Methods of generating mycelium materials with improved properties

Abstract

To provide improved mycelium materials with favorable mechanical properties, aesthetic properties, and other advantages, as well as materials and methods for making the improved mycelium materials.SOLUTION: A method for producing a composition of a mycelium material comprises: a. generating a cultivated mycelium material; b. contacting the cultivated mycelium material with a solution comprising one or more proteins to produce a composition comprising the cultivated mycelium material and one or more proteins, wherein the one or more proteins are derived from a species other than a fungal species from which the cultivated mycelium material is generated; and c. pressing the cultivated mycelium material. Also provided is an article of footwear comprising the mycelium material.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 767,433, filed November 14, 2018, and U.S. Provisional Application No. 62 / 782,277, filed December 19, 2018, the contents of which are incorporated by reference in their entirety. [Background technology]

[0002] background Mycelium is a next-generation sustainable material of growing interest due to its biological efficiency, strength and low environmental footprint. To this end, various methods of growing intertwined mycelial networks, both alone and as composites (i.e., intertwined particulate, fiber, or fiber networks), have been discussed for various applications. However, mycelial materials currently under development suffer from poor mechanical qualities, such as increased delamination and tearing under stress, and also from poor aesthetic qualities. Thus, what is needed are improved mycelial materials with favorable mechanical properties, aesthetic properties, and other advantages, as well as materials and methods for producing the improved mycelial materials. Summary of the Invention

[0003] overview In one aspect, provided herein is a composition comprising cultured mycelial material and one or more proteins, wherein the one or more proteins are derived from a species other than the fungal species from which the cultured mycelial material is produced.

[0004] In some embodiments, the one or more proteins are derived from a plant source.

[0005] In some embodiments, the plant source is a pea plant.

[0006] In some embodiments, the plant source is a soybean plant.

[0007] In some embodiments, the composition comprises a dye.

[0008] In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

[0009] In some embodiments, the composition comprises a plasticizer.

[0010] In some embodiments, the plasticizer is selected from the group consisting of oils, glycerin, and fat liquors.

[0011] In some embodiments, the composition is flexible.

[0012] In some embodiments, the one or more proteins are cross-linked.

[0013] In some embodiments, the one or more proteins are cross-linked with transglutaminase.

[0014] In some embodiments, the composition comprises an enzyme.

[0015] In some embodiments, the enzyme comprises transglutaminase.

[0016] In another aspect, provided herein is a composition comprising cultured mycelial material stained with a dye to develop a color, wherein the color of the cultured mycelial material is substantially uniform across one or more surfaces of the cultured mycelial material.

[0017] In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

[0018] In some embodiments, the composition comprises one or more proteins derived from a species other than the fungal species from which the cultured mycelial material is generated.

[0019] In some embodiments, the one or more proteins are derived from a plant source.

[0020] In some embodiments, the plant source is a pea plant.

[0021] In some embodiments, the plant source is a soybean plant.

[0022] In some embodiments, the dye permeates the entire interior of the composition.

[0023] In some embodiments, the composition comprises a plasticizer.

[0024] In some embodiments, the plasticizer is selected from the group consisting of oils, glycerin, and fatliquors.

[0025] In some embodiments, the composition is flexible.

[0026] In some embodiments, the composition comprises a tannin.

[0027] In some embodiments, the composition includes a finish applied to one or more surfaces of the composition.

[0028] In some embodiments, the finish is selected from the group consisting of a urethane, a wax, nitrocellulose, or a plasticizer.

[0029] In another aspect, provided herein is a method, comprising: generating cultured mycelium material; contacting the cultured mycelial material with a solution comprising one or more proteins to produce a composition comprising the cultured mycelial material and one or more proteins, the one or more proteins being derived from a species other than the fungal species from which the cultured mycelial material was produced; and compressing said cultured mycelium material.

[0030] In some embodiments, the contacting step comprises submerging the cultured mycelial material in the solution.

[0031] In some embodiments, the contacting step comprises contacting the cultured mycelial material with the solution in a single step.

[0032] In some embodiments, the contacting step comprises contacting the cultured mycelial material with the solution in one or more stages.

[0033] In some embodiments, the one or more proteins are derived from a plant source.

[0034] In some embodiments, the plant source is a pea plant.

[0035] In some embodiments, the plant source is a soybean plant.

[0036] In some embodiments, the solution comprises a dye.

[0037] In some embodiments, the composition is colored with a dye to develop a color, and the color of the cultured mycelial material is substantially uniform across one or more surfaces of the cultured mycelial material.

[0038] In some embodiments, the dye permeates the entire interior of the composition.

[0039] In some embodiments, the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

[0040] In some embodiments, the solution includes a plasticizer.

[0041] In some embodiments, the plasticizer is selected from the group consisting of oils, glycerin, and fatliquors.

[0042] In some embodiments, the composition is flexible.

[0043] In some embodiments, the one or more proteins are cross-linked.

[0044] In some embodiments, one or more proteins are cross-linked with transglutaminase.

[0045] In some embodiments, the solution comprises an enzyme.

[0046] In some embodiments, the enzyme comprises transglutaminase.

[0047] In some embodiments, the compacting step comprises compacting the cultured mycelium material to a thickness of between 0.1 inches and 0.5 inches.

[0048] In some embodiments, the compacting step comprises compacting the cultured mycelium material to a thickness of 0.25 inches.

[0049] In some embodiments, the compression step is repeated one or more times.

[0050] In some embodiments, the compacting step comprises compacting the cultured mycelium material to a thickness of 0.25 inches.

[0051] In some embodiments, the compacting step comprises compacting the cultured mycelium material with a roller.

[0052] In some embodiments, the solution comprises tannin.

[0053] In some embodiments, the method further comprises the step of incubating the composition.

[0054] In some embodiments, the incubating step comprises incubating the composition at a set temperature for a period of time.

[0055] In some embodiments, the set temperature is 40°C.

[0056] In some embodiments, the method further comprises drying the composition.

[0057] In some embodiments, the method further comprises applying a finish to one or more surfaces of the composition.

[0058] In some embodiments, the finish is selected from the group consisting of a urethane, a wax, nitrocellulose, or a plasticizer.

[0059] In another aspect, provided herein is an article of footwear that includes an upper, a lasting board attached to the upper and defining an interior cavity for receiving a foot, and an outsole coupled to the upper opposite the lasting board, wherein the upper includes at least a portion of mycelium material that includes one or more proteins derived from an organism other than mycelium.

[0060] In some embodiments, the upper comprises multiple sections of mycelium material, each of whose implementations has different physical properties.

[0061] In some embodiments, the different physical properties are selected to correlate with desired characteristics of corresponding locations of portions within the upper.

[0062] In some embodiments, one of the portions of the mycelial material includes a bump, and each implementation of the mycelial material has a relatively high flexibility compared to at least one of the portions.

[0063] In some embodiments, one of the sections of mycelial material comprises a heel counter, and each implementation of the mycelial material has a higher relative stiffness compared to at least one of the sections.

[0064] In some embodiments, the mycelium material is at least one of tanned and dyed to resemble leather.

[0065] In some embodiments, the article further includes a midsole attached to the lasting board, and an outsole attached to the midsole and coupled to the upper.

[0066] In some embodiments, said upper comprises a plurality of separate portions of said mycelium material.

[0067] In some embodiments, the portions are assembled using at least one of a topstitch, a felt stitch, and a backstitch construction.

[0068] In some embodiments, the parts are assembled using at least one of a solvent-based adhesive, a UV-curable adhesive, a heat-activated adhesive, and a water-based adhesive.

[0069] In some embodiments, at least one of the portions is split to resemble suede leather.

[0070] In some embodiments, at least one of the portions includes an edge that has been thinned by skiving.

[0071] In some embodiments, the portions are assembled using thermal bonding.

[0072] In some embodiments, the upper further comprises at least one additional portion of a textile material.

[0073] In some embodiments, the fabric material is thermoplastic and is attached to at least one of the portions of the mycelium material by thermal bonding.

[0074] In some embodiments, the upper includes an interlining assembled with a portion thereof.

[0075] In some embodiments, the perforations are parallel to a portion thereof.

[0076] In some embodiments, the perforations vary in at least one of size and relative spacing across an area of ​​the upper.

[0077] In some embodiments, the upper is laser etched along portions thereof.

[0078] In some embodiments, the upper includes at least one reinforcing portion injection molded thereon.

[0079] In some embodiments, the upper includes at least one 3D printed element fused thereto.

[0080] In some embodiments, at least a portion of the upper includes at least one portion that is molded into a three-dimensional shape.

[0081] In some embodiments, said upper is constructed from a single molded piece of said mycelium material.

[0082] In some embodiments, the mycelial material comprises multiple layers of the mycelial material bonded together, each of the packaging forms having different physical properties.

[0083] In some embodiments, at least one of said lasting board and said outsole comprises at least a portion of said mycelium material.

[0084] In another aspect, provided herein is an athletic sneaker comprising an upper comprising at least a portion of a mycelium material comprising one or more proteins derived from an organism other than mycelium, a lasting board attached to the upper and defining an interior cavity for receiving a foot, a midsole made of a foam material and attached to the lasting board, and an outsole made of a rubber material and attached to the midsole opposite the lasting board, wherein the mycelium material is at least one of tanned and dyed to resemble leather, and the upper is constructed and assembled to resemble leather athletic footwear.

[0085] In another aspect, provided herein is an athletic sneaker that includes an upper that includes at least a portion of a mycelium material that includes one or more proteins from an organism other than mycelium, a lasting board attached to the upper and defining an interior cavity for receiving a foot, a midsole made of a foam material and attached to the lasting board, and an outsole made of a rubber material and attached to the midsole opposite the lasting board, wherein the upper includes at least one portion that is molded into a three-dimensional shape. [The present invention 1001] a. producing cultured mycelium material; b. contacting the cultured mycelial material with a solution comprising one or more proteins to produce a composition comprising the cultured mycelial material and one or more proteins, the one or more proteins being derived from a species other than the fungal species from which the cultured mycelial material was produced; and c. compressing the cultured mycelium material A method comprising: [The present invention 1002] The method of claim 10, wherein said contacting step comprises submerging said cultured mycelial material in said solution. [The present invention 1003] The method of claim 1001, wherein said contacting step comprises contacting said cultured mycelial material with said solution in a single step. [The present invention 1004] The method of claim 10, wherein said contacting step comprises contacting said cultured mycelial material with said solution in one or more stages. [The present invention 1005] The method of any one of claims 1 to 10, wherein said one or more proteins are derived from a plant source. [The present invention 1006] The method of claim 1005, wherein the plant source is a pea plant. [The present invention 1007] The method of claim 1005, wherein the plant source is a soybean plant. [The present invention 1008] The method of claim 1001, wherein the solution comprises a dye. [The present invention 1009] The method of claim 1008, wherein said composition is colored with said dye to develop a color, and said color of said cultured mycelial material is substantially uniform across one or more surfaces of said cultured mycelial material. [The present invention 1010] The method of claim 1008, wherein the dye penetrates throughout the interior of the composition. [The present invention 1011] The method of claim 1008, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes. [The present invention 1012] The method of claim 1001, wherein the solution comprises a plasticizer. [The present invention 1013] The method of claim 10, wherein the plasticizer is selected from the group consisting of oil, glycerin, and fatliquoring agents. [The present invention 1014] The method of claim 1012, wherein the composition is flexible. [The present invention 1015] The method of any one of claims 1 to 10, wherein said one or more proteins are crosslinked. [The present invention 1016] The method of claim 10, wherein said one or more proteins are cross-linked with transglutaminase. [The present invention 1017] The method of claim 10, wherein the solution comprises an enzyme. [The present invention 1018] The method of claim 1017, wherein the enzyme comprises transglutaminase. [The present invention 1019] The method of claim 1001, wherein said compressing step comprises compressing said cultured mycelium material to a thickness of between 0.1 inches and 0.5 inches. [The present invention 1020] The method of claim 1019, wherein said compressing step comprises compressing said cultured mycelium material to a thickness of 0.25 inches. [The present invention 1021] 1001. The method of claim 1001, wherein the compressing step is repeated one or more times. [The present invention 1022] The method of claim 1001, wherein said compressing step comprises compressing said cultured mycelium material to a thickness of 0.25 inches. [The present invention 1023] The method of claim 1001, wherein said compacting step comprises compacting said cultured mycelium material with a roller. [The present invention 1024] The method of claim 1001, wherein the solution contains tannin. [The present invention 1025] The method of claim 1001, further comprising the step of incubating said composition. [The present invention 1026] The method of claim 1025, wherein said incubating step comprises incubating said composition at a set temperature for a period of time. [The present invention 1027] The method of claim 1026, wherein the set temperature is 40°C. [The present invention 1028] The method of claim 1001, further comprising the step of drying said composition. [The present invention 1029] The method of claim 1001, further comprising the step of applying a finish to one or more surfaces of said composition. [The present invention 1030] The method of claim 1029, wherein the finish is selected from the group consisting of urethane, wax, nitrocellulose, or a plasticizer. [The present invention 1031] 1. A composition comprising cultured mycelial material and one or more proteins, wherein the one or more proteins are derived from a species other than the fungal species from which the cultured mycelial material is produced. [The present invention 1032] The composition of the present invention 1030, wherein said one or more proteins are derived from a plant source. [The present invention 1033] The composition of claim 1032, wherein the plant source is a pea plant. [The present invention 1034] The composition of claim 1032, wherein the plant source is a soybean plant. [The present invention 1035] The composition of the present invention 1030, comprising a dye. [The present invention 1036] The composition of the present invention 1035, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes. [The present invention 1037] The composition of the present invention 1030, comprising a plasticizer. [The present invention 1038] The composition of the present invention 1037, wherein the plasticizer is selected from the group consisting of oil, glycerin and fatliquors. [The present invention 1039] The composition of the present invention 1037, which is flexible. [The present invention 1040] The composition of claim 1030, wherein said one or more proteins are crosslinked. [The present invention 1041] The composition of claim 1030, wherein said one or more proteins are cross-linked with transglutaminase. [The present invention 1042] The composition of the present invention 1030, comprising an enzyme. [The present invention 1043] The composition of claim 1042, wherein the enzyme comprises transglutaminase. [The present invention 1044] 1. A composition comprising cultured mycelial material that has been stained with a dye to develop a color, wherein the color of the cultured mycelial material is substantially uniform across one or more surfaces of the cultured mycelial material. [The present invention 1045] The composition of the present invention 1044, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes. [The present invention 1046] The composition of the present invention 1044, comprising one or more proteins derived from a species other than the fungal species from which the cultured mycelial material is produced. [The present invention 1047] The composition of claim 1046, wherein the one or more proteins are derived from a plant source. [The present invention 1048] The composition of claim 1047, wherein the plant source is a pea plant. [The present invention 1049] The composition of claim 1047, wherein the plant source is a soybean plant. [The present invention 1050] The composition of claim 1044, wherein the dye permeates the entire interior of the composition. [The present invention 1051] The composition of the present invention 1044, comprising a plasticizer. [The present invention 1052] The composition of the present invention wherein the plasticizer is selected from the group consisting of oil, glycerin, and fatliquoring agents. [The present invention 1053] The composition of the present invention 1051 is flexible. [The present invention 1054] The composition of the present invention 1044, comprising tannin. [The present invention 1055] The composition of the present invention further comprising a finish applied to one or more surfaces of said composition. [The present invention 1056] The composition of the present invention, wherein the finish is selected from the group consisting of urethane, wax, nitrocellulose, or plasticizer. [The present invention 1057] Footwear products, including: A.Upper, b. a lasting board attached to said upper and defining an interior cavity for receiving a foot; c. an outsole connected to the upper on the side opposite the lasting board; d. wherein said upper comprises at least a portion of mycelial material that comprises one or more proteins derived from an organism other than mycelium. [The present invention 1058] The article of footwear of the present invention 1057, wherein the upper comprises multiple portions of the mycelium material, each of the implementations having different physical properties. [The present invention 1059] The article of footwear of the present invention 1058, wherein the different physical properties are selected to correlate with desired characteristics of corresponding locations of portions within the upper. [The present invention 1060] The footwear article of claim 1059, wherein one of the portions of the mycelium material includes a bump, and each of the implementations of the mycelium material has a higher relative flexibility compared to at least one of the portions. [The present invention 1061] The footwear article of claim 1059, wherein one of the portions of mycelium material includes a heel counter, and each of the implementations of the mycelium material has a higher relative stiffness compared to at least one of the portions. [The present invention 1062] The footwear article of claim 1057, wherein the mycelium material is at least one of tanned and dyed to resemble leather. [The present invention 1063] The article of footwear of the present invention 1057 further comprises a midsole attached to the lasting board, and an outsole attached to the midsole and coupled to the upper. [The present invention 1064] The article of footwear of the present invention 1057, wherein the upper comprises a plurality of separate portions of the mycelium material. [The present invention 1065] The article of footwear of the present invention 1064, wherein the portions are constructed using at least one of a topstitch, a felt stitch, and a backstitch construction. [The present invention 1066] The article of footwear of the present invention 1064, wherein the portions are assembled using at least one of a solvent-based adhesive, a UV-curable adhesive, a heat-activated adhesive, and a water-based adhesive. [The present invention 1067] The article of footwear of the present invention, wherein at least one of the portions is split to resemble suede leather. [The present invention 1068] The footwear article of claim 1064, wherein at least one of said portions includes an edge that has been thinned by skiving. [The present invention 1069] The article of footwear of the present invention, wherein the portions are assembled using thermal bonding. [The present invention 1070] The article of footwear of the present invention, wherein the upper further comprises at least one additional portion of textile material. [The present invention 1071] The article of footwear of claim 1064, wherein the fabric material is thermoplastic and is attached to at least one of the portions of the mycelium material by thermal bonding. [The present invention 1072] The article of footwear of the present invention 1057, wherein the upper includes an interlining assembled with a portion thereof. [The present invention 1073] The article of footwear of the present invention 1057 includes perforations along a portion thereof. [The present invention 1074] The article of footwear of claim 1073, wherein the perforations vary in at least one of size and relative spacing across the area of ​​the upper. [The present invention 1075] The article of footwear of claim 1057, wherein the upper is laser etched along portions thereof. [The present invention 1076] The article of footwear of the present invention 1057, wherein the upper includes at least one reinforcing portion injection molded thereon. [The present invention 1077] The footwear article of the present invention 1057, wherein the upper includes at least one 3D printed element fused thereto. [The present invention 1078] The article of footwear of the present invention 1057, wherein at least a portion of the upper includes at least one portion molded into a three-dimensional shape. [The present invention 1079] The footwear article of claim 1057, wherein the upper is constructed from a single molded piece of the mycelium material. [The present invention 1080] The footwear article of claim 1057, wherein the mycelium material comprises multiple layers of the mycelium material bonded together, each layer having different physical properties. [The present invention 1081] The article of footwear of the present invention 1057, wherein at least one of the lasting board and the outsole comprises at least a portion of the mycelium material. [The present invention 1082] Athletic sneakers, including: a. an upper comprising at least a portion of mycelial material that contains one or more proteins derived from an organism other than mycelium; b. a lasting board attached to said upper and defining an interior cavity for receiving a foot; c. a midsole made of a foam material and attached to the lasting board; and d. an outsole made of a rubber material and attached to the midsole opposite the lasting board; e. wherein said mycelium material is at least one of tanned and dyed to resemble leather, and said upper is constructed and assembled to resemble leather athletic footwear. [The present invention 1083] Athletic sneakers, including: a. an upper comprising at least a portion of mycelial material that contains one or more proteins derived from an organism other than mycelium; b. a lasting board attached to said upper and defining an interior cavity for receiving a foot; c. a midsole made of a foam material and attached to the lasting board; and d. an outsole made of a rubber material and attached to the midsole opposite the lasting board; e. wherein said upper includes at least one portion that is molded into a three-dimensional shape. [Brief description of the drawings]

