 Shown are quantitative measurements of spunlace samples using a Konica Minolta CM-5 spectrophotometer.

[0292] Fig.85 Images of dyed spunlace samples after binder application and curing are shown at various temperatures and times. DETAILED DESCRIPTION

[0293] definition

[0294] The details of multiple embodiments of the present disclosure are set forth in the following description. Other features, purposes and advantages of the present disclosure will be apparent from the description. Unless otherwise defined herein, the scientific and technical terms used in conjunction with the present disclosure should have the meanings commonly understood by those skilled in the art. In addition, unless the context otherwise requires, singular terms should include plural, and plural terms should include singular. Unless otherwise indicated by the context, terms "a" and "an" include plural references. Typically, the nomenclature and technology associated with biochemistry, enzymology, molecule and cell biology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art.

[0295] Unless otherwise indicated, the following terms shall be understood to have the following meanings:

[0296] The term "hyphae" refers to the morphological structure of a fungus, characterized by a branched, filamentous shape.

[0297] The term "hyphal" refers to an object whose components include hyphae.

[0298] The term "mycelium" refers to a structure formed by one or more masses of branching hyphae. "Mass" refers to a quantity of matter. Mycelium is a distinct and separate structure from the fruiting body of a fungus or sporocarp.

[0299] The terms "cultivate" and "cultivated" mean the deliberate growth of fungi or other organisms using defined techniques.

[0300] The term "cultivated mycelial material" refers to a material comprising one or more masses of cultivated mycelium, or only cultivated mycelium. In some embodiments, the one or more masses of cultivated mycelium are disrupted as described herein. In most cases, the cultivated mycelial material has been produced on a solid substrate, as described below.

[0301] The term "composite mycelium material" refers to any material comprising a cultivated mycelium material, the cultivated mycelium material and another material, such as an adhesive or support material as described herein, such as a cross-linking agent, a natural adhesive or a synthetic adhesive combination. In some embodiments, the mycelium comprises a support material. Suitable support materials include, but are not limited to, a mass of continuous disordered fibers (e.g., non-woven fibers), a perforated material (e.g., a metal mesh fabric, a perforated plastic), a mass of discontinuous particles (e.g., wood chips) or any combination thereof. In a specific embodiment, the support material is selected from the group consisting of a mesh fabric, a gauze, a fabric, a knitted textile, a woven textile, and a non-woven textile. In some embodiments, the mycelium comprises a reinforcing material. A reinforcing material is a support material entangled in the mycelium or composite mycelium material. In some embodiments, the mycelium comprises a base material. A base material is a support material located on one or more surfaces of the mycelium or composite mycelium material.

[0302] The term "incorporate" refers to any substance that can be combined or contacted with another substance, such as a cultivated mycelial material, a composite mycelial material, or an adhesive. In a specific embodiment, the mycelial or composite mycelial material can be combined with, contacted with, or incorporated into a support material, such as woven, twisted, wrapped, folded, coiled, tangled, or interwoven together to produce a mycelial material that has been incorporated with the support material. In another embodiment, one or more adhesives can be incorporated into the cultivated mycelial material to be bonded in its disrupted or undisrupted state, such as embedded throughout the material, or added as a thin coating, such as by spraying, saturating, dipping, rolling, coating, etc., to produce a mycelial material.

[0303] As used herein, the term "disrupted" with respect to one or more masses of branched hyphae refers to one or more masses of branched hyphae to which one or more disruptions have been applied. As described herein, "disruption" can be mechanical or chemical, or a combination thereof. In some embodiments, the one or more masses of branched hyphae are disrupted by mechanical action. As used herein, "mechanical action" refers to manipulation of or associated with a machine or tool. Exemplary mechanical actions include, but are not limited to, blending, chopping, impacting, compacting, jumping, shredding, grinding, compression, high pressure, shearing, laser cutting, hammer milling, and water jet forces. In some embodiments, mechanical action may include, for example, applying physical forces in one or more directions so that at least some of the branched hyphae masses are aligned in parallel in one or more directions, wherein the physical forces are applied repeatedly. In some other embodiments, the one or more masses of branched hyphae are disrupted by chemical treatment. As used herein, "chemical treatment" refers to contacting the cultivated mycelial material or composite mycelial material with an amount of a chemical agent (e.g., an alkali or other chemical agent) sufficient to cause disruption. In various embodiments, a combination of mechanical action and chemical treatment may be used herein. The amount of mechanical action (e.g., the amount of pressure) and / or chemical agent applied, the period of time for which the mechanical action and / or chemical treatment is applied, and the temperature at which the mechanical action and / or chemical agent is applied depend in part on the components of the cultivated mycelial material or composite mycelial material and are selected to provide optimal disruption to the cultivated mycelial material or composite mycelial material.

[0304] As used herein, the term "plasticizer" refers to any molecule that interacts with a structure to increase the mobility of the structure.

[0305] As used herein, the term "processed mycelium material" refers to mycelium that has been post-processed by any combination of treatments using preservatives, plasticizers, finishes, dyes, and / or protein treatments.

[0306] As used herein, the term "web" refers to a mycelial material or composite mycelial material that has been disrupted, converted to a slurry, and aligned in a form (eg, a dry lay, an air lay, and / or a wet lay).

[0307] As used herein, the term "spunlace" refers to a mycelial material or composite mycelial material that has been disrupted and hydraulically entangled, wherein one or more clumps of branched hyphae are entangled using water jets or the like.

[0308] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of skill in the art to which the disclosed subject matter belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the disclosed subject matter, preferred methods and materials are now described. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials associated with the publications cited.

[0309] Where a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of that range (unless otherwise clearly indicated herein, to one-tenth of the unit of the lower limit) and any other stated or intervening values ​​in that stated range are encompassed within aspects of the invention. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges, and are also encompassed within aspects of the invention, subject to any specifically excluded limits in the stated ranges. Where a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also encompassed within aspects of the invention.

[0310] Certain ranges are set forth herein with numerical values ​​preceded by the term "about". The term "about" is used herein to provide literal support for the exact number that follows, as well as for numbers that are close to or approximately the number that follows the term. In determining whether a number is close to or approximately a specifically stated number, the close or approximately unstated number may be a number that, in the context in which the number is set forth, provides a substantial equivalent to the specifically stated number.

[0311] Exemplary methods and materials are described below, but methods and materials similar or equivalent to those described herein can also be used in the practice of the present invention and will be apparent to those skilled in the art. All publications and other references mentioned herein are incorporated by reference in their entirety. In the event of a conflict, the present specification (including definitions) shall prevail. Materials, methods and examples are illustrative only and are not intended to be limiting.

[0312] Mycelium compositions and methods of making

[0313] Provided herein are cultivated mycelial materials and composite mycelial materials and scalable methods for making the cultivated mycelial materials and composite mycelial materials. In some or most embodiments, the composite mycelial material comprises a cultivated mycelial material having one or more clumps of branched hyphae and a binder, wherein the one or more clumps of branched hyphae are disrupted. Also provided are methods for making the cultivated mycelial materials and composite mycelial materials.

[0314] Exemplary patents and applications discussing methods of growing mycelium include, but are not limited to: WIPO Patent Publication No. 1999 / 024555; British Patent No. 2,148,959; British Patent No. 2,165,865; U.S. Patent No. 5,854,056; U.S. Patent No. 2,850,841; U.S. Patent No. 3,616,246; U.S. Patent No. 9,485,917; U.S. Patent No. 9,879,219; U.S. Patent No. 9,469,838; U.S. Patent No. 9,914,906; U.S. Patent No. 9,555,395; U.S. Patent Publication No. 2015 / 0101509; U.S. Patent Publication No. 2015 / 0033620, all of which are incorporated herein by reference in their entirety. U.S. Patent Publication No. 2018 / 0282529, published on October 4, 2018, discusses various mechanisms for solution-based post-processing of mycelium materials to produce materials with favorable mechanical characteristics for processing into textiles or leather substitutes.

[0315] like Figure 1 As shown, an exemplary method of making mycelium material according to some embodiments described herein includes cultivating mycelium material, optionally disrupting the cultivated mycelium material, optionally adding a binder, optionally incorporating additional materials such as support materials, and combinations thereof. In various embodiments, conventional papermaking equipment can be adapted or used to perform some or all of the steps set forth herein. In the described embodiments, the mycelium material is manufactured using conventional papermaking equipment.

[0316] The description of the embodiment with several components connected to each other does not imply that all the components are needed. On the contrary, a variety of optional components can be described to illustrate multiple possible embodiments of one or more aspects of the present invention and to more fully illustrate one or more aspects of the present invention. Similarly, although process steps, method steps, algorithms, etc. can be described in a continuous order, unless specifically stated to the contrary, the process, method and algorithm can generally be configured to work in an alternative order. In other words, any order or sequence of the steps that can be described herein does not indicate that the steps are required to be performed in that order. The steps of the described process can be performed in any actual order. In addition, some steps can be performed simultaneously, even if described or implied as non-simultaneous occurrence (for example, because one step is described after another step). In addition, the process described in the figure does not imply that the illustrated process excludes other changes and modifications, does not imply that the illustrated process or any of its steps is necessary for one or more embodiments, and does not imply that the illustrated process is preferred. In addition, generally each embodiment describes a step once, but this does not mean that it must occur once, or it can only occur once when performing or implementing a process, method or algorithm each time. Some steps may be omitted in some embodiments or circumstances, or some steps may be performed more than once in a given embodiment or circumstances.

[0317] Cultivated mycelium material

[0318] Embodiments of the present disclosure include various types of cultivated mycelial materials. Depending on the particular embodiment and the requirements of the material being explored, various known methods of cultivating mycelium may be used. Any fungus that can be cultivated as mycelium may be used. Suitable fungal species for use include, but are not limited to: black mushroom; Agrocybe aegypti; ); Amylotrichum; Amylotrichum; Armillaria; Aspergillus nidulans; Aspergillus niger; Aspergillus oryzae; Ceriporia lacerata; Coprinus comatus; Soporus wilandii; Hepatic beef mushroom; Flammulina velutipes; Medicinal phyllotoxin; Ganoderma lucidum; Ganoderma pine; Hericium erinaceus; Smoky curtain mushroom; Sub-brick red weeping mushroom; Inonotus birchii; Salmon yellow lactarius; Tall ring mushroom; Black veined morel; Thermophilic myceliophthora; Neurospora crassa; Penicillium carmenii; Penicillium chrysogenum; Penicillium rubrum; Buxomyces rubrum; Red oyster mushroom; Oyster mushroom; Broad scale polypore; Aquatic small brittle stalk mushroom; Microsporus microsporus; Rhizopus oryzae; Schizophyllum; Streptomyces venezuelae; Stropharia; Terrestrial fusella; and Ustilago maydis. In some embodiments, the fungi used include Ganoderma lucidum, Neurospora crassa, and / or Buxomyces rubrum.

[0319] In some embodiments, strains or species of fungi can be bred to produce cultivated mycelial material with specific characteristics, such as dense hyphal networks, highly branched hyphal networks, hyphal fusions within the hyphal networks, and other characteristics that can alter the properties of the cultivated mycelial material. In some embodiments, strains or species of fungi can be genetically modified to produce cultivated mycelial material with specific characteristics.

[0320] In most embodiments, the cultivated mycelium can be grown by first inoculating a solid or liquid substrate with a mycelium inoculum from a selected fungal species. In some embodiments, the substrate is pasteurized or sterilized before inoculation to prevent contamination or competition with other organisms. For example, the standard method of cultivating mycelium includes inoculating a sterilized solid substrate (e.g., grain) with a mycelium inoculum. Other standard methods of cultivating mycelium include inoculating a sterilized liquid medium (e.g., liquid potato dextrose) with a mycelium inoculum or pure culture eggs. In some embodiments, the solid and / or liquid substrate will include lignocellulose as a carbon source for mycelium. In some embodiments, the solid and / or liquid substrate will contain a monosaccharide or a complex sugar as a carbon source for mycelium.

[0321] Reference Fig.13 , a method for making a mycelial material 100 is illustrated. The method 100 includes inoculating a nutrient source on a solid support 104, and incubating the mixture at 106 to grow mycelial biomass, collecting the cultivated mycelial biomass at 108, netting the mycelial biomass at 110 to form a mycelial network, and entangling mycelial branches in the mycelial network at 112.

[0322] At step 106, the inoculated nutrient source is incubated to promote the growth of mycelial biomass. The conditions of the nutrient source and the solid support can be selected to promote the growth of mycelial biomass having a plurality of hyphal branches having sufficient morphological characteristics for entanglement in downstream processes. Exemplary morphological characteristics include a minimum length of hyphal branches, a desired density of the hyphal network, a desired degree of branching of the hyphae, a desired aspect ratio, and / or a desired degree of hyphal fusion of the hyphal network. According to one aspect of the present disclosure, the conditions of the solid support in the incubation step at 106 are selected to promote the growth of mycelial biomass having a plurality of hyphal branches having a length of at least about 0.1 mm. For example, the hyphae may have a length of about 0.1 mm to about 5 mm, about 0.1 mm to about 4 mm, about 0.1 mm to about 3 mm, about 0.1 mm to about 2 mm, about 0.1 mm to about 1 mm, about 1 mm to about 5 mm, about 1 mm to about 4 mm, about 1 mm to about 3 mm, about 1 mm to about 2 mm, about 2 mm to about 5 mm, about 2 mm to about 4 mm, or about 2 mm to about 3 mm.

[0323] Incubation step 106 can occur under aerobic conditions in the presence of oxygen. Optionally, the solid support can be sealed to a chamber during all or part of the incubation step. In some embodiments, oxygen can be introduced into the chamber. The incubation temperature can be selected based on a specific fungal species. In some embodiments, the temperature of the chamber during incubation is about 20°C to about 40°C, about 25°C to about 40°C, about 30°C to about 40°C, about 35°C to about 40°C, about 20°C to about 35°C, about 25°C to about 35°C, about 30°C to about 35°C, about 20°C to about 30°C, or about 25°C to about 30°C.

[0324] Incubation step 106 is configured to promote the growth of mycelial biomass including a plurality of hyphal branches. Incubation step 106 may terminate when the cultivated mycelial biomass is collected at step 108. Incubation step 106 may terminate at a predetermined time or when a predetermined concentration of mycelial biomass is reached. After the cultivated biomass is collected at step 108, there may be some continued growth of mycelium. Optionally, the mycelial biomass may be treated to stop the growth of mycelium.

[0325] At step 108, collect the mycelium biomass of cultivation. The collected biomass can be made into slurry by adding dry mycelium biomass to aqueous solution. At step 108, the concentration of the collected mycelium biomass in such slurry can be adjusted based on the subsequent web-forming process at step 110. In certain embodiments, the mycelium biomass of cultivation is in slurry form. The concentration of mycelium biomass can be adjusted by increasing the volume of slurry or by concentrating mycelium biomass by removing at least a portion of liquid from slurry. In certain embodiments, the concentration of mycelium biomass can be adjusted to a concentration of about 10g / L to about 30g / L, about 10g / L to about 25g / L or about 10g / L to about 20g / L. In other examples, the mycelium biomass of cultivation can be collected and dried.

[0326] In some aspects, a binder can be optionally added to the cultivated mycelial biomass before, during, or after the web-forming process at step 110. The binder can be added before, during, or after collecting the cultivated mycelial biomass and / or adjusting the concentration of the cultivated mycelial biomass. The binder can include any of the adhesives, resins, cross-linking agents, or polymer matrix materials described herein, and combinations thereof.

[0327] In some aspects, the plurality of hyphae branches may be optionally disrupted before, during, or after the web-forming process at step 110. The plurality of hyphae branches may be disrupted according to any mechanical and / or chemical method described herein for disrupting the hyphae. For example, the hyphae may be mechanically disrupted by mechanical action such as blending, chopping, impacting, compacting, jumping, shredding, grinding, compressing, high pressure water spraying, or shearing force prior to the web-forming process at step 110. The hyphae may be disrupted before, during, or after adjusting the concentration of the cultivated mycelial biomass.

[0328] In some aspects, the collected mycelial biomass can be optionally combined with natural and / or synthetic fibers before, during, or after the web-forming process at step 110. In one aspect, the fibers can be combined with the mycelium before, during, or after disrupting the plurality of hyphal branches. The fibers can have any suitable size. Non-limiting examples of suitable fibers include cellulose fibers, cotton fibers, rayon fibers, lyocell fibers, TENCEL fibers, and polyester fibers. TM In one aspect, the fiber can have a length of less than about 25 mm, less than about 20 mm, less than about 15 mm, or less than about 10 mm. For example, the fiber can have a length of about 1 mm to about 25 mm, about 1 mm to about 20 mm, about 1 mm to about 15 mm, about 1 mm to about 10 mm, about 1 mm to about 5 mm, about 5 mm to about 25 mm, about 5 mm to about 20 mm, about 5 mm to about 15 mm, about 5 mm to about 10 mm, about 10 mm to about 25 mm, about 10 mm to about 20 mm, or about 10 mm to about 15 mm. The fiber can be combined with the mycelium at a desired concentration. In one embodiment, the fibers may be combined with the mycelium in an amount of about 1 wt % to about 25 wt %, about 1 wt % to about 20 wt %, about 1 wt % to about 15 wt %, about 1 wt % to about 10 wt %, about 1 wt % to about 5 wt %, about 5 wt % to about 25 wt %, about 5 wt % to about 20 wt %, about 5 wt % to about 15 wt %, about 5 wt % to about 10 wt %, about 10 wt % to about 25 wt %, about 10 wt % to about 20 wt %, or about 10 wt % to about 15 wt %.

[0329] At step 110, the mycelial biomass collected in step 108 may be processed according to a web-forming process to form a mycelial network. The web-forming process may include any of the wet-laying, dry-laying, or air-laying techniques described herein. The mycelium of the web formed in step 110 may be optionally chemically and / or thermally bonded using any of the adhesives described herein.

[0330] Optionally, the web-forming at step 110 may include laying the mycelial branches on a support material. As described herein, in some aspects, the support material is a reinforcing material. Non-limiting examples of suitable support materials include woven fibers, a continuous mass of disordered fibers (e.g., non-woven fibers), perforated materials (e.g., metal mesh fabrics or perforated plastics), a mass of discontinuous particles (e.g., wood chips), gauze, fabrics, knitted fibers, coarse cloth, and textiles. Mycelium can be combined with, contacted with, and / or incorporated into the support material. For example, in some aspects, mycelium can be woven, twisted, wound, folded, coiled, tangled, and / or interwoven with the support material to form a mycelium material, as described herein. In some aspects, the fibers can be laid on the support material before, during, and / or after adding a chemical adhesive. In some aspects, a reinforcing material can be combined with mycelial branches before, during, or after the web-forming step 110.

[0331] At step 112, the mycelial network formed at step 110 may undergo an entanglement process to entangle the plurality of mycelial branches in the mycelial network. The entanglement process may include needle punching (also referred to as felting) and / or hydroentanglement. When a support material is present, the entanglement process optionally includes entanglement of at least a portion of the plurality of mycelial branches with the support material. The entanglement process may form mechanical interactions between mycelia and optionally between mycelia and the support material (when present). In some embodiments, the mycelia are not entangled with the support material.

[0332] In some aspects, the entanglement at step 112 is achieved via a needle punch or needle felting process, wherein one or more needles are transferred into and out of the mycelial network. The needle movement entering and leaving the mycelial network promotes entangled mycelia and optionally oriented mycelia. A needle punching machine with a needle array can be used to pierce the mycelial network at multiple locations during each transfer of the needle array. The number of needles, the distance between needles, the shape of needles, and the size of needles (i.e., needle gauge) can be selected to provide the desired mycelial network entanglement degree. For example, the needle can be barbed and have any suitable shape, and its non-limiting examples include pointed blades, star blades, and conical blades. The number of needle punches and the penetration rate of each region can also be selected to provide the desired mycelial network entanglement degree. The parameters of the needle punch or needle felting process can be selected at least in part based on the fungal species, the morphology and size of the mycelia forming the mycelial network, the desired entanglement degree, and / or the end-use application of the mycelial material.

[0333] In some aspects, the entanglement at step 112 is achieved via a hydraulic entanglement process. The hydraulic entanglement process directs high pressure liquid jets into the mycelial network to promote entanglement of the mycelia. The liquid can be any suitable liquid, examples of which include water. The entanglement process can include a spinneret having an array of holes, the array of holes being configured to direct a stream of liquid at a specific location in the mycelial network. The diameter of the holes can be selected to provide a liquid jet having a desired diameter to be directed at the mycelial network. Additional aspects of the spinneret, such as the number of holes in the array and the spacing of the holes in the array, can be selected to provide a desired degree of entanglement of the mycelial network. The mycelial network and the spinneret can be moved relative to each other so that the liquid jets are directed at the mycelial network in a certain pattern. For example, the spinneret can be moved relative to the mycelial network in a generally "Z" or "N" shaped pattern to provide multiple passes of the spinneret over the mycelial network. The number of passes and the application pattern can be selected to provide a desired degree of entanglement of the mycelial network. The parameters of the hydroentanglement process may be selected based at least in part on the fungal species, the morphology and size of the hyphae forming the mycelial network, the desired degree of entanglement, and / or the end-use application of the mycelial material. In some embodiments, the hydroentanglement process occurs in stages where a portion of the mycelial material is webbed (e.g., a wet layup), the hydroentanglement process continues, and then a second portion of the mycelial material is webbed on top of the first portion and the hydroentanglement process is repeated. This process of webbing a portion of the mycelial material and hydroentangling the webbed portion may be repeated any number of times until a material of a final thickness is webbed.

[0334] The liquid pressure, the opening diameter in the spinneret and / or the flow rate of the liquid can be selected to provide the desired degree of entanglement of the mycelial network and optionally the entanglement of the mycelial network and the support material. For example, the liquid pressure in the hydraulic entanglement process can be at least 100 psi, at least 200 psi, at least 300 psi, at least 400 psi, at least 500 psi, at least 600 psi, at least 700 psi, at least 800 psi, at least 900 psi or at least 1000 psi. In some embodiments, the liquid jet pressure is from about 700 to about 900 psi. In some embodiments, the opening diameter in the spinneret is at least about 10 microns, at least about 30 microns, at least about 50 microns, at least about 70 microns, at least about 90 microns, at least about 110 microns, at least about 130 microns or at least about 150 microns. For example, the opening diameter in the spinneret can be about 10 microns to about 150 microns, 20 microns to about 70 microns, about 30 microns to about 80 microns, about 40 microns to about 90 microns, about 50 microns to about 100 microns, about 60 microns to about 110 microns, or about 70 microns to about 120 microns. In some embodiments, the opening has a diameter of about 50 microns. In some embodiments, the flow rate of the liquid can be about 100 mL / min to about 300 mL / min. In some embodiments, the belt speed during the entanglement process is about 1 meter / minute.

[0335] After the entanglement process is completed at 112, the mycelium material can be processed according to any post-processing methods and / or treatments described herein. Non-limiting examples of post-processing methods and treatments include treatment with a plasticizer, treatment with tannins and / or dyes, treatment with a preservative, treatment with a protein source, treatment with a coating and / or finish, a drying process, a rolling or pressing process, and treatment during an embossing process.

[0336] In a plurality of embodiments, the liquid or solid matrix can be supplemented with one or more different nutrient sources.The nutrient source can contain lignocellulose, monosaccharide (e.g., dextrose, glucose), complex sugar, agar, malt extract, nitrogen source (e.g., ammonium nitrate, ammonium chloride, amino acid) and other minerals (e.g., magnesium sulfate, phosphate).In some embodiments, one or more nutrient sources can be present in wood waste (e.g., wood chips, including from hardwood, beech and hickory) and / or agricultural waste (e.g., livestock manure, straw, corn stalks).Once the matrix has been inoculated and optionally supplemented with one or more different nutrient sources, the cultivated mycelium can be grown.The method for growing mycelium has been fully established in the art.Exemplary methods for growing mycelium include, but are not limited to, U.S. Patent No. 5,854,056; U.S. Patent No. 4,960,413; and U.S. Patent No. 7,951,388.

[0337] In some embodiments, the growth of the cultivated mycelium will be controlled to prevent the formation of fruiting bodies. A variety of methods for preventing the formation of fruiting bodies are discussed in detail in U.S. Patent Publication No. 2015 / 0033620; U.S. Patent No. 9,867,337; and U.S. Patent No. 7,951,388. In other embodiments, the cultivated mycelium can be grown so that it does not have any morphological or structural changes. Depending on the embodiment explored, growth conditions such as exposure to light (e.g., sunlight or growth lamps), temperature, carbon dioxide can be controlled during growth.

[0338] In some embodiments, the cultivated mycelium can be grown on an agar medium. Nutrients can be added to the agar / water base. Standard agar media commonly used for cultivating mycelial materials include, but are not limited to, fortified versions of malt extract agar (MEA), potato dextrose agar (PDA), oatmeal agar (OMA), and dog food agar (DFA).

[0339] In most embodiments, the cultivated mycelial material can be grown as a solid mass and can be subsequently disrupted. As described below, the disrupted cultivated mycelial material can be a live mat, preserved or otherwise treated to kill the mycelium (ie, stop mycelial growth).

[0340] In some embodiments, the cultivated mycelium material can be grown to include elongated hyphae, which define thin filaments that are interconnected with each other and can be further interconnected with a variety of support materials provided in the growth program, as further described below. The thin filaments can be analyzed using optical magnification or imaging devices to determine whether the growth length of the thin filaments is sufficient to support sufficient network interconnections between the thin filaments and a variety of additives. The thin filaments should not only have sufficient length, but also have flexibility to provide appropriate interconnections therebetween.

[0341] In some embodiments, the cultivated mycelium material can be processed using dry array, wet layer or air layer technology. In dry layer or dry array, the inert or growing mycelium network of branched mycelium can be pulled apart and disentangled to expand the volume of the network. Similarly, in wet layer technology, the inert or growing mycelium network of branched mycelium can be saturated in a liquid medium to disentangle and expand the volume of the network. In addition, in air layer technology, the inert or growing mycelium network of branched mycelium can be suspended in the air to establish a net that expands the volume of the network. After such technology, the expanded network can be compressed to provide a dense or tight network. The net can be densified to include a total density profile of at least 6g per cubic meter. The tight net can be embossed with a replica leather pattern to provide a leather substitute material.

[0342] In some embodiments, the method includes the step of webbing the collected mycelial biomass. In some embodiments, the step of webbing the collected mycelial biomass includes depositing the mycelial biomass on a support material.

