HK40118040A - Fungal textile materials and leather analogs - Google Patents

Description

Fungal fabric materials and leather analogues

This application is a divisional application of patent application No. 202080057866.X, filed on June 17, 2020, with the earliest priority date being June 18, 2019, and entitled "Fungal Fabric Materials and Leather Analogs".

Cross-reference to related applications

This application claims priority to the following U.S. provisional patent applications: 62/862,860, filed June 18, 2019; 62/951,332, filed December 20, 2019; and 62/966,525, filed January 27, 2020, all of which are incorporated herein by reference in their entirety.

Technical Field

This invention relates generally to fungal materials, and more specifically to materials derived from filamentous fungi that can be used as leather analogs and in other fabrics and cloths.

Background Technology

Many current textile materials, including but not limited to leather, generate environmental problems during manufacturing and may be difficult or impossible to dispose of or recycle in an environmentally safe manner during the product's lifespan. As a non-limiting example, leather production depends on cattle feeding (which in itself has significant environmental impacts and can cause animal welfare issues) and requires a tanning process that may use highly toxic chemicals such as chromium, formic acid, mercury, and various solvents. Furthermore, leather undergoes a slow biodegradation process over approximately 25–40 years. Many textile materials present similar environmental or ethical problems.

Therefore, there is a need in the art for textile materials that can be produced cost-effectively, have minimal environmental impact, and pose no animal welfare or other ethical concerns. Further advantageously, such materials should retain the various physical and/or mechanical properties of conventional textile materials such as leather, including tensile strength, tear strength, flexural stiffness, elasticity, texture, thermal properties, and sensory attributes.

Summary of the Invention

In one aspect of the invention, a method for preparing a durable sheet material comprising fungal biomass comprises (a) infiltrating a solution into inactivated fungal biomass, the solution comprising a solvent and a component selected from polymers, crosslinking agents, and combinations and mixtures thereof; and (b) curing the biomass to remove the solvent from the biomass and form the durable sheet material.

In various embodiments, the fungal biomass may comprise fungal mycelium.

In various embodiments, the inactivated fungal biomass may be crushed (size-reduced) prior to step (a), and step (a) may include blending the crushed inactivated fungal biomass with the solution to form a blend composition. This method may, but does not necessarily, further include casting the blend composition to form a cast sheet, and removing the solvent from the cast sheet in step (b). The crushed inactivated fungal biomass may, but does not necessarily, have an average particle size not greater than about 125 micrometers.

In various embodiments, the inactivated fungal biomass may comprise cohesive fungal biomass and (a) may include agitating the inactivated fungal biomass and the solution together for a period of time. The cohesive fungal biomass may, but does not necessarily, be produced by a surface fermentation process or a submerged solid-state surface fermentation process. The period may, but does not necessarily, be selected from at least about 4 hours, at least about 5 hours, at least about 10 hours, at least about 15 hours, at least about 20 hours, or at least about 25 hours. The period may, but does not necessarily, be about 10 hours to about 20 hours. The agitation may, but does not necessarily, be carried out at a pressure different from atmospheric pressure, which may be below or above atmospheric pressure. The method may, but does not necessarily, further comprise treating the inactivated fungal biomass with at least one chemical selected from calcium hydroxide and tannins.

In various embodiments, the inactivated fungal biomass may comprise a fungal paste produced through deep fermentation.

In various embodiments, the polymer may be selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations thereof.

In various embodiments, the polymer may be selected from the following amounts present in the durable sheet material: no more than about 25% by weight of the durable sheet material, no more than about 20% by weight of the durable sheet material, no more than about 15% by weight of the durable sheet material, no more than about 10% by weight of the durable sheet material, and no more than about 5% by weight of the durable sheet material.

In various embodiments, the crosslinking agent may be selected from citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted plant tannins, glyoxal, and combinations thereof.

In various embodiments, the solution may further comprise a plasticizer. The plasticizer may, but need not, be selected from glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols and their esters, epoxidized triglyceride vegetable oils, castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations thereof.

In various embodiments, the fungal biomass may comprise at least one filamentous fungus selected from the orders Ustilaginales, Russulales, Agaricales, Pezizales, and Hypocreales.

In various embodiments, the fungal biomass may comprise at least one filamentous fungus belonging to a family selected from the following: Ustilaginaceae, Hericiaceae, Polyporaceae, Grifolaceae, Lyophyllaceae, Strophariaceae, Lycoperdaceae, Agaricomaceae, Pleurotaceae, Physalacriaceae, Omphalotaceae, Tuberaceae, Morchellaceae, Sprassidaceae, Nectriaceae, and Cordycipitaceae.