[0086] The foregoing summary, as well as the following detailed description, will be better understood when read in conjunction with the accompanying drawings. For purposes of illustration, certain aspects of the disclosure are shown in the drawings. It will be understood, however, that the disclosure is not limited to the precise arrangements and instrumentalities shown. The drawings are not necessarily to scale. Certain features may be exaggerated in scale or shown in schematic form for clarity and conciseness.

[0087] [Figure 1] FIG. 1 is a front perspective view of an athletic sneaker according to an aspect of the present disclosure. [Diagram 2] FIG. 2 is a front perspective exploded view of an athletic sneaker. [Diagram 3] FIG. 2 is a front perspective exploded view of the upper of an athletic sneaker. [Figure 4] FIG. 2 is a front perspective view of an athletic sneaker according to another aspect of the present disclosure. [Diagram 5] FIG. 2 is a front perspective exploded view of an athletic sneaker. [Figure 6] FIG. 1 is a top view of a cut sheet of mycelium material that can be used to construct the upper of an athletic sneaker. [Figure 7] FIG. 2 is a front perspective view of an article of footwear according to another aspect of the present disclosure. [Figure 8] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 9] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 10] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 11] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 12] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 13] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 14] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 15] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 16] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 17] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 18] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 19] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 20] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 21] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 22] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 23] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 24] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Diagram 25] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 26] 1 shows cross-sections of exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 27] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 28]A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 29] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 30] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 31] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 32] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 33] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 34] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 35] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 36] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 37]A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 38] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 39] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 40] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 41] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 42] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 43] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 44] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 45] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 46]A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 47] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 48] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 49] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 50] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 51] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 52] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Diagram 53] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 54] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 55]A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 56] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 57] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 58] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 59] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 60] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 61] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 62] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 63] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 64] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 65] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 66] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 67] A shows a cross section of an exemplary cultured mycelium material after the indicated dyes and processing steps.B shows an exemplary cultured mycelium material after color fastness testing and the indicated dyes and processing steps. [Figure 68] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 69] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 70] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 71] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 72] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 73] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 74] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 75]A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 76] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 77] 2 shows an exemplary cultured mycelium material after Nitrocellulose and Protein Sandable Finish-Box Effect treatment. [Figure 78] 6 shows an exemplary cultured mycelium material after nitrocellulose finish-box effect treatment. [Figure 79] 1 shows an exemplary cultured mycelium material after a conventional polyurethane finishing treatment. [Figure 80] 1 shows an exemplary cultured mycelium material after antique effect finishing treatment. [Figure 81] 1 shows an exemplary cultured mycelium material after a distress finishing treatment. [Figure 82] 1 shows an exemplary cultured mycelium material after an embossed Luganil Olive Brown finish treatment. [Figure 83] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 84] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 85] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 86] A shows a cross section of an exemplary cultured mycelial material after the indicated dyes and processing steps.B shows an exemplary cultured mycelial material after the indicated dyes and processing steps. [Figure 87] FIG. 1 shows an exemplary mycelium material after pea protein finishing. [Figure 88]1 shows an exemplary mycelium material after unmixed soy protein finishing. [Figure 89] 1 shows an exemplary mycelium material after agitation soy protein finishing. [Figure 90] 1 shows an exemplary mycelium material after hemp protein finishing. [Figure 91] FIG. 1 shows an exemplary mycelium material after 50:50 pea protein to FI50 finishing. [Figure 92] 4 shows an exemplary mycelium material after 50:50 soy protein to FI50 finishing. [Figure 93] FIG. 1 shows an exemplary mycelium material after pea protein and cross-linker finishing. [Figure 94] 1 shows an exemplary mycelium material after Luganil Brown dye and carnauba flake wax finish. [Figure 95] 1 shows an exemplary mycelium material after Luganil Bordeaux dye, washing, and a carnauba flake wax finish. [Figure 96] 1 shows an exemplary mycelium material after Luganil Yellow dye, washing, and a Carnauba liquid wax finish. [Figure 97] Illustrated is an exemplary mycelium material after Luganil Brown dye, washing, and a Carnauba liquid wax finish. [Figure 98] 1 shows an exemplary mycelium material after waxy filler, water-based PU, and carnauba flake wax finish. [Figure 99] FIG. 1 shows an exemplary mycelium material after one coat of pea protein and crosslinker finish. [Figure 100] FIG. 1 shows an exemplary mycelium material after two coats of pea protein and crosslinker finish. [Figure 101] FIG. 1 shows an exemplary mycelium material after pea protein, cross-linking agent, and filler finishing without embossing. [Figure 102] FIG. 1 shows an exemplary mycelium material with embossing after pea protein, cross-linking agent, and filler finish. [Figure 103]Illustrated is an exemplary mycelium material after Luganil Red dye, washing, and pea protein and crosslinker finishing. [Figure 104] Illustrated is Luganil Brown dye, as well as exemplary mycelium material after glycerin soak, pea protein and crosslinker finish. [Figure 105] Illustrated is an exemplary mycelium material after Luganil Bordeaux dye and pea protein and crosslinker finish. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0088] Detailed Description Details of various embodiments are described in the following description. It should also be understood that the specific products, components, and steps shown in the accompanying drawings and described in the following specification are merely illustrative of the concepts defined in the appended claims. Thus, specific dimensions and other physical characteristics related to the embodiments disclosed herein should not be considered limiting unless the claims expressly state otherwise. Other features, objects, and advantages will become apparent from the description. Unless otherwise defined herein, scientific and technical terms shall have the meanings commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include plurals and plural terms shall include the singular. The terms "a" and "an" include plural referents unless the context dictates otherwise. In general, the nomenclature used in connection with, and techniques of, biochemistry, enzymology, molecular and cell biology, microbiology, genetics, protein, and nucleic acid chemistry, and hybridization described herein are well known and commonly used in the art.

[0089] The following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0090] The term "polynucleotide" or "nucleic acid molecule" refers to a polymeric form of nucleotides of at least 10 bases in length. The term includes DNA molecules (e.g., cDNA or genomic DNA or synthetic DNA) and RNA molecules (e.g., mRNA or synthetic RNA), as well as analogs of DNA or RNA that contain non-natural nucleotide analogs, non-natural internucleoside bonds, or both. The nucleic acid can be in any topological conformation. For example, the nucleic acid can be in a single-stranded, double-stranded, triple-stranded, quadruple-stranded, partially double-stranded, branched, hairpinned, circular, or padlock conformation.

[0091] Unless otherwise indicated, and as an example of all sequences described herein in the general format "SEQ ID NO:", a "nucleic acid comprising SEQ ID NO:1" refers to a nucleic acid, at least a portion of which has either (i) the sequence of SEQ ID NO:1, or (ii) a sequence complementary to SEQ ID NO:1. The choice between the two is determined by the context. For example, if the nucleic acid is used as a probe, the choice between the two is determined by the requirement that the probe be complementary to a desired target.

[0092] An "isolated" RNA, DNA or mixed polymer is one that is substantially separated from other cellular components which naturally accompany the natural polynucleotide in its natural host cell, such as the ribosomes, polymerases and genomic sequences with which it is naturally associated.

[0093] An "isolated" organic molecule (e.g., silk protein) is one that has been substantially separated from cellular components (membrane lipids, chromosomes, proteins) of the host cell from which it was derived, or from the medium in which the host cell was cultured. The term does not require that the biomolecule be separated from all other chemicals, although certain isolated biomolecules may be purified to near homogeneity.

[0094] The term "recombinant" refers to a biological molecule, such as a gene or protein, that is: (1) removed from its natural environment, (2) not associated with all or a portion of a polynucleotide with which the gene is found in nature, (3) operably linked to a polynucleotide with which it is not linked in nature, or (4) not occurring in nature. The term "recombinant" can be used in reference to cloned DNA isolates, chemically synthesized polynucleotide analogs, or polynucleotide analogs biologically synthesized by heterologous systems, as well as proteins and / or mRNAs encoded by such nucleic acids.

[0095] An endogenous nucleic acid sequence (or the protein product of the sequence encoded) in the genome of an organism is considered herein to be "recombinant" when a heterologous sequence is placed adjacent to the endogenous nucleic acid sequence and the expression of the endogenous nucleic acid sequence is altered. In this context, a heterologous sequence is a sequence that is not naturally adjacent to the endogenous nucleic acid sequence, regardless of whether the heterologous sequence itself is endogenous (originating from the same host cell or its progeny) or exogenous (originating from a different host cell or its progeny). As an example, a promoter sequence can replace the native promoter of a gene in the genome of a host cell (e.g., by homologous recombination) so that the gene has an altered expression pattern. The gene is "recombinant" because it is separated from at least a portion of the sequences that naturally flank it.

[0096] A nucleic acid is also considered to be "recombinant" if it contains any modification that does not naturally occur in the corresponding nucleic acid in a genome. For example, an endogenous coding sequence is considered to be "recombinant" if it contains an insertion, deletion, or point mutation introduced artificially, e.g., by human intervention. "Recombinant nucleic acid" also includes a nucleic acid integrated into a host cell chromosome at a heterologous site, and a nucleic acid construct that exists as an episome.

[0097] As used herein, the term "peptide" refers to short polypeptides, e.g., usually less than about 50 amino acids in length, more typically less than about 30 amino acids in length. The term, as used herein, encompasses analogs and mimetics that mimic structural and thus biological function.

[0098] The term "polypeptide" encompasses both naturally occurring and non-naturally occurring proteins, as well as fragments, variants, derivatives, and analogs thereof. Polypeptides may be monomeric or polymeric. Furthermore, a polypeptide may contain multiple distinct domains, each of which has one or more distinct activities.

[0099] The term "isolated protein" or "isolated polypeptide" refers to a protein or polypeptide that, based on its origin or source, (1) is not associated with naturally associated components that accompany it in its natural state, (2) is present in a purity not found in nature (e.g., free of other proteins of the same species), (3) is expressed by a cell of a different species, or (4) does not occur in nature (e.g., it is a fragment of a naturally found polypeptide or contains amino acid analogs or derivatives not found in nature, or bonds other than standard peptide bonds). Thus, a polypeptide that is chemically synthesized or synthesized in a cellular system different from the cell from which it is naturally derived is "isolated" from naturally associated components. A polypeptide or protein can also be rendered substantially free of naturally associated components by isolation using protein purification techniques well known in the art. When so defined, "isolated" does not necessarily require that the protein, polypeptide, peptide, or oligopeptide so described has been physically removed from its natural environment.

[0100] The term "polypeptide fragment" refers to a polypeptide having a deletion, e.g., an amino-terminal and / or carboxy-terminal deletion, compared to a full-length polypeptide. In a preferred embodiment, the polypeptide fragment is a contiguous sequence in which the amino acid sequence of the fragment is identical to the corresponding positions in the naturally occurring sequence. A fragment is usually at least 5, 6, 7, 8, 9, or 10 amino acids in length, preferably at least 12, 14, 16, or 18 amino acids in length, more preferably at least 20 amino acids in length, even more preferably at least 25, 30, 35, 40, or 45 amino acids in length, even more preferably at least 50 or 60 amino acids in length, even more preferably at least 70 amino acids in length.