[0343] In some embodiments, the support material comprises woven fibers, non-woven fibers, mesh fabric, perforated plastic, wood chips, gauze, fabric, knitted fibers, scrim, textile, or a combination thereof.

[0344] In some embodiments, entangling the plurality of hyphal branches comprises entangling at least a portion of the plurality of hyphal branches with a support material.

[0345] In some embodiments, the method further comprises combining the reinforcing material with the mycelial biomass, one of before the web-forming step, during the web-forming step, or after the web-forming step. In some embodiments, web-forming comprises wet-laying, air-laying, or dry-laying.

[0346] In some embodiments, the method further includes combining one of natural fibers, synthetic fibers, or a combination thereof with the mycelial biomass, one of before the web-forming step, during the web-forming step, or after the web-forming step.

[0347] In some embodiments, the fibers have a length of less than 25 millimeters.

[0348] Disrupted cultivated mycelial material

[0349] Various types of cultivated mycelial materials including one or more masses of branched hyphae can be disrupted at multiple points during the manufacturing process, thereby producing one or more masses of disrupted branched hyphae. In the embodiment, the cultivated mycelial material comprises one or more masses of disrupted branched hyphae. The cultivated mycelial material can be disrupted before or after the addition of the adhesive. In one aspect, the cultivated mycelial material can be disrupted simultaneously with the addition of the adhesive. Exemplary embodiments of disruption include, but are not limited to, mechanical action, chemical treatment, or a combination thereof. For example, the one or more masses of branched hyphae can be disrupted by both mechanical action and chemical treatment, mechanical action alone, or chemical treatment alone.

[0350] In some embodiments, the one or more masses of branched hyphae are disrupted by mechanical action. Mechanical action may include blending, chopping, impacting, compacting, jumping, shredding, grinding, compression, high pressure, water spraying and shearing force. In some embodiments, the mechanical action includes blending the one or more masses of branched hyphae. Exemplary methods for achieving such disruption include the use of blenders, grinders, hammer mills, drum combs, heat, pressure, liquids (such as water), mills, beaters and refiners. In an exemplary manufacturing process, the cultivated mycelial material is mechanically disrupted by conventional unit operations such as homogenization, grinding, coagulation, grinding, jet grinding, water spraying, etc.

[0351] According to another aspect, the mechanical action comprises applying a physical force to the one or more clumps of branched hyphae so that at least some of the clumps of branched hyphae are aligned in a particular form, such as in a parallel form or along or against a stress direction. The physical force may be applied to one or more layers of cultivated mycelium material or composite mycelium material. The disrupted mycelium material may typically be constructed into layers with different orientations. Exemplary physical forces include, but are not limited to, tension and alignment forces. Exemplary methods for achieving such disruption include the use of rollers and drafting devices. In some embodiments, the physical force is applied in one or more directions so that at least some of the clumps of branched hyphae are aligned in parallel in one or more directions, wherein the physical force is applied repeatedly. In the embodiment, the physical force may be applied at least twice, such as at least three times, at least four times, or at least five times.

[0352] In some other embodiments, the one or more masses of branched hyphae are disrupted by chemical treatment. In such embodiments, the chemical treatment comprises contacting the one or more masses of branched hyphae with a base or other chemical agent sufficient to cause disruption, including but not limited to alkaline peroxide, beta-glucanase, surfactants, acids, and bases such as sodium hydroxide and sodium carbonate (or soda ash). The pH of the mycelial material cultivated in the solution can be monitored for the purpose of maintaining an optimal pH.

[0353] In some embodiments, the disruption described herein produces one or more groups of disrupted branched hyphae, such as subnetworks. As used herein, "subnetwork" refers to individual branched hyphae groups produced after disruption (such as mechanical action or chemical treatment). Subnetworks can have a variety of shapes, such as spherical, square, rectangular, diamond and special-shaped subnetworks, etc., and each subnetwork can have a variable size. The cultivated mycelium material can be fully disrupted to produce one or more groups of disrupted branched hyphae (such as subnetworks), and its size is within the required range. In many cases, the disruption can be fully controlled to obtain both the size and size distribution of the subnetwork within the required range. In other embodiments where a more accurate size distribution of the subnetwork is required, the cultivated mycelium material of the disruption can be further processed or selected, such as by screening, aggregation, etc. to provide a desired size distribution. For example, the subnetwork can have a size represented by a length of, for example, about 0.1mm to about 5mm, including, for example, about 0.1mm to about 2mm, about 1mm to about 3mm, about 2mm to about 4mm and about 3mm to about 5mm. In some embodiments, a subnetwork may have a size represented by a length of about 2 mm. The "length" of a subnetwork is a measure of the distance equal to the maximum extended dimension of the subnetwork. Other measurable dimensions include, but are not limited to, length, width, height, area, and volume.

[0354] In various embodiments, physical forces can be used to create new physical interactions (i.e., re-entanglement) between one or more clumps of branched hyphae after disruption. A variety of known methods for creating entanglements between fibers can be used, including methods for creating nonwoven materials by creating mechanical interactions between fibers. In some embodiments described below, hydraulic entanglement can be used to create mechanical interactions between hyphae after the hyphae have been disrupted.

[0355] Preserved cultivated mycelial material

[0356] Once the cultivated mycelium material has grown, it may optionally be separated from the substrate in any manner known in the art, and optionally subjected to post-processing to prevent further growth by killing the mycelium and otherwise rendering the mycelium inert to decay, referred to herein as "preserved mycelium material." Suitable methods of producing preserved mycelium material may include drying or dehydrating the cultivated mycelium material (e.g., pressurizing the cultivated mycelium material to expel moisture) and / or heat treating the cultivated mycelium material.

[0357] In a specific embodiment, the cultivated mycelial material is pressurized to 0.25 inches at 190,000 pounds force for 30 minutes. The cultivated mycelial material may be pressurized by at least 100, 1000, 10,000, 100,000, 110,000, 120,000, 130,000, 140,000, 150,000, 160,000, 170,000, 180,000, 190,000, 200,000, or 300,000 pounds force or more. The cultivated mycelial material may be pressurized to at least 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.51, 0.52, 0.53, 0.54, 0.55 .54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.7, 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0.78, 0.79, 0.8, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.9, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.99 or 1 inch or more.The cultivated mycelial material may be pressurized to at least 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.5, 0.51, 0.52, 0.53, 0.54, 0.55 .54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.6, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.7, 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0.78, 0.79, 0.8, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.9, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.99 or 1 cm or more. The cultivated mycelial material may be pressurized for at least 1 min, 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min or 60 min or longer.

[0358] Suitable methods for drying organic matter so that it does not decay are well known in the art. In one specific embodiment, the cultivated mycelial material is dried in an oven at a temperature of 100°F or higher. In another specific embodiment, the cultivated mycelial material is heat pressed.

[0359] In other cases, one or more solutions are used to process living or dry cultivated mycelium materials, which are used to remove waste and moisture from mycelium. In some embodiments, the solution includes a solvent, such as ethanol, methanol or isopropanol. In some embodiments, the solution includes a salt, such as calcium chloride. Depending on the embodiment, the cultivated mycelium material can be immersed in a solution under pressure and without pressure for a variety of durations. In some embodiments, the cultivated mycelium material can be immersed in several solutions continuously. In a specific embodiment, the cultivated mycelium material can first be immersed in one or more first solutions including alcohol and salt, and then immersed in a second solution including alcohol. In another specific embodiment, the cultivated mycelium material can first be immersed in one or more first solutions including alcohol and salt, and then immersed in a second solution including water. After being treated with the solution, the cultivated mycelium material can be dried using a hot or cold method through pressurization and / or using a variety of methods including air drying and / or vacuum drying. These embodiments are described in detail in U.S. Patent Publication No. 2018 / 0282529, which is incorporated herein by reference in its entirety.

[0360] In one aspect, the cultivated mycelial material can be fixed by adjusting the pH using an acid such as formic acid. In specific embodiments, the pH will be at least 2, 3, 4, or 5. In some embodiments, the pH of the cultivated mycelial material will be adjusted to an acidic pH of 3 in order to fix the cultivated mycelial material using various agents such as formic acid. In specific embodiments, the pH will be adjusted to a pH of less than 6, 5, 4, or 3 in order to fix the cultivated mycelial material. In one embodiment, the pH will be adjusted to pH 5.5.

[0361] Adhesives

[0362] Aspects of the present disclosure include adhesives. As used herein, "adhesives" may include any suitable agent that provides increased strength and / or other properties (such as additional softness, strength, durability, and compatibility). The adhesive may be an agent that reacts with a portion of the cultivated mycelium material, enhances the treatment of the cultivated mycelium material, is co-treated with the cultivated mycelium material, or is treated independently, but becomes a network with the cultivated mycelium material to produce a composite mycelium material. In some aspects, the adhesive is added before disruption. In other aspects, the adhesive is added after disruption. In some other aspects, the adhesive is added when the sample is disrupted. The adhesive includes an adhesive, a resin, a cross-linking agent, and / or a matrix. The composite mycelium material described herein includes a cultivated mycelium material and an adhesive, which may be water-based, 100% solid, UV and moisture curable, a two-component reactive blend, pressure sensitive, self-crosslinking hot melt, etc.

[0363] In some embodiments, the adhesive is selected from the group comprising natural adhesive or synthetic adhesive. In the embodiment, the natural adhesive can include an adhesive based on natural latex. In a specific embodiment, the adhesive based on natural latex is leather glue or weld. The adhesive can include anionic, cationic and / or nonionic agents. On the one hand, the adhesive can include a crosslinking agent.

[0364] In some embodiments, the adhesive has a particle size of less than or equal to 1 μm, a sub-zero glass transition temperature, or a self-crosslinking function. In some embodiments, the adhesive has a particle size of less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function. In some embodiments, the adhesive has a particle size of less than or equal to 1 μm. In some embodiments, the adhesive has a sub-zero glass transition temperature. In some embodiments, the adhesive has a self-crosslinking function. In some embodiments, the adhesive has a particle size of less than or equal to 500 nanometers. Specific exemplary adhesives include vinyl acetate ethylene copolymers, such as Elite Plus and Elite 22.

[0365] In some embodiments, the adhesive has a glass transition temperature of -100--10°C, -100--90°C, -90--80°C, -80--70°C, -70--60°C, -60--50°C, -50--40°C, -40--30°C, -30--20°C, -20--10°C, -10--10°C, -30--25°C, -25--20°C, -20--15°C, -15--10°C, -10--5°C, -5--0°C, -90°C, -80°C, -70°C, -60°C, -50°C, -40°C, -35°C, -30°C, -25°C, -20°C, -15°C, -10°C, -5°C or 0°C. In some embodiments, the adhesive has a glass transition temperature of -15°C.

[0366] Other exemplary adhesives include, but are not limited to, transglutaminase, polyamide-epichlorohydrin resin (PAE), citric acid, genipin, alginate, vinyl acetate-ethylene copolymer and vinyl acetate-acrylic acid copolymer. In some embodiments, the adhesive is polyamide-epichlorohydrin resin (PAE). In some embodiments, the adhesive is vinyl acetate-ethylene copolymer. In some embodiments, the adhesive is vinyl acetate-acrylic acid copolymer.

[0367] In some embodiments, the adhesive includes one or more reactive groups. For example, the adhesive reacts with a group (such as an amine, a hydroxyl and a carboxyl) containing active hydrogen. In a specific embodiment, the adhesive cross-links one or more groups of branched hyphae via the one or more reactive groups. In some cases, amine is present on chitin, and hydroxyl and carboxyl are present on the polysaccharide and protein around chitin. In a specific embodiment, PAE includes a cationic azetidinium group. In the embodiment, the cationic azetidinium group on PAE acts as a reactive site in the polyamidoamine backbone, and reacts with a group (such as an amine, a hydroxyl and a carboxyl) containing active hydrogen in one or more mycelial branches.

[0368] Other examples of adhesive include but are not limited to citric acid combined with sodium hypophosphite or monosodium phosphate or sodium dichloroacetate, alginate combined with sodium hypophosphite or monosodium phosphate or sodium dichloroacetate, epoxidized soybean oil, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC), polyamide epichlorohydrin resin (PAE) and ammonium persulfate. Some examples of adhesive include epoxy resin, isocyanate, sulfur compound, aldehyde, anhydride, silane, aziridine and azetidinium compound and compound with all described functional groups. Possible formaldehyde-containing adhesive includes formaldehyde, phenol formaldehyde, urea formaldehyde, melamine urea formaldehyde, melamine formaldehyde, phenol resorcinol and any combination thereof.

[0369] Other examples of suitable adhesives include latex materials such as butadiene copolymers, acrylates, vinyl-acrylics, styrene-acrylics, styrene-butadiene, nitrile-butadiene, polyvinyl acetate, olefin-containing polymers (e.g., vinyl acetate-ethylene copolymers), vinyl ester copolymers, halogenated copolymers (e.g., vinylidene chloride polymers). Latex-based agents may contain functionality when used. Any type of latex may be used, including acrylics. Representative acrylics include those formed by ethyl acrylate, butyl acrylate (meth) methyl acrylate, carboxylated forms thereof, glycosylated forms thereof, self-crosslinked forms thereof (e.g., those including N-methylacrylamide), and copolymers and blends thereof (including copolymers formed with other monomers such as acrylonitrile). Natural polymers may also be used, such as starch, natural rubber latex, dextrin, lignin, cellulose polymers, sugar gums, etc. In addition, other synthetic polymers can also be used, such as epoxy resin, urethane, phenolic resin, chloroprene rubber, butyl rubber, polyolefin, polyamide, polypropylene, polyester, polyvinyl alcohol and polyester amide.As used herein, term " polypropylene " comprises the polymer of propylene or the polymer of propylene and other aliphatic polyolefins (such as ethylene, 1-butylene, 1-pentene, 3-methyl-1-butylene, 4-methyl-1-pentene, 4-methyl-1-hexene, 5-methyl-1-hexene and its mixture) polymerization.In particular embodiments, adhesive includes but is not limited to natural adhesive (for example based on the adhesive of natural latex, such as leather glue or weld, latex, based on the adhesive of soy protein), synthetic adhesive (polyurethane), chloroprene rubber (PCP), acrylic copolymer, styrene-butadiene copolymer, ethylene-vinyl acetate-b, nitrocellulose, polyvinyl acetate (PVA) and vinyl acetate ethylene (VAE).In other embodiments, the adhesive is VAE.

[0370] In one aspect, one or more adhesives may be incorporated into the cultivated mycelium material to be bonded in its disrupted or undisrupted state, for example embedded in the entire material, or added as a thin coating, such as by spraying, dipping, rolling, coating, etc., to produce a composite mycelium material. In another aspect, one or more adhesives may be incorporated while the disruption occurs. In accordance with the present disclosure, any suitable bonding method may be used. Bonding of the surfaces may occur when dry, and a strong cured bond may be developed. Bonding of one or more adhesives may include the use of open or closed cell foam materials, such as urethane, olefin rubber, and vinyl foam materials, as well as textiles, metals, and fabrics in a variety of laminated arrangements.

[0371] The bonded assembly (i.e., laminate) can be prepared by uniformly applying the aqueous adhesive to the cultivated mycelial material. In some embodiments, the thin layer comprises two continuous layers. In some embodiments, the thin layer comprises three continuous layers. A variety of coating methods can be used, such as spraying, roller coating, saturation, etc. The coated substrate can be dried before bonding.

[0372] The composite mycelium material can be chemically bonded by impregnating the composite mycelium material with a chemical binder to connect the fibers to each other, including connecting the cellulose fibers to each other. Non-limiting examples of suitable binders include gum arabic, vinyl acetate-ethylene (VAE), and adhesives. Examples of suitable adhesives include S-10 (available from USA Adhesives, USA) and Bish's Original Tear Mender Instant Fabric & Leather Adhesive (available from Tear Mender, USA). An example of a suitable VAE-based binder is Elite 22, which is available from Celanese Emulsions, USA. Another example of a suitable VAE-based binder is Elite Plus, which is available from Celanese Emulsions, USA. Another exemplary binder includes 2833, available from Celanese Emulsions, USA, and is described as a self-crosslinking vinyl acetate acrylic. In a web of interconnected hyphae, the chemical binder will have to saturate the web to diffuse through the web and reach the core of the network. Therefore, the composite mycelium material can be immersed in the binder solution to completely impregnate the material. Spraying of the chemical binder can also be provided to the composite mycelium material. Spraying of the chemical binder can be assisted by capillary action for dispersion, or can be assisted by vacuum coating to draw the chemical binder through the material. A coater can also be used to coat the composite mycelium material.

[0373] The composite mycelium material can be bonded using a thermal bonding technique, wherein an additive is provided with the composite mycelium material. This additive can be a "meltable" material that melts at a known heat level. The cellulosic material of the composite mycelium material does not melt, so that the composite mycelium material together with the additive can be heated to the melting point of the additive. When melted, the additive can be dispersed within the composite mycelium material and then cooled to harden the overall material.

[0374] The present disclosure is not limited to the above list of suitable adhesives. Other adhesives are known in the art. No matter how the type, the effect of adhesive is partly to provide a number of reactive sites per molecule. The type and amount of the adhesive used in the present disclosure depends on the desired properties. In a plurality of embodiments, an effective amount of adhesive can be used. As used herein, "effective amount" about adhesive refers to the amount of agent, and the amount is enough to provide increased intensity and / or other characteristics (such as other softness, strength, durability and compatibility).

[0375] The adhesive may be added to a cultivated mycelium material that has been pressurized, has one or more clumps of disrupted hyphae, and / or is hydroentangled. The adhesive may be added to the cultivated mycelium material before disrupting one or more clumps of branched hyphae or pressurizing. The adhesive may be added to the cultivated mycelium material during disrupting one or more clumps of branched hyphae or pressurizing. The adhesive may be added to the cultivated mycelium material after disrupting one or more clumps of branched hyphae or pressurizing.

[0376] In some embodiments, the pressurized cultivated mycelium material is contacted with an adhesive. In some embodiments, the disrupted cultivated mycelium material is contacted with an adhesive. In some embodiments, the adhesive is added before the branched mycelium mass is disrupted. In some embodiments, the adhesive is added during the disruption of one or more mass of branched mycelium. In some embodiments, the adhesive is added after the branched mycelium mass is disrupted. In some embodiments, the adhesive is added before the cultivated mycelium material is pressurized. In some embodiments, the adhesive is added during the pressurization of the cultivated mycelium material. In some embodiments, the adhesive is added after the cultivated mycelium material is pressurized.

[0377] Support Materials

[0378] According to one aspect, the cultivated mycelium material or composite mycelium material may further include a support material, for example to form a bonded assembly (i.e., a laminate). As used herein, the term "support material" refers to any material or combination of one or more materials that provides support to the cultivated mycelium material or composite mycelium material. In some embodiments, the support material is a scaffold. In some embodiments, the support material is a scrim.

[0379] In some embodiments, the support material is entangled in the cultivated mycelium material or composite mycelium material (e.g., reinforcing material). In some other embodiments, the support material is located on the surface of the cultivated mycelium material or composite mycelium material (e.g., base material). In some embodiments, the support material includes but is not limited to mesh fabrics, gauze, fabrics, multiple fibers, knitted textiles, woven textiles, non-woven textiles, knitted fibers, woven fibers, non-woven fibers, films, surface spray coatings, and fiber additives. In some embodiments, knitted textiles are knitted fibers. In some embodiments, woven textiles are woven fibers. In some embodiments, non-woven textiles are non-woven fibers. In some embodiments, the support material can be constructed in whole or in part by any combination of synthetic fibers, natural fibers (e.g., lignocellulosic fibers), metals, or plastics. The support material can be partially entangled in the cultivated mycelium material or composite mycelium material, for example, using known entanglement methods (e.g., felting or needle punching). In some aspects, the support material is not entangled in the cultivated mycelium material or composite mycelium material. A variety of methods known in the art can be used to form laminates as described herein. In some other embodiments, the support material includes a base material, which is applied, for example, to the top or bottom surface of the cultivated mycelium material or composite mycelium material. The support material can be attached by any means known in the art, including but not limited to chemical attachment, such as a suitable spray material, particularly a suitable adhesive, or alternatively, such as due to its inherent stickiness.

[0380] The laminate according to the present disclosure may include at least one support material. If more than one support material is used, the cultivated mycelium material or composite mycelium material may include an inner layer of a multi-layer sandwich structure, wherein the inner layer is, for example, a support material such as a knitted or woven fabric or a scaffold. In this case, the support material will be embedded in the cultivated mycelium material or composite mycelium material.

[0381] As used herein, the support material may include a scaffold or a textile. As used herein, a "scaffold" refers to any material known in the art that is different from the cultivated mycelium material and provides support to the cultivated mycelium material or the composite mycelium material. The "scaffold" may be embedded in the cultivated mycelium material or the composite mycelium material, or layered above, below, or within the cultivated mycelium material or the composite mycelium material. In the present disclosure, all kinds and types of scaffolds may be used, including but not limited to films, textiles, scrims, and polymers. As used herein, a "textile" refers to a type of scaffold that may be any woven, knitted, or non-woven fiber structure. In the case where multiple layers are included in the cultivated mycelium material or the composite mycelium material, such as Fig.11A and Fig. 11B As shown, two or more layers may include a support; or in other embodiments, two or more layers may include gauze. Useful supports include woven and non-woven supports, directional and non-directional supports, and orthogonal and non-orthogonal supports. Useful supports may include conventional supports, which include multiple yarns oriented longitudinally or along the length of the support, and multiple yarns oriented transversely or across the width of the support. These yarns may be referred to as warp yarns and weft yarns, respectively. A variety of yarns may be used, including but not limited to fiber materials and polymers. For example, yarns may include but are not limited to glass fiber, aluminum or aramid polymers. In one embodiment, the support includes glass fiber yarn. The support can be adhered together or locked in position using conventional adhesives (such as cross-linkable acrylic resins, polyvinyl alcohol or similar adhesives). The support can also be mechanically entangled by using techniques such as, but not limited to, needle punching. In another embodiment, the support can be locked in position by weaving. Combinations of support materials can be used according to the present disclosure.

[0382] In some embodiments, the support material can be incorporated into the cultivated mycelium material or composite mycelium material as described herein according to methods known in the art, including but not limited to the methods described in U.S. Patent No. 4,939,016 and U.S. Patent No. 6,942,711 (the patents are incorporated herein by reference in their entirety). For example, the support material can be incorporated into the cultivated mycelium material or composite mycelium material via hydraulic entanglement. In the embodiments, the support material can be incorporated into the cultivated mycelium material or composite mycelium material before or after adding an adhesive and / or a cross-linking agent. In some embodiments, a liquid (such as water) guided to the cultivated mycelium material or composite mycelium material via one or more holes for hydraulic entanglement can be passed through the cultivated mycelium material or composite mycelium material. In some embodiments, the liquid is a high-pressure liquid. In some embodiments, pressure and water flow can vary, depending in part on the type and pore size of the support material. In a plurality of embodiments, the water pressure is at least 100psi, for example at least 200psi, at least 300psi, at least 400psi, at least 500psi, at least 600psi, at least 700psi, at least 800psi, at least 900psi and at least 1000psi. In a plurality of embodiments, the water pressure is about 100psi to about 5000psi, including for example about 200psi to about 1000psi, about 300psi to about 2000psi, about 400psi to about 3000psi, about 500psi to about 4000psi and about 600psi to about 5000psi. In some embodiments, the water pressure is about 750psi. In a plurality of embodiments, the one or more holes have a diameter of at least 10 microns, for example at least 30 microns, at least 50 microns, at least 70 microns, at least 90 microns, at least 110 microns, at least 130 microns and at least 150 microns. In various embodiments, the one or more holes have a diameter of about 10 microns to about 150 microns, including, for example, about 20 microns to about 70 microns, about 30 microns to about 80 microns, about 40 microns to about 90 microns, about 50 microns to about 100 microns, about 60 microns to about 110 microns, and about 70 microns to about 120 microns. In some embodiments, the one or more holes have a diameter of about 50 microns.

[0383] The cultivated mycelium material or composite mycelium material may also include adjuvants for use in the foam material. Adjuvants or additives include crosslinking agents, processing aids (e.g., filter aids), dispersants, flocculants, viscosity reducers, flame retardants, dispersants, plasticizers, antioxidants, compatibilizers, fillers, pigments, UV protectants, fibers (such as abaca fibers), and the like. It is further contemplated that foaming agents may be used to introduce chemical binders into the composite mycelium material. Such foaming agents may make the mesh of the composite mycelium material more porous by introducing air into the mesh.

[0384] Plasticizers

[0385] A variety of plasticizers can be applied to cultivated mycelium materials or composite mycelium materials to change the mechanical properties of cultivated mycelium materials or composite mycelium materials. In the embodiment, the cultivated mycelium materials or composite mycelium materials further include plasticizers. U.S. Patent No. 9,555,395 discusses the addition of a variety of humectants and plasticizers. Specifically, U.S. Patent No. 9,555,395 discusses the use of glycerol, sorbitol, triglyceride plasticizers, oils (such as linseed oil, castor oil, drying oils), ions and / or nonionic glycols and polyethylene oxides. U.S. Patent Publication No. 2018 / 0282529 further discusses the use of plasticizers such as glycerol, sorbitol or another humectant to treat cultivated mycelium materials or composite mycelium materials to retain moisture and otherwise enhance the mechanical properties of cultivated mycelium materials or composite mycelium materials, such as the elasticity and flexibility of cultivated mycelium materials or composite mycelium materials. In such embodiments, the cultivated mycelium material or composite mycelium material is flexible.

[0386] Other similar plasticizers and humectants are well known in the art, such as polyethylene glycols and fatliquors, which are obtained by emulsifying natural oils with a liquid immiscible with the oil (e.g., water) so that droplets of the oil can penetrate the material. Various fatliquors contain emulsified oil in water to which other compounds, such as ionic and nonionic emulsifiers, surfactants, soaps, and sulfates, have been added. Fatliquors may include various types of oils, such as mineral, animal, and plant-based oils. Suitable fatliquors include, but are not limited to LEX fatliquor (Trumpler, Germany), DXV fatliquor (Trumpler, Germany), diethoxydimethylammonium chloride (DEEDMAC), Downy fabric softener, sorbitol, erythritol, Tween 20 and Tween 80.

[0387] Tannins and dyes

[0388] In various embodiments of the present disclosure, it may be desirable to impart color to cultivated mycelial material or composite mycelial material. As discussed in U.S. Patent Publication No. 2018 / 0282529, tannins may be used to impart color to cultivated mycelial material, composite mycelial material, or preserved composite mycelial material.