In various embodiments, the fungal biomass may comprise at least one filamentous fungus belonging to the genera selected from: Agaricus, Calocybe, Calvatia, Cordyceps, Disciotis, Fomes, Fusarium, Ganoderma, Grifola, Hericulum, Hypholoma, Hypsizygus, Morchella, Pholiota, Pleurotus, Polyporous, Sprassis, Stropharia, Tuber, and Ustilago.

In various embodiments, the fungal biomass may comprise at least one filamentous fungus selected from the following: *Ustilago esculenta*, *Hericulum erinaceus*, *Polyporous squamosus*, *Grifola fondosa*, *Hypsizygus marmoreus*, *Hypsizygus ulmarius*, *Calocybegambosa*, *Pholiota nameko*, *Calvatia gigantea*, *Agaricus bisporus*, *Stropharia rugosoannulata*, and *Hyphantria thunbergii*. The following fungi are listed: *Pleurotus eryngii*, *Pleurotus ostreatus*, *Pleurotus ostreatus var. columbinus*, *Tuber borchii*, *Morchella esculenta*, *Morchella conica*, *Morchella importuna*, *Sparassis crispa*, *Fusarium venenatum*, MK7 ATCC accession number PTA-10698, *Disciotis venosa*, and *Cordyceps militaris*.

In various embodiments, the solution may further comprise at least one of a pigment, a solubilizer, and a pH adjuster. The solubilizer may, but does not need to, be selected from hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations thereof. The pH adjuster may, but does not need to, be selected from hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations thereof.

In various embodiments, the durable sheet material may contain proteins cross-linked with isopeptide bonds.

In various embodiments, the method may further comprise, after step (b), at least one of the following: (i) adding a thermal dopant to the inactivated fungal biomass and (ii) adding a thermal dopant to the durable sheet material. The amount of the thermal dopant may, but does not need to, be selected from at least about 2.5% by weight, at least about 5% by weight, at least about 7.5% by weight, at least about 10% by weight, at least about 12.5% by weight, at least about 15% by weight, and at least about 17.5% by weight of the durable sheet material. The amount of the thermal dopant may, but is not required, be selected from no more than about 20% by weight, no more than about 17.5% by weight, no more than about 15% by weight, no more than about 12.5% by weight, no more than about 10% by weight, no more than about 7.5% by weight, and no more than about 5% by weight of the durable sheet material. The thermal dopant may, but is not required, be selected from ceramic materials, metallic materials, polymer materials, and combinations thereof. The thermal dopant may, but is not required, be selected from activated carbon, alumina, bentonite, diatomaceous earth, ethylene vinyl acetate, lignin, nano-silica, polycaprolactone, polylactic acid, organosilicon, and yttrium oxide.

In various embodiments, the inactivated fungal biomass is pulverized inactivated fungal biomass.

In various embodiments, the inactivated fungal biomass may comprise a biomat or a portion thereof produced by a surface fermentation process. The carbon-to-nitrogen molar ratio in the growth medium of the surface fermentation process may, but is not necessary, be about 5 to about 20, or about 7 to about 15.

In another aspect of the invention, the fabric composition comprises inactivated fungal biomass; and at least one component selected from plasticizers, polymers, crosslinking agents, and dyes, wherein the at least one component is distributed in the fungal mycelial biomass.

In various embodiments, the fabric composition may have a thickness of at least about 1 mm.

In various embodiments, the fabric composition may have a tear strength of at least about 30 N.

In various embodiments, the fabric composition may have a tear strength of at least about 10 N/mm.

In various embodiments, the fabric composition may have a flexural stiffness of no more than about 5 g-cm.

In various embodiments, the fabric composition may have a tensile strength of at least about 10 MPa.

In various embodiments, the fabric composition may have at least about 3 water spot gray levels.

In various embodiments, the fabric composition may have a light color fastness rating of at least about 4 for blue wool.

In various embodiments, the fabric composition may have a grayscale rating of at least about 3 for colorfastness to rubbing when dry.

In various embodiments, the fabric composition may have a grayscale rating of at least about 2 for colorfastness to rubbing when wet.