[0101] A protein has "homology" or is "homologous" to a second protein if the nucleic acid sequence encoding the protein has a similar sequence to the nucleic acid sequence encoding the second protein. Alternatively, a protein has homology to a second protein if the two proteins have "similar" amino acid sequences. (Thus, the term "homologous proteins" is defined to mean that two proteins have similar amino acid sequences.) As used herein, homology between two regions of amino acid sequence (especially with respect to predicted structural similarities) is taken to imply similarity in function.

[0102] When "homologous" is used in reference to proteins or peptides, it is recognized that non-identical residue positions often differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is replaced by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution does not substantially change the functional properties of a protein. When two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of homology may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well known to those of skill in the art. See, for example, Pearson, 1994, Methods Mol. Biol. 24:307-31 and 25:365-89 (incorporated herein by reference).

[0103] The twenty conventional amino acids and their abbreviations follow conventional usage. nd ed. 1991). Stereoisomers of the 20 conventional amino acids (e.g., D-amino acids), unnatural amino acids, such as α-,α-disubstituted amino acids, N-alkyl amino acids, and other unconventional amino acids may also be suitable components of the polypeptides described herein. Examples of unconventional amino acids include 4-hydroxyproline, γ-carboxyglutamic acid, ε-N,N,N-trimethyllysine, ε-N-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine, 3-methylhistidine, 5-hydroxylysine, N-methylarginine, and other similar amino acids and imino acids (e.g., 4-hydroxyproline). In the polypeptide notation used herein, the left-hand end corresponds to the amino terminus and the right-hand end corresponds to the carboxy terminus, in accordance with standard usage and convention.

[0104] The following six groups each contain amino acids that are conservative substitutions for one another: 1) serine (S), threonine (T), 2) aspartic acid (D), glutamic acid (E), 3) asparagine (N), glutamine (Q), 4) arginine (R), lysine (K), 5) isoleucine (I), leucine (L), methionine (M), alanine (A), valine (V), and 6) phenylalanine (F), tyrosine (Y), tryptophan (W).

[0105] Sequence homology of polypeptides, sometimes referred to as percent sequence identity, is usually measured using sequence analysis software. See, for example, the Sequence Analysis Software Package of the Genetics Computer Group (GCG), University of Wisconsin Biotechnology Center, 910 University Avenue, Madison, Wis. 53705. Protein analysis software matches similar sequences using measures of homology assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For example, GCG includes programs such as "Gap" and "Bestfit" that can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species, or between a wild-type protein and its mutant protein. See, for example, GCG version 6.1.

[0106] A useful algorithm for comparing a particular polypeptide sequence with a database containing a large number of sequences from different organisms is the computer program BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990); Gish and States, Nature Genet. 3:266-272 (1993); Madden et al., Meth. Enzymol. 266:131-141 (1996); Altschul et al., Nucleic Acids Res. 25:3389-3402 (1997); Zhang and Madden, Genome Res. 7:649-656 (1997)), in particular blastp or tblastn (Altschul et al., Nucleic Acids Res. 25:3389-3402 (1997)).

[0107] Preferred parameters for BLASTp are: expectation: 10 (default), filter: seg (default), gap opening cost: 11 (default), gap extension cost: 1 (default), maximum alignments: 100 (default), word size: 11 (default), number of displays: 100 (default), penalty matrix: BLOWSUM62.

[0108] Preferred parameters for BLASTp are: expectation: 10 (default), filter: seg (default), gap opening cost: 11 (default), gap extension cost: 1 (default), maximum alignment: 100 (default), word size: 11 (default), number of displays: 100 (default), penalty matrix: BLOWSUM62. The length of polypeptide sequences compared for homology is generally at least about 16 amino acid residues, usually at least about 20 residues, more usually at least about 24 residues, typically at least about 28 residues, and preferably more than about 35 residues. When searching a database containing sequences from multiple different organisms, it is preferable to compare amino acid sequences. Database searches using amino acid sequences can be measured by algorithms other than BLASTp known in the art. For example, polypeptide sequences can be compared using FASTA, a program in GCG version 6.1. FASTA provides alignments and percent sequence identity of the best overlapping regions between the query and search sequences. Pearson, Methods Enzymol. 183:63-98 (1990), incorporated herein by reference. For example, percent sequence identity between amino acid sequences can be determined using FASTA with its default parameters (word size 2, and scoring matrix PAM250) as provided in GCG version 6.1, incorporated herein by reference.

[0109] The terms "cultivate" and "cultured" refer to the use of defined techniques for the deliberate growth of fungi or other organisms.

[0110] The term "hyphae" refers to the morphological structure of fungi characterized by a branched, filamentous shape.

[0111] The term "mycelium" refers to the structure formed by one or more masses of branching hyphae. The mycelium is a separate structure distinct from the fruiting body or sporocarp of a fungus.

[0112] The term "cultured mycelial material" refers, in part, to material that includes one or more clumps of cultured mycelium, or that includes only cultured mycelium. As used herein, the term "cultured mycelial material" encompasses composite mycelial material, as defined below.

[0113] The term "composite mycelial material" refers to any mass of cultured mycelial material that has been grown to be intertwined with a second material. In some embodiments, the second material is embedded and / or intertwined within the composite mycelial material. In some embodiments, the second material is disposed on one or more surfaces of the composite mycelial material. Suitable second materials include, but are not limited to, fabrics, masses of continuous, disorganized fibers (e.g., nonwoven fibers), perforated materials (e.g., metal mesh, perforated plastic), masses of discontinuous particles (e.g., wood chips), or any combination thereof. In certain embodiments, the second material is selected from the group consisting of mesh, cheesecloth, cloth, knitted fibers, woven fibers, and nonwoven fibers.

[0114] As used herein, the term "plasticizer" refers to any molecule that interacts with a structure to increase the fluidity of the structure.

[0115] As used herein, the term "treated mycelial material" refers to mycelium that has been post-treated with any combination of treatments with preservatives, plasticizers, finishes, dyes, and / or protein treatments.

[0116] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the subject matter of this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the subject matter of this disclosure, the preferred methods and materials are described herein. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in which the publications are cited in connection therewith.

[0117] Where a range of values ​​is given, it is understood that each intervening value between the upper and lower endpoints of that range, to one-tenth of the unit of the lower endpoint, as well as any other stated or intervening value in that stated range, is encompassed herein unless the context clearly dictates otherwise. The upper and lower endpoints of these smaller ranges may be independently included within the smaller ranges, and are also encompassed herein, subject to any specifically excluded endpoints in the stated range. Where the stated range includes one or both endpoints, ranges excluding either or both included endpoints are also included herein.

[0118] In this specification, certain ranges are presented with numerical values ​​preceded by the term "about". The term "about" is used in this specification to provide literal support for the exact number it precedes, as well as a number that is close to or approximately the number it precedes. In determining whether a number is close to or approximately equal to a specifically stated number, the number that is close to or approximately equal to the unstated number may be a number that, in the context in which it is presented, provides a substantial equivalent to the specifically stated number.

[0119] Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used 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 case of conflict, the present specification, including definitions, will control. The materials, methods, and examples are illustrative only and are not intended to be limiting.

[0120] overview Provided herein are compositions and scalable methods for post-treating mycelial and / or composite mycelial materials. In some or most embodiments, the mycelial and / or composite mycelial materials are post-treated prior to processing to form preserved mycelial materials.

[0121] Exemplary patents and applications discussing methods of growing mycelium include: WIPO Patent Publication No. 1999 / 024555, GB Patent No. 2,148,959, GB 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. Additionally, U.S. Patent Publication No. 2018 / 0282529, filed October 4, 2018, discusses various mechanisms of solution-based post-treatment of mycelial material to produce materials with favorable mechanical properties for processing into fabrics or leather substitutes.

[0122] The description of an embodiment with several components in communication with each other does not imply that all such components are required. Conversely, various optional components may be described to illustrate a wide variety of possible embodiments and to more fully describe one or more aspects. Similarly, although process steps, method steps, algorithms, and the like may be described in sequence, such processes, methods, and algorithms may generally be configured to occur in another order unless otherwise specified. In other words, any sequence or order of steps that may be described herein does not in itself indicate a requirement that the steps occur in that order. The described process steps may be performed in any practical order. Furthermore, some steps may occur simultaneously, even though they are described or implied not to occur simultaneously (e.g., even if one step is described after another step). Furthermore, the description of steps in the drawings by their depiction does not imply that the described steps are exclusive of other variations and modifications, does not imply that the described steps or any of their steps are required for one or more embodiments, and does not imply that the described steps are preferred. Also, although steps are generally described once in an embodiment, this does not mean that they must or can only be performed once each time the process, method, or algorithm is performed or executed. Some steps may be omitted in some embodiments or some occurrences, and some steps may be performed multiple times in a given embodiment or occurrence.

[0123] Cultivation of mycelial material The embodiments of the present disclosure include various compositions of cultured mycelial material and methods for its manufacture. Depending on the particular embodiment and the requirements of the desired material, various known methods of culturing mycelium may be used. Any fungus that can be cultured as a mycelium may be used. Suitable fungi for use include Pleurotus ostreatus, Agrocybe brasiliensis, Polyporus squamosus, Rhizopus microspores, Schizophyllum commune, Flammulina velutipes, Hypholoma capnoides, Hypholoma sublaterium, Morchella angusticeps, Macrolepiota procera, Coprinus comatus, Agaricus arvensis, Ganoderma lucidum, and others. Examples of suitable fungi include, but are not limited to, Ganoderma tsugae, Ganoderma sessile, and Inonotus obliquus.

[0124] In some embodiments, fungal strains or species can be bred to produce mycelium with specific characteristics, such as a dense network of hyphae, a highly branched network of hyphae, hyphal fusion within the hyphal network, and other characteristics that can alter the material properties of the cultured mycelial material. In some embodiments, fungal strains or species can be genetically engineered to produce mycelium with specific characteristics.

[0125] In most embodiments, the cultured mycelium material may be grown by first inoculating a solid or liquid substrate with an inoculum of mycelium from a selected fungal species. In some embodiments, the substrate is pasteurized or sterilized prior to inoculation to prevent contamination or competition with other organisms. For example, a standard method for culturing mycelium includes inoculating a sterilized solid substrate (e.g., cereal) with an inoculum of mycelium. Another standard method for culturing mycelium includes inoculating a sterilized liquid medium (e.g., liquid potato dextrose) with an inoculum of mycelium. In some embodiments, the solid and / or liquid substrate includes lignocellulose as a carbon source for the mycelium. In some embodiments, the solid and / or liquid substrate includes simple or complex sugars as a carbon source for the mycelium.

[0126] In various embodiments, the liquid or solid substrate may be supplemented with one or more different nutrient sources. The nutrient sources may include lignocellulose, simple sugars (e.g., dextrose, glucose), complex sugars, agar, malt extract, nitrogen sources (e.g., ammonium nitrate, ammonium chloride, amino acids), and other minerals (e.g., magnesium sulfate, phosphate). In some embodiments, one or more nutrient sources may be included in wood waste (e.g., sawdust) and / or agricultural waste (e.g., livestock manure, straw, corn stover).

[0127] After the substrate is inoculated and optionally supplemented with one or more different nutrient sources, the cultured mycelial material and / or composite mycelial material may grow to some extent. In embodiments producing composite mycelial material, the inoculated substrate may form part of the composite material, such as the particles described in U.S. Pat. No. 9,485,917. In some embodiments, the cultured mycelial material may grow through a second material that intertwines with the mycelium to form a composite material. Various methods of growing a network of cultured mycelial material that intertwines with another material to form a composite material are disclosed in U.S. Pat. No. 9,485,917, U.S. Patent Publication No. US2016 / 0302365, and U.S. Patent Publication No. US2013 / 0263500, the entireties of which are incorporated herein by reference.

[0128] In various embodiments, the cultured mycelium material can grow alone without a second material. In some embodiments, the growth of the mycelium material is controlled to prevent fruit body formation. Various methods of preventing fruit body formation are discussed in detail in US Patent Publication No. US2015 / 0033620, which is incorporated by reference in its entirety. In other embodiments, the cultured mycelium material can be grown such that there is no morphological or structural change in the cultured mycelium material. Depending on the embodiment required, growth conditions, such as exposure to light (e.g., sunlight or growth lamp), temperature, carbon dioxide, can be controlled during the course of growth.

[0129] In some embodiments, the cultured mycelium material can be grown on agar medium. Nutrients can be added to the agar / water base. Standard agar media commonly used for culturing mycelium include, but are not limited to, enriched malt extract agar (MEA), potato dextrose agar (PDA), oatmeal agar (OMA), and dog food agar (DFA).

[0130] Preservation of mycelial material After the cultured mycelium material has grown, it may be separated from the substrate and optionally post-treated to kill the mycelium or to render the mycelium non-rotting, thereby preventing further growth (referred to herein as "preserved mycelium material"). Suitable methods for producing preserved mycelium material may include drying or dehydrating the cultured mycelium material (e.g., compressing the cultured mycelium material to remove water) and / or heat treating the cultured mycelium material. In a particular embodiment, the cultured mycelium material is compressed at 190,000 pounds force for 30 minutes to 0.25 inches. In another embodiment, the cultured mycelium material is compressed for 5 minutes to 0.25 inches. Suitable methods for drying organic matter to render it non-rotting are well known in the art. In one particular embodiment, the cultured mycelium material is dried in an oven at a temperature of 100° F. or higher. In another particular embodiment, the cultured mycelium material is heat compressed. Various post-treatment methods, including heat and pressure, are disclosed in U.S. Patent Publication Nos. 2017 / 0028600 and 2016 / 0202365, which are incorporated by reference in their entireties.

[0131] In some cases, to produce preserved mycelial material, the cultured mycelial material is treated with one or more agents known to convert the chitin contained in the mycelium to chitosan and / or to add functional groups to the chitin. In various embodiments, the chitin contained in the mycelium (or the chitin that has been converted to chitosan) may be treated with an alkaline solution, an epoxide reagent, an aldehyde reagent, a cyclodextrin reagent, graft polymerization, chelating chemistry, a carboxymethyl reagent, an epoxide reagent, a hydroxyl alkyl reagent, or any combination thereof. Specific examples of these chemistries are disclosed in U.S. Pat. No. 9,555,395, the majority of which is incorporated herein by reference. After functionalization of the chitin, various agents may be used to crosslink the chitin. Depending on the functionalization of the chitin groups, conventional tanning agents, including chromium, vegetable tannins, tanning oils, epoxies, aldehydes, and syntans, may be used to attach functional groups. Due to toxicity and environmental concerns of chromium, other minerals used in tanning, such as aluminum, titanium, zirconium, iron and combinations thereof with or without chromium, may also be used.