[0389] Since the cultivated mycelial material and / or the composite mycelial material partially comprises chitin, it lacks the functional sites that are abundant in protein-based materials. Therefore, it may be necessary to functionalize the chitin in the cultivated mycelial material or the composite mycelial material in order to establish binding sites for acidic and direct dyes. Methods for functionalizing chitin are discussed above.

[0390] A variety of dyes can be used to impart color to the cultivated mycelium material or composite mycelium material, such as acid dyes, direct dyes, disperse dyes, sulfur dyes, synthetic dyes, reactive dyes, pigments (e.g., black iron oxide and cobalt blue) and natural dyes. In some embodiments, the cultivated mycelium material or composite mycelium material is immersed in an alkaline solution before applying a dye solution to promote dye absorption and penetration into the material. In some embodiments, the cultivated mycelium material or composite mycelium material is pre-soaked in ammonium chloride, ammonium hydroxide and / or formic acid before applying a dye solution to promote dye absorption and penetration into the material. In some embodiments, tannins may be added to the dye solution. In a plurality of embodiments, the cultivated mycelium material or composite mycelium material can be preserved as discussed above before dye treatment or pretreatment.

[0391] Depending on the embodiment, the dye solution can be applied to the cultivated mycelium material or composite mycelium material using different application techniques. In some embodiments, the dye solution can be applied to one or more outer surfaces of the cultivated mycelium material or composite mycelium material. In other embodiments, the cultivated mycelium material or composite mycelium material can be immersed in the dye solution.

[0392] In addition to pre-soaking with a variety of solutions, agents can also be added to the dye solution to promote dye absorption and penetration into the material. In some embodiments, ammonium hydroxide and / or formic acid promote dye absorption and penetration into the material together with acidic or direct dyes. In some embodiments, ethoxylated fatty amines are used to promote dye absorption and penetration into the processed material.

[0393] In various embodiments, a plasticizer is added after or during the addition of the dye. In various embodiments, the plasticizer may be added with the dye solution. In certain embodiments, the plasticizer may be coconut oil, vegetable glycerin, or a sulfited or sulfated fatliquor.

[0394] In some embodiments, the dye solution can be maintained at an alkaline pH using a base such as ammonium hydroxide. In specific embodiments, the pH will be at least 9, 10, 11 or 12. In some embodiments, the pH of the dye solution will be adjusted to an acidic pH in order to use a variety of agents (such as formic acid) to fix the dye. In specific embodiments, the pH will be adjusted to a pH less than 6, 5, 4 or 3 in order to fix the dye.

[0395] In various methods, the cultivated mycelial material, composite mycelial material and / or preserved composite mycelial material can be subjected to mechanical processing or agitation when the dye solution is applied to promote dye absorption and penetration into the material. In some embodiments, subjecting the cultivated mycelial material, composite mycelial material and / or preserved composite mycelial material to squeezing or other forms of pressure in the dye solution enhances dye absorption and penetration. In some embodiments, the cultivated mycelial material, composite mycelial material and / or preserved composite mycelial material can be subjected to sonication.

[0396] Using the methods described herein, the cultivated mycelium material or composite mycelium material can be dyed or colored so that the color of the cultivated mycelium material or composite mycelium material processed is substantially uniform. In some embodiments, the cultivated mycelium material or composite mycelium material is colored with a dye, and on one or more surfaces of the cultivated mycelium material or composite mycelium material, the color of the cultivated mycelium material or composite mycelium material is substantially uniform. Using the above method, the cultivated mycelium material or composite mycelium material can be dyed or colored so that the dye and color are not only present in the surface of the cultivated mycelium material or composite mycelium material, but also penetrate into the kernel of the material via the surface. In the embodiment, the dye is present in the entire interior of the cultivated mycelium material or composite mycelium material.

[0397] In various embodiments of the present disclosure, the cultivated mycelium material or composite mycelium material can be dyed so that the cultivated mycelium material or composite mycelium material is colorfast. A variety of techniques can be used to measure colorfastness, such as ISO 11640:2012: Colorfastness Test-Colorfastness Test-Colorfastness of Resistance to Periodic Reciprocating Friction or ISO 11640:2018 (which is an update of ISO11640:2012). In a specific embodiment, colorfastness will be measured according to the grayscale used as a metric to determine friction fastness and sample changes. In some embodiments, the cultivated mycelium material or composite mycelium material will demonstrate strong colorfastness, indicated by a grayscale of at least 3, at least 4, or at least 5.

[0398] Protein sources

[0399] In a number of embodiments, it may be beneficial to optionally process the cultivated mycelial material or composite mycelial material with one or more protein sources (i.e., exogenous protein sources) that are not naturally present in the cultivated mycelial material or composite mycelial material. In some embodiments, the one or more proteins are from species other than fungal species, from which the cultivated mycelial material is produced. In some embodiments, the cultivated mycelial material or composite mycelial material may be optionally processed with a plant protein source (such as pea protein, rice protein, hemp protein, and soy protein). In some embodiments, the protein source will be animal protein, such as insect protein or mammal protein. In some embodiments, the protein will be a recombinant protein produced by a microorganism. In some embodiments, the protein will be a fibrous protein, such as silk or collagen. In some embodiments, the protein will be an elastomeric protein, such as elastin or elastin. In some embodiments, the protein will have one or more chitin binding domains. Exemplary proteins with chitin binding domains include elastin and a variety of bacterial chitin binding proteins. In some embodiments, the protein will be an engineered or fusion protein that includes one or more chitin binding domains.Depending on the embodiment, the cultivated mycelial material or composite mycelial material may be preserved as described above prior to treatment or may be treated without prior preservation.

[0400] In a specific embodiment of the present disclosure, the cultivated mycelial material or composite mycelial material is immersed in a solution comprising a protein source. In a specific embodiment, the solution comprising a protein source is an aqueous solution. In other embodiments, the solution comprising a protein source comprises a buffer, such as phosphate buffered saline.

[0401] In some embodiments, the solution comprising a protein source will include an agent for cross-linking the protein source. Depending on the embodiment, a variety of known agents that interact with amino acid functional groups can be used. In a specific embodiment, the agent for cross-linking the protein source is transglutaminase. Other suitable agents for cross-linking amino acid functional groups include tyrosinase, genipin, sodium borate and lactase. In other embodiments, traditional tanning agents can be used to cross-link proteins, including chromium, vegetable tannins, tanning oils, epoxy resins, aldehydes and synthetic tanning agents. As discussed above, due to the toxicity and environmental concerns of chromium, PAE other minerals, such as aluminum, titanium, zirconium, iron and combinations thereof, with and without chromium can be used.

[0402] In a plurality of embodiments, the processing using a protein source can occur before, after or in parallel with preserving the cultivated mycelial material or composite mycelial material, plasticizing the cultivated mycelial material or composite mycelial material and / or dyeing the cultivated mycelial material or composite mycelial material. In some embodiments, the processing using a protein source can occur before or during the use of a solution comprising alcohol and salt to preserve the cultivated mycelial material or composite mycelial material. In some embodiments, the processing using a protein source occurs before or in parallel with dyeing the cultivated mycelial material or composite mycelial material. In some of these embodiments, the protein source is dissolved in the dye solution. In a specific embodiment, the protein source will be dissolved in the alkaline dye solution that optionally includes one or more agents that promote dye absorption.

[0403] In some embodiments, a plasticizer will be added to the dye solution comprising a dissolved protein source to plasticize the cultivated mycelial material or composite mycelial material in parallel. In a specific embodiment, the plasticizer can be a fatting agent. In a specific embodiment, the plasticizer will be added to the protein source, which is dissolved in the alkaline dye solution comprising one or more agents that promote dye absorption.

[0404] Coatings and Finishes

[0405] After the cultivated mycelium material or composite mycelium material has been processed using any combination of methods as described above, the cultivated mycelium material or composite mycelium material can be treated with a finishing agent or coating. A variety of finishing agents common in the leather industry can be used, such as proteins in binder solutions, nitrocellulose, synthetic waxes, natural waxes, waxes with protein dispersions, oils, polyurethanes, acrylic polymers, acrylic resins, emulsion polymers, water-resistant polymers and their various combinations. In a specific embodiment, a finishing agent including nitrocellulose can be applied to the cultivated mycelium material or composite mycelium material. In another specific embodiment, a finishing agent including conventional polyurethane finishing agents will be applied to the cultivated mycelium material or composite mycelium material. In a plurality of embodiments, one or more finishing agents will be applied to the cultivated mycelium material or composite mycelium material in sequence. In some cases, the finishing agent will be combined with a dye or pigment. In some cases, the finishing agent will be combined with a feel modifier (i.e., a sensory modifier or tactile), including one or more of natural and synthetic waxes, silicones, paraffins, saponified fatty substances, fatty acid amides, amide esters, stearamides, emulsions thereof, and any combination of the foregoing. In some cases, the finishing agent will be combined with a defoamer. In some embodiments, an external element or force is applied to the cultivated mycelium material or composite mycelium material. In the embodiments, the external element or force comprises heating and / or pressurization. In some embodiments, the external element or force is hot pressing. In some embodiments, an external force is applied to the cultivated mycelium material or composite mycelium material. In the embodiments, the external force comprises heating and / or pressurization. In some embodiments, the external force is hot pressing.

[0406] Processed mycelium material

[0407] In various embodiments of the present disclosure, the cultivated mycelium material or composite mycelium material is sonicated, perforated or vacuum processed. The perforation may include needle puncture, air puncture or water puncture.

[0408] In various embodiments of the present disclosure, the cultivated mycelial material or composite mycelial material can be mechanically processed and / or chemically processed in various ways in a solution (i.e., a dye solution, a protein solution, or a plasticizer) and after the cultivated mycelial material or composite mycelial material has been removed from the solution. In the embodiments, the methods include mechanically processing and / or chemically processing the cultivated mycelial material or composite mycelial material, wherein a processed mycelial material is produced.

[0409] While the cultivated mycelium material or composite mycelium material is in solution or dispersion, it may be stirred, sonicated, squeezed or pressurized to ensure absorption of the solution. The extent of mechanical processing will depend on the specific treatment applied and the level of brittleness of the cultivated mycelium material or composite mycelium material at its processing stage. The squeezing or pressurizing of the cultivated mycelium material or composite mycelium material may be achieved by hand wringing, mechanical wringing, flat presses, felt rollers or calendar rollers.

[0410] Likewise, as discussed above, the cultivated mycelial material or composite mycelial material can be pressurized or otherwise processed to remove the solution from the composite mycelial material after it is removed from the solution. The treatment with the solution and pressurizing the material can be repeated several times. In some embodiments, the material is pressurized at least twice, at least three times, at least four times, or at least five times.

[0411] Once the cultivated mycelium material or composite mycelium material is sufficiently dried (e.g., using the heat, pressure or other drying techniques described above), the cultivated mycelium material or composite mycelium material can be subjected to additional mechanical and / or chemical processing. Depending on the technique used to treat the cultivated mycelium material or composite mycelium material and the resulting toughness of the cultivated mycelium material or composite mycelium material, different types of mechanical processing can be applied, including but not limited to sanding, brushing, electroplating, riveting, tumbling, vibrating and cross-rolling.

[0412] In some embodiments, the cultivated mycelial material or composite mycelial material may be embossed with any heat source or by applying chemicals. In some embodiments, the cultivated mycelial material or composite mycelial material in solution may be subjected to additional chemical processing, such as maintaining at an alkaline pH using a base such as ammonium hydroxide. In specific embodiments, the pH will be at least 9, 10, 11 or 12. In some embodiments, the pH of the cultivated mycelial material or composite mycelial material in solution will be adjusted to an acidic pH in order to fix the composite mycelial material using a variety of agents (such as formic acid). In specific embodiments, the pH will be adjusted to a pH of less than 6, 5, 4 or 3 in order to fix the cultivated mycelial material or composite mycelial material.

[0413] Finishing, coating and other steps may be performed after or before mechanical and / or chemical processing of the dried cultivated mycelium material or composite mycelium material. Likewise, final pressing steps, including decorative steps such as embossing or engraving, may be performed after or before mechanical and / or chemical processing of the dried cultivated mycelium material or composite mycelium material.

[0414] Fig.14 Flow chart illustrating a method 200 for converting a raw mycelium material into a crusting material that may be processed according to a desired finishing process (e.g., finishing coating, decorative step, final pressing step) based on the end-use application of the material. The raw mycelium material may be a dried, refrigerated, or frozen material manufactured according to any of the methods described herein. The raw material may optionally be slit on the top and / or bottom to provide a mycelium sheet of desired thickness. Slitting may also provide a smoother surface at the cut. The crusting material may be dyed, plasticized, dried, and / or otherwise post-processed as described herein.

[0415] Still refer to Fig.14 At step 202, a pre-finishing treatment solution may be prepared based on the size and mass of the mycelial material. In one embodiment, the pre-finishing treatment solution may be prepared in a volume of about 6 mL per gram of wet mycelial material or 20 mL per gram of dry mycelial material. The pre-finishing treatment solution may include one or more dyes, tannins, and / or plasticizers (e.g., fatliquors) in a suitable solvent (such as water). In one embodiment, the pre-finishing treatment solution includes one or more dyes and / or tannins and one or more fatliquors. The amount of dye added may be based on the specific type of dye and the desired color of the resulting product. An exemplary pre-finishing treatment solution includes: one or more acid dyes at a concentration that produces a desired color; about 25 g / L plant tannins; about 6.25 g / L LEX fatliquor (Trumpler, Germany); and about 18 g / L to about 19 g / L DXV fatliquor (Trumpler, Germany).

[0416] At step 204, the pre-finishing treatment solution may be applied to the mycelium material by a combination of soaking and pressurizing processes. In one embodiment, the material is soaked in the pre-finishing treatment solution for a predetermined period of time (e.g., 1 minute) and then moved through a pressurizing system. An example of a suitable pressurizing system includes moving the soaked material through a pair of rollers that are spaced apart to provide the desired degree of pressurization to the material with each pass between the rollers. The material may be pushed and / or pulled through the rollers. The rate at which the material passes through the rollers may vary. According to one aspect of the present disclosure, the soaking and pressurizing process at step 204 may be repeated one or more times (e.g., 1, 2, 3, 4, 5 times or more).

[0417] After the pre-finishing treatment application at 204, the material can proceed to a fixing process 206. The fixing process 206 includes adjusting the pH of the pre-finishing treatment solution to a pH that can be used to fix the dye. In one embodiment, the fixing process is an acid fixing process, which includes reducing the pH of the pre-finishing treatment solution. Non-limiting examples of acids that can be used for acid fixing include acetic acid and formic acid. For example, acetic acid can be used to reduce the pH of the above exemplary pre-finishing treatment solution to a pH of 3.15±1.0.

[0418] At step 210, the mycelium material may be soaked in a pH adjusted pre-finishing treatment solution and flattened in a manner similar to that described above with respect to step 204. The soaking and pressing process at step 210 may be repeated one or more times (e.g., 1, 2, 3, 4, 5, or more times).

[0419] Step 212 includes a final, extended soak of the material in a pH adjusted pre-finishing treatment solution. The material may be inverted about halfway through the extended soak period. The extended soak period may be about 30 minutes to 1 hour or more. When the extended soak period is complete, the material may be processed through a final pressurization process at 214. The final pressurization process may be the same or different than that described above with respect to steps 204 and 210.

[0420] After the fixing process 206, at step 216, the material can be dried with or without heating. During the drying step 216, the material can be held substantially vertically, horizontally, or in any orientation therebetween. During the drying step, the material can optionally be restrained. For example, one or more clamps can be used to restrain all or a portion of the material during drying. In some embodiments, the drying step 216 is performed under ambient conditions.

[0421] Mechanical properties of composite mycelium materials

[0422] The various methods of the present disclosure can be combined to provide processed cultivated or composite mycelium materials having various mechanical properties. In the embodiments, the mycelium material includes mechanical properties such as wet tensile strength, initial modulus, elongation at break, thickness, and / or crack tear strength. Other mechanical properties include, but are not limited to, elasticity, stiffness, yield strength, ultimate tensile strength, ductility, hardness, toughness, creep resistance, and other mechanical properties known in the art.

[0423] In various embodiments, the processed mycelium material can have a thickness of less than 1 inch, less than 1 / 2 inch, less than 1 / 4 inch, or less than 1 / 8 inch. In some embodiments, the composite mycelium material has a thickness of about 0.5 mm to about 3.5 mm, including, for example, about 0.5 mm to about 1.5 mm, about 1 mm to about 2.5 mm, and about 1.5 mm to about 3.5 mm. The thickness of the material within a given piece of material can have a varying coefficient of variation. In some embodiments, the thickness is substantially uniform to produce a minimum coefficient of variation.

[0424] In some embodiments, the mycelium material may have an initial modulus of at least 20 MPa, at least 25 MPa, at least 30 MPa, at least 40 MPa, at least 50 MPa, at least 60 MPa, at least 70 MPa, at least 80 MPa, at least 90 MPa, at least 100 MPa, at least 110 MPa, at least 120 MPa, at least 150 MPa, at least 175 MPa, at least 200 MPa, at least 225 MPa, at least 250 MPa, at least 275 MPa, or at least 300 MPa. In some embodiments, the mycelium material can have an initial modulus of about 0.5 MPa to about 300 MPa, including, for example, about 0.5 MPa to about 10 MPa, about 1 MPa to about 20 MPa, about 10 MPa to about 30 MPa, about 20 MPa to about 40 MPa, about 30 MPa to about 50 MPa, about 40 MPa to about 60 MPa, about 50 MPa to about 70 MPa, about 60 MPa to about 80 MPa, about 70 MPa to about 90 MPa, about 80 MPa to about 100 MPa, about 90 MPa to about 150 MPa, about 100 MPa to about 200 MPa, and about 150 MPa to about 300 MPa. In a specific embodiment, the mycelium material has an initial modulus of 0.8 MPa. In one aspect, the mycelium material has an initial modulus of 1.6 MPa. In another aspect, the mycelium material has an initial modulus of 97 MPa.

[0425] In some embodiments, the mycelium material can have a wet tensile strength of about 0.05 MPa to about 50 MPa, including, for example, about 1 MPa to about 5 MPa, about 5 MPa to about 20 MPa, about 10 MPa to about 30 MPa, about 15 MPa to about 40 MPa, and about 20 MPa to about 50 MPa. In a specific embodiment, the mycelium material can have a wet tensile strength of about 5 MPa to about 20 MPa. In one aspect, the mycelium material has a wet tensile strength of about 7 MPa. In a specific embodiment, the wet tensile strength will be measured by ASTM D638.

[0426] In some embodiments, the mycelium material may have a breaking strength ("ultimate tensile strength") of at least 1.1 MPa, at least 6.25 MPa, at least 10 MPa, at least 12 MPa, at least 15 MPa, at least 20 MPa, at least 25 MPa, at least 30 MPa, at least 35 MPa, at least 40 MPa, at least 45 MPa, or at least 50 MPa.

[0427] In some embodiments, the mycelium material has an elongation at break of less than 2%, less than 3%, less than 5%, less than 20%, less than 25%, less than 50%, less than 77.6%, or less than 200%. For example, the mycelium material can have an elongation at break of about 1% to about 200%, including, for example, about 1% to about 25%, about 10% to about 50%, about 20% to about 75%, about 30% to about 100%, about 40% to about 125%, about 50% to about 150%, about 60% to about 175%, and about 70% to about 200%.

[0428] In some embodiments, the initial modulus, ultimate tensile strength, and elongation at break are measured using ASTM D2209 or ASTM D638. In a specific embodiment, the initial modulus, ultimate tensile strength, and elongation at break are measured using a modified version of ASTM D638 that uses the same sample size as ASTM D638 and the strain rate of ASTM D2209.

[0429] In some embodiments, the mycelium material may have a single needle tear strength of at least 15 N, at least 20 N, at least 25 N, at least 30 N, at least 35 N, at least 40 N, at least 50 N, at least 60 N, at least 70 N, at least 80 N, at least 90 N, at least 100 N, at least 125 N, at least 150 N, at least 175 N, or at least 200 N. In a particular embodiment, the notched tear strength will be measured by ASTM D4786.

[0430] In some embodiments, the mycelium material may have a double needle tear strength of at least 20 N, at least 40 N, at least 60 N, at least 80 N, at least 100 N, at least 120 N, at least 140 N, at least 160 N, at least 180 N, or at least 200 N. In a particular embodiment, the notched tear strength will be measured by ASTM D4705.

[0431] In some embodiments, the mycelium material may have a notch tear strength (also referred to as slit tear strength) of at least 1.8 N, at least 15 N, at least 25 N, at least 35 N, at least 50 N, at least 75 N, at least at least 100 N, at least 150 N, or at least 200 N, as measured by ISO-3377. In a specific embodiment, the notch tear strength will be measured by ASTM D4704. In some embodiments, the mycelium material may have a slit tear strength of at least 1 N, at least 20 N, at least 40 N, at least 60 N, at least 80 N, at least 100 N, at least 120 N, at least 140 N, at least 160 N, at least 180 N, or at least 200 N, as measured by ISO-3377-2. In one aspect, the mycelium material has a split tear strength of about 1 N to about 200 N, including, for example, about 10 N to about 30 N, about 20 N to about 40 N, about 30 N to about 50 N, about 40 N to about 60 N, about 50 N to about 70 N, about 60 N to about 80 N, about 70 N to about 90 N, about 80 N to about 100 N, about 90 N to about 110 N, about 100 N to about 120 N, about 110 N to about 130 N, about 120 N to about 140 N, about 130 N to about 150 N, about 140 N to about 160 N, about 150 N to about 170 N, about 160 N to about 180 N, about 170 N to about 190 N, and about 180 N to about 200 N, as measured by ISO-3377-2.

[0432] In some embodiments, the mycelium material has a flexural modulus (flexure) of at least 0.2 MPa, at least 1 MPa, at least 5 MPa, at least 20 MPa, at least 30 MPa, at least 50 MPa, at least 80 MPa, at least 100 MPa, at least 120 MPa, at least 140 MPa, at least 160 MPa, at least 200 MPa, at least 250 MPa, at least 300 MPa, at least 350 MPa, at least 380 MPa. In a specific embodiment, compression will be measured by ASTM D695. In some embodiments, the mycelium material has a flexural modulus of about 5-10 MPa. In some embodiments, the mycelium material has a flexural modulus of about 10-20 MPa. In some embodiments, the mycelium material has a flexural modulus of about 20-30 MPa. In some embodiments, the mycelium material has a flexural modulus of about 30-40 MPa. In some embodiments, the mycelium material has a flexural modulus of about 10-11 MPa. In some embodiments, the mycelium material has a flexural modulus of about 10 MPa.

[0433] In various embodiments of the present disclosure, the mycelium material has different absorption characteristics, which are measured as the percentage increase in mass after immersion in water. In some embodiments, the percentage increase in mass after immersion in water for 1 hour is less than 1%, less than 5%, less than 25%, less than 50%, less than 74%, or less than 92%. In a specific embodiment, ASTM D6015 is used to measure the percentage increase in mass after immersion in water for 1 hour.

[0434] Method for producing mycelium material

[0435] A method of making a mycelium material as described herein is also provided. According to one embodiment of the present disclosure, the mycelium material can be made by producing a cultivated mycelium material comprising one or more masses of branched hyphae; disrupting the cultivated mycelium material comprising the one or more masses of branched hyphae; and adding an adhesive to the cultivated mycelium material (e.g., by contacting the disrupted cultivated mycelium material with a solution comprising an adhesive); thereby producing a composite mycelium material. In some embodiments, the cultivated mycelium material comprises one or more masses of disrupted branched hyphae. In some embodiments, the one or more masses of disrupted branched hyphae have a length. In the embodiment, the one or more masses of disrupted branched hyphae have a length of about 0.1 mm to about 5 mm.

[0436] On the other hand, the mycelium material can be manufactured by producing a cultivated mycelium material; pressurizing the cultivated mycelium material; and adding a binder to the cultivated mycelium material (e.g., by contacting the pressurized cultivated mycelium material with a solution containing the binder), thereby manufacturing a composite mycelium material.

[0437] In some embodiments, the production includes producing a cultivated mycelium material on a solid substrate. In some embodiments, the method further includes incorporating a support material into the mycelium material. In some embodiments, the support material is a reinforcing material. In some embodiments, the support material is a base material. In some embodiments, the disruption includes disrupting the one or more clumps of branched hyphae by mechanical action. In some embodiments, the method further includes adding one or more proteins from species other than the fungal species that produces the cultivated mycelium material. In some embodiments, the method further includes adding a dye to the cultivated mycelium material or the mycelium material. In some embodiments, the method further includes adding a plasticizer to the cultivated mycelium material or the mycelium material. In some embodiments, the method further includes adding tannin to the cultivated mycelium material or the mycelium material. In some embodiments, the method further includes adding a finishing agent to the mycelium material. In some embodiments, the method further comprises determining the mechanical properties of the mycelium material, wherein the mechanical properties include, but are not limited to, wet tensile strength, initial modulus, elongation at break, thickness, tear strength, elasticity, stiffness, yield strength, ultimate tensile strength, ductility, hardness, toughness, creep resistance, etc. For example, the mycelium material has a wet tensile strength of about 0.05 MPa to about 50 MPa, an initial modulus of about 0.5 MPa to about 300 MPa, an elongation at break of about 1% to about 200%, a thickness of about 0.5 mm to about 3.5 mm, and / or a tear strength of about 1 N to about 200 N.

[0438] In some embodiments, conventional papermaking equipment is used to make the cultivated mycelium material or composite mycelium material.

[0439] Example

[0440] The following examples are presented to provide those skilled in the art with a complete disclosure and description of how to make and use the invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the following experiments are all or the only experiments performed. Every effort has been made to ensure accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should be considered. Unless otherwise indicated, parts are parts by weight, molecular weights are weight average molecular weights, temperatures are expressed in degrees Celsius, and pressures are at or near atmospheric pressure. Standard abbreviations may be used, such as bp, base pairs; kb, kilobases; pl, picoliters; s or sec, seconds; min, minutes; h or hr, hours; aa, amino acids; kb, kilobases; bp, base pairs; nt, nucleotides; im, intramuscular; ip, intraperitoneal; sc, subcutaneous; etc.

[0441] Materials and methods

[0442] The following materials and methods were used in the examples.

[0443] Composite mycelium material samples

[0444] For each sample described below, the components were blended together in a blender (Vitamix or Blendtec). The resulting slurry was poured into a mold resting on a papermaking screen, or a sizing cloth through which water was passed. After waiting for about 1-15 minutes, the mold was removed from the slurry. The material was then pressed to about 0.25 inches via a hand press. The resulting material was then removed from the screen and dried in front of a fan. The sample was dried and then pressurized in a hot press. A scaffold is optionally included in the composite mycelium material, as described below.