In various embodiments, the fabric composition may further comprise at least one backing layer of a non-fungal fabric material. This non-fungal fabric material may, but need not, be selected from acrylic fabrics, alpaca fabrics, angora fabrics, cashmere fabrics, coir fabrics, cotton fabrics, eisengarn fabrics, hemp fabrics, jute fabrics, Kevlar fabrics, flax fabrics, microfiber fabrics, mohair fabrics, nylon fabrics, olefin fabrics, pashmina fabrics, polyester fabrics, pineapple fiber fabrics, ramie fabrics, rayon fabrics, seasilk fabrics, silk fabrics, sisal fabrics, spandex fabrics, spider silk fabrics, wool fabrics, and combinations and mixtures thereof.

In various embodiments, the fabric composition may further comprise a thermal dopant. The thermal dopant may, but not necessarily, be selected from ceramic materials, metallic materials, polymeric materials, and combinations and mixtures thereof. The thermal dopant may, but not necessarily, be selected from activated carbon, alumina, bentonite, diatomaceous earth, ethylene vinyl acetate, lignin, nano-silica, polycaprolactone, polylactic acid, organosilicon, and yttrium oxide. The thermal properties of the fabric composition may, but not necessarily, differ from the same thermal properties of the fabric composition without the thermal dopant, wherein the thermal properties are selected from thermal effusivity, thermal conductivity, heat capacity, and combinations thereof.

In another aspect of the invention, the manufactured article comprises a fabric composition as described herein, wherein the manufactured article is selected from articles of clothing, accessory items, and furniture.

In another aspect of the invention, a method for manufacturing a durable sheet material includes (a) contacting inactivated fungal biomass with an aqueous solution containing calcium hydroxide to form limed inactivated fungal biomass; (b) contacting the limed inactivated fungal biomass with an aqueous solution containing ammonium sulfate to form delimed inactivated fungal biomass; (c) contacting the delimed inactivated fungal biomass with an aqueous solution containing a polymer to form pickled inactivated fungal biomass; (d) contacting the pickled inactivated fungal biomass with an aqueous solution containing a crosslinking agent to form tanned inactivated fungal biomass; (e) contacting the tanned inactivated fungal biomass with an aqueous solution containing a plasticizer to form plasticized inactivated fungal biomass; (f) drying the plasticized inactivated fungal biomass to form dried inactivated fungal biomass; and (g) hot-pressing the dried inactivated fungal biomass to form the durable sheet material.

In various embodiments, the method may further include rinsing the inactivated fungal biomass with water to remove residual aqueous solution between any pair of steps selected from steps (a) and (b), steps (b) and (c), steps (c) and (d), and steps (d) and (e).

In various embodiments, at least one of steps (a) to (e) may include agitating the inactivated fungal biomass together with the aqueous solution.

In various embodiments, the aqueous solution of at least one of steps (a) to (c) may further contain a surfactant or solubilizer. The surfactant or solubilizer may, but does not need to, be selected from polysorbates, hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations and mixtures thereof.

In various embodiments, the polymer may be selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations and mixtures thereof.

In various embodiments, the aqueous solution of step (c) may further comprise a plasticizer selected from: glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols and their esters, epoxidized triglyceride vegetable oil, castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof.

In various embodiments, the aqueous solution of step (c) may further contain an alkali metal halide. This alkali metal halide may, but does not need to, be sodium chloride.

In various embodiments, the crosslinking agent may be selected from citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations and mixtures thereof.

In various embodiments, the plasticizer may be selected from glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols and their esters, epoxidized triglyceride vegetable oil, castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof.

In another aspect of the invention, a method of manufacturing a durable sheet material includes (a) inactivating the fungal biomass by boiling it in water; (b) contacting the inactivated fungal biomass with an aqueous solution containing calcium hydroxide to form lime-treated inactivated fungal biomass; (c) contacting the lime-treated inactivated fungal biomass with an aqueous solution containing ammonium sulfate to form delime-treated inactivated fungal biomass; (d) contacting the delime-treated inactivated fungal biomass with an aqueous solution containing an alkali metal halide to form impregnated inactivated fungal biomass; (e) contacting the impregnated inactivated fungal biomass with a first crosslinking agent to form tanned inactivated fungal biomass; and (f) contacting the tanned inactivated fungal biomass with at least one of a second crosslinking agent and a polymer. (g) Contacting the retanned inactivated fungal biomass with an aqueous solution to form retanned inactivated fungal biomass; (h) Contacting the retanned inactivated fungal biomass with a fatliquoring oil to form fatliquoring inactivated fungal biomass; (i) Adhering a non-fungal fabric backing to the inactivated fungal biomass to form backed inactivated fungal biomass; (j) Hot-pressing the backed inactivated fungal biomass to form hot-pressed inactivated fungal biomass; (k) Drying the hot-pressed inactivated fungal biomass to form dried inactivated fungal biomass; and (d) Applying at least one of finishing wax, finishing oil, and nitrocellulose to the dried inactivated fungal biomass to form the durable sheet material.