[0132] In other examples, fresh or dried cultured mycelium material is treated with one or more solutions that function to remove waste and water from the mycelium. In some embodiments, the solution includes a solvent such as ethanol, methanol, or isopropyl alcohol. In some embodiments, the solution includes a salt such as calcium chloride. In some embodiments, the cultured mycelium material may be submerged in the solution for various periods of time, with or without pressure. In some embodiments, the cultured mycelium material may be submerged in several solutions sequentially. In certain embodiments, the cultured mycelium material may be submerged first in one or more first solutions including alcohol and salt, and then in a second solution including alcohol. In another particular embodiment, the cultured mycelium material may be submerged first in one or more first solutions including alcohol and salt, and then in a second solution including water. After treatment with the solutions, the cultured mycelium material may be compressed using a high or low temperature process, and / or dried using various methods, including air drying and / or vacuum drying. U.S. Patent Publication No. 2018 / 0282529, which is incorporated by reference in its entirety, describes these embodiments in detail.

[0133] Plasticization of cultured mycelial material Various plasticizers may be applied to cultured mycelium material to change the mechanical properties of the cultured mycelium material. US Patent No. 9,555,395 discusses the addition of various humectants and plasticizers. Specifically, US Patent No. 9,555,395 discusses the use of glycerol, sorbitol, triglyceride plasticizers, oils such as linseed oil, drying oils, ionic and / or non-ionic glycols. US Patent Publication No. 2018 / 0282529 further discusses treating solution-processed mycelium material with plasticizers such as glycerol, sorbitol or other humectants to retain moisture and to increase the mechanical properties of the cultured mycelium material, such as the elasticity and flexibility of the cultured mycelium material.

[0134] Fatliquors are well known in the art, which are obtained by emulsifying other similar plasticizers and humectants, such as polyethylene glycol, and natural oils with an oil-immiscible liquid (such as water) so that microdroplets of the oil penetrate the material. Various fatliquors contain oil emulsified in water, with other compounds added, such as ionic and nonionic emulsifiers, surfactants, soaps, and sulfates. Fatliquors can contain various types of oils, such as mineral oils, animal oils, and vegetable oils.

[0135] Tanning and dyeing of cultured mycelium material In various embodiments, it may be desirable to impart color to cultured or preserved mycelial material. As described in U.S. Patent Publication No. 2018 / 0282529, tannins may be used to impart color to cultured or preserved mycelial material.

[0136] Because cultured mycelial material contains some chitin, it lacks functional sites that are abundant in protein-based materials. Therefore, it may be necessary to functionalize the chitin in the cultured mycelial material to create binding sites for acid and direct dyes. Methods for functionalizing chitin are discussed above.

[0137] A variety of dyes may be used to impart color to the cultured mycelium material, including acid dyes, direct dyes, disperse dyes, sulfur dyes, synthetic dyes, pigments, and natural dyes. In some embodiments, the cultured mycelium material is submerged in an alkaline solution prior to application of the dye solution to promote exhaustion of the dye and penetration into the material. In some embodiments, the cultured mycelium material is pre-soaked in ammonium chloride, ammonium hydroxide, and / or formic acid prior to application of the dye solution to promote exhaustion of the dye and penetration into the material. In some embodiments, tannins may be added to the dye solution. In various embodiments, the cultured mycelium material may be optionally stored as discussed above prior to dye treatment or pretreatment.

[0138] In some embodiments, the dye solution may be applied to the cultured mycelial material using different application techniques. In some embodiments, the dye solution may be applied to one or more exterior surfaces of the cultured mycelial material. In other embodiments, the cultured mycelial material may be submerged in the dye solution.

[0139] In addition to presoaking with various solutions, agents may be added to the dye solution to promote dye exhaustion and penetration into the material. In some embodiments, ammonium hydroxide and / or formic acid along with acid or direct dyes promote dye exhaustion and penetration into the material. In some embodiments, ethyloxylated fatty amines are used to promote dye exhaustion and penetration into the treated material.

[0140] In various embodiments, a plasticizer is added after or during the addition of the dye. In various embodiments, the plasticizer can be added together with the dye solution. In certain embodiments, the plasticizer can be coconut oil, vegetable glycerin, or a sulfited or sulfated fatliquor.

[0141] In some embodiments, the dye solution may be maintained at a basic pH using a base such as ammonium hydroxide. In certain embodiments, the pH is at least 9, 10, 11, or 12. In some embodiments, the pH of the dye solution is adjusted to an acidic pH using various agents such as formic acid to fix the dye. In certain embodiments, the pH is adjusted to less than pH 6, 5, 4, or 3 to fix the dye.

[0142] In various methods, the cultured and / or preserved mycelial material may be subjected to mechanical processing or agitation while the dye solution is being applied to enhance dye exhaustion and penetration into the material. In some embodiments, the cultured and / or preserved mycelial material may be subjected to squeezing or other forms of pressure while in the dye solution to enhance dye exhaustion and penetration. In some embodiments, the cultured and / or preserved mycelial material may be subjected to ultrasonic treatment.

[0143] Using the methods described herein, the cultured mycelial material may be dyed or colored such that the color of the treated mycelial material is substantially uniform. Using the methods described above, the cultured mycelial material may be dyed or colored such that the dye and color are not only present on the surface of the cultured mycelial material, but instead penetrate through the surface to the core of the treated mycelial material.

[0144] In various embodiments, the cultured mycelium material can be dyed so that the cultured mycelium material does not discolor. Color fastness can be measured using various techniques, such as ISO 11640:2012: Color Fastness Test, i.e., color fastness to cycles of double rubbing or ISO 11640:2018, which is a revision of ISO 11640:2012. In certain embodiments, color fastness is measured using a grayscale rating as a metric to identify rub fastness and sample changes, as described above. In some embodiments, the mycelium exhibits high color fastness, as indicated by a grayscale rating of at least 3, at least 4, or at least 5.

[0145] Treatment of cultured mycelial material with a protein source In various embodiments, it may be beneficial to treat the cultured mycelial material with one or more protein sources that are not naturally present in the mycelium (i.e., exogenous protein sources). In some embodiments, the one or more proteins are from a species other than the fungal species from which the cultured mycelial material is generated. In some embodiments, the cultured mycelial material may be treated with a plant protein source, such as pea protein, rice protein, hemp protein, and soy protein. In some embodiments, the protein source is an animal protein, such as an insect protein or a mammalian protein. In some embodiments, the protein is a recombinant protein produced by a microorganism. In some embodiments, the protein is a fibrous protein, such as silk or collagen. In some embodiments, the protein is an elastic protein, such as elastin or resilin. In some embodiments, the protein has one or more chitin-binding domains. Exemplary proteins with chitin-binding domains include resilin and various bacterial chitin-binding proteins. In some embodiments, the protein is a modified or fusion protein with one or more chitin-binding domains. In some embodiments, the cultured mycelial material may be stored as described above prior to treatment, or may be treated without prior storage.

[0146] In certain embodiments, the cultured mycelium material is submerged in a solution comprising the protein source. In certain embodiments, the solution comprising the protein source is aqueous. In other embodiments, the solution comprising the protein source comprises a buffer, such as phosphate buffered saline.

[0147] In some embodiments, the solution containing the protein source includes an agent that functions to cross-link the protein source. In some embodiments, various known agents that interact with amino acid functional groups can be used. In certain embodiments, the agent that functions to cross-link the protein source is transglutaminase. Other suitable agents that cross-link amino acid functional groups include tyrosinase, genipin, sodium borate, and lactase. In other embodiments, proteins can be cross-linked using traditional tanning agents, including chromium, vegetable tannins, tanning oils, epoxies, aldehydes, and syntans. As mentioned above, due to the toxicity and environmental concerns of chromium, other inorganics, such as aluminum, titanium, zirconium, iron, and combinations thereof with or without chromium, may be used.

[0148] In various embodiments, the treatment with a protein source may occur before, after, or simultaneously with the storage of the cultured mycelial material, the plasticization of the cultured mycelial material, and / or the dyeing of the cultured mycelial material. In some embodiments, the treatment with a protein source may occur before or during the storage of the cultured mycelial material using a solution containing alcohol and salt. In some embodiments, the treatment with a protein source occurs before or simultaneously with the dyeing of the cultured mycelial material. In some of these embodiments, the protein source is dissolved in the dye solution. In certain embodiments, the protein source is dissolved in a basic dye solution that includes one or more agents that promote dye exhaustion.

[0149] In some embodiments, a plasticizer is added to the dye solution containing the dissolved protein source, and simultaneously plasticizes the treated mycelium material. In certain embodiments, the plasticizer can be a fatliquor. In certain embodiments, the plasticizer is added to the protein source dissolved in the basic dye solution containing one or more agents that promote dye exhaustion.

[0150] Coating and finishing of cultured mycelium material After the cultured mycelium material is treated using any combination of the above plasticization, protein treatment, preservation and tanning, the cultured mycelium material may be treated with a finish or coating. Various finishes common to the leather industry may be used, such as binder solutions containing proteins, nitrocellulose, synthetic waxes, natural waxes, waxes containing protein dispersions, oils, polyurethanes, acrylic polymers, acrylic resins, emulsion polymers, water-resistant polymers and various combinations thereof. In a particular embodiment, a finish containing nitrocellulose may be applied to the cultured mycelium material. In another particular embodiment, a finish containing a conventional polyurethane finish may be applied to the cultured mycelium material. In various embodiments, one or more finishes are applied to the cultured mycelium material in sequence. In some cases, the finish is mixed with a dye or pigment. Optionally, the finish is mixed with a hand feel modifier (i.e., feel modifier or touch) including one or more of natural and synthetic waxes, silicones, paraffins, saponified fatty substances, amides of fatty acids, amide esters, stearamides, emulsions thereof, and any combination of the foregoing. Optionally, the finish is mixed with a defoamer.

[0151] Mechanical processing of materials in solution and after post-treatment In various embodiments, the cultured mycelium material can be mechanically processed in different ways both while in solution (i.e., dye solution, protein solution, or plasticizer) and after the cultured mycelium material is removed from the solution.

[0152] While the cultured mycelium material is in solution, it may be agitated, sonicated, squeezed or compressed to ensure uptake of the solution. The degree of mechanical processing depends on the specific treatment being applied and the level of friability of the cultured mycelium material at that stage of processing. Squeezing or compressing the cultured mycelium material may be done by hand squeezing, mechanical squeezing, platen pressing, lino rollers or calendar rollers.

[0153] Similarly, as noted above, the cultured mycelium material may be compressed or otherwise processed to remove solution from the cultured mycelium material after it has been removed from the solution. Treatment of the material with solution and compression may be repeated several times.

[0154] After the cultured mycelium material is sufficiently dried (e.g., by using heat, compression or other dehydration techniques as mentioned above), the cultured mycelium material may be subjected to further mechanical processing. Depending on the technique used to process the cultured mycelium material and the toughness of the cultured mycelium material obtained, different types of mechanical processing may be applied, including but not limited to sanding, brushing, plating, staking, tumbling, vibrating and cross-rolling. The cultured mycelium material may be embossed with any heat source or through the application of chemicals.

[0155] In some embodiments, the composite mycelial material may be embossed with any heat source or through the application of chemicals. In some embodiments, the composite mycelial material in solution may be subjected to further chemical treatment, for example, may be maintained at a basic pH using a base such as ammonium hydroxide. In certain embodiments, the pH is at least 9, 10, 11 or 12. In some embodiments, the pH of the solution containing the composite mycelial material is adjusted to an acidic pH using various agents such as formic acid to fix the composite mycelial material. In certain embodiments, the pH is adjusted to less than pH 6, 5, 4 or 3 to fix the composite mycelial material.

[0156] Finishing, coating and other steps may occur after or before mechanical processing of the dried cultured mycelium material. Similarly, final compression steps, including embossing steps, may occur after or before mechanical processing of the dried cultured mycelium material.

[0157] Mechanical properties of post-treated mycelial materials The various methods described herein may be combined to provide treated mycelial materials with different mechanical properties.

[0158] In various embodiments, the treated mycelium material may have a thickness of less than 1 inch, less than ½ inch, less than ¼ inch, or less than ⅛ inch. The thickness of the material within a given piece of material may have various coefficients of variation. In some embodiments, the thickness is substantially uniform, resulting in a minimal coefficient of variation.

[0159] In some embodiments, the treated mycelial 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 treated mycelial 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. In some embodiments, the treated 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%. In some embodiments, the initial modulus, ultimate tensile strength, and elongation at break are measured using ASTM D2209 or ASTM D638. In certain embodiments, the initial modulus, ultimate tensile strength, and elongation at break are measured using a modified ASTM D638 using the same sample dimensions as ASTM D638 at the strain rate of ASTM D2209.

[0160] In some embodiments, the treated mycelium material may have a double stitch 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 certain embodiments, the tongue tear strength is measured by ASTM D4705.

[0161] In some embodiments, the treated mycelium material may have a single stitch 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 certain embodiments, the tongue tear strength is measured by ASTM D4786.

[0162] In some embodiments, the treated mycelial material may have a tongue 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 100 N, at least 150 N, or at least 200 N. In certain embodiments, the tongue tear strength is measured by ASTM D4704.

[0163] In some embodiments, the treated mycelium material may have a flexural modulus (deflection) 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 certain embodiments, the compression is measured by ASTM D695.

[0164] In various embodiments, the treated mycelium material has different absorption properties measured as a percentage of weight gain after immersion in water. In some embodiments, the weight gain % 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 certain embodiments, the weight gain % after immersion in water for 1 hour is measured using ASTM D6015.

[0165] Method for producing cultured mycelium Provided herein are methods that include: generating cultured mycelium material; contacting the cultured mycelial material with a solution comprising one or more proteins to produce a composition comprising the cultured mycelial material and one or more proteins, the one or more proteins being derived from a species other than the fungal species from which the cultured mycelial material was produced; and compressing said cultured mycelium material.

[0166] In some embodiments, the method comprises submerging the cultured mycelial material in the solution, hi some embodiments, the contacting step comprises contacting the cultured mycelial material with the solution in a single step.

[0167] Exemplary Products Using Mycelial Material It is to be understood that the growing, processing, and processing steps applied to the above mycelium in various combinations (including those specifically discussed above and those that may be apparent or derived based on the above) are generally derived or adapted to produce materials that resemble leather. To that end, such processing steps may be applied to produce specific mycelium-based materials that have characteristics or properties similar to those of leather, including various types of leather (such as tactile, visual, and physical, as described in more detail herein), or that have various known characteristics or attributes. In this manner, the mycelium-based materials can be cultured, preserved, plasticized, tanned, dyed, protein-treated, coated, finished, or post-treated according to the steps and variations thereof described herein, and in various combinations, to produce raw materials that can be manufactured or assembled into different products that are typically, or in various forms, primarily leather or that otherwise characterize or include leather. In certain forms and compositions, the mycelium-based material may result in products or articles that meet or exceed consumer, retailer, or manufacturer expectations for similar leather or leather-containing products, including being amenable to or usable with the same or similar processing, assembly, and manufacturing techniques used in working with leather. In other aspects, the methods of cultivating and protein treating mycelium material may allow, among other things, the breeding, modification, or selection of fungi and the use of specific liquid and solid substrates, nutrient sources, entanglement materials, and the like, and the proteins for treatment may allow for the controlled production of mycelium with specific properties that provide improved workability or manufacturability over conventional leather, including being amenable to further assembly, manufacturing, or finishing techniques. In this manner, such products comprised of, using, or incorporating various types of mycelium material that may be produced according to or in light of the above description may provide consumers and manufacturers with benefits beyond those possible with conventional leather, in addition to the ecological, environmental, and humanitarian benefits that may be realized by substituting the mycelium material described herein for leather.