[0445] The following samples were used:

[0446] HM1-1-1: 15 g of dried cultivated mycelial material, 375 mL of water, and 3 g of pea protein (Nutribiotic) were blended together. 3.75 g of transglutaminase from BDF (BDF TG) was added. The blend was mixed with a spatula and incubated at room temperature (RT) for 30 min, and then poured into a 6×6 inch mold, pressed to 1 / 4" thick, dried, and labeled HM1-1-1. One third of this material was rubbed with 3 g of epoxidized soybean oil and then the sample was pressed at 120° C. at 1 metric ton pressure for 1 min and labeled HM1-1-11_120p.

[0447] HM1-1-7: 15 g of dried cultivated mycelial material, 375 mL of water, 3 g of pea protein (Nutribiotic), and 3 g of leather glue were blended together. 3.75 g of BDF TG was added. The blend was mixed with a spatula and incubated at RT for 30 min, and then poured into a 6×6 inch mold, pressed to 1 / 4" thick, dried, and labeled as HM1-1-7.

[0448] HM1-1-9: 2.5 g of dried cultivated mycelial material, 75 mL of water, and 0.5 g of pea protein (Nutribiotic) were blended together and poured into a 6×6 inch mold, pressed to 1 / 4" thick, dried, and labeled HM1-1-9.

[0449] HM1-1-11: 10 g of dried cultivated mycelial material, 400 mL of water, 4 g of pea protein (Nutribiotic), and 7.5 g of epoxidized soybean oil were blended together and poured into a 6×6 inch mold, pressed to 1 / 4" thick, dried, and labeled HM1-1-11. Half of the sample was then pressed at 120° C. at 1 metric ton pressure for 1 min.

[0450] HMO refers to a blended sample made of 15 g of dry cultivated mycelial material, 3 g of pea protein, and 5% glycerol in 400 mL of water.

[0451] HM25: 5 g of dried cultivated mycelial material, 125 mL of water, 125 mL of 40 mM phosphate buffer (pH=7) containing 1.5% polyamide epichlorohydrin resin (PAE resin, Polycup 9200 from Solenis), and 1 g of pea protein were blended together. Two 2×2 inch squares were made. One was heated at 105° C. for 5 minutes (labeled: HM25_5min) and one was heated at 105° C. for 10 minutes (labeled: HM25_10).

[0452] HM1-3-1: 5 g of cultivated mycelial material, 125 mL of 50 mM phosphate buffer (pH=7.4) containing 1.5% PAE, and 1 g of pea protein were blended together. Two 2×2 inch pads were made. The pads were heated at 105° C. for 5 min; after placing the pads, the oven took 5 min to reach 105° C. Next, one pad was soaked in 5% glycerol for 10 minutes and dried in a fume hood, and the other pad was subjected to wet tensile testing as is.

[0453] HM1-3-2: 5 g of cultivated mycelial material, 125 mL of 50 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 2.5 g of of leather glue and 1g of pea protein were blended together. Two 2×2 inch pads were made and heated at 105°C for 5 min; after placing the pads, the oven took 5 min to reach 105°C. Next, one pad was soaked in 5% glycerol for 10 minutes and dried in a fume hood, and the other pad was subjected to wet tensile testing as is.

[0454] HM1-3-3: 15 g of cultivated mycelial material, 400 mL of 50 mM phosphate buffer (pH=7.4) containing 1.5% PAE, and 3 g of pea protein were blended together. After drying, salt crystals formed on the outside of the homogenized mycelial plate. A 6×6 inch pad was made and heated at 105° C. for 5 min; after placing the pad, the oven took 5 min to reach 105° C. Next, the pad was soaked in 5% glycerol for 10 minutes and dried in a fume hood.

[0455] HM1-3-4: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH=7.4) containing 1.5% PAE, and 1 g of pea protein were blended together. Two 2×2 inch pads were made; the pads were poured into a 2×2 inch mold and then rolled unidirectionally with baking pins between two papermaking screens. The orientation of the pins was parallel to the longer sides of the rectangular plate.

[0456] HM1-3-5: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH=7.4) containing 3.0% PAE, and 1 g of pea protein were blended together. Two 2×2 inch pads were made. The pads were heated at 105° C. for 5 min; after placing the pads, the oven took 5 min to reach 105° C.

[0457] HM1-3-6: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 2.5 g of of leather glue and 1g of pea protein were blended together. A cotton textile support (support 2) was incorporated into the center of two 2×2 inch pads. After waiting 5 minutes for the oven to reach 105° C., one plate was pressed to 1 metric ton at 105° C. for 2 min and the other plate was heated at 105° C. for 5 min. A 1.54 mm spacer was used to limit the extent to which the plates were pressed.

[0458] HM1-3-7: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 5 g of of leather glue and 1g of pea protein were blended together. A cotton textile support (support 2) was incorporated into the center of two 2×2 inch pads. After waiting 5 minutes for the oven to reach 105° C., one plate was pressed to 1 metric ton at 105° C. for 2 min and the other plate was heated at 105° C. for 5 min. A 1.54 mm spacer was used to limit the extent to which the plates were pressed.

[0459] HM1-3-8: 5 g of cultivated mycelial material and 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE were blended together. Two 2×2 inch pads were made. The pads were heated at 105° C. for 5 min; after placing the pads, the oven took 5 min to reach 105° C.

[0460] HM1-3-9: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH=7.4) containing 1.5% PAE, and 1 g of pea protein were blended together. Two 2×2 inch pads were made. The pads were pressed at 105° C. at 1 metric ton for 2 minutes to a height of 1.45 mm.

[0461] HM1-3-10: 5 g of cultivated mycelium material, 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, and 1 g of pea protein were blended together. The textile scaffold (scaffold 2) was incorporated into a 2×2 inch pad. The scaffold was coated in dry cultivated mycelium material, which had been poured onto the scaffold in a thin slurry the day before and allowed to dry. The plate was then pressed to 1.5 mm at 1 metric ton pressure for 2 min at 105°C.

[0462] HM1-3-11: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH=7.4) containing 1.5% PAE, and 1 g of pea protein were blended together. Two 2×2 inch pads were made. The pads and a cotton textile support (support) with 1 / 8 inch holes were coated with Weldwood contact glue and pressurized with 2 L of water in a beaker for 2.5 h at room temperature. Next, the material was pressurized to 2.54 mm at 1 metric ton pressure for 4 min at 105°C.

[0463] HM1-3-12: 2.5 g of cultivated mycelial material, 62.5 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, and 0.5 g of pea protein were blended together. A paper scaffold (scaffold 3, black, non-woven, plastic) was incorporated into a 2×2 inch pad. The plate was then pressed to 1.5 mm at 1 metric ton pressure for 2 min at 105°C.

[0464] HM1-3-13: 2.5 g of cultivated mycelial material, 62.5 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, and 0.5 g of pea protein were blended together. A 2×2 inch pad was made, which incorporated the support 4 inside, onto which the mycelial slurry had been poured the night before. The plate was then pressed to 1.5 mm at 1 metric ton pressure for 2 min at 105°C.

[0465] HM1-3-14: 2.5 g of cultivated mycelial material, 62.5 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, and 0.5 g of pea protein were blended together. A 2×2 inch pad with a clean support 4 was incorporated inside. The plate was then pressed to 1.5 mm at 1 metric ton pressure for 2 min at 105°C.

[0466] HM1-3-15: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, and 1 g of pea protein were blended together. Two 2×2 inch pads were made. The pads and a cotton textile support with 1 / 8 inch holes (support 4) were coated with leather adhesive from Springfield Leather Company and pressurized with 2 L of water in a beaker for 2.5 h at room temperature. Next, the material was pressurized to 2.54 mm at 1 metric ton pressure for 4 min at 105°C.

[0467] HM1-4-1: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 1 g of pea protein, 1 g of iron (III) oxide black or 1 g of cobalt blue, and 5% glycerol were blended together. Cotton textile support (support 4) was incorporated internally. Two 2×2 inch pads were made. The pads were pressed and heated at 105° C. under 1 metric ton pressure for 2 min.

[0468] HM1-4-2: 5 g of cultivated mycelial material, 125 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 1 g of pea protein, 0.125 g of brown acid dye, and 5% glycerol were blended together. A cotton textile support (support 4) was incorporated internally. Two 2×2 inch pads were made. The pads were pressed and heated at 105° C. under 1 metric ton pressure for 2 min.

[0469] HM1-4-3: 15 g of cultivated mycelial material, 400 mL of 25 mM phosphate buffer (pH = 7.4) containing 1.5% PAE, 3 g of pea protein, 5 g of iron (III) oxide black, and 5% glycerol were blended together. Two 6×6 inch pads were made. The pads were pressed and heated at 105° C. under 1 metric ton pressure for 2 min.

[0470] HM1-4-4: Same as HM1-4-3.

[0471] HM1-4-5: Same as HM1-4-3 and HM1-4-4, except 8g from The leather glue is also mixed in.

[0472] HM3: 15 g of dry cultivated mycelium material, 500-600 mL of water, and 3 g of pea protein (Nutribiotic) were blended together. 3.75 g of BDF TG was added, mixed with a spatula, half of the mold was poured into a 6×6 inch mold, pre-wetted Scaffold 1 was pressed into the material, and the other half of the material was poured into the mold. The mixture was incubated for 30 min, then pressed to 1 / 4" thick, and dried. This sample was cut in half, and 3 g of epoxidized soybean oil was rubbed into half of the sample. The sample was then hot pressed at 120°C under 1 metric ton pressure for 2 min.

[0473] HM22: 15 g of dry cultivated mycelium material, 550 mL of water and 3 g of pea protein were blended together. A strip of gauze (Scaffold 1) was incorporated inside by hook needle needle technique. Scaffold 1 was not evenly placed in the middle of the material.

[0474] Wet tensile test

[0475] The standard test method for tensile testing of composite mycelium materials is performed according to the ASTM D638 protocol. The samples are conditioned for 24 hours at 65 ± 2% RH. In some embodiments, the samples are soaked in water for 1 hour at room temperature before testing. The samples are punched using an ASTM standard die (such as ASTM D638 Type IV dog bone). The thickness, width, and mass of each sample are measured. The appropriate tensile test method is then run on a universal testing machine from Zwick (zwikiLine Materials Testing Machine Z5.0TH).

[0476] Crack tear test

[0477] The standard test method for crack tear testing of composite mycelium materials was performed using a universal testing system from Zwick according to ISO 3377-2 protocol. The samples were conditioned at 65±2% RH for 24 h. In some embodiments, the samples were equilibrated at 65% relative humidity for 16 h at room temperature prior to testing. 1" x 2" specimens with a center crack were die cut using ISO 3377-2. The thickness and mass of each specimen were measured. The appropriate crack tear test method was then run on a universal mechanical tester from Zwick.

[0478] Drawing of mycelial material

[0479] The mycelial hyphae are aligned by manually stretching the sheet of material in the direction. The stretching force applied to the material does not exceed the breaking force.

[0480] Scanning electron microscopy (SEM) imaging and Fourier transform (FT) analysis

[0481] Scanning electron microscopy (SEM) uses a focused electron beam to evaluate the morphology of materials through secondary electrons. The electron beam is scanned in a raster pattern to collect micrographs at scales between 1 mm and 10 nm or between 10X and 100,000X magnification. The SEM method uses a low vacuum (1 to 10 Torr), avoiding the need for dehydration or sputter coating of biological samples.

[0482] The SEM micrographs were then cropped to square size and analyzed using Fourier Transform (FT). The FT of the image represents the sum of the complex exponentials of the magnitude (i.e., intensity), frequency, and phase angle of the variation. The resulting frequency domain reveals the image periodicity that varies with the angle. Because aligned fibers induce periodicity that is orthogonal to the fiber axis, the frequency domain is used to quantify preferential fiber alignment. The polar coordinate frequency domain image is then converted to Cartesian coordinates to extract the pattern of the azimuthal distribution. The azimuthal distribution is then fitted with a Gaussian peak to calculate the full amplitude at half maximum and the maximum angular position.

[0483] Polarized Fourier Transform Infrared (FTIR) Spectroscopy

[0484] Fourier transform infrared (FTIR) spectroscopy was used to evaluate the secondary and tertiary structures of the composite mycelium materials. Depending on the embodiment, different wave numbers (cm -1 ) to evaluate the different chemical functionalities present in the chitin of mycelial hyphae. The wavenumber of the methyl deformation mode corresponding to N-acetyl was found to be around 1410 cm -1 , while the ether vibration mode was found to be around 950 cm -1 .

[0485] In the attenuated total reflection mode, the infrared beam is internally reflected inside the internal reflection component. The light absorption is caused by the attenuation of the evanescent wave at the interface.

[0486] In polarized FTIR, the light is polarized with s (perpendicular to the plane of reflection). The sample is positioned obliquely along the polarized light s vector (0 degrees) or perpendicular (90 degrees). The ratio of the absorbance at 0 and 90 degrees defines the dichroic ratio (R), from which it is possible to calculate the second order Legendre parameters <p2>=(R-1) / (R-2).

[0487] Example 1: Tensile properties of composite mycelium materials

[0488] As shown in the following results, by incorporating an adhesive (such as a cross-linking agent polyamide-epichlorohydrin (PAE)) and an optional textile support, the wet tensile strength and tear strength of the material are dramatically improved. PAE is a cross-linking agent traditionally used in the paper industry, but is also found in sausage casings. The performance of the harder textile support in the composite mycelium material is better than that of the less hard, more elastic support. It is not intended to be bound by any particular theory, and it is advocated that the less hard support ultimately does not bear any load when incorporated into the material. In addition, compared with the harder support, the less hard support is more likely to be delaminated when broken. Water-based latex adhesives provide further benefits to the material in terms of strength and plastic deformation.

[0489] In some samples including PAE, the composite mycelium material had higher wet tensile strength and higher rip tear than the intact cultivated mycelium material. In some samples with PAE, the composite mycelium material had lower elongation at break (plastic deformation) than the intact cultivated mycelium material.

[0490] Adding adhesive

[0491] Table 1 depicts the wet tensile strength (MPa), initial modulus (MPa), and elongation at break (%) of various composite mycelium materials.

[0492] Table 1

[0493]

[0494]

[0495] The addition of a binder (such as PAE or polyurethane) and in some samples a textile scaffold dramatically improved the tensile properties of the composite mycelium material. The wet tensile strength increased from 0.11 MPa to at least 0.86 MPa, and as high as 7.38 MPa. The initial modulus increased from 1.97 MPa to at least 3.48 MPa, and as high as 96.98 MPa. The elongation at break increased from 8.20% to at least 10.39%, and as high as 278.5%. The thickness of the composite mycelium material ranged from 0.5 mm to 3.5 mm. The sub-network size (e.g., length) of the disrupted mycelium material ranged from 0.5 mm to 2 mm.

[0496] PAE cross-linking

[0497] The presence of pea protein did not affect PAE crosslinking. Sample HM1-3-8 was crosslinked with 1.5% PAE and without pea protein (Table 2). Increasing the PAE concentration from 1.5% to 3% did not significantly increase the wet tensile strength (Table 2). Figure 2 Illustrated are stress-strain curves for hot pressed mycelium sheets containing PAE and Scaffold 3 (HM1-3-12) (dashed line) and hot pressed mycelium sheets containing PAE and Scaffold 4 (HM1-3-13) (solid line).

[0498] Table 2 depicts the wet tensile strength (MPa), initial modulus (MPa), and elongation at break (%) of various composite mycelium materials. Samples were prepared to compare PAE crosslinked samples at 1.5% dry weight percentage with PAE crosslinked samples at 3% dry weight percentage in a board, and to compare PAE+pea protein crosslinked samples with samples crosslinked in the absence of pea protein.

[0499] Table 2

[0500]

[0501]

[0502] Additive Enhancement

[0503] Latex adhesive (hide glue) improved wet tensile strength and elongation at break and did not affect the ability of PAE to cross-link the mycelium (Table 3).

[0504] Table 3 depicts the wet tensile strength (MPa), initial modulus (MPa), and elongation at break (%) of various composite mycelium materials, comparing mycelium samples cross-linked by PAE in the absence of glue to mycelium samples cross-linked by PAE in the presence of glue.

[0505] Table 3

[0506]

[0507]

[0508] Samples HM1-4-1 to HM1-4-5 included a dye, a plasticizer, and a scaffold, and were all compressed to approximately 1.4 mm.

[0509] Hot Pressing

[0510] Hot pressing the samples at 105°C instead of crosslinking the samples in an oven at 105°C resulted in a two-fold increase in wet tensile strength (Table 4).

[0511] Table 4 depicts the wet tensile strength (MPa), initial modulus (MPa), and elongation at break (%) of various composite mycelium materials. Samples were prepared to compare samples hot pressed at 105°C (HM1-3-9p) with samples crosslinked in an oven at 105°C.

[0512] Table 4

[0513]

[0514] Incorporating support material

[0515] The incorporated support material increases the wet tensile strength of the composite mycelium material, wherein a stiffer support material (such as a scaffold) produces a higher initial modulus than a less stiff support material. In some samples, the incorporated support material increases the wet tensile strength of the superheated and pressed PAE samples by about 2 to 5 times. Figure 3 Shows the different support materials incorporated into the composite mycelium material used herein. From left to right, Figure 3 Depicted are a gauze support having pores slightly smaller than 1 / 16 inch (support 1); a cotton textile support having pores smaller than 1 / 32 inch (support 2); a non-textile support having pores 1 / 16 inch in size (support 3); and a cotton textile support having larger pores 1 / 8 inch in size (support 4). Figure 4 Depicted is bracket 4 with Weldwood adhesive after wet tensile testing.

[0516] Table 5 depicts exemplary mechanical properties of four scaffolds used herein. Mechanical properties were tested on a Zwick system.

[0517] Table 5

[0518] n＝？ sample Tensile strength(MPa) Initial modulus (MPa) Elongation at break (%) 1 Bracket 1 7.32 3.44 20.6 1 Bracket 2 2.53 0.1 49 1 Bracket 3 51.7 1110 4.96 1 Bracket 4 8.7 2.32 31.1

[0519] The incorporated support material increases the wet tensile strength of one or more composite mycelium material samples, wherein a stiffer support material (such as a scaffold) bears more load before the disrupted mycelium breaks compared to a less stiff scaffold (e.g. Scaffold 3, non-textile, compared to cotton textile Scaffold 2). Since the one or more composite mycelium materials are relatively stiff, the elongation at break is quite low, so the relatively stiff scaffold proves to be more effective in producing tensile property values ​​comparable to cow leather. It is desirable for the support material to have a higher initial modulus and optionally a lower elongation at break compared to the composite mycelium material so that the scaffold will initially bear the strain from any tension and then break before the composite mycelium material breaks.

[0520] Scaffold 3 (non-textile scaffold) meets these desired requirements. Scaffold 2 has a low initial modulus and high elongation at break. Scaffold 4 is made of natural material (cotton) and has appropriate tensile strength and initial modulus. Compared to Scaffold 1, it is more difficult to tear Scaffold 4.

[0521] Table 6 depicts the mechanical properties of pressurized, cross-linked composite mycelium materials with and without incorporated support materials (such as scaffolds).The mechanical properties of upholstery leather were used for comparison.

[0522] Table 6

[0523]

[0524] Example 2: Crack tear strength of composite mycelium material

[0525] The crack tear strength of the composite mycelium material was compared with that of the intact cultivated mycelium material.

[0526] Table 7 depicts the split tear strength (N) and thickness (mm) of various composite mycelium materials.

[0527] Table 7

[0528]

[0529] The split tear strength of various composite mycelium materials ranges from about 7N to about 50N. Figure 5 A graph depicting the tear strength of various composite mycelium materials versus thickness, including stressed samples (HM1-4-3 and HM1-1-11_120p) and unstressed samples. The tear strength of the stressed samples is much stronger than that of the unstressed samples. HM1-4-3 was stressed in the presence of 1.5% PAE and HM1-1-11_120p was stressed in the presence of epoxidized soybean oil. The tear strength of the unstressed samples in the absence of PAE is linear with thickness.

[0530] Example 3: Alignment of mycelial hyphae from Ganoderma lucidum

[0531] Next, the cultivated mycelial material or composite mycelial material is disrupted by physically aligning the branching hyphae in one or more directions. Figure 6 A stress-strain curve is shown, which plots the through-thickness drawing stress as a function of strain for an aligned mycelium. Strain cycles were completed in 10% increments from 10% to 80% before drawing to maximum elongation. The force was measured while the branched mycelial mass was aligned. The curve illustrates the proportional limit, followed by a maximum in the curve where necking occurs.

[0532] Table 8 describes Figure 6 The maximum drawing stress and elongation range for full thickness drawing described in .

[0533] Table 8

[0534]

[0535]

[0536] Fig. 7A and Figure 7B Shown before stretching ( Fig. 7A ) and after stretching ( Figure 7B ). In this embodiment, the fibers are aligned along the stress direction. The thin layer comprises three consecutive layers.

[0537] Figure 8 Fourier transforms of SEM images of mycelium before (black squares) and after (grey circles) drawing are shown. The graphs illustrate the normalized grey scale (%) as a function of the fiber alignment angle. Fig. 9 Polarized FTIR spectra of aligned mycelial hyphae are shown along with polarization (0 degrees) and perpendicular to the polarization (90 degrees). The spectrum of pure chitin is shown for comparison. Fig.10 The second-order Legendre parameters ( <p2>). The graph demonstrates the presence of alignment of hyphae at specific frequencies. Fig.11A and Fig. 11B Shown at 150X( Fig.11A ) and 500X( Fig. 11B ) SEM micrograph of two aligned mycelium thin layers bonded with a polyurethane hot melt adhesive. The surface of the layers was measured. Fig. 12A and Fig. 12B Shown in dry state ( Fig. 12A ) and wet state ( Fig. 12B Stress-strain curves of aligned mycelia tested after acclimation at 65% RH under 37°C and aligned mycelia bonded with polyurethane hot melt adhesive.

[0538] Table 9 depicts the tensile properties of aligned mycelium and aligned mycelium bonded with a polyurethane (PU) hot melt adhesive tested after acclimation at 65% relative humidity (RH) and after one hour of water immersion.

[0539] Table 9

[0540]

[0541] The dry tensile strength of the composite mycelium materials was measured for wet tensile strength. For example, the stretched mycelium PU laminates produced dry tensile strengths of 5.750 MPa to 8.804 MPa and wet tensile strengths of 3.298 MPa to 4.549 MPa. It was observed that the initial modulus dropped due to wetting. There was also a large reduction in the value that occurred for the non-adhesive laminates as compared to the bonded laminates. In the stretched mycelium thin layer samples without polyurethane, the absence of adhesive did not change the tensile strength. In addition, the use of polyurethane produced a material that had four times the strength when wet. Without intending to be bound by any particular theory, the tensile strength properties may depend on the specific manipulation type of the mycelium.

[0542] Example 4: Hydroentanglement of composite mycelium materials

[0543] The components for the material are disrupted in a blender and the resulting slurry includes at least the cultivated mycelial material and water. The water is directed through a hole having a diameter of about 50 microns at about 750-1000 psi in the mycelial slurry. The mycelial slurry is immersed in a solution including one or more adhesives. Without intending to be bound by any particular theory, it is proposed that one or more clumps of branched hyphae of the composite mycelial material are effectively entangled via hydraulic entanglement, resulting in improvements in certain mechanical properties of performance, such as wet tensile strength, initial modulus, elongation at break, thickness and / or crack tear strength. The results are included herein, for example, Example 15.

[0544] Example 5: Infiltration of the mycelial material with a processing solution

[0545] Mycelial sheets are porous materials made of tangled hyphae. The hyphae are tangled to form a 3D network containing open pores between the hyphae structures, such as Fig.15 As shown. The pore diameter of these hyphae can range from hundreds of nanometers to several microns. When making a mycelium material, it can be beneficial to use a compound that can bind the mycelium hyphae together, which helps to improve the structural integrity and lamination of the mycelium material. However, the compound must also be able to penetrate the mycelium material in order to effectively act as a material binder.

[0546] The pore size distribution inside the mycelial material was characterized using a capillary flow porosimeter (Porous Materials Inc., Ithaca NY) according to ASTM standard F316-03. Fig.16 It is shown that most pore diameters fall within the range between 0.36 and 0.84 microns, with a sharp peak of very high pore size distribution between 0.72 and 0.78 microns. The maximum pore size is approximately 2.2 microns, as determined by the bubble point, which is the pressure at which the first continuous bubble is detected. Therefore, solutions with particle sizes outside this range may be retained or blocked by the surface of the mycelial material and will not penetrate into the center of the mycelial material.

[0547] Particle size of mimosa tannin, fatliquor and dye solution

[0548] Next, the particle size of various processing solutions was determined. A Malvern Zetasizer Nano-S was used to measure the particle size in selected solutions and emulsions. This device can measure particles between 0.1 nanometers and 5 micrometers. Exemplary solutions include mimosa tannin, fatliquor, and dye. The particle size of a 5% aqueous solution of mimosa tannin is between Fig.17 The particle size of the 5 wt% aqueous solution of the fatliquor (Trupon DVX:Truposol LEX=3:1) is shown in Fig.18 Shown in.

[0549] In the Mimosa tannin solution, most particles were between 100 nm and 1 μm, whereas in the fatliquor solution, most particles were between 50 and 500 nm. Therefore, it was expected that Mimosa tannin and fatliquor particles penetrated through the mycelial material.

[0550] Particle size of gum arabic, tannin and fatliquor solution

[0551] Gum Arabic, a natural gum consisting of hardened tree sap and useful as a binder material, has been considered as a candidate for mycelial treatment. However, processing of gum Arabic with materials showed that only surface impregnation occurred. Measurements of the particle size distribution in gum Arabic solutions showed that the particle size of gum Arabic in aqueous solution was relatively large, likely resulting in particles filtering through the mycelium and being located at the mycelial surface, creating a crusty surface layer. The particle size of gum Arabic in a 10% wt / wt aqueous solution ( Fig.19 ), and the particle size of gum arabic (10% wt / wt) with tannin and fatliquor in crust solution ( Fig. 20 ).