In various embodiments, the method may further include rinsing the inactivated fungal biomass with water to remove residual aqueous solution between any pair of steps selected from steps (b) and (c), steps (c) and (d), and steps (e) and (f).

In various embodiments, at least one of steps (a) to (g) may include agitating the inactivated fungal biomass together with the aqueous solution.

In various embodiments, the aqueous solution of at least one of steps (b) and (c) may further contain a surfactant or solubilizer. The surfactant or solubilizer may, but does not need to, be selected from polysorbates, hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations and mixtures thereof.

In various embodiments, the polymer may, but need not, be selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations and mixtures thereof.

In various embodiments, the alkali metal halide can be sodium chloride.

In various embodiments, the aqueous solution of at least one of steps (d) to (f) may contain a pH adjuster. This pH adjuster may, but does not necessarily, contain hydrochloric acid, acetic acid, formic acid, lactic acid, or combinations or mixtures thereof, or a metal hydroxide.

In various embodiments, the first crosslinking agent may comprise an aluminum salt, a chromium salt, a titanium salt, an aldehyde, or a combination or mixture thereof. The first crosslinking agent may, but does not need to be, an aluminosilicate.

In various embodiments, the second crosslinking agent may be selected from citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations and mixtures thereof.

In various embodiments, the polymer may be selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations and mixtures thereof.

In various embodiments, the aqueous solution of step (f) may further contain anionic dyes.

In various embodiments, the fattening oil may be selected from sulfated castor oil, beeswax, coconut oil, vegetable oil, olive oil, linseed oil, oleic acid, and combinations and mixtures thereof.

In various embodiments, the fattening oil may comprise an emulsion and the method may further include, between steps (g) and (h), contacting the fattening oil with an acid to dissociate the emulsion.

In various embodiments, the coating wax may be selected from carnauba wax, candelilla wax, and combinations and mixtures thereof.

Various embodiments of the present invention generally relate to the production of durable sheet materials comprising fungal biomass. In some embodiments, the durable sheet material may have controlled, modified, and/or adjusted thermal properties. As a first non-limiting example, the thermal properties of the durable sheet material of the present invention can be controlled, modified, and/or adjusted by controlling the size, number, and/or spatial distribution of bubbles in the durable sheet material. As a second non-limiting example, the thermal properties of the durable sheet material of the present invention can be controlled, modified, and/or adjusted by adding a thermal dopant having desired thermal properties (e.g., heat capacity, thermal conductivity, heat storage coefficient, and combinations thereof) and thus changing the same thermal properties of the durable sheet material as a whole. As a third non-limiting example, the thermal properties of the durable sheet material of the present invention can be controlled, modified, and/or adjusted by controlling the mass, volume, thickness, spatial distribution, etc., of the thermal dopant included in the durable sheet material, thereby providing a modified or designed spatial pattern for heat exchange within and through the durable sheet material.

Various embodiments of the present invention provide fungal fabric materials, and particularly fungal leather analogues, made from whole, cohesive fungal biomass (e.g., fungal biomats produced by surface fermentation or any other suitable process), crushed or homogenized fungal biomass, or any other physical form of fungal biomass, especially filamentous fungal biomass. The materials of the present invention typically comprise both inactivated fungal biomass and a component selected from polymers, plasticizers, crosslinking agents, and dyes, and the methods of the present invention allow for the introduction of one or more such components into the inactivated fungal biomass to produce materials having desired chemical, physical, and/or thermal properties. The materials of the present invention can generally be provided as durable sheet materials suitable for use in the same or similar applications as conventional fabrics.

Attached Figure Description

Figure 1 is a schematic overview of a method for manufacturing fungal fabric materials according to various embodiments of the present invention.

Figure 2 is a schematic diagram outlining a method for manufacturing fungal fabric materials according to various embodiments of the present invention.

Figure 3 is a schematic diagram outlining a method for manufacturing fungal fabric materials according to various embodiments of the present invention.