[0168] Use of mycelium materials in footwear According to the above description, in one example, the mycelium material described herein can be used in various types and forms of footwear, including as a replacement for leather, which is used in various forms for substantially all parts of at least some types of footwear. In various forms, the mycelium material described herein can be used in all or part of the upper of many types of shoes. Furthermore, dress shoes and the like usually include an insole (e.g., at the top, foot-contacting, surface) that is made only of leather or includes leather, and in some applications, the welt, midsole, and outsole (including at least the forefoot portion) may also be made of leather. In any of these cases, the leather can be replaced with a specific implementation of the mycelium material described herein that has the required characteristics and is assembled or manufactured accordingly into the desired form. Similarly, either or both of the outsole and upper of various types of slippers may be made with the mycelium material of the present invention, for example, replacing leather, and either or all of the upper, outsole, laces, and at least some stitching of moccasins or deck shoes may be made with the mycelium material of the present invention.

[0169] With reference to the embodiment shown in FIG. 1, reference numeral 10 generally denotes a shoe, particularly in the form of an athletic sneaker. In particular, the terms "athletic" and "sneaker," as discussed herein, whether used alone or in combination with a particular type or style of footwear, in no way imply or require that such footwear is strictly used or usable for any type of athletic activity or athletic competition. In this regard, a footwear product may be merely of or reminiscent of the style or construction of athletic footwear, so as to encompass such footwear, whether or not used or intended for athletic activity (e.g., athleisure or fashion footwear shaped like or similar to athletic sneakers, or other variations of athletic footwear, as described below). Furthermore, the description made herein, including with reference to the drawings, is merely exemplary with respect to the footwear described and illustrated, and furthermore, modifications may be made to the footwear described herein for purposes of style and fit, and / or to create footwear based on the principles and constructions described herein suitable for various purposes or conditions. Additionally, although construction and manufacturing techniques may be discussed herein with respect to a particular style of footwear (e.g., athletic sneakers), such construction and manufacturing techniques discussed with respect to one type of footwear may be acceptable alternatives for equivalent construction and manufacturing techniques discussed herein with respect to other types of footwear (e.g., hiking boots, sandals (including sport sandals), etc.).

[0170] 1, the illustrated athletic sneaker 10 is illustrative of a typical structure for an athletic sneaker and includes an upper 12, a midsole 14, and an outsole 16, with the upper 12 generally defining an interior 18 adapted to receive a wearer's foot, and the outsole 16 forming the portion of the athletic sneaker 12 that contacts the ground beneath the wearer's foot. In this regard, the structure of the depicted athletic sneaker 10 is generally specific to other types of footwear, although it is noted that the integrated midsole 14 and outsole 16 may be collectively referred to as the "outer" of the footwear, and other than the depicted midsole 14 and outsole 16 may be used in a variety of forms. In one example, the outer may be comprised of a midsole material (e.g., compression molded ethyl vinyl acetate ("EVA") that exhibits both acceptable cushioning and resilience) that may be formed into the midsole material, at least in the portion of the ground contact surface that is typically included in a separate outsole. Similarly, the outer can include a rubber outsole 16 and can be used alone, without a cushioned midsole, for applications such as athletic footwear commonly referred to as "barefoot" style running shoes. Such variations are considered to be within the scope of this disclosure, and the depicted examples are modified accordingly. As shown in the example of FIG. 1, the midsole 16 is disposed between the upper 12 and the outsole 16, and serves to support and cushion the sole of the foot, particularly against impact with the ground provided by the outsole 16. As shown in FIG. 2, the interior 18 of the upper 12 is generally surrounded at its lower portion by a lasting board 24, and the upper 12 is attached to the lasting board 24 around or adjacent to the lower perimeter 22 of the upper 12 (depending on the particular construction method, as discussed further below). The lasting board 24 and / or the portion of the upper 12 adjacent to the perimeter 22 are then attached to the midsole 14 by placing the lasting board 24 on the midsole 14. As shown in FIG. 3, an insole 24 can be disposed within the interior 18, over the lasting board 24.The insole 20 may be at least somewhat cushioned to provide additional comfort to the user and to cover the stitching used to attach the lasting board 24 around the perimeter 22. In one embodiment, the insole 20 may also include mycelium material. This may be done by constructing the insole 20 entirely from mycelium material, or by covering a cushioned layer of foam with a thin layer of mycelium material, such that the top, foot-contacting surface of the insole 20 is made of mycelium material.

[0171] As seen in Figures 1 and 2, the described athletic sneaker 10 is an example of a sneaker, and in particular an upper 12, manufactured using a "cut and sew" process in which the upper 12 is assembled from several individual pieces of raw material corresponding to the various portions of the upper 12. In particular, the individual pieces are cut from a flat two-dimensional shape of raw material as necessary according to the desired final form of the upper 12, and are sewn together along various seams that at least partially give the upper 12 a desired three-dimensional shape. Such sewing may be augmented by the use of various adhesives along the seams, and may be performed on the entire shoe last or on a portion corresponding to the desired shape of the interior 18 of the upper 12. In particular, the lasting board 24 is typically sewn to the upper 12 on the last, and for the conventional construction of the depicted athletic sneaker 10, and similar footwear, is finished using a "strobel" stitch using a special machine that joins the material portions of the upper 12 that define the perimeter 22 with the lasting board 24 at adjacent edge-to-edge seams. The resulting "strobel sock" including the assembled upper 12 and lasting board 24 is then attached to the midsole 14. In most cases, this is done using adhesives or the like. In some constructions, the attachment between the lasting board 24 and the midsole 14 may be augmented or finished using seams such as break stitches or seams along specific areas of the upper 12 related to features associated with the midsole 14, as discussed further below. Generally, the midsole 14 is made of a foam material and may be made of a number of different foam materials, including various densities of EVA, or various inserts including plastics and the like. The outsole 16 may be formed from one or more sections of rubber (such as various synthetic rubbers) that are glued, bonded, or otherwise joined to the midsole 14, at least in areas where ground contact is made and / or where grip or durability is desired.

[0172] With regard to the assembly of the cut-and-sew upper 12, the pieces and portions of the upper 12 generally correspond to the particular regions of the upper 12 described above, but may vary depending on their particular shape and arrangement according to the desired style appearance of the athletic sneaker 10, as well as the desired fit, flexibility, and support of the athletic sneaker 10, which may be influenced or dictated by the intended use of the athletic sneaker. In the exemplary depiction of Figures 1 and 2, the various portions of the upper 12 may include a toe tip 26 and a vamp 28 that extends upwardly from the toe tip 26 to a throat 30 of the athletic sneaker 10. A tongue 32 extends upwardly from the vamp 28 along the throat 30, and opposing medial and lateral quarters 34a and 34b extend rearwardly from the toe tip 26 and define portions of the lower perimeter 22 along each side of the upper 20 and downwardly from the throat 30. A heel counter 36 extends around the rear of the upper and connects between the two quarters 34a and 34b to surround the heel of the wearer. Additionally, medial and lateral collar portions 38a and 38b may extend upwardly from the heel counter 36 and rearwardly from the respective medial and lateral quarters 34 to define respective portions of a topline 40 of the upper 12. A heel tab 42 is disposed above the heel counter and connects between the rearmost ends of the respective collar portions 38a and 38b to define the rear of the topline 40. An inner liner 44 (FIG. 3) may extend through all or a portion of the upper 12 to define the interior 18 thereof and may be attached to the respective lateral portion of the upper 12 through which it extends.

[0173] As noted above, the shapes and configurations of the above portions of the upper are merely exemplary and may be modified to achieve different appearances, as well as different fit and performance characteristics (flexibility, support, weight, etc.). In one embodiment, all or a portion of the depicted collar portions 38a and 38b may be integral with the respective quarters 34a and 34b. Additionally, the toe tip 26 may be integral with one or both of the quarters 34a and 34b, or may itself be formed into one or more portions (e.g., extending separately from the respective quarters 34a and 34b), and the vamp 24 may also be integral with the toe tip 26. In such a construction, additional portions may be assembled with the vamp 24 and / or toe tip 26 (e.g., foxing) to cover various seams and / or provide additional support, protection, or style effects (e.g., bicycle toe, etc.). In still further variations, collar portions 38a and 38b and / or heel tab 42 may extend upward relative to the depicted features (or additional portions may be added over existing portions), thereby raising the topline 40 to the level of a mid-top or high-top sneaker (i.e., at or above the wearer's ankles) for further support or protection of the wearer and / or for aesthetic purposes.

[0174] 3, additional components may be added between the exterior portion of the upper 12 and the liner 44. In particular, a collar lining 46 (which may include or be bonded with padding) may be attached to portions of the collar portions 38a and 38b, the heel tab 42, and the quarters 34a and 34b (if applicable) and may be wrapped inwardly over portions of the liner 44 to provide a finished appearance while also providing padding or grip around the topline 40 that may be beneficial to the wearer. Similarly, additional lining or padding may be added to the interior of the tongue 32 to more evenly distribute the force of the laces 50 used to tie the quarters 34a and 34b together to close the throat 30 on the foot.

[0175] In accordance with the above, the upper 12 may be made entirely or in part using one or more specific implementations of the mycelium material described above. As described above, the cultivation, preservation, plasticization, tanning, dyeing, protein treatment, coating, finishing, and post-treatment steps may be individually tailored and combined together in various ways to achieve properties that are particularly suitable for use in the athletic sneaker 10 depicted and described. In some respects, such properties may enable the mycelium material to mimic or meet the expectations of the leather material from which sneakers of the depicted type were originally manufactured and from which the construction and assembly techniques of such sneakers originate, as described above. In particular, in many cases, leather is already increasingly being replaced by other materials, including woven or knitted fabrics, synthetic leather or suede, various polymeric sheet materials, and combinations thereof. The use of such materials may provide certain cost advantages over leather (including due to availability), as well as various manufacturing advantages, including the ability to create the upper, or portions thereof, in a more seamless manner using material properties or available manufacturing techniques (e.g., so-called three-dimensional weaving or knitting techniques, which allow for variations in materials and patterns, as well as shapes). Some synthetic materials may also be formable or adaptable in ways that traditional leather is not. In other aspects, synthetic and textile (including synthetic and natural textile) materials may compromise or reduce the support or durability of sneakers made from such materials compared to those made from leather. Additionally, the appearance and tactile qualities of leather may be preferred by consumers in many athletic sneaker (and other footwear) implementations. In this manner, the mycelium material of the present invention can be used in place of leather, and even in place of synthetic materials and fabrics (in whole or in part), to address the various availability (and potentially cost) and ecological issues that exist with respect to leather, as well as the preference, support, and durability of synthetic and fabric materials, particularly when used in the manufacture of sneakers, including the depicted athletic sneaker 10.This may, in some cases, make the mycelium material of the present invention suitable for use in the manufacture of so-called "retro" sneakers that may recall or be directly based on certain sneaker designs of traditional leather. Similarly, implementations of the mycelium material of the present invention may be used in other applications where the properties of leather are preferred, including for activities where the durability and support of leather are advantageous or where the appearance of leather is also desired.

[0176] Thus, in one example, the athletic sneaker 10 shown in Figures 1-3 may be such that the upper 12 is manufactured, in whole or in major part, from the mycelium material of the present invention using construction and assembly techniques used to manufacture shoe uppers made of leather or primarily made of leather. In this regard, the various portions of the upper 12 described above, including the toe tip 26, vamp 28, quarters 34a and 34b, heel counter 36, collar portions 38a and 38b, and heel tab 42 (all of which are depicted in Figures 1-3 and modified for the above purposes within the scope of this disclosure), may be cut into desired shapes from flat sheets of raw mycelium material of desired composition and sewn together along seam lines at the interfaces between adjacent portions of the cut-out pieces of material to give the upper 12 its desired shape. In particular, the cut-out mycelium material may be joined by top stitching 52 along seams defined by overlapping portions of the material. At the location of the topstitch 52, the raw edge of the mycelium material generally meets along the respective cuts of the top / outermost pieces, and the topstitch 52 meets as well, which may be doubled or tripled along at least a portion of the seam for added durability or decorative effect. It is noted that in areas where additional tolerances or tolerances are required, a felt seam secured by one or more topstitches (such as a lap or top felt seam) may be used. Where both the raw edge and the seam are to be hidden (or adjacent parts are to be joined adjacently), a backstitch seam 54 may be used. As shown, the inner and outer quarters 34a and 34b may be joined by a backstitch seam. Similarly, the collar portions 38a and 38b and the heel tab 42 (and optionally the quarters 34a and 34b) may be joined to the collar lining 46 by a backstitch seam. Additionally, the tongue 32 can be made from the mycelium material of the present invention and can be joined to the lining by a backstitch seam, where the tongue 32 and tongue lining can be sewn together along their outer edges and top ends (and optionally any additional padding on the outside of the liner) with the desired outer surfaces facing each other.The assembled tongue 32 and liner 48 can then be inverted to expose the outer surface and encased in padding before being assembled with the vamp and / or quarters 34a and 34b (if applicable) using topstitching 52.

[0177] In some respects, the properties of mycelium, which are generally comparable to leather, allow the assembly to be completed using techniques with parameters and equipment similar or comparable to those used to assemble sneaker uppers made of leather, resulting in a similar appearance and efficiency using established techniques and existing machinery. In this manner, the cut pieces of mycelium material may have further processing steps performed on them to reduce the thickness of the material before stitching, resulting in a cleaner appearance and easier completion of the backstitch seam 54, or the incorporation of any felt stitches into the upper 12. Such skiving may include compressing or cutting the material at the desired seam edges, and may be completed using machinery used to skive the edges of leather. In addition to the typical assembly stitches 52 and 54 shown in Figures 1-3, the pieces of mycelium material may be embroidered before or after assembly of the upper 12. In one example, eyelets 56 on the upper through which the laces pass may benefit from the additional reinforcement that may be provided by such embroidery 58 around or through eyelets 56a. Additional structural embroidery (such as to surround or attach additional structural members such as metal or plastic strips) may also be used along quarters 34a and 34b, and decorative embroidery (i.e., stitching not associated with a seam), including logos or other identifying or identifying information, may be provided elsewhere on the upper, including, but not limited to, heel tab 42, tongue 32, heel counter 36, and quarters 34a and 34b.

[0178] Similarly, the mycelium material may be suitable for other processing and assembly techniques used for leather, which may be useful in the manufacture of the athletic sneaker 10 of the present invention. In particular, during or after the tanning process described above, the mycelium material may be split and portions of the leather corresponding to the "top grain" may be removed to provide a mycelium material that resembles suede, exhibiting comparable tactile and material properties, a more supple yet rough feel, and greater flexibility than leather. Similarly, the mycelium material may be sanded, buffed, or stamped to resemble nubuck leather (in appearance and various material properties), or tanned or dyed with soluble materials to resemble aniline leather. In various examples, the vamp 28, outer quarter 34a, and collar portions 38a and 38b may be made from split mycelium material that resembles suede, providing increased flexibility and comfort in areas where less support may be needed. Similarly, the tongue liner 48 and collar liner 46 may be made from a split mycelium material that resembles suede to improve grip and / or flexibility. In another example, the plasticization process can be tailored and applied to split the mycelium material (and optionally embossed) to produce a material similar to bicast leather (or additional applications of polyurethane or vinyl can be applied). This may be used in parts of the upper, including the heel counter 36, that may benefit from the added stiffness that such a material provides.