[0552] The particles of the gum arabic solution are between 1 μm and more than 5 μm ( Fig.19 ). Since the Malvern equipment measures particles less than 5 μm, the peak at the right edge of the graph is truncated. Unexpectedly, the gum arabic / tannin / fatting agent combination solution does not have particles between 50nm and 1000nm, as would be expected from the particles of the mimosa tannins and fatting agent. Instead, a peak at about 2 μm is observed next to the gum arabic signal peak. Since the fatting agent is a mixture of softening oils, surfactants, defoamers and other ingredients, the addition of mimosa tannins and gum arabic, which contain a large amount of hydroxyl groups, may affect the surface energy. In such solutions, this can change the particle size, especially the micellar particle size.

[0553] Polymer solution particle size

[0554] The particle sizes of various self-crosslinking copolymers were also evaluated. Vinyl acetate-acrylic and vinyl acetate-ethylene are self-crosslinking polymers that can bind to the mycelial material and itself, and thus act as both a binder and a crosslinker. Additional particle size test results showed that vinyl acetate-ethylene (VAE) from different manufacturers had micelle particle sizes that were 10 times different from those in the emulsion (0.23 μm vs. 2.2 μm). Fig.21 10wt% is shown Particle size distribution of 2833 emulsion (Celanese). Fig. 22 Particle size distribution of VAE S-10 emulsion (US Adhesive) is shown. Thus, even similar polymer solutions show a range of particle sizes, depending on the manufacturer source.

[0555] Example 6: Treatment of mycelium with a cross-linking agent and gum arabic binder

[0556] PAE and APS crosslinkers

[0557] The polymer crosslinkers PAE and APS were studied to improve the mechanical strength of mycelial materials. These crosslinkers are used to crosslink mycelial hyphae and can be used with another polymer binder (such as gum arabic or latex). The crosslinkers used are polyamide epichlorohydrin (PAE) and ammonium persulfate (APS). Polyamide epichlorohydrin resins are water-soluble, cationic resins with azetidinium groups that can be used to crosslink polysaccharides (such as cellulose) with carboxyl groups.

[0558] Anionic radicals generated by the decomposition of APS can cross-link polymer chains containing hydroxyl groups.Mycelium is mainly composed of polysaccharides, which have multiple hydroxyl groups on their side chains.

[0559] PAE crosslinker

[0560] About PAE samples, two collections of four kinds of 10% (w / w) mycelium slurries in water were prepared. In each collection, two samples were mixed with PAE to a final 1.5% (w / w) PAE concentration, one sample was mixed with PAE to a final 3% (w / w) PAE concentration, and one sample was not mixed with PAE as a control. The resulting mixture was allowed to stand for 10 minutes, filtered and pressed into a 0.25 inch pad, and then dried at room temperature. The control sample, a 1.5% PAE sample, and a 3% PAE sample were heated at 105°C for 10 minutes. Another 1.5% PAE sample remained unheated. One sample collection was tested to be dry, and the other was wet. ISO 3377-2 double edge tearing, ASTM D2209 tensile and ASTM D4704 notch tearing tests were then performed on a ZwickiLine material testing machine Z5.0 to determine the relative strength of each PAE-processed sample compared to the control sample heated only.

[0561] The tensile strength results of PAE cross-linked samples are Fig.23 The 3% PAE and heat treated samples had the highest stress intensity and the 1.5% PAE and heat treated samples had the second highest stress intensity under both wet and dry conditions. Under wet conditions, the 1.5% PAE and no heat sample had the third highest strength, while under dry conditions, the 1.5% PAE and no heat sample had similar strength to the heated only sample. Therefore, adding PAE and heating the samples increased the strength of the mycelial material in a concentration-dependent manner, and the full effectiveness of PAE cross-linking required the application of heat. Adding PAE and heating to cross-link the mycelial material improved the mechanical quality of the mycelial board.

[0562] APS crosslinker

[0563] For APS samples, the dried mycelium mat was soaked in a 1% (w / w) solution of ammonium persulfate in deionized water or phosphate buffered saline (PBS) and brought to 80°C for 3.5 hours with gentle agitation. The resulting mat was rinsed in deionized water and dried at 40°C for 3 hours. ISO3377-2 double edge tear, ASTM D2209 tensile and ASTM D4704 notched tear tests were then performed on a ZwickiLine material testing machine Z5.0 to determine the relative strength relative to the control sample.

[0564] The results of the tear and tensile strength tests are shown in Tables 13 and 14.

[0565] Table 13. Wet tensile strength, initial modulus and elongation at break.

[0566]

[0567] Table 14. Crack tear test

[0568]

[0569] As compared to the control samples, the APS treated samples had significantly increased wet tensile strength, initial modulus, elongation, and slit tear strength. Thus, adding APS to crosslink the mycelium material improves the mechanical quality of the mycelium board.

[0570] Gum Arabic and Latex or APS

[0571] The ability of gum arabic to improve the mechanical strength of mycelium was also investigated. Additionally, ammonium persulfate (APS) was added to crosslink the gum arabic and mycelium.

[0572] Three samples of mycelium and gum arabic were tested. In the first two, gum arabic (AEP Colloids Division, Sarcom Inc) was crosslinked using 3% wt / wt and 5% wt / wt APS: gum arabic mass. In the third sample, latex was mixed with gum arabic in a 30:70 ratio. The latex contains monomers with unsaturated C=C bonds and can self-polymerize and / or crosslink. The samples were also treated with mimosa tannins and a fatliquor.

[0573] Table 15 provides details of the samples and processing conditions.

[0574]

[0575] All three samples were processed by soaking in the solution for 1 minute followed by a rolling step. The soaking and rolling cycle was performed four times. The processed mycelium plates were dried in a convection oven at 90°C for 2 hours. The plates were removed from the oven and dried overnight at room temperature in a fume hood. The plates were cut and TM The cross sections were imaged using a -3030Plus environmental scanning electron microscope at an accelerating voltage of 15 kV.

[0576] In the cross-sectional SEM image of gum arabic with latex, two dense layers ( Fig.24 ). The gum arabic adhesive penetrated approximately 100 to 150 μm from the surface.

[0577] Fig.25 The samples treated with gum arabic and latex are shown to have similar stiffness to the samples treated with gum arabic and APS, and are all slightly stiffer than the mycelium plates treated with only fatliquor (2.5% w / w in aqueous solution), as indicated by the higher flexural modulus. The higher flexural modulus indicates an increase in stiffness and a decrease in flexibility of the material. This increase in stiffness may be caused by a denser surface layer on the gum arabic / latex treated samples, which has been observed in the SEM images. The data are normalized to the unit sample thickness of the untreated sample, which is calculated by force divided by thickness, with units of N / mm.

[0578] The normalized crack tearing maximum strength of the mycelium boards treated with gum arabic and APS was slightly higher than that of the untreated boards ( Fig.26 ). The gum arabic and latex treated panels showed significantly higher crack tear strength than the gum arabic and APS treated samples. The latex treated mycelium samples showed high crack tear strength, indicating that 30% latex in the binder mixture helps improve the crack tear strength. However, gum arabic and latex have low penetration of solution particles and increased flexural modulus in the material.

[0579] Example 7: Improving mycelium mechanical properties using a self-crosslinking polymer binder

[0580] Infiltration of mycelial material with polymer binder emulsion

[0581] Mycelial mats were incubated with solutions containing various concentrations of polymer emulsion binders to evaluate the effects on mechanical properties. Vinyl acetate-ethylene and vinyl acetate-acrylic acid are self-crosslinking copolymers. The copolymers can act as a binder polymer and bind to the mycelial material and / or itself, allowing a single molecule to act as both a binder and a crosslinker.

[0582] First, an aqueous solution containing vegetable tannin, fatliquor and dye was prepared according to the composition described in Table 17. The amount of water used to prepare the solution was 20 times the mass of the input mycelium sample.

[0583]

[0584] Binders were then added to the solution to achieve a specific binder solids content. The binders and concentration ranges for each binder are described in Table 18. Tear mender latex was used as a control.

[0585]

[0586] 1 The glass transition temperature of natural rubber latex is -70°C.

[0587] 2 Approximate glass transition temperature.

[0588] The dried mycelium samples were immersed in each solution for one minute and then passed through rollers to force the solution through the material. The immersion and rolling process was repeated until the sample appeared to be uniformly saturated with the solution. The samples were dried under ambient conditions and then calendered to a final thickness of 1.3-1.7 mm. The sample mass was recorded before and after processing to determine the mass change relative to its starting mass and final mycelium content.

[0589] For some experiments, tannins, fatliquors and dyes were omitted to understand the effect of individual binder concentration on mass uptake in the final sample. Fig. 27 Shown are the mycelium content of the samples as a function of the binder concentration in the solutions of the samples impregnated with natural rubber latex (Tear Mender) and S-10 vinyl acetate-ethylene.

[0590] like Fig. 27 As shown, the binder absorption in the final sample is proportional to the binder concentration in the solution and inversely proportional to the final mycelium content in the sample. Fig. 27 The decrease in mycelium content in the final sample and the increase in the binder solution concentration indicate that the final sample total mass contains more binder. Therefore, in order to achieve the desired final mycelium content, the binder concentration in the solution can be adjusted according to the relationship shown.

[0591] The surface and interior of the impregnated mycelium samples were examined using scanning electron microscopy (SEM) to qualitatively assess the extent of impregnation. SEM was performed using a Hitachi TM-3030Plus environmental scanning electron microscope at an accelerating voltage of 15 kV. Fig.30 Representative scanning electron micrographs of untreated mycelium samples and mycelium samples impregnated with natural rubber latex or S-10 vinyl acetate-ethylene are shown. Fig.30 As shown, the binders S-10 vinyl acetate-ethylene and natural rubber latex were effectively impregnated into the mycelial network, as evidenced by the filled voids in the cross-section and torn surfaces (Figs. b, c, d, f), compared to the void spaces between the hyphae in the untreated samples (Figs. a and e).

[0592] Evaluation of mechanical properties

[0593] To evaluate the delamination resistance of the binder impregnated mycelium samples, a T-peel test was performed according to ASTM D 1876 using a Zwick Proline universal testing system with a 500 N load cell. Prior to testing, the samples were conditioned at 65% relative humidity until their mass stabilized within 0.25 wt % for one hour.

[0594] Fig.28 The maximum T-peel force of the mycelium sample impregnated with Tear mender natural rubber latex is shown. Fig.29 Shown are the maximum T-peel forces for mycelium samples impregnated with a range of S-10 vinyl acetate-ethylene concentrations. Points represent individual T-peel tests. For these samples, fatliquor, dye and tannin were omitted.

[0595] To evaluate the crack tear resistance of the binder impregnated mycelium samples, crack tear tests were performed using the Zwickiline Universal Test System according to ISO 3377-2. Prior to testing, the samples were conditioned at 65% relative humidity until their mass stabilized within 0.25 wt% for one hour. Fig.31 The maximum force of crack tearing of samples impregnated with various binders at various concentrations is shown, normalized to the sample thickness. Specifically, the maximum force is divided by the sample thickness, in units of N / mm

[0596] Fig.32 Shows the impregnation with the same binder concentration Maximum T-peel force of mycelium samples impregnated with 2833 vinyl acetate-acrylic or Dur-O-Set Elite 22 vinyl acetate-ethylene. Maximum T-peel force for mycelium samples of 2833 vinyl acetate-acrylic or Dur-O-Set Elite 22 vinyl acetate-ethylene. Points represent individual T-peel tests. Diamonds represent upper and lower 95% confidence intervals for the mean maximum T-peel force.

[0597] like Fig.31 and Fig.32 As shown, the delamination resistance of the mycelium can be affected by the choice of binder used for impregnation and its concentration. In general, for both binders studied, the average maximum T-peel force of the impregnated samples was greater than that of the non-impregnated samples.

[0598] Therefore, the crack tear resistance of the mycelium can be influenced by the choice of binder used for impregnation and its concentration.

[0599] Example 8: Processing method for improving mycelium penetration efficiency

[0600] Sonication-assisted solution infiltration process with optional heating

[0601] Sonication is a mechanical method that aids solution infiltration, and thus can make the impregnation process more efficient. The mycelium sheet (wet or dry) is soaked in a bag with a processing solution (a "crusting" solution, such as a dye, fatliquor and / or tannin solution, and an optional binder) and rolled with a roller to help the solution penetrate the mycelium material. Next, the bagged mycelium material and processing solution are sonicated in a bath sonicator or with a probe sonicator. The mycelium material is rolled again in the processing solution, removed from the solution, and dried. The mycelium is finally mechanically softened by additional rolling to smooth and soften the material.

[0602] To investigate the effectiveness of sonication, three solutions representing different viscosity levels were used: a low viscosity crust solution, a medium viscosity crust solution with 7 wt% latex, and a high viscosity crust solution with 10 wt% gum arabic. Water alone was used as a baseline control. The control samples were soaked in the test solutions for 24 hours. Sonication was performed in a bath sonicator. The sample weight change from dry intact mycelium plates to infiltrated plates after drying was Fig.33 As shown in Figure 2, sonication of the mycelium panels with low, medium, and high viscosity crusting solutions resulted in a significant increase in the weight of the mycelium panels as compared to soaking alone. The medium viscosity crusting solution with 7% wt latex resulted in the greatest increase in weight. Thus, sonication improved the penetration of the processing solution in each solution condition tested.

[0603] A modified sonic bath with heating may also be used because (1) the heated solution helps the particles move faster due to Brownian motion; (2) the heated solution also strengthens the mycelial material, and (3) sonicating the mycelium directly in the processing solution in the sonic bath can skip the bagging step.

[0604] Sonication and perforation penetration

[0605] Mechanical perforation of mycelial materials was investigated to determine if it improves solution penetration, for example by creating full thickness microchannels. Two penetration media were evaluated: needle and water. Air penetration can also be used. For the needle penetration method, a roller with a 3 mm needle length (perforation depth) was used. For the water perforation method, high pressure water with a 50 μm diameter spinneret was used. In addition to the mechanical perforation step, sonication was also used. The percent weight change of a crust solution with a 5% gum arabic concentration as described in Table 17 was compared to a crust solution alone using only sonication. The mycelial plates were weighed before and after soaking, and the weight gain of the final plate is shown as the percent weight change of the starting plate weight.

[0606] Fig.34 Shown is the percent change in weight of the binder-infiltrated mycelium panels in the samples perforated by the needle perforation method and sonicated as compared to sonication only. Fig.35 Shown is the percent change in weight of binder penetrated mycelium sheets in samples perforated with water perforation method and sonicated as compared to sonication only. Mechanical perforation on the mycelium sheet surface was found to improve solution wetting for solutions containing gum arabic.

[0607] Vacuum Assisted Solution Infiltration

[0608] Due to the high porosity of the mycelium board (up to 95 vol%), vacuum was used to remove air prior to solution infiltration. This allowed determination of the ability of vacuum compacting the porous mycelium board in order to minimize air volume and increase solution transfer by vacuum force. A vacuum machine setup was constructed from a solution tank, a single-sided mold, a vacuum bag, a solution collector, and a vacuum pump. The boards were incubated with various solutions under vacuum, removed from vacuum and allowed to dry, and then mechanically softened.

[0609] Three solutions representing different viscosity levels were used: a crusting solution (low viscosity), a crusting solution with 7 wt% latex (medium viscosity), and a crusting solution with 10 wt% cold soluble Starch A (high viscosity) (Cargill, USA) as described in Table 17. The mycelial plates were weighed before and after soaking in the crusting solutions, and the weight gain of the final plates is shown as the percentage weight change of the starting plate weight.

[0610] The change in sample weight from the dried intact mycelium plate to the infiltrated plate after drying is shown in Fig.36 The effectiveness of the vacuum-assisted infiltration process depends on the vacuum conditions, as shown by higher vacuum levels, resulting in higher sample weight gain in the crust solution with 7 wt % latex sample.

[0611] Example 9: Lamination of mycelium materials with binders and adhesives

[0612] In addition to improving the internal bond strength of mycelium materials, binders can also be used as adhesives to build mycelium-based composites. Lamination using binders and adhesives can be used to make mycelium materials with improved mechanical properties.

[0613] The tear and tensile properties of the mycelium material can also be improved by adding a textile scrim without sacrificing the flexibility of the material. Depending on the type of textile scrim used (e.g., fabric or plastic scrim), the tear and tensile properties can be adjusted to match a given specification. The scrim can be added to the back of the mycelium material by using an adhesive or binder (such as latex) or it can be laminated between two layers.

[0614] Laminated and Scrims

[0615] First, mycelium samples were prepared as described in Example 7. Briefly, aqueous solutions containing vegetable tannins, fatliquor, and dye were prepared according to the compositions described in Table 17 and the mycelium materials were soaked and rolled as previously described. After drying, the samples were laminated by spreading a thin layer of Tear Mender natural rubber latex on the surfaces of the two mycelium samples, pressed together, and then passed through rollers to improve adhesion and remove excess solution. This produced a laminated double layer mycelium material. In some samples, woven scrim was laminated between the two layers. A piece of woven scrim material cut to the same size as the crust sample was placed between the two samples, and the laminate was then passed through rollers. Prior to testing, the samples were dried at ambient conditions for 96 hours.

[0616] The crack tear force was tested as previously described. Fig.37 The maximum rip tear force is shown for mycelium samples laminated with and without cheesecloth. Laminated bilayers of mycelium only and mycelium bilayers with latex were also tested. Cheesecloth coated in latex was tested as a control. Dots represent individual rip tear tests. Diamonds represent upper and lower 95% confidence intervals for the mean maximum rip tear force.

[0617] like Fig.37 As shown, lamination can be used to substantially increase the maximum rip tear force of the mycelium composite. Each additional lamination step (doubling and pressing, adding a latex layer, adding scrim) increases the rip tear force required to tear the material.

[0618] Fig.38 Representative crack tear force-travel curves are shown for various mycelium samples and composites: single layer mycelium sample; two layers of mycelium tested simultaneously; two layers of mycelium laminated with natural rubber latex; two layers of mycelium laminated with natural rubber latex with a woven scrim in between.

[0619] Fig.37 and Fig.38 It is shown that while lamination alone is a useful method for making higher strength materials, incorporating textile scrim during lamination provides two advantages over lamination alone. First, the maximum force can be increased to that of the scrim, rather than relying on the strength of the mycelium and adhesive alone. Second, the scrim inhibits tear propagation through the material, as seen by the continuous peak in force after the yield point, which is due to the continuous loading and rupture of the scrim yarn. In contrast, in samples not laminated with the scrim, tear propagation proceeds uninhibited until the mycelium is completely torn.

[0620] To construct another type of mycelium-based composite, lamination was performed by pressing two mycelium samples together with a layer of latex after the first soaking cycle and completing the remainder of the process with the samples stuck together, in parallel with the solution infiltration process as described in Example 7. Unlaminated samples (single layer + latex) were prepared as described in Example 7; a single sheet of mycelium material was soaked in a crusting solution containing latex tear mender as a binder and rolled through a pasta sheeter for a total of 4 soaking and rolling cycles.

[0621] Fig.39 The maximum crack tear force of the mycelium samples laminated in parallel and impregnated is shown, compared to the unlaminated (single layer + latex) and unimpregnated (single layer) samples. The double layer and latex laminated samples have higher crack tear force compared to the unlaminated and unimpregnated samples. The dots represent individual crack tear tests. The diamonds represent the upper and lower 95% confidence intervals for the average maximum crack tear force. Therefore, improved crack tear resistance can be achieved by performing the lamination process in parallel with the impregnation process, rather than performing the lamination as a separate process.

[0622] In addition to improving tear resistance, the design and processing of the composite can also be selected to adjust the flexibility of the material. The "Scrim + Latex Bond" and "Scrim + Latex Adhesion" samples were prepared as previously described in Examples 7 and 8 above. "Scrim + Latex Bond" refers to the process in which the scrim is added during the lamination / crusting process. "Scrim + Latex Adhesion" refers to the process in which the scrim is added after the crusting process is complete and the sample is dry. As previously described in Example 6, the gum arabic and APS samples were prepared with 10% gum arabic and 3% APS. To prepare the semi-crusted samples with scrim, the dried mycelium samples were first cut to approximately 0.5 inches thick and then impregnated with the crusting solution as described in Table 17, using the same soaking and rolling process described in Example 7, but without the addition of a binder.

[0623] To measure the flexural initial modulus, samples were subjected to a three-point bending test according to ASTM D790, but with the strain rate increased from 1% / min to 10% / min. Fig.40 The flexural modulus is shown for a double layer with latex, a double layer with scrim and latex, a double layer treated with gum arabic and APS, and a single layer with scrim attached to one side. The double layer treated with the gum arabic and APS samples was made by taking a mycelium single layer, cutting it longitudinally in half, and then bringing the two halves together. Fig.40 As shown, different material designs and processing methods can be selected to adjust the flexural initial modulus by an order of magnitude, which can be used to design products with varying feel and aesthetics. In this case, the flexibility of the material is based on whether the material is impregnated with a latex emulsion or with a gum arabic solution; the latex binder is impregnated throughout the thickness of the material or is used only as a binder; and is controlled by controlling the thickness of the input mycelium material. As a comparison, A1 and A2 cowhide handbag samples were used. The double layer with latex was the stiffest at about 110 MPa, while the single layer with the scrim adhered had a flexural modulus similar to that of the A1 handbag leather benchmark at about 15 MPa. Therefore, the choice of lamination, binder, and scrim can be selected to provide greater tear resistance or greater or less flexibility of the resulting material.

[0624] Hot pressing of mycelium materials

[0625] The mechanical properties of mycelium materials, such as tear resistance and delamination resistance, can be improved by adding binders and by heat pressing. Heat pressing increases the areal density and fuses the mycelium hyphae together, which results in improved mechanical properties.

[0626] 20 mm thick mycelium panels were treated with the crusting solution as described in Table 17 and hot pressed at 70°C for 2 min to achieve 1.5 mm panel thickness. Samples were conditioned at 65% relative humidity for 24 hours prior to testing. Parallel samples were also incubated with 0.5% S-10 vinyl acetate-ethylene prior to hot pressing. T-peel testing was performed according to ASTM D1876 and crack tear properties were tested using a universal tensile tester according to ISO3377-2.

[0627] like Fig.41 As shown, delamination resistance (T-peel maximum force) is increased by heat pressing. Further improvement in T-peel maximum force is achieved by combining heat pressing with the addition of S-10 vinyl acetate-ethylene adhesive.

[0628] Fig.42 It was shown that hot pressing significantly increased the maximum force of crack tearing of the treated mycelium panels.The combination of hot pressing and S-10 vinyl acetate-ethylene binder further improved the tear resistance.

[0629] Example 10: Treatment of mycelial material with a plasticizer

[0630] Fatliquor, fabric softener or sorbitol

[0631] Materials and methods

[0632] Several different types of chemicals were tested to improve the softness of the processed boards after treatment with fatliquors, binders and / or crosslinkers. Softeners were also evaluated for changes in the mechanical strength of the boards, as a reduction in mechanical strength is a disadvantage.

[0633] The mycelium board was treated with a crusting solution containing a total of 2.5 wt% fatliquor with a ratio of 3:1 of Trupon DXV to Truposol LEX (Trumpler Gmbh) and dried. The dried board was then soaked in deionized water for 1 minute and then passed through a dip roller with a gap setting of 1. The soaking and rolling steps were repeated 8 times to ensure that the mycelium board was completely wetted and that any soluble debris or chemicals were removed. Next, a variety of plasticizers shown in Table 19 were diluted in water.

[0634] Table 19: Plasticizer types and concentrations in aqueous solutions

[0635]

[0636] *Diethoxydimethylammonium chloride (DEEDMAC) is the active ingredient in Downy fabric softener.

[0637] The samples were treated with 200 mL of the plasticizer solution shown in Table 19 for about 10 min to 1 hour, depending on the volume of water. Control samples were incubated with water only. After treatment, the samples were dried at ambient conditions for 24 hours, then conditioned overnight at 65% relative humidity (calculated by dividing the partial pressure of water vapor by the equilibrium vapor pressure of water at a given temperature) before mechanical testing.

[0638] result

[0639] The material thickness was constant in all treated samples ( Fig.43 The final density of the treated samples was also estimated by dividing the mass by the volume. The density change indicates plasticizer absorption. The sorbitol 2X sample had a higher final density as compared to the other samples, which can be attributed to the higher sorbitol concentration in the solution ( Fig.44 ).

[0640] Determine the flexural modulus of each sample ( Fig.45 ). All plasticizers tested resulted in similar ranges of mycelium board flexural modulus, and significant reductions in flexural modulus, as compared to water-only treated mycelium. Samples treated with Downy fabric softener and sorbitol had a slightly harder feel than samples treated with fatliquor and sorbitol 2x. Samples treated with sorbitol 2X and fatliquor had the same feel.

[0641] The crack tear of the treated panels was also determined as previously described. The maximum crack treatment strength for each panel was Fig.46 The fatliquor produced the lowest tear strength under all conditions. The sorbitol 2X sample had tear strength comparable to that of the untreated mycelium board, even though the board was as flexible as the fatliquor sample. The maximum tear force was also normalized to the thickness of each board ( Fig.47 The normalized (by force divided by sample thickness, in N / mm) maximum slit tear strengths showed the same tear strength trend for the samples as compared to the original maximum strength force, indicating that the tear strength improvements observed in the sorbitol 2X samples were not simply due to increased material thickness or density.

[0642] Tween 20 and Tween 80

[0643] Sorbitol is a sugar alcohol that can be used as a plasticizer or softener for mycelial sheets to reduce the flexural modulus while maintaining crack tear strength. Polysorbates (Tween 20, Tween 80) are oily liquids derived from ethoxylated sorbitan (a derivative of sorbitol) esterified with fatty acids. Polysorbates are nonionic surfactants with a variety of well-understood molecular structures. The ability of polysorbates to act as plasticizers was evaluated.

[0644] Untreated mycelium plates were pre-weighed and cut so that each plate had the same 3.2 g dry weight. Tween 20 and Tween 80 solutions were prepared by dissolving 0.5 g or 1.5 g weight of Tween 20 or Tween 80 in 20 ml of DI water. The amount and concentration of the Tween solution are shown in Table 20. Three drops of food coloring were also added to the solution to help identify solution penetration.

[0645] Table 20. Plasticizer type and amount

[0646]

[0647] Each plate was soaked in deionized water and compressed by passing through a dip roller with a gap setting of 1. The soaking and rolling steps were repeated a total of 9 times to ensure that the mycelium plates were completely wetted and to ensure that any soluble debris or chemicals were removed. The mycelium plates were then dried overnight under ambient conditions inside a fume hood.

[0648] For plasticizer solution treatment, the compressed and dried mycelium plates were manually massaged in Tween solution. Solution penetration was determined by observing the food coloring level inside the mycelium plates and massaging the plates until the color penetrated through the cut cross-sections. The samples were allowed to dry overnight in a fume hood. All samples were conditioned overnight at 65% RH prior to testing.