Figure 4 is a schematic diagram outlining a method for manufacturing fungal fabric materials according to various embodiments of the present invention.

Figure 5 is a schematic diagram outlining a method for manufacturing fungal fabric materials according to various embodiments of the present invention.

Figure 6 is a schematic diagram outlining a method for manufacturing fungal fabric materials according to various embodiments of the present invention.

Figure 7 is a graph showing the tensile strength of MK7 leather analog material as a function of glycerol content according to various embodiments of the present invention.

Figure 8 is a graph showing the fracture strain of MK7 leather analog material as a function of glycerol content according to various embodiments of the present invention.

Figure 9 is a graph showing the degree of swelling of MK7 leather analog material as a function of glycerol content according to various embodiments of the present invention.

Figure 10 is a graph showing the immersion mass loss of MK7 leather analog material as a function of glycerol content according to various embodiments of the present invention.

Figure 11 is a graph showing the tensile strength of MK7 leather analog material as a function of loading ratio according to various embodiments of the present invention.

Figure 12 is a graph showing the fracture strain of MK7 leather analog material as a function of loading ratio according to various embodiments of the present invention.

Figure 13 is a graph showing the degree of swelling of MK7 leather analog material as a function of loading ratio according to various embodiments of the present invention.

Figure 14 is a graph showing the post-immersion mass loss of MK7 leather analog material as a function of loading ratio according to various embodiments of the present invention.

Figure 15 is a graph showing the tensile strength of MK7 leather analog material as a function of the polyvinyl alcohol:chitosan ratio according to various embodiments of the present invention.

Figure 16 is a graph showing the fracture strain of MK7 leather analog material as a function of the polyvinyl alcohol:chitosan ratio according to various embodiments of the present invention.

Figure 17 is a graph showing the degree of swelling of MK7 leather analog material as a function of the polyvinyl alcohol:chitosan ratio according to various embodiments of the present invention.

Figure 18 is a graph showing the post-immersion mass loss of MK7 leather analog material as a function of the polyvinyl alcohol:chitosan ratio according to various embodiments of the present invention.

Figures 19A, 19B, 19C, and 19D are histograms of broken fungal particles after blending in a conventional household blender for 10, 20, 40, and 60 seconds, respectively, according to various embodiments of the present invention.

Figure 20 is a graph showing the blending overrun, heating overrun, total overrun, and density of a solution of fungal particles in water as a function of loading ratio according to various embodiments of the present invention.

Detailed Implementation

As used herein, unless otherwise specifically stated, the term "biodegradable" refers to a material that biodegrades more rapidly than "genuine" (i.e., animal) leather under a given set of conditions (e.g., the conditions detailed in ISO 20136:2017, "Leather—determination of degradability by micro-organisms").

As used herein, unless otherwise specified, the term "degree of swelling" refers to the relative amount of change in mass of a solid article when the solid is saturated with a liquid. As a non-limiting example, a solid article having a mass of 200 g when dry and 300 g when saturated with water has a degree of swelling of 50%, or 0.5%, in water. When the term "degree of swelling" is used herein without explicitly specifying the liquid, the liquid may be assumed to be water.

As used herein, unless otherwise specifically stated, the term "durable" means a material having at least one of the following: a tear strength of at least about 5 N/mm, a tear force of at least about 5 N, and a tensile strength of at least about 1.5 MPa.

As used herein, unless otherwise specifically stated, the term "fungal biomass" refers to a substance in which fungi have been cultured, fermented, or grown by any suitable process. It should be clearly understood that fungal biomass can be produced by any of the many methods known in the art and disclosed herein, including but not limited to surface fermentation methods, submerged fermentation methods, solid-substrate submerged fermentation (SSSF) methods, and methods disclosed in PCT application publication WO2019/099474 ("474 Publication") (the entire contents of which are incorporated herein by reference).

As used herein, unless otherwise specified, the terms “hide leather” and “genuine leather” are interchangeable and each refers to a durable, flexible material produced by tanning the raw hide or skin of an animal.

As used herein, unless otherwise specifically stated, the term "inactivated" refers to fungal biomass that has been killed or otherwise prevented from active growth by suitable inactivation means such as boiling, steaming, rinsing, irradiation, freezing, treatment with an aqueous solution of at least 70% ethanol, treatment with ethanol vapor, treatment with alkali or otherwise raising the pH (with or without heating), treatment with acid or otherwise lowering the pH (with or without heating), or mechanical disturbance or destruction (e.g., by blending or otherwise crushing). It should be clearly understood that fungal biomass may be inactivated during, in combination with, and or as a result of, additional process steps (e.g., crushing, lime treatment, or deliquifying).