[0179] In one embodiment, the process by which the currently used mycelium material is produced can be tailored for the further processing and to provide desirable characteristics derived therefrom. In one example, the mycelium material can be cultured to provide a structure where the "middle" split resembles the type of tanned leather preferred for the production of traditional suede (e.g., lamb, goat, calf, etc.). This has a tighter network of fibers, resulting in less "hairy" fuzz on the exposed surface of the resulting material. Such modifications can be made to provide a variety of different specific leather-like mycelium materials for different parts of the upper 12, i.e., softer or stiffer materials for the parts that may utilize or benefit from such properties, etc.

[0180] Additionally, the material may be perforated as is or after cutting to enhance flexibility or breathability in desired areas. The size and shape of the perforations 60 may vary between different portions and within a particular perforation area. In one example, the vamp 28 may be laser cut perforated in an extended pattern 60 after the outer quarter is cut from the stock (or during the process in which the vamp 28 and / or other portions of the upper 12 are laser cut from the stock) to enhance flexibility and breathability in areas where less support or stiffness is required. Similarly, laser etching may be used to thin the mycelium material in various areas (without cutting it completely) or to provide decoration, including selectively removing top grain. In one example, the mycelium material may be manufactured to facilitate perforation or improve the quality of the perforations, for example, by controlling or plasticizing the fiber network formation to reduce degradation or pilling of the material within the perforations 60 (which may also improve the quality and elasticity of the raw edges adjacent the topstitches 52). In other examples, a plasticization step can be performed to provide raw edges, including within perforations, that are "self-repairing" during the laser cutting process or that are more suitable for laser cutting or laser etching (e.g., requiring lower power or being less susceptible to fuels) compared to leather.

[0181] As noted above, adhesives may be used to improve the strength of various seams between portions of the upper 12, including both topstitched seams 52, backstitched seams 54, and felted seams that may be used in the construction of the upper 12. Additionally, adhesives may be used alone to attach the combined upper 12 and lasting board 24 to the midsole 14. Solvent-based adhesives (also called cements) have been used for such purposes, including the attachment of the midsole 14, and are generally recognized to be relatively low cost, have short fixing times, and are easy to work with. Such solvent-based adhesives and cements may be used in parts or portions of the upper 12 of the mycelium material of the present invention, such as to secure seams of overlapping portions of mycelium material in the same manner that they may be used in leather, and / or to help secure the mycelium forming the portion of the upper 12 adjacent the lower perimeter 22 (or the insole 20, which, as noted above, may also be made from mycelium material) to the midsole 14. In further embodiments, such adhesives may be used to attach the outsole 16 to the midsole 14 or to attach additional elements together with the upper 12, including the depicted heel stabilizer 62, which is secured between the rear of both the upper 12 and the lasting board 24 and the midsole 14.

[0182] In some circumstances, ultraviolet ("UV") light cured or activated adhesives may be used to replace solvent-based adhesives in whole or in part. Such UV cured or UV activated adhesives may include acrylic cements or modified epoxy materials. In either case, the compound includes a photoinitiator that undergoes a chemical reaction upon exposure to UV light, causing the release of by-products to the reaction. These by-products may interact with the remaining compound and cause the compound to harden, or may initiate a reaction that results in hardening. The incorporation and reliance on a photoinitiator allows the cement or adhesive to cure "on demand" rather than within a short time of application (e.g., exposure to air in the case of acrylic cements or mixing in the case of epoxies). This may allow various portions of the upper 12 and / or midsole 14 to be coated along those portions that correspond to the seams 52, 54, or may allow attachment to another element upon cutting, e.g., with the adhesive portion of each piece activated when it is ready to be attached to the other piece or element desired. Various heat activated adhesives may be used in a similar manner. Generally, such adhesives may be made to cure upon application of heat above a certain threshold temperature or may utilize heat as a curing catalyst (e.g., in the case of epoxies). In one example, the heat-activated adhesive is applied prior to stitching of an assembled upper 12 and / or assembled athletic sneaker 10, which are then passed through a heat tunnel to initiate or amplify the curing of the adhesive, resulting in a finished part or product. In some applications, the adhesive may initially exhibit a relatively low level of adhesion such that the pieces or components can be assembled without stitching and then heat applied to cure the heat-activated adhesive.

[0183] Furthermore, because solvents often contain volatile organic compounds ("VOCs") or other polluting chemicals (which may also be flammable), water-based adhesives and cements have been developed to serve as an alternative to solvent-based compounds. In one example, for example, polyurethane adhesives may have water as their primary "solvent" since the adhesive's curing requires water to evaporate from the compound. Thus, heat may be used to accelerate or even cause the adhesive to cure. Additionally, preheating the material to be attached may also accelerate the curing process. Water-based adhesives may provide certain characteristics that make them advantageous for use in shoe construction, including assembling the athletic sneaker 10 of the present invention with the above-mentioned sections made of the mycelium material of the present invention. In particular, the crosslinking of the compound during drying may be less susceptible to ambient humidity (addition of a curing agent may further improve moisture resistance, as well as initial bond strength, heat resistance, and water degradation resistance performance). Water-based adhesives may reduce the stiffness of the material and may be less prone to disrupt stitching. Additionally, they may be made with a relatively high viscosity to prevent absorption into the material before curing, while still being sufficiently sprayable. Thus, in the same manner as above, a water-based adhesive can be used to help secure the seams 52, 54 and / or to attach additional elements to the upper 12 or to secure the upper 12 and lasting board 24 to the midsole 14.

[0184] Additionally, as shown in FIG. 3, the upper 12 may include additional structural elements in the form of various interlining elements. In particular, a heel counter interlining layer 64 may be disposed between the heel counter 36 and the underlying portions of the quarters 34a and 34b and / or collar portions 38a and 38b. Similarly, the medial and lateral quarters 34a and 34b may include eyelet interlining 66 along their edges adjacent the throat 30. In both cases, the interlining 64, 66 may be comprised of a relatively stiff fabric or a relatively soft polymeric sheet material, so that the use of the interlining 64, 66 provides additional support to the upper 12 in the area in which it is used. In particular, the heel counter interlining 64 (which may be smaller than the heel counter 36 to avoid interference with the stitching 52 and to keep the interlining 64 hidden) may provide additional stability to the heel of the wearer. Similarly, the eyelet interlining may provide additional support to the quarters 34a and 34b in the area of ​​the eyelets 56, prevent tightening of the lace 50 from damaging the quarters 34a and 34b, and / or allow the eyelets 56 to be positioned closer to the throat 30. The interlinings 64 and 66 may, at least initially, be attached to the heel counter 36 and the quarters 34a and 34b with an adhesive, including any of the adhesives described above.

[0185] In one aspect, the ability to control the material properties of the mycelium material of the present invention can also make it more suitable for adhesives than traditional leathers, making it easier to assemble using existing techniques and equipment and assemblies, and providing the athletic sneaker 10 of the present invention, and variations thereof, with improved strength and resilience. In various examples, the mycelium material of the present invention can be specifically manufactured to increase surface roughness and reduce overall porosity to improve adhesion with various adhesives. Additionally, adjustments can be made to increase heat resistance and / or heat absorption, allowing for higher preheating of the material for use with water-based adhesives.

[0186] Furthermore, further properties of the mycelial material of the present invention may lead to the use of further assembly techniques and may facilitate the implementation of different types of overall structures with different functional and aesthetic characteristics. In one example, the plasticization process described above may impart a degree of thermoplasticity to the mycelial material. Most notably, the thermoplasticity of the mycelial material allows it to be molded and bonded using heat. The particular level of such thermoplasticity exhibited by the material may be controlled by applying different ones of the plasticization process depending on the various parameters described above, as well as the particular characteristics of the culturing, tanning, and dyeing processes that may affect the outcome of the plasticization process.

[0187] In one example, the mycelium material may be manufactured to be securely assembled by adhesive, so that stitches 52 and 54 shown in Figures 1-3 may be eliminated. This may be made possible by the thermoplastic nature of the mycelium material, which may further facilitate heat-activated bonding between various portions of the upper 12. For example, when using a water-based adhesive, the mycelium pieces may be fused directly by heating the material to facilitate drying of the adhesive. Furthermore, by using specialized equipment, various portions of the upper 12, or the entire upper 12, may be fused using heat and pressure without the use of threaded seams or adhesives. Similarly, portions of the upper, such as the upper collar insert 68 or the vamp 28, may be assembled with fabrics to add flexibility to the upper 12. In one application, various thermoplastic fabrics may be used, which may be heat-bonded to adjacent portions of the upper 12 as well. Heat may also be used in joining the assembled upper 12 and insole 20 to the midsole, particularly in applications where the insole 20 is made of the mycelium material of the present invention.

[0188] In another example shown in Figures 4-6, an athletic sneaker variant 110 of the present disclosure can include an upper 112 assembled from a single piece of mycelium material that can be cut into a shape including toe tip 126, vamp 128, tongue 132, quarters 134a and 134b, heel counter 136, and portions thereof corresponding to counter portions 138a and 138b. As will be appreciated, cut-and-sew constructions, such as those of athletic sneaker 10 described above, rely on the construction of seams and the relative placement of individual parts to provide the three-dimensional shape of the assembled upper 12. The one-piece upper 112 shown in Figure 4 is similarly cut from a flat stock of the mycelium material, but since there are no seams for the relative placement of parts (except for the joining of heel counter portion 136 and collar portion 138a), thermoforming can be used to contribute to the desired three-dimensional shape of the upper 112. In this manner, a sheet of material 170 shown in Figure 6 is heated before being molded onto a shoe last and assembled with an insole 120. This heating causes the sheet of material 170 to pliable so that it imparts a three-dimensional shape as it is molded over a shoe last. The assembled upper 112 and insole 120 can then be bonded to the midsole 114 and outsole 116, as shown in Figure 5. In another example, the sheet of material can be roughly formed into the desired shape, such as by first attaching the heel counter 136 and collar portion 138a using an adhesive (such as the heat-activated adhesive described above), and placed into a special mold where heat is applied to the sheet 170 and pressure from the mold imparts the desired three-dimensional shape to the upper 112.

[0189] As further shown, additional features such as a collar lining 146 can be assembled prior to bonding the upper 112 and insole 120 to the midsole, which can be done using adhesives, heat bonding, or traditional stitching. In a variant, the collar lining 146 can be made of the mycelium material, which can be placed in a mold for direct bonding, for example, along with a sheet material 170, while the upper 112 is molded. Additional elements such as an external heel counter reinforcement can be assembled with the implementation of the mycelium material of the present invention and bonded to the upper 112 using adhesives and / or heat. In one application, the thermoplastic nature of the mycelium material can facilitate overmolding plastic directly onto the upper 112, including by injection molding or the like. In this way, the depicted variations of the heel counter reinforcement 172, as well as the quarter band 174 and eyelet reinforcement 178, can be added to the upper 112 after formation by a further step, where the upper 112 is placed in a subsequent mold with cavities for the heel counter reinforcement 172 and the quarter band 174, so that these features are molded directly into the upper 112 from a flexible plastic or thermoplastic elastomer material. In a further variation, such features can be 3-D printed directly into the upper 112, for example, by filament deposition, where the heat used to extrude a filament of material promotes fusion with the mycelium material. In certain embodiments, the features can be 3-D printed into the sheet of material 170 prior to further shaping. Alternatively, the features can be 3-D printed onto the formed upper 112 using specially adapted equipment. Additionally, fabric portions, such as counter insert 176 depicted in Figures 3-6, can be assembled with sheet 170, including by using an adhesive, or, if a thermoplastic fabric is used, by thermal bonding as described above.

[0190] In further variations, a single sheet of mycelium material 170 may be formed from different specific implementations or types of mycelium material, which may be bonded together, either in a slit sheet of material or after individual portions of the sheet have been cut separately. In one embodiment, the material may be bonded in separate layers, such that different outer layers may be bonded to one inner layer, providing different material properties in different areas of the upper 112 (e.g., less stiff material for the vamp area 128 or in the collar portions 138a and 138b). In this manner, a generally "seamless" upper 112 may be constructed with different portions of mycelium material having different properties or characteristics. Additionally, additional layers, including waterproof layers, other laminates, and the like, may be added in a similar process (as may be done in conjunction with the materials used to form the individual pieces of the upper 12). In one example, a collar insert 168 and a collar lining 146 may be included in the sheet 170 and may comprise bonded portions of the sheet 170 that have greater flexibility and / or grip.

[0191] In any of the above embodiments of athletic sneakers 10 and 110 described herein, various designs, logos, and the like can be added to the sneakers 10, 110 using techniques similar to those used in connection with existing sneakers and other footwear. In various examples, the various mycelium material areas of the uppers 12 and 112 can be printed, such as by pad printing or screen printing. The mycelium material of the present invention can also be printed using a sublimation process, where a special ink is printed onto a special sheet and pressed against the uppers 12 and 112 at high temperatures, causing the ink to sublimate and penetrate the surface of the mycelium material, before returning to a solid state and becoming a generally permanent part of the mycelium material. Additionally, the thermoplastic nature of the mycelium material of the present invention can allow for the embossing of graphics or other functional elements using heat and pressure.

[0192] It should be understood that the techniques and assembly methods using the mycelium material can also be used to assemble other types of footwear, including the various types described above (slippers, sandals, moccasins, deck shoes), using techniques generally similar to those used to make such footwear from leather, while taking advantage of many additional properties of the mycelium material and providing additional advantages to such footwear and its construction, according to the principles and variations described above. In this manner, various styles of dress shoes, boots, and the like can also be made from the mycelium material of the present invention, using various of the processes and techniques described above. In one example, the dress shoe 210 depicted in FIG. 7 can be made from the mycelium material of the present invention, which allows its toe 226 to be molded using heat, without the need to stretch leather to the shape, which can make the shoe 210 easier and cheaper to manufacture. Additional parts of the depicted dress shoe 210 may be generally similar to those of the athletic sneakers 10 and 110 described above, and are similarly numbered. In a variant, the dress shoe 210 or boot of similar construction may include a single outer as described above, including a midsole 214 material suitable for providing a surface that may contact the ground in place of the depicted outer 216. In one application, the dress shoe 210 or boot may include a "crepe sole" made of crepe rubber or a suitable replica or substitute, which exhibits a durometer low enough to provide cushioning with a layer of rubber of a thickness comparable to that of the midsole and outsole combined. Other similar applications are also possible.

[0193] Those skilled in the art will appreciate that the construction of the invention and other components described herein is not limited to any particular materials, and other exemplary embodiments of the invention disclosed herein may be formed from a wide variety of materials, unless otherwise stated herein.

[0194] For purposes of this disclosure, the term "coupled" (in all of its forms, i.e., couple, coupling, coupled, etc.) generally means directly or indirectly coupling two components to one another. Such coupling may be fixed in nature or movable in nature. Such coupling may be achieved by two components and an optional additional intermediate member that is integrally molded with one another or with the two components as a single unitary structure (e.g., the upper may be coupled directly to the outsole or via a midsole in between). Such coupling may be permanent in nature or removable or releasable in nature, unless otherwise specified.