[0649] result

[0650] Fig.48 The sample thickness after plasticizer treatment is shown. Fig.49 The density of the samples after plasticizer treatment is shown. The comparative thickness and density of the samples treated with the previously described fatliquor, Downy and sorbitol are also shown. As compared to the control sample, the Tween plasticizer did not change the thickness of the mycelial material ( Fig.48 However, Tween 20 and Tween 80 increased the density of the samples in a dose-dependent manner as compared to the control samples ( Fig.49 Increasing the concentration of Tween 20 or Tween 80 resulted in a comparable increase in sample density, as seen by the higher density of the 1.5 g treated sample compared to the 0.5 g treated sample.

[0651] Fig.50 The highest flexural modulus for each sample is shown. Fig.51 The average flexural modulus for each sample is shown. Comparative flexural moduli for the previously described fatliquor, Downy, and sorbitol treated samples are also shown. Tween surfactants have a plasticizing effect, as seen by the reduction in the total flexural modulus and the average modulus ( Fig.50 and Fig.51 ). The absolute values ​​of the Tween flexural modulus are in a similar range to fatliquor and sorbitol. However, the plasticizer concentration does not affect the flexural modulus. Both concentrations of Tween 20 and Tween 80 produce similar flexural moduli, but the 25 mg / ml (0.5 g) Tween 20 sample has a larger standard deviation of the measured flexural modulus.

[0652] Fig.52 The highest split tear strength for each sample is shown. Fig.53 The normalized tear strength for each sample is shown. The comparative flexural modulus of the previously described fatliquor, Downy and sorbitol treated samples is also shown. The water control sample also shows a lower tear strength than the previous control sample, which can be attributed to the different application methods, or due to the non-uniformity of the starting board. The tear strength does not appear to be significantly reduced by the polysorbate solution treatment, or affected by the Tween type or Tween concentration. 0.5g Tween into a 3.2g mycelium board (about 15.1% weight absorption) may be sufficient.

[0653] The surface color uniformity (data not shown) of the sample processed by tween is also assessed. Color is due to the food dye included in the Tween solution. The sample processed with Tween has a more uniform coloration, even after drying and adaptation. Therefore, using Tween as a surfactant can be used to improve liquid penetration and dye levelness in the mycelial material.

[0654] Erythritol and fat-liquoring agent content

[0655] Mycelium sheets were cut into 3" x 5" sizes. Each sheet was soaked in water and compressed by passing through a dip roller with a gap setting of 1. The soaking and rolling steps were repeated a total of 5 times. The mycelium sheets were then dried in a convection oven at 40°C overnight.

[0656] Two different plasticizer application methods were used. The plasticizer solutions are described in Table 21. The conventional soaking and rolling method was used to treat the fatliquor control (treated with 100 ml of 2.5% w / w fatliquor aqueous solution) and the erythritol sample. The amount of fatliquor used was based on the previous example. In the parallel samples with fatliquor, the fatliquor was pre-weighed to 15% or 30% of the mycelium board weight, diluted in a small amount of water (4 times the board weight), and applied to the mycelium board using gentle manual massage. After the plasticizer was applied, the mycelium board was dried overnight under ambient conditions in a fume hood. All mycelium samples were weighed before and after the plasticizer application to make it possible to determine the percentage of the board weight increase due to plasticizer penetration. Table 21 provides the board starting and ending weights, and the calculated percentage of weight increase due to plasticizer absorption.

[0657] Table 21. Plasticizer types and solution concentrations

[0658] Processing Description Solution concentration Water control No treatment Fatliquor 2.5% DXV:LEX=3:1, 2.5% in 100ml aqueous solution Erythritol 5% in 100ml aqueous solution Fatliquor 15% DXV:LEX=3:1, 0.45g diluted in 12ml water Fatliquor 30% DXV:LEX=3:1, 0.83g diluted in 11ml water

[0659] Table 22. Plasticizer Type and Percentage Weight Gain on Plates

[0660] describe Weight before treatment (g) Weight after treatment (g) Board weight increase (%) Fatliquor 2.5% 2.89 3.18 7.75 5% Erythritol 3.05 3.45 13.01 Fatliquor 15% 3.02 3.41 13.00 Fatliquor 30% 2.75 3.47 26.13

[0661] The 2.5% fatliquor concentration resulted in a 7.75% board weight increase for the mycelial board. The mycelial board treated with 5% erythritol had a 13% weight increase. Regarding the 15% and 30% fatliquor samples applied by massage, the mass of both boards increased by almost equal percentages (13% and 26%, respectively), indicating that most of the fatliquor matrix was absorbed into the mycelial board and only had a small amount of loss during processing.

[0662] The flexural modulus of the mycelium panels was tested and Fig.54 In the embodiment of the present invention, the flexural modulus of the 15% fatliquor sample is shown in Table 22. Increasing the fatliquor content in the mycelium board from 8% to 13% significantly reduces the flexural modulus (see the % increase in board weight of the 2.5% fatliquor sample compared to the 15% fatliquor sample shown in Table 22). The fatliquor 15% sample also feels softer than the fatliquor control sample. The fatliquor 30% sample has a slightly softer feel than the fatliquor 15% sample. However, the flexural modulus of the two samples are almost the same, indicating that the 13% fatliquor absorption in the 15% fatliquor sample is sufficient. Therefore, additional fatliquor loading may not improve material softness.

[0663] Treatment with erythritol solution (5%) also reduced the flexural modulus, but was not as effective as the fatliquor ( Fig.54 ). The soaking and rolling treatment using 5% erythritol showed similar flexural modulus as the 2.5% fatliquor solution treatment.

[0664] Example 11: Acid dyes and reactive dyes

[0665] After being treated for tear resistance and softness, the mycelium boards were dyed to color them. Previous dye development used acid dyes to color mycelium boards. Acid dyes fully penetrate into the mycelium boards. However, the colorfastness of acid dyes in the mycelium boards was unsatisfactory, so alternative dyes were studied. Known reactive dyes react with cellulose and polysaccharides containing hydroxyl groups and form covalent bonds, thereby producing preferred colorfastness. Jacquard Procion MX brown reactive dye was tested for colorfastness in mycelium materials and compared with Acid Brown 14 dye.

[0666] Regarding the acid dyeing process, the preparation of acid dyes (such as Acid Brown M (TCI America TM , CIAcid Brown 14) or Lanasyn Black (CIAcid Black 194), vegetable tannin and fatting agent aqueous solution and as previously described via four rounds of soaking and rolling applied to mycelial material. pH 3.15 solution is prepared with formic acid or acetic acid. The plate is soaked in an acidic solution and rolled 3 times for 1 minute, then soaked in a fresh pH 3.15 fixative solution for 1 hour. The plate is rinsed with water and dried in a fume hood.

[0667] About reactive dye dyeing process, preparation is less than the aqueous solution of 10g / L reactive dye (Jacquard Procion MX) and 5-100g / L NaCl and is applied to mycelium material via four rounds of immersion lasting 30min and rolling as previously mentioned.Sodium carbonate is added in the described solution with the ultimate concentration of 20g / L and the described plate experiences six rounds of 10 minutes of immersion / rolling circulation lasting a total of 1 hour. Rinse the described plate with water and roll four times and dry in a fume hood. Handle two mycelium samples with acid dyes and handle three mycelium samples with reactive dyes. Each of the described five samples is all cut into small slices and is immersed in water and lasts about one hour. Repeat the described soaking step, until visually hardly observing that dyestuff leaches into the soaking solution. About the acid dye sample, this needs five soaking steps and about the reactive dye sample, this needs three soaking steps. Collect water after each soaking step and use the full visible spectrum (360-740nm) to measure the UV / vis absorbance of the described solution to evaluate the color of the sample. Fig.55A -E shows the maximum absorbance of water containing dye leached from dyed mycelium samples after five rounds of progressive immersion in water. The two samples on the left are mycelium dyed with acid dyes, and the three samples on the right are mycelium dyed with reactive dyes. Fig.55A Shown are the absorbances after 1 round of soaking. Fig.55B Shown are the absorbances after 2 rounds of soaking. Fig.55C Shown are the absorbances after 3 rounds of soaking. Fig.55D Shown are the absorbances after 4 rounds of soaking. Fig.55E Shown are the absorbances after 5 rounds of soaking.

[0668] Small sections of washed mycelium were then air dried to see how much color remained. The acid dye samples lost most of their color, while the reactive dye samples retained their color well for multiple rounds of washing (data not shown). The reactive dye treated mycelium plates had much better colorfastness than the acid dyes.

[0669] Example 12: Optimization of crust solution of mycelial material

[0670] The processing solution ("crusting solution") is also optimized to produce material of acceptable quality for downstream finishing procedures.

[0671] Table 23 provides a list of the variables, the ranges tested, and comments about the variables

[0672]

[0673] Mycelial material plates from 5 different batches were used in the test. The length, width, thickness and mass of each original starting sample were collected. Therefore, wet density and starting thickness may be used as additional input variables for the final analysis. Each sample was processed by preparing an aqueous solution of dye, vegetable tannin, fatliquor, pea protein and TG in water. The pH of the solution was adjusted to 3.5, 5.5 or 7 using formic acid or acetic acid. The mycelial material was soaked in the crust solution for 1 minute and then passed through an Italian pasta press 4 or 10 times. The soaked and rolled plates were then incubated at 40°C for 0, 4 or 18 hours and dried.

[0674] Finished panels were evaluated for the following properties: crack tear strength (mechanical test—maximum tear force, normalized tear strength); dye penetration, via visual grading 1-5; color fastness, using an electronic crockmeter (SDL Atlas Company, USA), grading 1-5; and surface color analysis, using a light box and ImageJ grayscale rating.

[0675] result

[0676] The board starting thickness or density has the greatest impact on crack tear strength. Fig.56 The maximum force of the crack tear is shown as compared to the starting thickness (cm) of the panel. Fig.57 The maximum force to tear at the crack is shown as compared to the final thickness (cm) of the board. Thicker starting or final boards have higher maximum force to tear at the crack.

[0677] The soaking solution pH also has one of the most significant effects as it affects the final coloration and dye penetration of the sample. Fig.58 The coloration averages as compared to the soaking pH are shown, with lower coloration values ​​indicating darker colors. Fig.59 Shown is dye penetration as compared to soaking pH. Fig.60 Shown is colorfastness as compared to soaking pH.

[0678] The main trends observed were: higher starting density resulted in higher final density; higher starting thickness resulted in higher final thickness; roll setting affected final thickness as wide roll spacers resulted in thicker final material; higher starting thickness resulted in higher maximum tear force; low pH resulted in poorer dye penetration; more pea protein resulted in lighter coloration.

[0679] Additional slight trends observed were: TG increased final density; roll setting affected maximum tear force and normalized tear force, as material processed with smaller roll spacing resulted in lower tear force; longer incubation resulted in higher normalized tear strength; more pea protein or TG resulted in poorer dye penetration; higher solution temperature resulted in higher dye penetration; and higher pH resulted in higher colorfastness.

[0680] Thus, higher thickness and / or density in the starting material results in higher slit tear strength. For example, to achieve a product specification of 20N, an exemplary starting board should be at least 3.4 cm thick. However, if the product specification force is higher, then simply increasing the board thickness may not result in excellent slit tear properties. Therefore, additional techniques such as composites (e.g., adding scaffolds, scrims, adhesives, etc.) or improved growth structures such as hydroentanglement should be explored.

[0681] Additionally, low pH solutions adversely affect the plates in the presence of pea proteins and TG. Therefore, if a low pH fixation step is used, it should be a separate step.

[0682] Example 13: Treatment of mycelium material with Elite-plus binder and other additives

[0683] Mechanical properties of mycelium material with Elite-plus binder

[0684] Materials and methods

[0685] Mechanically break between 27 and 35 g of dried mycelium into small cubes. Add 1.5-1.75 L of water and use a low shear rotary blender to disrupt the slurry. Then dewater the slurry before redispersing it in another 1.5-1.75 L of water. Dewater it a second time before redispersing it in another 1.5-1.75 L of water. Add the prescribed amount of Dur-o-Set Elite Plus (Celanese Emulsions, USA) to the slurry dispersion and mix until uniform. Then wet-ply the resulting slurry on a sizing cloth and dewater to form a board. Dry this board in a convection oven at 45°C. Place the resulting board in a 7.5-15% fatliquor solution and allow it to soak for 30 minutes. Dry the board again in a convection oven at 45°C and submit for testing.

[0686] result

[0687] A range of different leather products were tested and the average peel force varied from 2.5 N / cm to more than 23 N / cm.

[0688] Table 24 provides a tabulation of the average peel forces for the different leather samples.

[0689] Leather samples Average peel force (N / cm) 1 2.5 2 14 3 >23

[0690] Table 25 provides mechanical data for samples using the Elite-Plus adhesive.

[0691]

[0692] Fig.61 The average peel force (N / cm) is shown as a binary fit to the estimated value (%) of the binder. As the concentration of the binder increases, the average maximum peel force also increases. The panels were also subjected to Taber testing. For example, panel P1015 was subjected to 7000 cycles under Taber abrasion using a 1000g weight and a CS-10 abrasive disc. The samples showed uniform wear with a mass loss of approximately 6%. The binder samples also withstood 30,000 cycles of Bally flex with minimal signs of cracking or damage.

[0693] Table 26 provides dry tensile strength data for samples using the Elite-Plus binder.

[0694]

[0695] Mechanical properties of mycelium material with Elite-22 binder and abaca fiber

[0696] Materials and methods

[0697] The dried and disrupted mycelium was crusted in 10% Elite 22 with 7.5% (1:3LEX:DXV) fatliquor solution by soaking for 20-30 sec and abaca fiber was added before passing through a pasta press. Soaking and pasta press were performed 4 times and the mycelium was dried in an oven at 113°F for 4-5 hours and hot pressed at 80°C for 1 min to smooth the surface. The mycelium was conditioned overnight in a humidity chamber at 50% moisture content before testing.

[0698] result

[0699] The samples were crusted through the above procedure. The tearing of the samples increased with the increase in the percentage of abaca fiber in the samples. Fig.62 The standard force values ​​for samples with various concentrations of abaca fiber are illustrated.

[0700] Table 27 provides thickness and rip tear data for various samples having abaca fibers.

[0701]

[0702] Example 14: Treatment of mycelial material with other additives

[0703] APS, EDC, sodium periodate, ionic liquids and PAE resins

[0704] Ammonium persulfate (APS), ethyl-3(3-dimethylaminopropyl)carbodiimide (EDC), sodium periodate, ionic liquids and PAE resins were studied to improve the mechanical strength of mycelial materials. These crosslinking agents were used to crosslink mycelial hyphae and can be used with additional polymeric binders. Hot pressing was also used in some experiments. Mycelial mats were treated with the APS crosslinking agent according to the method described in Example 6.

[0705] Materials and methods

[0706] APS crosslinker

[0707] For APS samples, the dried mycelium mats were soaked in a 1% (w / w) solution of ammonium persulfate in deionized water or phosphate buffered saline (PBS) and brought to 80°C for 3.5 hours with gentle agitation. In some samples, a 1% (w / w) solution of lignin in deionized water was also added. The resulting mats were rinsed in deionized water and dried at 40°C for 3 hours. ISO 3377-2 double edge tear, ASTM D2209 tensile and ASTM D4704 notched tear tests were then performed on a ZwickiLine material testing machine Z5.0 to determine the relative strength relative to the control sample.

[0708] APS crosslinker + hot pressing

[0709] For APS samples, dried mycelium mats with or without binder were soaked in a 1% (w / w) solution of ammonium persulfate in deionized water and brought to 80°C for 1 hour with gentle agitation. The mats were then transferred to an 80°C hot press where they were pressed to 1.4-2.0 mm for 2.5 hours. The resulting mats were rinsed in deionized water and dried at 40°C for 3 hours. ISO 3377-2 double edge tear, ASTM D2209 tensile and ASTM D4704 notched tear tests were then performed on a ZwickiLine material testing machine Z5.0 to determine the relative strength relative to the control sample.

[0710] EDC crosslinker

[0711] Dried mycelium mats with or without a binder were soaked in a 1% (w / w) solution of 1-ethyl-3 (3-dimethylaminopropyl) carbodiimide in deionized water at buffer pH 5.5, 7.4, 8.3 or 9.5 at room temperature for six hours. In some samples, a 1% (w / w) solution of lignin in deionized water was also added. The resulting mats were rinsed in deionized water and dried at 40°C for 3 hours. ISO 3377-2 double edge tear, ASTM D2209 tensile and ASTM D4704 notched tear tests were then performed on a ZwickiLine material testing machine Z5.0 to determine the relative strength relative to the control sample.

[0712] Sodium periodate

[0713] The dried mycelium mat was soaked in a 1% (w / w) solution of sodium periodate in deionized water. The solution was brought to 80°C for 2 hours, then rinsed and dried at 40°C for 3 hours. ASTM D2209 tensile testing was then performed on a ZwickiLine material testing machine Z5.0 to determine relative strength compared to a control sample.

[0714] Ionic Liquids

[0715] The dried mycelium mat was soaked in the ionic liquid solution at room temperature for 5 min to 16 hours. The sample was then pressed at 80°C for 5 min. Wet tensile testing was then performed via ASTM D2209.

[0716] PAE resin

[0717] The samples were heated at 105° C. for 5 minutes or 10 minutes. The samples were at pH=5 (HM24) or pH=7 (HM25).

[0718] result

[0719] The results of mechanical properties tests are Fig.63 , Fig.64 , Fig.65 , Fig.66 , as shown in Table 28 and Table 29.

[0720] Table 28. Initial modulus, wet tensile strength, and elongation at break.

[0721]

[0722]

[0723] The treated samples had significantly increased wet tensile strength, initial modulus as compared to the control samples.Thus, adding APS to crosslink the mycelium material improves the mechanical quality of the mycelium board.

[0724] Table 29. Wet tensile strength and elongation at break.

[0725]

[0726] As compared to the control sample, the sodium periodate and EDC treated samples had significantly increased wet tensile strength.Thus, adding sodium periodate and / or EDC to crosslink the mycelial material improves at least one mechanical parameter of the mycelial board.

[0727] Fig.65 The tensile strength results for ionic liquid treated samples are shown as compared to control samples. IL-P0.75mm-16h indicates that the sample was soaked in ionic liquid for 16 hours at room temperature and then pressed to 0.75 mm for 5 min at 80°C. For the crosslinked samples, the elongation at break was about 50%.

[0728] Fig.66 The tensile strength results for the PAE resin treated samples are shown as compared to the control samples. HM18_105 does not contain PAE resin, but it does have epoxidized soybean oil and was heated at 105°C for 4 hours, indicating that heat treatment alone does not result in increased wet tensile strength.

[0729] Example 15: Lamination of mycelium material, scrim addition, heat pressing and hydroentanglement

[0730] Materials and methods

[0731] Slurry preparation

[0732] Mycelium slurry was prepared by blending dry mycelium in tap water using a Blendtec Pro 800 blender. The concentration of the slurry ranged from 0.5-2.5 w / v%, depending on the experiment. The mycelium was blended to make a uniform slurry, typically for 90 seconds at setting 5. After blending, the slurry was sieved on a 500 micron sieve to remove fines and soluble components. The sieved mycelium was then resuspended back to 1 w / v%, manually stirred to disperse, and sieved / resuspended two more times. After the final resuspension, a solution of sodium dodecyl sulfate (SDS) in DI water was added and the slurry was blended at setting 5 for 10 seconds to produce foam. The final concentration of sodium dodecyl sulfate in the slurry was typically 0.002 w / v%. The foam remains trapped in the slurry and helps create porosity in the web after wet layup.

[0733] Wet cloth layer

[0734] A laboratory scale wet lamination apparatus was constructed to make webs from slurries. The apparatus consisted of a 150 mm diameter Buchner funnel attached to a 4 L vacuum flask, which was lined with a sizing cloth. The vacuum flask was attached to a vacuum pump (Vacuubrand VARIO PC 3001 Select) which allowed the vacuum set point to be controlled with an accuracy of 1 Torr. A three-way valve between the vacuum pump and the flask controlled whether the flask was under vacuum or vented to atmosphere. Wet lamination was achieved by pouring the slurry into the Buchner funnel and then vacuum filtering at a determined vacuum level (typically 600 Torr) until no more water was observed dripping from the funnel (typically 90 seconds). The resulting web had a moisture content between 75-80 wt%. The web was then manually peeled from the sizing cloth and dried at 45°C under convection until its mass reached a steady state (typically several hours).

[0735] Hard crust

[0736] A planetary centrifugal mixer (THINKY ARE-400 TWIN) was used to prepare a crusting solution with binder emulsion and fatliquor, the typical composition of which is listed in Table 30. 100 ml of solution was sufficient to perform the impregnation of a single web formed on a 150 mm diameter Buchner funnel as described above.

[0737] Table 30. Composition of crusting solution for dipping.

[0738] Components Concentration (wt%) DUR-O-SET Elite 22VAE 10(Based on the solid content of binder emulsion) Trupon DXV fatliquor 5.625 Truposol LEX Fatliquor 1.875 water The rest

[0739] To perform the impregnation, the dry web is immersed in the crust solution for at least 1 minute to wet it, and then passed between two rollers with a spacing of about 2 mm to remove air and force the solution through the material. The immersion and rolling process is repeated until the web is uniformly saturated with the crust solution, typically 4 iterations. Thereafter, the crust is dried at 45°C under convection until its quality reaches a steady state (typically several hours).

[0740] Lamination

[0741] Lamination, scrim incorporation and heat pressing may be performed in parallel to compress the materials and smooth the surface. To laminate the two crusted webs together, first apply a thin layer of DUR-O-SET Elite 22VAE to one side of each web using a brush or roller. A typical coating is approximately 9 mg / cm on a wet basis. 2 After coating, the adhesive is exposed to ambient conditions for about 30 seconds, during which the adhesive becomes tacky. The webs are then pressed together (adhesive side inward) at 80°C using a manual hot press (Carver 4120) for 1 minute to a final thickness in the range of 1-3 mm (controlled by placing a spacer between the press plates). Cheese may be incorporated into the material by placing cheesecloth between the two webs before pressing (typically 95gsm woven cotton). After pressing, the laminate is dried at 45°C under convection until its mass reaches a steady state (typically several hours) to remove residual water from the adhesive before curing.

[0742] Curing

[0743] DUR-O-SET Elite 22VAE contains self-crosslinking N-methylol acrylamide side chains which, once crosslinked, increase the polymer's initial modulus, ultimate tensile strength, and water resistance (especially at high temperatures that may be encountered during aqueous dyeing processes). Before curing the crust or laminate, the sample is dried to remove residual water to prevent expanding vapors from delaminating the material. Curing is achieved by placing the sample in an oven between 90 and 135°C for 0.5 to 1 hour.

[0744] Hydroentanglement

[0745] Hydroentanglement of the mycelial webs was performed according to the method described in Example 4. The samples were hydroentangled with 50 micron water jets at 1000 psi. Each sample was hydroentangled with multiple passes in both vertical and horizontal directions.

[0746] result

[0747] Effect of coarse cloth in mycelium material

[0748] The mechanical properties of typical samples are shown in Table 31. Note that these samples were constructed with scrim. The area density was calculated by dividing the mass of the sample by the sample area. The apparent bulk density was calculated by dividing the mass of the sample by the product of the sample thickness and the sample area as measured according to ASTM D1813.

[0749] Table 31. Mechanical properties and dimensional characteristics of typical samples.

[0750]

[0751]

[0752] Effects of heat pressing on mycelium materials

[0753] Pressurization of the mycelium material is performed at or above ambient temperature to control final thickness and surface roughness, which in turn affects downstream operations such as finishing and embossing, as well as the flexibility and feel of the final product.

[0754] To study the effect of press time, temperature and pressure on the final thickness of the material, samples were prepared according to the above process, but without the curing step, and then pressed at different points in the time-temperature-pressure parameter space as shown in Table 32. The thickness of the material before pressing was 2.2 ± 0.1 mm. The thickness of the material was measured according to ASTM D1813 before and after pressing.

[0755] Table 32. Time-temperature-pressure parameter space studied for hot pressing experiments.

[0756] factor level Pressurization time(s) 5、60 Pressurization temperature (℃) 22、50、80 Pressurization pressure (MPa) 0.34、1.7

[0757] Fig.67 It is shown that the pressing temperature has the greatest impact on the final thickness, and the pressing time and pressure have smaller effect sizes of comparable magnitude. The surface roughness is qualitatively proportional to the final thickness. The results of this model may be used to adjust the final thickness of the material as required for downstream operations.

[0758] Effect of wet layering process parameters on mesh porosity

[0759] The porosity of the web affects several downstream properties, such as areal and bulk density, compressibility, flexibility, through-thickness strength, and ability to be impregnated with solutions. Therefore, understanding which factors affect web porosity and controlling them helps to tailor the behavior of the intermediate material and the properties of the final product.

[0760] To study the effect of slurry volume, concentration, and vacuum treatment on web porosity, a slurry was prepared as described in this example, but the sieving step, resuspension step, and addition of sodium lauryl sulfate were omitted. The slurry was then wet-layered using a laboratory-scale web-forming apparatus at various points in the parameter space described in Table 33. After drying as previously described, a sample of the web was cut using a scalpel to allow observation of a web cross-section by scanning electron microscopy. Scanning electron microscopy was performed using a Hitachi TM The 3030Plus scanning electron microscope was operated at an accelerating voltage of 15 kV.

[0761] Table 33. Parameter space investigated to determine the effect of wet layup parameters on web porosity.

[0762] factor level Slurry volume (L) 0.5、1 Slurry concentration (w / v%) 1、2.5 Vacuum Set Point (Torr) 35、600

[0763] Fig.68 Representative scanning electron micrographs of meshes produced at various points in the parameter space described in Table 33 are shown, along with their corresponding area densities. The relationships between the factors studied and the thickness, qualitative porosity, and qualitative compressibility of the meshes are summarized in Table 34.

[0764] Table 34. Relationship between the studied wet layup parameters and the thickness, porosity and compressibility of the resulting webs.

[0765]

[0766] The results of this experimentation can generally be used to adjust the thickness, porosity, and compressibility of the material, which in turn affects downstream processing and final properties.