As used herein, unless otherwise specifically stated, the term "penetration" refers to the penetration of a solution into and/or saturation of a solid material, such that the solution or a portion thereof is distributed within the solid material. For example, and without limitation, a polymer solution penetrates the interstitial spaces of a fungal biomass composed of mycelium. It is not desired to be theoretically constrained to penetrate fungal mycelium biomass with a solution containing, for example, polymers and plasticizers, resulting in the distribution of such components within the fabric material after solvent removal through curing. Such distribution may be substantially uniform or non-uniform.

As used herein, unless otherwise specified, the term “load ratio” refers to the weight ratio of fungal biomass to polymer in a fungal fabric composition.

As used herein, unless otherwise specified, the term "loss of mass after immersion" refers to the relative amount of mass lost by a solid article after immersion in a liquid, without regard to the mass of liquid absorbed by the solid article. As a non-limiting example, a solid article having a mass of 100 grams when dry and a mass of 95 grams after immersion in water (not considering the mass of absorbed liquid) has a 5% loss of mass after immersion in water. When the term "loss of mass after immersion" is used herein without explicitly specifying the liquid, the liquid may be assumed to be water.

As used herein, unless otherwise specified, the term "sheet" refers to a layer of solid material having a generally flat or planar shape and a high surface area to thickness ratio.

As used herein, unless otherwise specified, the term "tannin" generally refers to any molecule that forms a strong bond with protein structures, and more specifically, when applied to raw hides and leathers, it binds strongly to the protein portions of the collagen structure in the skin to improve the strength and resistance to degradation of the leather. The most commonly used types of tannins are vegetative tannins, i.e., tannins extracted from trees and plants, and chromium tannins such as chromium(III) sulfate. Other examples of tannins when the term is used herein include modified naturally derived polymers, biopolymers, and salts of metals other than chromium, such as aluminosilicates (sodium aluminum silicate, potassium aluminum silicate, etc.).

Referring now to FIG1, one embodiment of a method 100 for manufacturing a fungal fabric material is shown. In the first step 110 of the method 100 shown in FIG1, fungal biomass is produced by any of several suitable methods, including but not limited to those described below: PCT application PCT/US2017/020050 (“050 application”), filed February 28, 2017; PCT application PCT/US2018/048626 (“626 application”), filed August 29, 2018; U.S. Provisional Patent Application 62/811,421 (“421 application”), filed February 27, 2019; and 474, the entire contents of which are hereby incorporated by reference. As described in the 050, 626, and 421 applications, the fungal biomass can be grown by surface fermentation in an artificial culture medium to form a cohesive structure of interwoven or interconnected mycelia called a biomat. According to the methods described in applications '050, '626, and '421, in various embodiments, it is desirable to control the oil and/or lipid content of the fungal biomass by providing a growth medium having a preselected carbon-to-nitrogen ratio. Specifically, the production of certain lipids or oils, or the amount thereof, by the fungal biomass can result in the fungal fabric material possessing certain desired material properties, such as improved water resistance, reduced conditioning requirements, etc.; such properties can be easily controlled, modified, or adjusted by providing a preselected carbon-to-nitrogen molar ratio in the fungal growth medium (which in various embodiments may be about 5 to about 20, or about 7 to about 15). In some embodiments, the production of certain lipids or oils by the fungal biomass, such as oleic acid, linoleic acid, eicosenoic acid, palmitic acid, stearic acid, arachidic acid, sorbic acid, etc., allows for the use of certain polymers, solvents, etc., which might otherwise be unsuitable in the practice of this invention, thereby providing properties that the fungal fabric material might otherwise not obtain, or providing additional or synergistic effects of this kind.

In the optional second step 120 of method 100 shown in Figure 1, the fungal biomass can be crushed by any suitable method. As a non-limiting example, the method may include processing (e.g., in a mixer, food processor, or similar crushing device), compression (e.g., by moving jaws, rollers, rotary cones, or similar compression devices), impact (e.g., by hammers, high-speed jets of material, rollers, or similar impact devices), spray drying, etc. This crushing step can be carried out for any suitable duration (e.g., two minutes) in any suitable apparatus (e.g., a mixer). During this crushing step, at least a portion of the cohesive, interconnected, or interwoven mycelial network of the fungal biomass can be disturbed or destroyed.