[0195] It is also important to note that in the above examples, the configuration and arrangement of the elements of the product shown are merely illustrative. Although only a few embodiments of the innovation have been described in detail in this disclosure, those skilled in the art who review this disclosure will readily appreciate that many modifications (e.g., size, dimensions, structure, shape, and proportions of various elements, parameter values, attachment methods, use of materials, colors, orientations, etc.) are possible without substantially departing from the novel teachings and advantages of the described subject matter. For example, an element shown as integrally molded may be constructed from multiple parts, an element shown as multiple parts may be integrally molded, the operation of interfaces may be reversed or changed, the structure and / or length or width of members or connectors or other elements of the system may be changed, and the nature or number of adjustment positions provided between the elements may be changed. Accordingly, all such modifications are intended to be included within the scope of the innovation. Other substitutions, modifications, changes, and omissions may be made in the design, operating conditions, and arrangement of the desired and other exemplary embodiments without departing from the spirit of the innovation.

[0196] It will be understood that any described process or step within a described process may be combined with other disclosed processes or steps to form structures within the scope of the present invention. The exemplary structures and processes disclosed herein are for illustrative purposes and should not be construed as limiting.

[0197] It is to be understood that changes and modifications can be made in the structures and methods described above without departing from the concepts of the present invention, which concepts are intended to be covered by the following claims unless expressly stated otherwise by their language.

[0198] The above description is to be considered as being only a description of the illustrated embodiments. Modifications of the invention will occur to those skilled in the art and to those who make or use the invention. Accordingly, the above examples shown in the drawings are for illustrative purposes only and are not intended to limit the scope of the products defined by the following claims, which are to be interpreted in accordance with the principles of patent law, including the doctrine of equivalents. EXAMPLES

[0199] Example 1 - Plasticization of preserved mycelial material The effect of various methods of preserving the material prior to tanning and plasticization of the material was investigated. As a first step, Ganoderma sessile was cultured to form substantially homogenous (i.e., no fruiting bodies or substantial morphological changes) mats of cultured mycelial material approximately 21 inches long, 14 inches wide, and 2 inches thick. These mats of cultured mycelial material were then separated from the substrate on which they were grown and treated with two different treatment regimes.

[0200] For the first treatment regimen ("Treatment A"), mats of cultured mycelial material were submerged in a solution of methanol and 15% by weight calcium chloride (CaCl2) for 7 days. The solution was then replaced with clean solvent, and the mats were then submerged in the same solution for an additional 7 days. The solution was again replaced with clean solvent, and the mats were then submerged in the same solution for an additional 7 days, for a total of 21 days. The mats of cultured mycelial material were then compressed on a platen press for 5 minutes to a thickness of ½ inch. The mats were then submerged in methanol for 3 days to rinse the mats, and compressed again on a platen press for 30 minutes to a thickness of ¼ inch. The mats were then dried on the platen press for 1 day.

[0201] For the second treatment regimen ("Treatment B"), the mats of cultured mycelial material were first compressed on a platen press for 5 minutes to a thickness of ¼ inch. The compressed mats were then submerged in a solution of methanol and 15% by weight calcium chloride (CaCl2) for 14 days. The mats of cultured mycelial material were then submerged in water for 3 days to rinse the mats and compressed again on a platen press for 30 minutes to a thickness of ¼ inch. The mats of cultured mycelial material were then dried on the platen press for 1 day.

[0202] Both mats of cultured mycelial material that were treated were solution tanned in a tea solution and then plasticized by applying a 20% by weight aqueous solution of glycerin to the mats. These mats of cultured mycelial material were then compressed in a calendar press and an aqueous solution containing 10% by weight of a non-sulfated fatliquoring agent to a final width of 0.1 inches.

[0203] Various tests were performed on the mats from Treatment A and Treatment B to see if there were any differences between the treatments. These are summarized in Table 1 below along with the ASTM standards used to test the materials, where applicable. ASTM D638 was modified to set the strain rate at 10 inches per minute. ASTM D6015 was modified to use a smaller sample size of 0.25 x 1.0 inches.

[0204] Table 1. Test results from plasticized preserved mycelium material TIFF2025032176000002.tif189165

[0205] Example 2 - Protein solution immersion and cross-linking The effect of treating mycelial material with protein was investigated. As a first step, Ganoderma sessile was cultured to form substantially homogenous (i.e., no fruiting bodies or substantial morphological changes) mats of cultured mycelial material approximately 21 inches long, 14 inches wide, and 2 inches thick. These mats of cultured mycelial material were then separated from the substrate on which they were grown.

[0206] These mats of cultured mycelial material were then cut into 5 inch by 5 inch squares and compressed in a platen press for 5 minutes until they were ¼ inch thick. Each square of cultured mycelial material was immersed in one of four different solutions for one hour: 1) A 0.5% by weight pea protein solution in water ("0.5% pea protein solution in water, without TG"), 2) a 0.5% by weight aqueous pea protein solution containing about 0.25% transglutaminase ("0.5% aqueous pea protein solution + TG"); 3) 0.5% by weight pea protein in phosphate buffered saline containing 0.25% transglutaminase ("0.5% pea protein in PBS + TG"); 4) 0.25% by weight hemp protein solution in water containing 0.25% transglutaminase ("10% hemp protein solution in water + TG"); 5) An aqueous solution containing 0.25% transglutaminase ("water + TG").

[0207] After soaking in the protein and transglutaminase solutions for 1 hour, the squares of cultured mycelial material were again pressed in a platen press to a thickness of ¼ inch for 5 minutes and incubated for 16 hours at 37° C. After incubation, the squares of cultured mycelial material were exposed to 62° C. for 2 hours to inactivate the transglutaminase. The squares were then air-dried for 2 days.

[0208] To test the efficacy of transglutaminase, the same squares of mycelium mat were cut into smaller 0.5" x 0.5" squares and submerged in water to determine the % mass gain after immersion in water after 1 hour. Table 2 below summarizes the % mass gain for the various types of plant protein treatments.

[0209] (Table 2) Mass increase (%) after immersion in water for 1 hour TIFF2025032176000003.tif49153

[0210] To investigate the different effects, table 3 below summarizes the % mass gain after 1 hour of soaking in water for the different types of pea protein treatments.

[0211] (Table 3) Mass increase (%) after immersion in water for 1 hour TIFF2025032176000004.tif55158

[0212] Example 3 - Treatment of cultured mycelial material with dye solution A variety of different dyeing conditions were used to identify the optimal conditions for staining mats of cultured mycelial material preserved using Treatment A described in Example 1. Various combinations of acid and direct dyes were used to evaluate the penetration of the dyes into the cultured mycelial material under different conditions: Direct Red dye (DR37), Acid Green dye (AG68:1), Direct Black dye (DB168), Spirulina Blue dye, Anthraquinone, Natural Yellow 3, Acid Brown dyes (AB425 and AB322) were evaluated for penetration into the cultured mycelial material.

[0213] In various tests, cultured mycelial material was first treated with a presoak consisting of ammonium chloride with or without surfactant, after which the dye solution was applied. In some tests, ammonium hydroxide was added to the dye solution. In some tests, ethyloxylated fatty amines were added to the dye solution. In some tests, formic acid was added to the dye solution. In some tests, oxiranes were added to the dye solution. In some tests, sulfated fatliquors were added to the solution. The effect of pH was also examined by adjusting the amount of formic acid and / or ammonium hydroxide in the solution.

[0214] The specific penetration screening test conditions and results for each of Tests 1, 2, 3, 4, and 5 are shown in Table 4. The corresponding dye penetration images are shown in Figures 8, 9, 10, 11, and 12, as indicated. All images are cross-sections of the material taken at 32x magnification. Dye penetration into the cultured mycelial material was examined by microscope and visual inspection over different treatment time intervals. Dye penetration into the cultured mycelial material varied between different experimental conditions, with complete dye penetration into the mycelium being observed under some experimental conditions. In general, ammonium hydroxide was observed to promote dye penetration and exhaustion.

[0215] (Table 4) Dye penetration screening test TIFF2025032176000005.tif254169TIFF2025032176000006.tif177169

[0216] The mycelium swelled rapidly when submerged in solutions, especially mixtures involving ammonia and surfactants. Pressure was required to collapse the structure and remove the dye, producing a mat approximately 1-2 mm thick. For a mycelium mat measuring approximately 300 x 450 mm, a pressure of 190,000 lbsf was used.

[0217] Additionally, samples of different substrates were dyed using the same combination of direct and acid dyes to evaluate the variability of the dyeing process. The specific substrate screening test conditions and results for each of Tests 6, 7, 8, 9, and 10 are shown in Table 5. Images of the corresponding dye penetration are shown in Figures 13, 14, 15, and 16, as indicated. All images are cross-sections of the materials taken at 32x magnification.

[0218] (Table 5) TIFF2025032176000007.tif164169TIFF2025032176000008.tif158169

[0219] Further testing was performed using alternative dyes such as Direct Black 168 (CB168), Spirulina Blue, Natural Yellow 3, Anthraquinone, Acid Brown 322 (AB322), and Acid Brown 425 (AB425). Further penetration screening test conditions and results for each of Tests 10, 11, 12, 13, and 14 are shown in Table 6. Corresponding dye penetration images are shown in Figures 17, 18, 19, 20, and 21, as indicated. All images are cross-sections of the material taken at 32x magnification.

[0220] (Table 6) TIFF2025032176000009.tif186168TIFF2025032176000010.tif104168

[0221] These results indicate that standard synthetic dyes with known formulations, higher concentrations, and known penetrating performance are able to successfully penetrate this cultured mycelium material.

[0222] Cultured mycelium material was incubated with dye with and without agitation to evaluate the effect of agitation on dye penetration. Agitation test conditions and results for Tests 15 and 16 are shown in Table 7. Corresponding dye penetration images are shown in Figures 22 and 23, as indicated. All images are cross-sections of the material taken at 32x magnification.

[0223] (Table 7) TIFF2025032176000011.tif124169

[0224] Agitation of the cultured mycelial material aided in the exhaustion and penetration of the dye.

[0225] Cultured mycelium material was incubated with dyes at different pH to evaluate the effect of pH on dye penetration. The agitation test conditions and results for tests 17, 18, and 19 are shown in Table 8. The corresponding dye penetration images are shown in Figures 24, 25, and 26, as indicated. All images are cross-sections of the material taken at 32x magnification.

[0226] (Table 8) TIFF2025032176000012.tif177163

[0227] Increasing the pH improved dye penetration of the cultured mycelial material.

[0228] Color fastness was also evaluated in a rub test. Cultured mycelium material was dyed with various treatments and then rubbed using a Veslic device. Color fastness was graded after this rub test. Color fastness test conditions and results for tests 20, 21, and 22, which used larger amounts of cultured mycelium material, are shown in Table 9. Test 23 also used agitation, tests 24, 25, and 26 also used a post-dyeing wash step, and test 27 used a low dye concentration and a post-dyeing wash and squeeze step. Images of the corresponding dye penetration are shown in Figures 27A and 27B, 28A and 28B, 29A and 29B, 30A and 30B, 31A and 31B, 32A and 32B, 33A and 33B, and 34A and 34B, as indicated. All images are cross-sections of the material taken at 32x magnification. In each of Figures 27-34, panel A shows dye penetration and panel B shows color fastness.

[0229] (Table 9) TIFF2025032176000013.tif231168TIFF2025032176000014.tif212169TIFF2025032176000015.tif202169TIFF2025032176000016.tif222167

[0230] Test 27 shows that a squeezing action allowed for more rapid exhaustion of the dye compared to gentle agitation. When the dyed mycelium material was placed in water after the dyeing process, the material did not release the dye. Instead, pressure was required to release the dye from the mycelium material after dyeing.

[0231] These results indicate that the use of ammonia aided in dye penetration, and that an alkaline pH resulted in better dye penetration.

[0232] Example 4 - Treatment of cultured mycelial material with dye solution, protein solution and plasticizer Mats of cultured mycelial material preserved using treatment A described in Example 1 were treated with several different staining solutions in combination with plant proteins (soybean protein and pea protein) to determine the effect of protein treatment on the staining of cultured mycelial material. Briefly, 5.5g or 11g of protein (soybean or pea, both from Pulsin) was added to 500ml of water and sonicated at 40°C for 60 minutes. Samples of mycelial material were cut to 150mm x 35mm and incubated in the protein solution. While in the protein solution, the mycelial material was rolled (squeezed) five times with a lino roller, incubated for 15 minutes, rolled another five times, and then soaked for another 60 minutes. For staining, 2.5g of Acid Brown 425 (BASF) was added to 500ml of 50°C water and the pH was adjusted to 10 using ammonia solution. In some tests, a plasticizer was added to the dye solution. These samples were removed from the protein solution and placed in the dye solution. The samples were rolled 15 times, incubated for 15 minutes, and rolled on the reverse side for another 15 minutes. The samples were incubated overnight in the dye solution. Excess dye was removed by washing with water and gently squeezing for approximately 5 minutes. The samples were dried at room temperature. All tests were subjected to wet rub fastness testing using the protocol of BS EN ISO 11640:2012 for testing colour fastness of leather. 20 cycles of wet rubs were performed and graded using the Grey Scale Rating (GSR) system.

[0233] In most trials, the dye solution was maintained at a basic pH (pH 10) during dyeing, and then the pH was lowered to an acidic pH (pH 4-6) to fix the dye. In some trials, a plasticizer was added to the dye solution, such as fatliquors (e.g., Trupon® AMC and DXV from Trumpler), vegetable glycerin, or coconut oil. In some trials, a lino roller was used to squeeze the cultured mycelium material in the protein solution and / or dye solution. Control samples without plasticizers showed poor flexibility. Various amounts of protein were used, and it was found that excess protein produced undesirable results. In some trials, a mildewcide was added to the dye solution.

[0234] In some trials, tannins were used in combination with various dyes to treat cultured mycelial material with and without added proteins. In some trials, a plasticization step was performed after the dyeing step and fungicides were added to the plasticizer.

[0235] In addition to visual inspection of dye penetration, the hand of the samples was evaluated for softness and flexibility. Uniform distribution of dye (or color) across the surface of the cultured mycelium material was observed in some experimental conditions. In some conditions, penetration of the dye into the cultured mycelium material was observed. In many conditions, a soft and flexible material was produced. Some samples were evaluated by appearance and the rub fastness of the dye was evaluated by staining and change to the sample using Grey Scale Rating (GSR) as the metric. In some tests, a biocide was added to the dye solution.

[0236] The results of these tests are included as shown in Tables 10-16 and Figures 35A and 35B through 54A and 54B. All cross-sectional microscopic images were taken at 25.6x magnification. For each test, a cross-section of the stained mycelium material is shown in Panel A and the rub fastness is shown in Panel B.

[0237] Table 10: Protein fixation and staining test TIFF2025032176000017.tif235169TIFF2025032176000018.tif231169TIFF20250321760 00019.tif242169TIFF2025032176000020.tif248170TIFF2025032176000021.tif245169

[0238] Samples 13-16 had increased protein. All three samples performed poorly in the rub fastness test and had limited dye penetration. Without wishing to be bound by theory, this may be due to excess protein being present on the surface creating a barrier and preventing the dye from migrating into the structure of the material. Samples 17-20 did not contain plasticizer and represent the control samples for Samples 1-16. All these control samples (17-20) showed poor stiffness and flexibility. Samples 17-19 had poor rub fastness results. However, Sample 20 had improved rub fastness results and dye penetration. This difference may be due to the fact that Sample 20 was produced in the first series of tests, where the performance observations differed from later samples due to the length of time the sample was left in the dye solution.

[0239] The samples were similarly stained and then washed or not washed. The results of these tests are included as Table 11.