[0767] Adding a foaming agent to the slurry can also be used to adjust the porosity of the web. To study the effect of a foaming agent, slurries were prepared as described in this example at the same concentration (0.75 w / v%) with or without 0.05 wt% sodium dodecyl sulfate (SDS) incorporated into the slurry. The slurries were then wet-layered using a lab-scale wet-layering apparatus as described in this example, but at a vacuum set point of 450 Torr. Fig.69 Scanning electron micrographs of two webs and their corresponding dimensional data are shown.

[0768] Adding a foaming agent such as SDS to the slurry significantly increases porosity and reduces the apparent bulk density of the web. By adding SDS to the slurry and then blending, the resulting foam is trapped in the slurry and creates additional porosity after dehydration and drying.

[0769] In-situ binder addition

[0770] Table 35. This table shows wet end Elite-Plus hardskin data. This table demonstrates how web bulk density changes based on the presence or absence of vacuum during wet plying and the presence or absence of chemical defoamer. This change in web bulk density affects T-peel strength.

[0771] Table 35. In situ binder addition during web formation.

[0772]

[0773]

[0774] In addition to adding a binder solution in the slurry or impregnating the dry web with a binder solution, in-situ binder addition during web formation is another method for incorporating binders. A possible advantage of this method compared to other methods is that the amount of required binder can be less. Possible advantages relative to dry impregnation include not having to first dry the web before impregnation, and the binder load in the material can be better controlled and is not limited by the ability of the binder to penetrate the dry web. Adjusting the binder content in the final material can affect the mechanical performance of the material as well as its feel and other aesthetic properties.

[0775] In order to study the effect of the process parameters of the in-situ binder addition, the slurry was first prepared as described in this embodiment. Then the laboratory scale wet layering device was modified by adding a valve between the Buchner funnel rod and the vacuum flask, so that it was possible to control the volume of liquid discharged from the slurry. The binder solution was prepared by mixing DUR-O-SET Elite 22VAE with water to obtain a final solid content of 2.5 or 5wt%. Then the slurry was wet layered in the modified device, while the amount of water removed from the slurry before the binder solution was added, the amount of the binder solution added and the concentration of the binder solution added changed. After the binder addition and vacuum filtration were completed, the net was dried and the mass absorption relative to the input mycelium mass was recorded. In addition, Fourier transform infrared spectroscopy (FTIR) was used to semi-quantitatively evaluate the binder: mycelium ratio, which varied with the material for the Z position (through-thickness direction) of each point in the volume-concentration parameter space. FTIR was performed using a Bruker Alpha II attenuated total reflection (ATR) FTIR equipped with a diamond ATR assembly. The mesh sample was cut with a scalpel and its top, bottom and core surfaces were placed on the internal reflection element (IRE) of the instrument. -1 Resolution from 4000–600 cm -1 The spectrum was recorded by averaging 32 scans. The spectrum was then baseline corrected over the entire wavenumber region and the wavelength range was 1760-1700 cm -1 (corresponding to the C=O bond of the acetate group in DUR-O-SET Elite 22VAE) and 1180-880 cm -1 A straight baseline was drawn (corresponding to CO bonds in sugars, considered representative of mycelial content). The area was then integrated and the ratio of the peak areas reported.

[0776] Table 36 summarizes the parameter space studied in this experiment.

[0777] factor level Amount of water removed from the slurry before applying the binder solution (ml) 300、600 Volume of binder solution added (ml) 100、200、400 Concentration of binder solution (wt%) 2.5、5

[0778] Fig.70 A model is shown that relates the mass % of mycelium in a dry-bonded web to the volume of binder solution added to the web and the concentration of the binder solution. The points represent individual measurements. The curve represents the mean and the shaded area represents the 95% confidence interval of the mean. 2 = 0.97. The mass % calculations assumed a 25 wt% mass loss relative to the input mycelium mass due to fines and soluble losses during the sieving operation.

[0779] Fig.71 The model shown in describes how the binder content of a bonded web can be adjusted to a desired value by controlling the amount of binder solution added to the web and the concentration of that solution. In this experiment, the amount of water removed from the slurry before the binder solution was added did not significantly affect the final binder content.

[0780] Fig.71 A model is shown that describes the average binder:mycelium peak area ratio over the concentration-volume-Z position parameter space. Lines and curves represent the average for each reaction. Shaded areas and error bars represent the 95% confidence interval for the mean. R 2 =0.77.

[0781] Fig.72 It is shown that for low and high binder addition volumes, the binder: mycelium peak ratio varies with the Z position in the net. The error bars represent the 95% confidence interval of the mean value. Binder: mycelium ratio increases with both the volume and concentration of the added binder solution. The model also predicts that as the added volume increases, the benefits will decrease. In addition, the model shows that on average, the top of the bonded net (wherein the binder is added) has a higher binder content than the core or bottom. Finally, the model shows that in the case of low binder addition volumes, the difference in binder content between the top of the net and the core / bottom will intensify. Ensuring that the binder is evenly distributed over the entire thickness of the material helps to alleviate internal stratification.

[0782] Effects of hydroentanglement on mycelial materials

[0783] Fig.73 The effect of hydroentanglement on the wet tensile curve of mycelial webs is shown. The wet tensile results were determined from mycelial webs without binder fatliquors or heat pressing. Hydroentanglement reduces the initial modulus of the mycelial web and increases the elongation at break.

[0784] Example 16: Process Chemicals and Their Effects on Mechanical Properties of Mycelium Materials

[0785] Adhesive Dur-O-set Elite 22 Curing Conditions

[0786] Materials and methods

[0787] Experiments were conducted on mycelium webs that had been hydroentangled or as "spunlace". The spunlace was treated with a binder baseline solution, 10% Elite 22 solution, soaked for 15 minutes, rolled twice with a hand roller to remove air bubbles inside the spunlace, and then soaked for another 15 minutes. After the binder application, the spunlace was dried overnight at 45°C. After drying the spunlace, a fatliquor was applied using a 7.5wt% DXV / LEX solution (3:1 ratio DXV:LEX) and subjected to soaking for 15 minutes, one hand roller, and another soaking for 15 minutes. After the fatliquor application, the spunlace was dried again at 45°C overnight. The spunlace was then post-cured under various temperature and time conditions.

[0788] result

[0789] The results of the curing test are Fig.74 , Fig.75 and as shown in Table 37.

[0790] Table 37. Sample descriptions for curing experiments.

[0791]

[0792] like Fig.74 As shown, the flexural modulus increases with increasing curing temperature, indicating that the spunlace becomes stiffer during curing. When the temperature reaches 110°C for 30 minutes or 135°C for 15 minutes, the average flexural modulus becomes stable.

[0793] However, if Fig.75 As shown, the tear strength increases with increasing temperature and reaches a maximum when curing is continued at 135°C for 30 minutes. After that, additional curing temperatures result in a decrease in tear strength. The Elite 22 binder cures (crosslinks) above 130°C, so the mechanical strength increases as a result. The mechanical results show that crosslinking begins around 110°C. When curing more than 30 mixtures at 135°C, the tear strength decreases, which can be attributed to some damage to the mycelium material itself.

[0794] Mechanical strength of mycelium materials cured at different temperatures

[0795] Materials and methods

[0796] Samples C116, C133, and C134 are spunlace fabrics produced using the methods described herein. Each spunlace fabric contained 30% Tencel fiber (8 mm length) biomass, was wet-laid and hydroentangled. A baseline (soak-roll-soak) method was used to use a 10% Elite 22 binder solution, followed by drying each spunlace fabric at 45°C overnight. Sample C116 was then cured at 135°C for 2 hours. After curing, sample C116 was dyed, a fatliquor was added, and the spunlace fabric was dried. Samples C133 and C134 were dyed first, and then dried. Sample C133 was then cured at 135°C for 2 hours, followed by the addition of a fatliquor and the spunlace fabric was dried. A fatliquor was added to sample C134 without curing at 135°C, and the spunlace fabric was dried after the fatliquor was added.

[0797] result

[0798] Table 38. Crack tear strength of mycelium materials cured at different temperatures

[0799] Sample ID Thickness(mm) Crack Tear (N) 95% CI Number of tests C116(135C) 2.6 21.3 ±2.02 6 C133(135C) 2.14 26.19 ±2.20 8 C134(70C) 2.06 15.52 ±1.95 8

[0800] Sample C116 ( Fig.76 ) and C133( Fig.77 ), which shows a similar trend to the uncured sample C134 ( Fig.78 ). However, sample C116 encountered problems during dyeing, and the binder that had been cured at 135°C appeared to be stained by the reactive dye, leaving an uneven dyeing pattern on the crust surface. Samples C133 and C134 did not have the uneven dyeing problem. Sample C133 showed slightly more color variation as compared to sample C134, but dyed evenly. It is suggested that the mycelium material with the binder needs to be dyed first, then fully cured at 135°C.

[0801] Adhesive application using sonication and warm solution soaking

[0802] Different binder application methods were tested on wet-laid mycelium webs to identify steady state binder loadings.

[0803] Materials and methods

[0804] For the warm solution soaking method, the webs were subjected to a soak-roll-soak method using a 60°C water bath, and then the webs were sonicated at 60°C. Specifically, each web was sealed in a Ziploc bag with a 10% Elite 22 binder solution, which was then immersed in a 60°C water bath heated by a sous vide cooker. The webs, binder solution, and Ziploc bag were heated for 1 hour, and during that time, the wet weight of the web was measured every 15 minutes to monitor the binder solution soaking. After the warm soaking for one hour, the web was pressed or rolled using a hand roller, and then the bubbles were pushed out of the web. The web was then soaked in the warm binder solution in the Ziploc bag in the water bath for several more minutes to reach a stable state.

[0805] For the sonication method, the binder solution was directly placed in an ultrasonic cleaner and heated to 60° C. The web was then immersed in the 60° C. binder solution under ultrasonic treatment. The wet weight of the web was measured every 15 minutes, and the web was rolled after immersion for 1 hour.

[0806] result

[0807] Fig.79 Shown is the leaching of binder solution onto the web over time using the sonication and warm soak methods.

[0808] After 1.5 hours of processing, the sonication process reached a steady state, and rolling did not change the wet weight of the web, indicating that there were almost no bubbles in the web. The web that underwent warm soaking increased in wet weight after rolling, indicating that more bubbles were squeezed out. However, after rolling and soaking for an additional 30 minutes, the two methods reached similar binder loadings.

[0809] Table 39. Binder leaching of mycelial webs using sonication and warm soaking methods.

[0810]

[0811] Example 17: Salt Concentration and Dye Penetration

[0812] Reactive dye protocols provided by manufacturers typically call for the addition of high concentrations of sodium sulfate (30-100 g / L: higher concentrations for darker shades) during the exhaustion phase of dyeing. This salt addition greatly hinders the penetration of dyes into the mycelial wet-laid fabric or spunlace. In addition, sodium carbonate is added to the dye bath in order to increase the pH and promote fixation. Manufacturers generally recommend concentrations in the range of 8-20 g / L. This sodium carbonate addition can also affect dye penetration. A concentration of about 1 g / L raises the pH to about 10, thereby allowing proper fixation without preventing dye penetration.

[0813] Sodium sulfate concentration

[0814] Three spunlace samples were dyed with 2 g / L blue dye in Procion MX and varying amounts of sodium sulfate (0, 25, 50 g / L) at 60°C for 2 h. These samples were not subjected to fixing or rinsing in order to focus on the amount of dye that penetrated into the samples. Fig.80 As shown, increasing salt concentrations result in darker surface colors, but almost completely prevent any dye penetration. The spunlace sample with 0 g / L sodium sulfate produces good penetration, but the sample with 50 g / L sodium sulfate produces poor penetration. Higher concentrations of sodium sulfate result in poorly dyed spunlace.

[0815] Sodium carbonate concentration

[0816] Since the concentration of sodium sulfate reduces dye penetration, it was investigated whether reducing the sodium carbonate concentration during fixation would affect fixation and penetration. It was found that a 1 g / L sodium carbonate solution had a pH of 10, which should be within the appropriate range for dye fixation to fibers. Two spunlace samples were dyed with Navy HF dye for 90 min and then fixed with 0.5 and 1 g / L sodium carbonate at 60°C for 60 min. Fig.81 (0.5g / L sodium carbonate) and Fig.82 (1 g / L sodium carbonate), these samples had good penetration and the dye remained fixed to the matrix.

[0817] Effect of dyeing temperature and time

[0818] Use 2g / L at 22, 40 or 60℃ Six spunlace samples were dyed with Navy HF dye for 1 hour or 3 hours and then fixed by adding 1 g / L sodium carbonate. Fixation was performed at the same temperature as the dye exhaustion. Fig.83 As shown, it can be qualitatively seen that longer times and higher temperatures result in darker colors on the surface and longer times result in preferred dye penetration.

[0819] In addition, quantitative measurements were performed using a Konica Minolta CM-5. Fig.84 As shown, the spectrophotometer shows the same trend in reflectance mode. In these measurements, lower L* values ​​correspond to darker colors.

[0820] Effect of adhesive curing temperature and time

[0821] 10 wt% Elite 22 binder solution was applied to spunlace samples and then cured at different temperatures (70, 90, 110, 130°C) and times (15-45 min) before dyeing. Navy HF dye stains the sample for 2 hours. Fig.85 As shown, curing at 90°C and above results in poor dyeing of the spunlace. This indicates that any dyeing procedure should be performed before the final curing operation.

[0822] Additional aspects of the present disclosure include:

[0823] According to a first aspect of the present disclosure, a composite mycelium material includes: a cultivated mycelium material including one or more masses of branched hyphae, wherein the one or more masses of branched hyphae are disrupted; and a binder.

[0824] According to a second aspect of the present disclosure, the composite mycelium material of aspect 1, wherein the cultivated mycelium material has been produced on a solid substrate.

[0825] According to a third aspect of the present disclosure, the composite mycelium material of aspect 1 or 2, wherein the cultivated mycelium material comprises one or more masses of disrupted branched hyphae.

[0826] According to a fourth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 3, wherein the one or more masses of disrupted branched hyphae have a length of 0.1 mm to 5 mm.

[0827] According to a fifth aspect of the present disclosure, the composite mycelium material of aspect 4, wherein the one or more masses of disrupted branched hyphae have a length of 2 mm.

[0828] According to a sixth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 5, wherein the composite mycelium material further comprises a supporting material.

[0829] According to a seventh aspect of the present disclosure, the composite mycelium material of aspect 6, wherein the support material has a pore size of 1 / 16 inch.

[0830] According to an eighth aspect of the present disclosure, the composite mycelium material of aspect 6, wherein the supporting material comprises a reinforcing material.

[0831] According to a ninth aspect of the present disclosure, the composite mycelium material of aspect 8, wherein the reinforcement material is entangled within the composite mycelium material.

[0832] According to a tenth aspect of the present disclosure, the composite mycelium material of aspect 6, wherein the support material comprises a base material.

[0833] According to an eleventh aspect of the present disclosure, the composite mycelium material of aspect 10, wherein the base material is located on one or more surfaces of the composite mycelium material.

[0834] According to a twelfth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 11, wherein the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted fiber, woven fiber, non-woven fiber, knitted textile, woven textile and non-woven textile.

[0835] According to a thirteenth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 12, wherein the one or more masses of branched hyphae are disrupted by mechanical action.

[0836] According to a fourteenth aspect of the present disclosure, the composite mycelium material of aspect 13, wherein the mechanical action comprises blending the one or more masses of branched hyphae.

[0837] According to a fifteenth aspect of the present disclosure, the composite mycelium material of aspect 13, wherein the mechanical action comprises applying a physical force to the one or more masses of branched hyphae so that at least some of the branched hyphae masses are aligned in a parallel form.

[0838] According to a sixteenth aspect of the present disclosure, the composite mycelium material of aspect 15, wherein the physical force is a tensile force.

[0839] According to a seventeenth aspect of the present disclosure, the composite mycelium material of aspect 15, wherein the mechanical action comprises applying the physical force along one or more directions so that at least some of the branched hyphae masses are aligned in parallel along one or more directions, wherein the physical force is applied repeatedly.

[0840] According to an eighteenth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 17, wherein the one or more masses of branched hyphae are disrupted by chemical treatment.

[0841] According to a nineteenth aspect of the present disclosure, the composite mycelium material of aspect 18, wherein the chemical treatment comprises contacting the one or more masses of branched hyphae with an amount of alkali or other chemical agent sufficient to cause disruption.

[0842] According to the twentieth aspect of the present disclosure, the composite mycelium material of aspect 19, wherein the base comprises an alkaline peroxide.

[0843] According to a twenty-first aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 20, wherein the binder comprises one or more reactive groups.

[0844] According to a twenty-second aspect of the present disclosure, the composite mycelium material of aspect 21, wherein the one or more reactive groups react with active hydrogen-containing groups.

[0845] According to a twenty-third aspect of the present disclosure, the composite mycelium material of aspect 22, wherein the active hydrogen-containing groups include amine, hydroxyl and carboxyl groups.

[0846] According to a twenty-fourth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 23, wherein the binder comprises an adhesive, a resin, a cross-linking agent and / or a matrix.

[0847] According to a twenty-fifth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 23, wherein the binder is selected from the group consisting of transglutaminase, polyamide-epichlorohydrin resin (PAE), citric acid, genipin, alginate, natural adhesives and synthetic adhesives.

[0848] According to a twenty-sixth aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 23, wherein the binder is PAE.

[0849] According to the twenty-seventh aspect of the present disclosure, the composite mycelium material of aspect 26, wherein the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups including amine, hydroxyl and carboxyl groups in one or more branches of the hyphae.

[0850] According to a twenty-eighth aspect of the present disclosure, the composite mycelium material of aspect 25, wherein the natural adhesive comprises a natural latex-based adhesive.

[0851] According to a twenty-ninth aspect of the present disclosure, the composite mycelium material of aspect 28, wherein the natural latex-based adhesive is leather glue or weld.

[0852] According to a thirtieth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 29, wherein the composite mycelium material comprises one or more proteins from species other than the fungal species that produced the cultivated mycelium material.

[0853] According to the thirty-first aspect of the present disclosure, the composite mycelium material of aspect 30, wherein the one or more proteins are from a plant source.

[0854] According to the thirty-second aspect of the present disclosure, the composite mycelium material of aspect 31, wherein the plant source is a pea plant.

[0855] According to a thirty-third aspect of the present disclosure, the composite mycelium material of aspect 31, wherein the plant source is a soybean plant.

[0856] According to a thirty-fourth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 33, wherein the composite mycelium material further comprises a dye.

[0857] According to a thirty-fifth aspect of the present disclosure, the composite mycelium material of aspect 34, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes and reactive dyes.

[0858] According to a thirty-sixth aspect of the present disclosure, the composite mycelium material of aspect 34, wherein the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform on one or more surfaces of the composite mycelium material.

[0859] According to a thirty-seventh aspect of the present disclosure, the composite mycelium material of aspect 34, wherein the dye is present throughout the interior of the composite mycelium material.

[0860] According to a thirty-eighth aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 37, wherein the composite mycelium material further comprises a plasticizer.

[0861] According to the thirty-ninth aspect of the present disclosure, the composite mycelium material of aspect 38, wherein the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, water, ethylene glycol, triethyl citrate, water, acetylated monoglyceride and epoxidized soybean oil.

[0862] According to the fortieth aspect of the present disclosure, the composite mycelium material of aspect 38, wherein the composite mycelium material is flexible.

[0863] According to a 41st aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 40, wherein an external element is applied to the cultivated mycelium material.

[0864] According to a forty-second aspect of the present disclosure, the composite mycelium material of aspect 41, wherein the external element is applied via heating and / or pressure.

[0865] According to a 43rd aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 42, wherein the composite mycelium material further comprises tannin.

[0866] According to a 44th aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 43, wherein the composite mycelium material further comprises a finishing agent.

[0867] According to a forty-fifth aspect of the present disclosure, the composite mycelium material of aspect 44, wherein the finishing agent is selected from the group consisting of urethane, wax, nitrocellulose and a plasticizer.

[0868] According to a forty-sixth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 45, wherein the composite mycelium material comprises mechanical properties.

[0869] According to a forty-seventh aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the mechanical properties include wet tensile strength, initial modulus, elongation at break, thickness and / or slit tear strength.

[0870] According to a 48th aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 46, wherein the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

[0871] According to a forty-ninth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

[0872] According to a fiftieth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the composite mycelium material has a wet tensile strength of 7 MPa.

[0873] According to the fifty-first aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 46, wherein the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

[0874] According to a fifty-second aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the composite mycelium material has an elongation at break of 1% to 25%.

[0875] According to a fifty-third aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

[0876] According to a fifty-fourth aspect of the present disclosure, the composite mycelium material of any one of Aspects 1 to 46, wherein the composite mycelium material has a thickness of 2 mm.

[0877] According to a fifty-fifth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the composite mycelium material has a crack tear strength of 5N to 100N.

[0878] According to a fifty-sixth aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 46, wherein the composite mycelium material has a crack tear strength of 50N.

[0879] According to a fifty-seventh aspect of the present disclosure, the composite mycelium material of any one of aspects 1 to 56, wherein the composite mycelium material is manufactured using conventional papermaking equipment.

[0880] According to the fifty-eighth aspect of the present disclosure, a method for manufacturing a composite mycelium material comprises: producing a cultivated mycelium material comprising one or more clumps of branched hyphae; disrupting the cultivated mycelium material comprising the one or more clumps of branched hyphae; and adding an adhesive to the cultivated mycelium material; thereby manufacturing the composite mycelium material.

[0881] According to a fifty-ninth aspect of the present disclosure, the method of aspect 58, wherein the producing comprises producing the cultivated mycelial material on a solid substrate.

[0882] According to the sixtieth aspect of the present disclosure, the method of aspect 58 or 59, wherein the cultivated mycelium material comprises one or more masses of disrupted branched hyphae.

[0883] According to the sixty-first aspect of the present disclosure, the method of any one of aspects 58 to 60, wherein the one or more clumps of disrupted branched hyphae have a length of 0.1 mm to 5 mm.

[0884] According to a sixty-second aspect of the present disclosure, the method of aspect 61, wherein the one or more clumps of disrupted branched hyphae have a length of 2 mm.

[0885] According to a sixty-third aspect of the present disclosure, the method of any one of aspects 58 to 62, further comprising incorporating a support material into the composite mycelium material.

[0886] According to a sixty-fourth aspect of the present disclosure, the method of aspect 63, wherein the support material has a pore size of 1 / 16 inch.

[0887] According to a sixty-fifth aspect of the present disclosure, the method of aspect 63, wherein the supporting material comprises a reinforcing material.

[0888] According to a sixty-sixth aspect of the present disclosure, the method of aspect 65, wherein the reinforcement material is entangled within the composite mycelium material.

[0889] According to the sixty-seventh aspect of the present disclosure, the method of aspect 63, wherein the supporting material comprises a base material.

[0890] According to a sixty-eighth aspect of the present disclosure, the method of aspect 67, wherein the base material is located on one or more surfaces of the composite mycelium material.

[0891] According to the sixty-ninth aspect of the present disclosure, the method of any one of aspects 58 to 68, wherein the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted fiber, woven fiber, non-woven fiber, knitted textile, woven textile and non-woven textile.

[0892] According to the seventieth aspect of the present disclosure, the method of any one of aspects 58 to 69, wherein the disrupting comprises disrupting the one or more masses of branched hyphae by mechanical action.

[0893] According to the seventy-first aspect of the present disclosure, the method of aspect 70, wherein the mechanical action comprises blending the one or more masses of branched hyphae.

[0894] According to the seventy-second aspect of the present disclosure, the method of aspect 70, wherein the mechanical action comprises applying a physical force to the one or more masses of branched hyphae so that at least some of the mass of branched hyphae are aligned in a parallel form.

[0895] According to the seventy-third aspect of the present disclosure, the method of aspect 72, wherein the physical force is a tensile force.

[0896] According to the seventy-fourth aspect of the present disclosure, the method of aspect 72, wherein the mechanical action comprises applying the physical force along one or more directions so that at least some of the branched hyphae masses are aligned in parallel along one or more directions, wherein the physical force is applied repeatedly.

[0897] According to the seventy-fifth aspect of the present disclosure, the method of any one of aspects 58 to 74, wherein the one or more masses of branched hyphae are disrupted by chemical treatment.

[0898] According to a seventy-sixth aspect of the present disclosure, the method of aspect 75, wherein the chemical treatment comprises contacting the one or more masses of branched hyphae with an amount of alkali or other chemical agent sufficient to cause disruption.

[0899] According to the seventy-seventh aspect of the present disclosure, the method of aspect 76, wherein the base comprises an alkaline peroxide.

[0900] According to the seventy-eighth aspect of the present disclosure, the method of any one of aspects 58 to 77, wherein the binder comprises one or more reactive groups.

[0901] According to the seventy-ninth aspect of the present disclosure, the method of aspect 78, wherein the one or more reactive groups react with an active hydrogen-containing group.

[0902] According to the eightieth aspect of the present disclosure, the method of aspect 79, wherein the active hydrogen-containing group comprises an amine, a hydroxyl group, and a carboxyl group.

[0903] According to the eighty-first aspect of the present disclosure, the method of any one of aspects 58 to 80, wherein the adhesive comprises a sticker, a resin, a cross-linking agent and / or a matrix.

[0904] According to the eighty-second aspect of the present disclosure, the method of any one of aspects 58 to 80, wherein the adhesive is selected from the group consisting of transglutaminase, polyamide-epichlorohydrin resin (PAE), citric acid, genipin, alginate, natural adhesives and synthetic adhesives.

[0905] According to an eighty-third aspect of the present disclosure, the method of any one of aspects 58 to 80, wherein the binder is PAE.

[0906] According to the eighty-fourth aspect of the present disclosure, the method of aspect 83, wherein the PAE comprises a cationic azetidinium group, and the cationic azetidinium group reacts with active hydrogen-containing groups including amine, hydroxyl and carboxyl groups in one or more branches of hyphae.

[0907] According to the eighty-fifth aspect of the present disclosure, the method of aspect 82, wherein the natural adhesive comprises a natural latex-based adhesive.

[0908] According to the eighty-sixth aspect of the present disclosure, the method of aspect 85, wherein the natural latex-based adhesive is leather glue or weld.

[0909] According to an eighty-seventh aspect of the present disclosure, the method of any one of aspects 58 to 86, further comprising adding one or more proteins from a species other than the fungal species that produced the cultivated mycelial material.

[0910] According to an eighty-eighth aspect of the present disclosure, the method of aspect 87, wherein the one or more proteins are from a plant source.

[0911] According to the eighty-ninth aspect of the present disclosure, the method of aspect 88, wherein the plant source is a pea plant.