In the third step 130 of method 100 shown in Figure 1, the fungal biomass is mixed with a solution of a synthetic polymer and/or a biopolymer. The synthetic polymer may be any synthetic polymer soluble in a selected solvent, which may, but does not necessarily, be water; as a non-limiting example, the synthetic polymer may be polyvinyl alcohol, polyethylene glycol, polysiloxane, polyphosphazene, low- and/or high-density polyethylene, polypropylene, polyvinyl chloride, polystyrene, nylon, polytetrafluoroethylene, thermoplastic polyurethane, polychlorotrifluoroethylene, polycaprolactone, polyacrylic acid, and/or any one or more synthetic polymers sold under various brand names (e.g., Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora, Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.). The biopolymer may be any polymeric molecule naturally produced by animals, plants or fungi, including, as non-limiting examples, cellulose, chitin, chitosan, collagen, silk protein, hyaluronic acid, keratin, alginate, starch, and combinations thereof. In various embodiments, the solution (or another solution of the bio-pad and its combination in the same, preceding, or subsequent steps) may also contain additional components, such as, as non-limiting examples, plasticizers (e.g., glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g., mannitol, sorbitol) and their esters, epoxidized triglyceride vegetable oils (e.g., from soybean oil), castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof) and/or crosslinking agents (e.g., homo-bifunctional crosslinking agents, hetero-bifunctional crosslinking agents, photoreactive crosslinking agents, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations thereof). It should be clearly understood that the crushing step (if any) and the mixing step may be performed simultaneously or sequentially in any order.

In the fourth step 140 of method 100 shown in Figure 1, the biomass/solution mixture is stirred, typically at an elevated temperature (about 90°C to about 100°C, as a non-limiting example). After stirring, the biomass/solution mixture may optionally be further mixed with a dye to provide the desired color to the fungal fabric material. In some embodiments, the dye may be added early in the process.

In the fifth step 150 of method 100 shown in Figure 1, the bio-pad/solution mixture is cured, optionally after being cast into the desired shape. This curing step may involve drying or initiating a chemical reaction that may remove the solvent from the solution.

In the sixth step 160 of method 100 shown in Figure 1, the cured material is hot-pressed to form the desired fungal fabric material. In various embodiments, the fungal fabric material may have at least one physical, mechanical, and/or aesthetic property that mimics or closely resembles the physical, mechanical, and/or aesthetic properties of conventional fabric materials such as leather.

In some embodiments of the method of the present invention, the step of crushing the fungal biomass (e.g., the second step 120 shown in FIG. 1) may be omitted. In some such embodiments, the biomass (e.g., a biomat produced according to the methods described in '050, '626, and/or '421 applications) may have been or may not have been pre-crushed. In other embodiments, the biomass used may be biomass that does not require crushing, such as a fungal paste produced by a submerged fermentation method as known and described in the art.

Referring now to FIG2, another embodiment of a method 200 for manufacturing fungal fabric materials is shown. In the first step 210 of method 200 shown in FIG2, fungal biomass is produced and processed by any of several suitable methods, including but not limited to those described in applications '050, '626, '421, and '474. The biomass may be boiled, rinsed, irradiated, and/or pressed to inactivate the organism and/or remove excess water and/or other liquids. The biomass may also be frozen, particularly when it is desired or necessary to preserve the biomass for a period of time before proceeding to subsequent steps and thus extend the usable "shelf life" (storage period) of the biomass.

In the second step 220 of method 200 shown in Figure 2, the fungal biomass is thawed (if pre-frozen); crushed by any suitable method, which, as a non-limiting example, may include processing in a mixer, food processor, mill, ultrasonic generator, or similar crushing device; and blended or otherwise homogenized with water and optionally a pigment to provide the desired color to the fungal fabric material. This crushing sub-step can be carried out for any suitable duration (e.g., two minutes) in any suitable device (e.g., a mixer). This blending/homogenization sub-step produces a viscous, substantially homogeneous fungal paste. It should be clearly understood that the crushing sub-step and the blending/homogenization sub-step can be carried out simultaneously and sequentially in the same container, or sequentially in different containers; as a non-limiting example, water and optional pigment may be added to the mixer along with the fungal biomass prior to the crushing sub-step, and these components may be blended simultaneously in the mixer, thereby carrying out the crushing and blending/homogenization sub-steps simultaneously in the same container. In some embodiments, breaking down the fungal biomass can also lead to the inactivation of the fungal biomass (e.g., by disrupting the cellular structure of the fungus).