[0240] (Table 11) TIFF2025032176000022.tif240169TIFF2025032176000023.tif122168

[0241] Further studies were performed with increased sample size. Batch 2044 was used in Study 27. The results are shown in Table 12.

[0242] (Table 12) TIFF2025032176000024.tif93153

[0243] Tests were conducted at lower dye concentrations to evaluate the possibility of eliminating the wash step. Batch 2373 was used in these tests. The results are shown in Table 13.

[0244] (Table 13) TIFF2025032176000025.tif130168

[0245] The cultured mycelium material was then stained using vegetable tannin (mimosa). Batch 2342 was used in these tests. The results are shown in Table 14.

[0246] (Table 14) TIFF2025032176000026.tif254166

[0247] The presence of vegetable tannins increased the dye exhaustion and gave the material a more robust structure. Higher concentrations of vegetable tannins made the mycelium material feel stiffer and less flexible. This is similar to vegetable tanned leathers that are typically used for firm soles, belts, bridle leather, etc. Therefore, the concentration of vegetable tannins used should be reduced to increase the flexibility of the dyed mycelium material.

[0248] Exemplary dyes and vegetable tanning procedures followed are shown in Table 15 below.

[0249] TIFF2025032176000027.tif242169TIFF2025032176000028.tif75168

[0250] After drying, the mycelium material was rinsed again with water and then dried with paper towels. The material was then soaked in water at 100 kg / cm at 50°C. 2 This resulted in a less intense color, but a much improved finish.

[0251] Further tests were performed with additional color dyes: Luganil Bordeaux B, Luganil Red EB, Luganil Yellow G, and Luganil Olive Brown N. The results are shown in Table 16.

[0252] (Table 16) TIFF2025032176000029.tif240169TIFF2025032176000030.tif84169

[0253] Example 5 - Treatment of cultured mycelial material with dye and protein solutions Mats of cultured mycelial material preserved using Treatment A described in Example 1 were treated with several different finishes. The various finishes were applied in different sequences and evaluated for aesthetic and functional appearance of the cultured mycelial material. The finishing protocol is shown in Table 17. Various finishes were applied including: A nitrocellulose (LW65-370-Stahl) and protein sandable finish was applied after the application of a vegetable glycerol plasticizer, then the cultured mycelium material was rolled vertically to create a "box grain" effect. Nitrocellulose (LW65-370-Stahl) was mixed with hand feel modifier (HM-4896) and water in equal weight ratios. A conventional polyurethane finish containing pigments, wax emulsion, cationic wax, acrylic resin, water-based urethane resin and a water-based base coat. The base coat was followed by a top coat containing water lacquer, water-based lacquer and a hand modifier. Antique effect finish comprising a conventional polyurethane finish as above, with an antique effect coat applied between the base coat and top coat. The cultured mycelium material was also surface buffed. A distressed finish which includes the antique effect finish, an additional buffing step and an additional top coat. Contains cultured mycelium material stained with Luganil olive brown dye, which is then cooled to 100 kg / cm at 50°C. 3Below, an embossed sample embossed using a silicone mat. Various vegetable protein finishing agents, alone or with crosslinkers · Carnauba wax, alone or in combination with other known finishing agents.

[0254] An example of a Nitrocellulose and Protein Sandable Finish - Box Effect is shown in Figure 77. An example of a Nitrocellulose Finish - Box Effect is shown in Figure 78. An example of a Traditional Polyurethane Finish is shown in Figure 79. An example of an Antique Effect is shown in Figure 80. An example of a Distressed Effect is shown in Figure 81. An example of an Embossed Luganil Olive Brown is shown in Figure 82.

[0255] Table 17. Finishing Protocol TIFF2025032176000031.tif240169TIFF2025032176000032.tif247169

[0256] The protocol for the oil-based lift-off topcoat is shown in Table 18.

[0257] The results of these tests are shown in Table 19. The finishes were evaluated primarily on the basis of aesthetic appearance and hand. Many of the finishes produced the desired appearance and hand. Microscopic images of cross sections of each material are shown in Panel A of each figure, and macroscopic photographs of the materials are shown in Panel B of each figure.

[0258] (Table 19) TIFF2025032176000033.tif223169TIFF2025032176000034.tif77165

[0259] Various protein and wax finishes were also applied to the dyed cultured mycelium material. Figure 87 shows an exemplary mycelium material after a pea protein finish. Figure 88 shows an exemplary mycelium material after an unstirred soy protein finish. Figure 89 shows an exemplary mycelium material after an stirred soy protein finish. Figure 90 shows an exemplary mycelium material after a hemp protein finish. Figure 91 shows an exemplary mycelium material after a 50:50 pea protein to FI50 finish. Figure 92 shows an exemplary mycelium material after a 50:50 soy protein to FI50 finish. Figure 93 shows an exemplary mycelium material after a pea protein and crosslinker finish. Figure 94 shows an exemplary mycelium material after a Luganil Brown dye and a carnauba flake wax finish. Figure 95 shows an exemplary mycelium material after a Luganil Bordeaux dye, washing, and a carnauba flake wax finish. FIG. 96 shows an exemplary mycelium material after Luganil Yellow dye, washing, and a carnauba liquid wax finish. FIG. 97 shows an exemplary mycelium material after Luganil Brown dye, washing, and a carnauba liquid wax finish. FIG. 98 shows an exemplary mycelium material after a waxy filler, water-based PU, and a carnauba flake wax finish. FIG. 99 shows an exemplary mycelium material after one coat of a pea protein and crosslinker finish. FIG. 100 shows an exemplary mycelium material after two coats of a pea protein and crosslinker finish. FIG. 101 shows an exemplary mycelium material without embossing after a pea protein, crosslinker, and filler finish. FIG. 102 shows an exemplary mycelium material with embossing after a pea protein, crosslinker, and filler finish. FIG. 103 shows an exemplary mycelium material after Luganil Red dye, washing, and a pea protein and crosslinker finish. Figure 104 shows an exemplary mycelium material after Luganil Brown dye, washing, and glycerin soaking, pea protein and crosslinker finishing. Figure 105 shows an exemplary mycelium material after Luganil Bordeaux dye, and pea protein and crosslinker finishing.

Claims

1. a. Producing cultured mycelial material; b. contacting the cultured mycelial material with a solution comprising one or more proteins to produce a composition comprising the cultured mycelial material and one or more proteins, the one or more proteins being derived from a species other than the fungal species from which the cultured mycelial material was produced; and c. compressing the cultured mycelium material A method comprising:

2. The method of claim 1 , wherein said contacting step comprises submerging said cultured mycelial material in said solution.

3. 2. The method of claim 1, wherein said contacting step comprises contacting said cultured mycelial material with said solution in a single step.

4. 2. The method of claim 1, wherein said contacting step comprises contacting said cultured mycelial material with said solution in one or more stages.

5. The method of claim 1 , wherein the one or more proteins are derived from a plant source.

6. The method of claim 5 , wherein the plant source is a pea plant.

7. The method of claim 5 , wherein the plant source is a soybean plant.

8. The method of claim 1 , wherein the solution comprises a dye.

9. 9. The method of claim 8, wherein the composition is colored with the dye to develop a color, and the color of the cultured mycelial material is substantially uniform across one or more surfaces of the cultured mycelial material.

10. The method of claim 8 , wherein the dye penetrates throughout the interior of the composition.

11. 9. The method of claim 8, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

12. The method of claim 1 , wherein the solution comprises a plasticizer.

13. The method of claim 12, wherein the plasticizer is selected from the group consisting of oils, glycerin, and fatliquors.

14. The method of claim 12, wherein the composition is flexible.

15. The method of claim 1 , wherein the one or more proteins are cross-linked.

16. The method of claim 1 , wherein the one or more proteins are cross-linked with transglutaminase.

17. The method of claim 1 , wherein the solution comprises an enzyme.

18. 18. The method of claim 17, wherein the enzyme comprises transglutaminase.

19. 10. The method of claim 1, wherein said compacting step comprises compacting said cultured mycelium material to a thickness of between 0.1 inches and 0.5 inches.

20. 20. The method of claim 19, wherein the compacting step comprises compacting the cultured mycelium material to a thickness of 0.25 inches.

21. The method of claim 1 , wherein the compressing step is repeated one or more times.

22. 10. The method of claim 1, wherein said compacting step comprises compacting said cultured mycelium material to a thickness of 0.25 inches.

23. 10. The method of claim 1, wherein the compacting step comprises compacting the cultured mycelium material with a roller.

24. The method of claim 1 , wherein the solution comprises tannin.

25. The method of claim 1 , further comprising the step of incubating the composition.

26. 26. The method of claim 25, wherein the incubating step comprises incubating the composition at a set temperature for a period of time.

27. 27. The method of claim 26, wherein the set temperature is 40°C.

28. The method of claim 1 further comprising the step of drying the composition.

29. The method of claim 1 further comprising the step of applying a finish to one or more surfaces of the composition.

30. 30. The method of claim 29, wherein the finish is selected from the group consisting of a urethane, a wax, nitrocellulose, or a plasticizer.

31. 1. A composition comprising cultured mycelial material and one or more proteins, wherein the one or more proteins are derived from a species other than the fungal species from which the cultured mycelial material is produced.

32. 31. The composition of claim 30, wherein the one or more proteins are derived from a plant source.

33. 33. The composition of claim 32, wherein the plant source is a pea plant.

34. 33. The composition of claim 32, wherein the plant source is a soybean plant.

35. The composition of claim 30 comprising a dye.

36. 36. The composition 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. 31. The composition of claim 30 comprising a plasticizer.

38. 38. The composition of claim 37, wherein the plasticizer is selected from the group consisting of oils, glycerin, and fatliquors.

39. 38. The composition of claim 37 which is flexible.

40. The composition of claim 30 , wherein the one or more proteins are cross-linked.

41. 31. The composition of claim 30, wherein the one or more proteins are cross-linked with transglutaminase.

42. 31. The composition of claim 30 comprising an enzyme.

43. 43. The composition of claim 42, wherein the enzyme comprises transglutaminase.

44. 1. A composition comprising cultured mycelial material stained with a dye to develop color, wherein the color of the cultured mycelial material is substantially uniform across one or more surfaces of the cultured mycelial material.

45. 45. The composition of claim 44, wherein the dye is selected from the group consisting of acid dyes, direct dyes, synthetic dyes, natural dyes, and reactive dyes.

46. 45. The composition of claim 44, comprising one or more proteins derived from a species other than the fungal species from which the cultured mycelial material is produced.

47. 47. The composition of claim 46, wherein the one or more proteins are derived from a plant source.

48. 48. The composition of claim 47, wherein the plant source is a pea plant.

49. 48. The composition of claim 47, wherein the plant source is a soybean plant.

50. 45. The composition of claim 44, wherein the dye permeates throughout the interior of the composition.

51. 45. The composition of claim 44 comprising a plasticizer.

52. 52. The composition of claim 51, wherein the plasticizer is selected from the group consisting of oils, glycerin, and fatliquors.

53. 52. The composition of claim 51, which is flexible.

54. 45. The composition of claim 44, comprising a tannin.

55. 45. The composition of claim 44, comprising a finish applied to one or more surfaces of the composition.

56. 56. The composition of claim 55, wherein the finish is selected from the group consisting of a urethane, a wax, nitrocellulose, or a plasticizer.

57. Footwear products, including: a. Upper, b. a lasting board attached to the upper and defining an interior cavity for receiving a foot; c. an outsole connected to the upper on the side opposite the lasting board; d), wherein said upper comprises at least a portion of mycelial material that includes one or more proteins derived from an organism other than mycelium.

58. 58. The article of footwear of claim 57, wherein the upper comprises multiple portions of the mycelium material in their respective implementations having different physical properties.

59. 59. The article of footwear of claim 58, wherein the different physical properties are selected to correlate with desired characteristics of corresponding locations of portions within the upper.

60. 60. The article of footwear of claim 59, wherein one of the sections of mycelium material includes a bump, and the respective implementation of the mycelium material has a higher relative flexibility as compared to at least one of the sections.

61. 60. The article of footwear of claim 59, wherein one of the sections of mycelium material includes a heel counter, and the respective implementation of the mycelium material has a higher relative stiffness as compared to at least one of the sections.

62. 58. The article of footwear of claim 57, wherein the mycelium material is at least one of tanned and dyed to resemble leather.

63. 58. The article of footwear of claim 57, further comprising a midsole attached to the lasting board, and an outsole attached to the midsole and coupled to the upper.

64. 58. The article of footwear of claim 57, wherein the upper comprises a plurality of separate portions of the mycelium material.

65. 65. The article of footwear of claim 64, wherein the portions are constructed using at least one of a topstitch, a felt stitch, and a backstitch construction.

66. 65. The article of footwear of claim 64, wherein the portions are assembled using at least one of a solvent-based adhesive, a UV-curable adhesive, a heat-activated adhesive, and a water-based adhesive.

67. 65. The article of footwear of claim 64, wherein at least one of the portions is split to resemble suede leather.

68. 65. The article of footwear of claim 64, wherein at least one of the portions includes an edge that has been thinned by skiving.

69. 65. The article of footwear of claim 64, wherein the portions are assembled using thermal bonding.

70. 65. The article of footwear of claim 64, wherein the upper further comprises at least one additional portion of textile material.

71. 65. The article of footwear of claim 64, wherein the fabric material is thermoplastic and is attached to at least one of the portions of the mycelium material by thermal bonding.

72. 58. The article of footwear of claim 57, wherein the upper includes an interlining assembled with a portion thereof.

73. 58. The article of footwear of claim 57, including perforations along a portion thereof.

74. 74. The article of footwear of claim 73, wherein the perforations vary in at least one of size and relative spacing across areas of the upper.

75. 58. The article of footwear of claim 57, wherein the upper is laser etched along portions thereof.

76. 58. The article of footwear of claim 57, wherein the upper includes at least one reinforcing portion injection molded thereon.

77. 58. The article of footwear of claim 57, wherein the upper includes at least one 3D printed element fused thereto.

78. 58. The article of footwear of claim 57, wherein at least a portion of the upper includes at least one portion that is molded into a three-dimensional shape.

79. 58. The article of footwear of claim 57, wherein the upper is constructed from a single molded piece of the mycelium material.

80. 58. The article of footwear of claim 57, wherein the mycelium material comprises multiple layers of the mycelium material bonded together, each of the implementations having different physical properties.

81. 58. The article of footwear of claim 57, wherein at least one of the lasting board and the outsole includes at least a portion of the mycelium material.

82. Athletic sneakers, including: a. an upper comprising at least a portion of mycelial material that contains one or more proteins derived from an organism other than mycelium; b. a lasting board attached to the upper and defining an interior cavity for receiving a foot; c. a midsole made of a foam material and attached to the lasting board; and d. an outsole made of a rubber material and attached to the midsole opposite the lasting board; e. wherein said mycelium material is at least one of tanned and dyed to resemble leather, and said upper is constructed and assembled to resemble leather athletic footwear.

83. Athletic sneakers, including: a. an upper comprising at least a portion of mycelial material that contains one or more proteins derived from an organism other than mycelium; b. a lasting board attached to the upper and defining an interior cavity for receiving a foot; c. a midsole made of a foam material and attached to the lasting board; and d. an outsole made of a rubber material and attached to the midsole opposite the lasting board; e. wherein said upper includes at least one portion that is molded into a three-dimensional shape.

Images