[0912] According to the ninetieth aspect of the present disclosure, the method of aspect 88, wherein the plant source is a soybean plant.

[0913] According to a ninety-first aspect of the present disclosure, the method of any one of aspects 58 to 90, further comprising adding a dye to the cultivated mycelium material or the composite mycelium material.

[0914] According to the ninety-second aspect of the present disclosure, the method of aspect 91, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes and reactive dyes.

[0915] According to a ninety-third aspect of the present disclosure, the method of aspect 91, wherein the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

[0916] According to a ninety-fourth aspect of the present disclosure, the method of aspect 91, wherein the dye is present throughout the interior of the composite mycelium material.

[0917] According to a ninety-fifth aspect of the present disclosure, the method of any one of aspects 58 to 94, further comprising adding a plasticizer to the cultivated mycelium material or the composite mycelium material.

[0918] According to the ninety-sixth aspect of the present disclosure, the method of aspect 95, wherein the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, water, ethylene glycol, triethyl citrate, water, acetylated monoglyceride and epoxidized soybean oil.

[0919] According to a ninety-seventh aspect of the present disclosure, the method of aspect 95, wherein the composite mycelium material is flexible.

[0920] According to a ninety-eighth aspect of the present disclosure, the method of any one of aspects 58 to 97, further comprising applying an external element to the cultivated mycelium material.

[0921] According to a ninety-ninth aspect of the present disclosure, the method of aspect 98, wherein the external element is applied via heating and / or pressure.

[0922] According to the hundredth aspect of the present disclosure, the method of any one of aspects 58 to 99, further comprising adding tannin to the cultivated mycelium material or the composite mycelium material.

[0923] According to the one hundred and first aspect of the present disclosure, the method of any one of aspects 58 to 100 further includes adding a finishing agent to the composite mycelium material.

[0924] According to the one-hundred and second aspect of the present disclosure, the method of aspect 101, wherein the finishing agent is selected from the group consisting of urethane, wax, nitrocellulose and plasticizer.

[0925] According to the one-hundred and third aspect of the present disclosure, the method of any one of aspects 58 to 102 further comprises determining the mechanical properties of the composite mycelium material.

[0926] According to the one-hundred and fourth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the mechanical properties include wet tensile strength, initial modulus, elongation at break, thickness and / or slit tear strength.

[0927] According to the one-hundred and fifth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

[0928] According to the one-hundred and sixth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

[0929] According to the one-hundred and seventh aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a wet tensile strength of 7 MPa.

[0930] According to the one-hundred and eighth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

[0931] According to the one-hundred and ninth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has an elongation at break of 1% to 25%.

[0932] According to the one-hundred-tenth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

[0933] According to a one hundred eleventh aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a thickness of 2 mm.

[0934] According to the one hundred and twelfth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a split tear strength of 5N to 100N.

[0935] According to a thirteenth aspect of the present disclosure, the method of any one of aspects 58 to 103, wherein the composite mycelium material has a split tear strength of 50N.

[0936] According to a fourteenth aspect of the present disclosure, the method of any one of aspects 58 to 113, wherein conventional papermaking equipment is used to make the composite mycelium material.

[0937] Those skilled in the art will appreciate that the construction of the disclosed and other components described is not limited to any particular material. Unless otherwise described herein, other exemplary embodiments of the disclosed disclosure disclosed herein may be formed from a variety of materials.

[0938] It is also important to note that the construction and arrangement of the components of the present disclosure as shown in the exemplary embodiments are only illustrative. Although only some embodiments have been described in detail in the present disclosure, it will be readily understood by those skilled in the art who review the present disclosure that multiple modifications are possible (e.g., the size, dimensions, structure, shape and ratio, parameter values, installation arrangements, use of materials, colors, orientations, etc. of the various components), without essentially departing from the novel teachings and advantages of the stated subject matter. Therefore, all of the modifications are intended to be included within the scope of the present disclosure. Without departing from the spirit of the present disclosure, other substitutions, modifications, changes, and omissions may be made in the design, operating conditions, and arrangements of desired and other exemplary embodiments.

[0939] It should be understood that any of the processes or steps within the processes described herein can be combined with other disclosed processes or steps to form structures within the scope of the present disclosure. The exemplary structures and processes disclosed herein are for illustrative purposes and should not be interpreted as limiting.

Claims

1. A composite mycelium material, comprising: a. a cultivated mycelial material comprising one or more masses of branched hyphae, wherein said one or more masses of branched hyphae are disrupted; and b. an adhesive selected from the group consisting of vinyl acetate-ethylene (VAE) copolymers, vinyl acetate-acrylic copolymers, polyamide-epichlorohydrin resins (PAE), copolymers, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives, and synthetic adhesives.

2. The composite mycelial material of claim 1, wherein the cultivated mycelial material has been produced on a solid substrate.

3. The composite mycelium material of claim 1, wherein the cultivated mycelium material comprises one or more masses of disrupted branched hyphae.

4. The composite mycelium material of claim 3, wherein the one or more masses of disrupted branched hyphae have a length of 0.1 mm to 5 mm.

5. The composite mycelium material of claim 4, wherein the one or more masses of disrupted branched hyphae have a length of 2 mm.

6. The composite mycelium material of claim 1, wherein the one or more masses of branched hyphae are entangled, wherein entangling the hyphae comprises needle punching, felting, or hydroentangling.

7. The composite mycelium material of claim 6, wherein the entangled mycelium is hydroentangled.

8. The composite mycelium material of claim 1, wherein the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

9. The composite mycelium material of claim 1, wherein the binder is vinyl acetate-ethylene (VAE) copolymer.

10. The composite mycelium material of claim 1, wherein the binder is a vinyl acetate-acrylic acid copolymer.

11. The composite mycelium material of claim 1, wherein the binder is PAE.

12. The composite mycelium material of claim 11, wherein the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of the hyphae.

13. The composite mycelium material of claim 1, wherein the natural adhesive comprises a natural latex based adhesive.

14. The composite mycelium material according to claim 13, wherein the natural latex-based adhesive is leather glue or weld.

15. The composite mycelium material of claim 1, wherein the composite mycelium material further comprises a support material.

16. The composite mycelium material of claim 15, wherein the support material has a pore size of 1 / 16 inch.

17. The composite mycelium material of claim 15, wherein the support material comprises a reinforcement material.

18. The composite mycelium material of claim 17, wherein the reinforcement material is entangled within the composite mycelium material.

19. The composite mycelium material of claim 15, wherein the support material comprises a base material.

20. The composite mycelium material of claim 19, wherein the base material is located on one or more surfaces of the composite mycelium material.

21. The composite mycelium material of claim 15, wherein the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted textile, woven textile, and non-woven textile.

22. The composite mycelium material of claim 1, wherein the one or more masses of branched hyphae are disrupted by mechanical action.

23. The composite mycelium material of claim 22, wherein the mechanical action comprises intermixing the one or more masses of branched hyphae.

24. The composite mycelium material of claim 22, wherein the mechanical action comprises breaking the one or more clumps of branched hyphae.

25. The composite mycelium material of claim 22, wherein the mechanical action comprises applying a physical force to the one or more masses of branched hyphae such that at least some of the mass of branched hyphae are aligned in a parallel fashion.

26. The composite mycelium material of claim 25, wherein the physical force is a tensile force.

27. The composite mycelium material of claim 25, wherein the mechanical action comprises applying the physical force in one or more directions such that at least some of the branched hyphae masses are aligned parallel in one or more directions, wherein the physical force is applied repeatedly.

28. The composite mycelium material of claim 1, wherein the one or more masses of branched hyphae are disrupted by chemical treatment.

29. The composite mycelium material of claim 28, wherein the chemical treatment comprises contacting the one or more masses of branching hyphae with an amount of alkali or other chemical agent sufficient to cause disruption.

30. The composite mycelium material of claim 29, wherein the base comprises an alkaline peroxide.

31. The composite mycelium material of claim 1, wherein the composite mycelium material comprises one or more proteins from a species other than the fungal species from which the cultivated mycelium material was produced.

32. The composite mycelium material of claim 31, wherein the one or more proteins are from a plant source.

33. The composite mycelium material of claim 32, wherein the plant source is a pea plant.

34. The composite mycelium material of claim 32, wherein the plant source is a soybean plant.

35. The composite mycelium material of claim 1, wherein the composite mycelium material further comprises a dye.

36. The composite mycelium material of claim 35, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

37. The composite mycelium material of claim 36, wherein the dye is a reactive dye.

38. The composite mycelium material of claim 35, wherein the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

39. The composite mycelium material of claim 35, wherein the dye is present throughout the interior of the composite mycelium material.

40. The composite mycelium material of claim 1, wherein the composite mycelium material further comprises a plasticizer.

41. The composite mycelium material of claim 40, wherein the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

42. The composite mycelium material of claim 41, wherein the plasticizer is a fatliquor.

43. The composite mycelium material of claim 40, wherein the composite mycelium material is flexible.

44. The composite mycelium material of claim 1, wherein an external element is applied to the cultivated mycelium material.

45. The composite mycelium material of claim 44, wherein the external element is applied via heat and / or pressure.

46. ​​The composite mycelium material of claim 44, wherein the external element is a heat press.

47. The composite mycelium material of claim 1, wherein the composite mycelium material further comprises tannin.

48. The composite mycelium material of claim 1, wherein the composite mycelium material further comprises a finish.

49. The composite mycelium material of claim 48, wherein the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

50. The composite mycelium material of any one of claims 1-49, wherein the composite mycelium material comprises mechanical properties.

51. The composite mycelium material of claim 50, wherein the mechanical properties comprise wet tensile strength, initial modulus, elongation at break, thickness and / or slit tear strength.

52. The composite mycelium material of claim 50, wherein the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

53. The composite mycelium material of claim 50, wherein the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

54. The composite mycelium material of claim 50, wherein the composite mycelium material has a wet tensile strength of 7 MPa.

55. The composite mycelium material of claim 50, wherein the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

56. The composite mycelium material of claim 50, wherein the composite mycelium material has an elongation at break of 1% to 25%.

57. The composite mycelium material of claim 50, wherein the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

58. The composite mycelium material of claim 50, wherein the composite mycelium material has a thickness of 2 mm.

59. The composite mycelium material of claim 50, wherein the composite mycelium material has a split tear strength of 5N to 100N.

60. The composite mycelium material of claim 50, wherein the composite mycelium material has a split tear strength of 50N.

61. The composite mycelium material of any one of claims 1-49, wherein the composite mycelium material is made using conventional papermaking equipment.

62. A composite mycelium material comprising: a. a cultivated mycelial material comprising one or more masses of branched hyphae, wherein the one or more masses of branched hyphae are disrupted; b. an adhesive comprising a vinyl acetate-ethylene copolymer; and c. Reactive dyes.

63. A method of making a composite mycelium material, the method comprising: a. producing a cultivated mycelial material comprising one or more masses of branched hyphae; b. disrupting the cultivated mycelial material comprising one or more clumps of branched hyphae; and c. adding a binder to the cultivated mycelium material; thereby producing the composite mycelium material, wherein the binder is selected from the group consisting of vinyl acetate-ethylene copolymer, vinyl acetate-acrylic copolymer, polyamide-epichlorohydrin resin (PAE), copolymer, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives and synthetic adhesives.

64. The method of claim 63, wherein the binder is added before the branched hyphae mass is disrupted, during the disruption of the branched hyphae mass, or after the branched hyphae mass is disrupted.

65. The method of claim 63, wherein said producing comprises producing the cultivated mycelial material on a solid substrate.

66. The method of claim 63, wherein the cultivated mycelial material comprises one or more masses of disrupted branching hyphae.

67. The method of claim 66, wherein the one or more clumps of disrupted branched hyphae have a length of 0.1 mm to 5 mm.

68. The method of claim 67, wherein the one or more clumps of disrupted branched hyphae have a length of 2 mm.

69. The method of claim 63, further comprising entangling the one or more masses of branched hyphae, wherein entangling the hyphae comprises needling, felting, or hydroentangling.

70. The method of claim 69, wherein entangling the hyphae comprises hydroentangling.

71. The method of claim 69, wherein hydroentangling the one or more masses of branched hyphae comprises hydroentangling using liquid jets configured to spray liquid at a pressure of 700 psi to 1000 psi.

72. The method of claim 69, wherein the hydroentangling comprises using liquid jets configured to eject liquid at a flow rate of 100 mL / min to 300 mL / min.

73. The method of claim 63, wherein said disrupting comprises disrupting said one or more clumps of branched hyphae by mechanical action.

74. The method of claim 73, wherein the mechanical action comprises blending the one or more masses of branched hyphae.

75. The method of claim 73, wherein the mechanical action comprises breaking the one or more clumps of branched hyphae.

76. The method of claim 73, wherein the mechanical action comprises applying a physical force to the one or more masses of branched hyphae such that at least some of the masses of branched hyphae are aligned in a parallel fashion.

77. The method of claim 76, wherein the physical force is a tensile force.

78. The method of claim 76, wherein the mechanical action comprises applying the physical force in one or more directions such that at least some of the branched hyphae masses are aligned parallel in one or more directions, wherein the physical force is applied repeatedly.

79. The method of claim 63, wherein the one or more clumps of branching hyphae are disrupted by chemical treatment.

80. The method of claim 79, wherein the chemical treatment comprises contacting the one or more masses of branching hyphae with an amount of alkali or other chemical agent sufficient to cause disruption.

81. The method of claim 80, wherein the base comprises a basic peroxide.

82. The method of claim 63, wherein the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

83. The method of claim 63, wherein the binder is vinyl acetate-ethylene copolymer.

84. The method of claim 63, wherein the adhesive is a vinyl acetate-acrylic copolymer.

85. The method of claim 63, wherein the binder is PAE.

86. The method of claim 85, wherein the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of hyphae.

87. The method of claim 63, wherein the natural adhesive comprises a natural latex based adhesive.

88. The method of claim 87, wherein the natural latex-based adhesive is leather glue or weld.

89. The method of claim 63, further comprising incorporating a support material into the composite mycelium material.

90. The method of claim 89, wherein the support material has a pore size of 1 / 16 inch.

91. The method of claim 89, wherein the support material comprises a reinforcing material.

92. The method of claim 91, wherein the reinforcement material is entangled within the composite mycelium material.

93. The method of claim 89, wherein the support material comprises a base material.

94. The method of claim 93, wherein the base material is located on one or more surfaces of the composite mycelium material.

95. The method of claim 89, wherein the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted fibers, woven fibers, and non-woven fibers.

96. The method of claim 63, further comprising perforating, sonicating, and / or vacuum processing the cultivated mycelial material.

97. The method of claim 96, comprising perforating and sonicating the cultivated mycelial material.

98. The method of claim 63, further comprising adding one or more proteins from a species other than the fungal species that produced the cultivated mycelial material.

99. The method of claim 98, wherein the one or more proteins are from a plant source.

100. The method of claim 99, wherein the plant source is a pea plant.

101. The method of claim 99, wherein the plant source is a soybean plant.

102. The method of claim 63, further comprising adding a dye to the cultivated mycelial material or the composite mycelial material.

103. The method of claim 102, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

104. The method of claim 103, wherein the dye is a reactive dye.

105. The method of claim 102, wherein the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

106. The method of claim 102, wherein the dye is present throughout the interior of the composite mycelium material.

107. The method of claim 63, further comprising adding a plasticizer to the cultivated mycelium material or the composite mycelium material.

108. The method of claim 107, wherein the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

109. The method of claim 108, wherein the plasticizer is a fatliquor.

110. The method of claim 107, wherein the composite mycelium material is flexible.

111. The method of claim 63, further comprising applying an external element to the cultivated mycelial material.

112. The method of claim 111, wherein the external element is applied via heat and / or pressure.

113. The method of claim 111, wherein the external element is a heat press.

114. The method of claim 63, further comprising adding tannin to the cultivated mycelial material or the composite mycelial material.

115. The method of claim 63, further comprising adding a finish to the composite mycelium material.

116. The method of claim 115, wherein the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

117. The method of any one of claims 63-116, further comprising determining a mechanical property of the composite mycelium material.

118. The method of claim 117, wherein the mechanical properties comprise wet tensile strength, initial modulus, elongation at break, thickness and / or slit tear strength.

119. The method of claim 117, wherein the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

120. The method of claim 117, wherein the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

121. The method of claim 117, wherein the composite mycelium material has a wet tensile strength of 7 MPa.

122. The method of claim 117, wherein the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

123. The method of claim 117, wherein the composite mycelium material has an elongation at break of 1% to 25%.

124. The method of claim 117, wherein the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

125. The method of claim 117, wherein the composite mycelium material has a thickness of 2 mm.

126. The method of claim 117, wherein the composite mycelium material has a split tear strength of 5N to 100N.

127. The method of claim 117, wherein the composite mycelium material has a split tear strength of 50N.

128. The method of any one of claims 63-116, wherein conventional papermaking equipment is used to make the composite mycelium material.

129. A method of making a composite mycelium material, comprising: a. Produce cultivated mycelial material; b. pressurizing the cultivated mycelium material; and c. contacting the pressurized cultivated mycelium material with a solution comprising a binder, thereby producing the composite mycelium material, wherein the binder is selected from the group consisting of vinyl acetate-ethylene copolymer, vinyl acetate-acrylic copolymer, polyamide-epichlorohydrin resin (PAE), copolymer, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives and synthetic adhesives.

130. The method of claim 129, wherein the adhesive is added before the pressurizing step, during the pressurizing step, or after the pressurizing step.

131. The method of claim 129, wherein said producing comprises producing the cultivated mycelial material on a solid substrate.

132. The method of claim 129, further comprising incorporating a support material into the composite mycelium material.

133. The method of claim 132, wherein the support material has a pore size of 1 / 16 inch.

134. The method of claim 132, wherein the support material comprises a reinforcing material.

135. The method of claim 134, wherein the reinforcement material is entangled within the composite mycelium material.

136. The method of claim 135, wherein the support material comprises a base material.

137. The method of claim 136, wherein the base material is located on one or more surfaces of the composite mycelium material.

138. The method of claim 132, wherein the support material is selected from the group consisting of mesh fabric, gauze, fabric, knitted fibers, woven fibers, and non-woven fibers.

139. The method of claim 129, wherein the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

140. The method of claim 129, wherein the binder is vinyl acetate-ethylene copolymer.

141. The method of claim 129, wherein the adhesive is a vinyl acetate-acrylic copolymer.

142. The method of claim 129, wherein the adhesive is PAE.

143. The method of claim 142, wherein the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of hyphae.

144. The method of claim 143, wherein the natural adhesive comprises a natural latex based adhesive.

145. The method of claim 144, wherein the natural latex-based adhesive is leather glue or weld.

146. The method of claim 129, further comprising perforating, sonicating, and / or vacuum processing the cultivated mycelial material.

147. The method of claim 146, comprising perforating and sonicating the cultivated mycelial material.

148. The method of claim 129, further comprising adding one or more proteins from a species other than the fungal species that produced the cultivated mycelial material.

149. The method of claim 148, wherein the one or more proteins are from a plant source.

150. The method of claim 149, wherein the plant source is a pea plant.

151. The method of claim 149, wherein the plant source is a soybean plant.

152. The method of claim 129, further comprising adding a dye to the cultivated mycelial material or the composite mycelial material.

153. The method of claim 152, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

154. The method of claim 153, wherein the dye is a reactive dye.

155. The method of claim 152, wherein the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

156. The method of claim 152, wherein the dye is present throughout the interior of the composite mycelium material.

157. The method of claim 129, further comprising adding a plasticizer to the cultivated mycelium material or the composite mycelium material.

158. The method of claim 157, wherein the plasticizer is selected from the group consisting of oil, glycerol, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

159. The method of claim 158, wherein the plasticizer is a fatliquor.

160. The method of claim 157, wherein the composite mycelium material is flexible.

161. The method of claim 129, further comprising applying an external element to the cultivated mycelial material.

162. The method of claim 161, wherein the external element is applied via heat and / or pressure.

163. The method of claim 161, wherein the external element is a heat press.

164. The method of claim 129, further comprising adding tannin to the cultivated mycelial material or the composite mycelial material.

165. The method of claim 129, further comprising adding a finish to the composite mycelium material.

166. The method of claim 165, wherein the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

167. The method of any one of claims 129-166, further comprising determining a mechanical property of the composite mycelium material.

168. The method of claim 167, wherein the mechanical properties comprise wet tensile strength, initial modulus, elongation at break, thickness and / or slit tear strength.

169. The method of claim 168, wherein the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

170. The method of claim 168, wherein the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

171. The method of claim 168, wherein the composite mycelium material has a wet tensile strength of 7 MPa.

172. The method of claim 168, wherein the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

173. The method of claim 168, wherein the composite mycelium material has an elongation at break of 1% to 25%.

174. The method of claim 168, wherein the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

175. The method of claim 168, wherein the composite mycelium material has a thickness of 2 mm.

176. The method of claim 168, wherein the composite mycelium material has a split tear strength of 5N to 100N.

177. The method of claim 168, wherein the composite mycelium material has a split tear strength of 50N.

178. The method of any one of claims 129-166, wherein conventional papermaking equipment is used to make the composite mycelium material.

179. A composite mycelium material comprising: a. Pressurized cultivated mycelial material; and b. an adhesive selected from the group consisting of vinyl acetate-ethylene (VAE) copolymers, vinyl acetate-acrylic copolymers, polyamide-epichlorohydrin resins (PAE), copolymers, transglutaminase, citric acid, genipin, alginate, gum arabic, latex, natural adhesives, and synthetic adhesives.

180. The composite mycelial material of claim 179, wherein the cultivated mycelial material has been produced on a solid substrate.

181. The composite mycelium material of claim 179, wherein the binder is a copolymer having properties selected from the group consisting of: a particle size less than or equal to 1 μm, a sub-zero glass transition temperature, and a self-crosslinking function.

182. The composite mycelium material of claim 179, wherein the binder is a vinyl acetate-ethylene (VAE) copolymer.

183. The composite mycelium material of claim 179, wherein the binder is a vinyl acetate-acrylic acid copolymer.

184. The composite mycelium material of claim 179, wherein the binder is PAE.

185. The composite mycelium material of claim 184, wherein the PAE comprises a cationic azetidinium group that reacts with active hydrogen-containing groups comprising amine, hydroxyl, and carboxyl groups in one or more branches of the hyphae.

186. The composite mycelium material of claim 179, wherein the natural adhesive comprises a natural latex based adhesive.

187. The composite mycelium material of claim 186, wherein the natural latex-based adhesive is leather glue or weld.

188. The composite mycelium material of claim 179, wherein the composite mycelium material further comprises a support material.

189. The composite mycelium material of claim 188, wherein the support material has a pore size of 1 / 16 inch.

190. The composite mycelium material of claim 188, wherein the support material comprises a reinforcement material.

191. The composite mycelium material of claim 190, wherein the reinforcement material is entangled within the composite mycelium material.

192. The composite mycelium material of claim 188, wherein the support material comprises a base material.

193. The composite mycelium material of claim 192, wherein the base material is located on one or more surfaces of the composite mycelium material.

194. The composite mycelium material of claim 188, wherein the support material is selected from the group consisting of mesh, gauze, fabric, knitted fibers, woven fibers, and non-woven fibers.

195. The composite mycelium material of claim 179, wherein the composite mycelium material comprises one or more proteins from a species other than the fungal species that produced the cultivated mycelium material.

196. The composite mycelium material of claim 195, wherein the one or more proteins are from a plant source.

197. The composite mycelium material of claim 196, wherein the plant source is a pea plant.

198. The composite mycelium material of claim 196, wherein the plant source is a soybean plant.

199. The composite mycelium material of claim 179, wherein the composite mycelium material further comprises a dye.

200. The composite mycelium material of claim 199, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

201. The composite mycelium material of claim 200, wherein the dye is a reactive dye.

202. The composite mycelium material of claim 199, wherein the composite mycelium material is colored with the dye and the color of the composite mycelium material is substantially uniform across one or more surfaces of the composite mycelium material.

203. The composite mycelium material of claim 199, wherein the dye is present throughout the interior of the composite mycelium material.

204. The composite mycelium material of claim 179, wherein the composite mycelium material further comprises a plasticizer.

205. The composite mycelium material of claim 204, wherein the plasticizer is selected from the group consisting of oil, glycerin, fatliquor, sorbitol, diethoxydimethylammonium chloride, Tween 20, Tween 80, erythritol, water, ethylene glycol, triethyl citrate, water, acetylated monoglycerides, and epoxidized soybean oil.

206. The composite mycelium material of claim 205, wherein the plasticizer is a fatliquor.

207. The composite mycelium material of claim 206, wherein the composite mycelium material is flexible.

208. The composite mycelium material of claim 179, wherein an external element is applied to the cultivated mycelium material.

209. The composite mycelium material of claim 208, wherein the external element is applied via heat and / or pressure.

210. The composite mycelium material of claim 209, wherein the external element is a heat press.

211. The composite mycelium material of claim 179, wherein the composite mycelium material further comprises tannin.

212. The composite mycelium material of claim 179, wherein the composite mycelium material further comprises a finish.

213. The composite mycelium material of claim 212, wherein the finish is selected from the group consisting of urethane, wax, nitrocellulose, and plasticizers.

214. The composite mycelium material of any one of claims 179-213, wherein the composite mycelium material comprises mechanical properties.

215. The composite mycelium material of claim 214, wherein the mechanical properties comprise wet tensile strength, initial modulus, elongation at break, thickness and / or slit tear strength.

216. The composite mycelium material of claim 215, wherein the composite mycelium material has a wet tensile strength of 0.05 MPa to 10 MPa.

217. The composite mycelium material of claim 215, wherein the composite mycelium material has a wet tensile strength of 5 MPa to 20 MPa.

218. The composite mycelium material of claim 215, wherein the composite mycelium material has a wet tensile strength of 7 MPa.

219. The composite mycelium material of claim 215, wherein the composite mycelium material has an initial modulus of 1 MPa to 100 MPa.

220. The composite mycelium material of claim 215, wherein the composite mycelium material has an elongation at break of 1% to 25%.

221. The composite mycelium material of claim 215, wherein the composite mycelium material has a thickness of 0.5 mm to 3.5 mm.

222. The composite mycelium material of claim 215, wherein the composite mycelium material has a thickness of 2 mm.

223. The composite mycelium material of claim 215, wherein the composite mycelium material has a split tear strength of 5N to 100N.

224. The composite mycelium material of claim 215, wherein the composite mycelium material has a split tear strength of 50N.

225. The composite mycelium material of any one of claims 179-213, wherein the composite mycelium material is made using conventional papermaking equipment.

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