In an optional third step 230 of method 200 shown in Figure 2, the viscous, substantially homogeneous paste is degassed by any suitable method, which, as a non-limiting example, may include one or more of agitation and vacuum treatment. Degasing the fungal material can provide improved quality to the finished fungal fabric product, including, but not limited to, a more aesthetically pleasing and/or more similar texture or "feel" to the replicated material (e.g., genuine leather) for the user. In some embodiments, degasing may be omitted; in particular, in some embodiments, it is desirable to allow at least some air bubbles or cavities to remain in the fungal paste, as this can impart certain desired thermal or insulating properties to the finished fungal fabric material.

In the fourth step 240 of method 200 shown in Figure 2, the fungal paste is mixed with a solution of a polymer in a selected solvent. The solvent may, but does not, be water. The polymer may, but does not, be a biopolymer, i.e., any polymeric molecule naturally produced by animals, plants, or fungi, including, as non-limiting examples, cellulose, chitin, chitosan, collagen, silk fibroin, hyaluronic acid, keratin, alginate, starch, and combinations thereof. In various embodiments, the solution (or another solution in combination with the biopadded material in the same, preceding, or subsequent steps) may, in addition to the biopolymer, contain, or contain as a substitute for, the following synthetic polymers soluble in the solvent (e.g., polyvinyl alcohol, polyethylene glycol, polysiloxane, polyphosphazene, low- and/or high-density polyethylene, polypropylene, polyvinyl chloride, polystyrene, nylon, polytetrafluoroethylene, thermoplastic polyurethane, polychlorotrifluoroethylene, polycaprolactone, polyacrylic acid, and/or products sold under various brand names (e.g., Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora, Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.) Any one or more synthetic polymers. In various further embodiments, the solution may contain one or more additional components, such as plasticizers (e.g., glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g., mannitol, sorbitol) and their esters, epoxidized triglyceride vegetable oils (e.g., from soybean oil), castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof), crosslinking agents (e.g., homo-bifunctional crosslinking agents, hetero-bifunctional crosslinking agents, photoreactive crosslinking agents, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations thereof), solubilizers (e.g., hydrochloric acid, acetic acid, formic acid, lactic acid, etc.), and/or pH adjusters (e.g., hydrochloric acid, acetic acid, formic acid, lactic acid, etc.).

The solution can be prepared by combining the polymer and the solvent, along with optional one or more additional components, in a container, and heating the combination while stirring. In various embodiments where the solution includes a solubilizer and/or a pH adjuster, either or both of these can be added to the solution after heating and stirring the other components. Preferably, the polymer (biopolymer, synthetic polymer, or a combination thereof) is completely dissolved in the solvent, and then the solution is mixed with an optionally degassed fungal paste. The mixture can be heated (e.g., to about 90°C and/or to boiling) and/or stirred sufficiently to ensure that the mixture is substantially homogeneous, for example, about 30 minutes to about 45 minutes.

In an optional fifth step 250 of method 200 shown in Figure 2, the mixture produced in the fourth step is degassed by any suitable method, which, as a non-limiting example, may include one or more of agitation and vacuum treatment. Degasing the mixture can provide improved quality to the finished fungal fabric product, including, but not limited to, a more aesthetically pleasing and/or more similar texture or "feel" to the replicated material (e.g., genuine leather) for the user. In some embodiments, degasing may be omitted; in particular, in some embodiments, it is desirable to allow at least some air bubbles or cavities to remain in the mixture, as this can impart certain desired thermal or insulating properties to the finished fungal fabric material.

In the sixth step 260 of method 200 shown in Figure 2, the fungal mixture is cured, optionally after being cast into a desired shape (e.g., a flat or textured mold). This curing step may or may not involve curing or initiating a chemical reaction and may or may not remove the solvent from the solution. The curing step may be carried out at room temperature in ambient air. The curing may be permitted to continue for a duration sufficient to provide the desired quality (e.g., about 20% of the mass before drying/curing) and/or moisture content of the cured material.

In an optional seventh step 270 of method 200 shown in Figure 2, the cured material may be hot-pressed to form a desired fungal fabric material. In various embodiments, the fungal fabric material may have at least one physical, mechanical, and/or aesthetic property that mimics or closely resembles the physical, mechanical, and/or aesthetic properties of conventional fabric materials such as leather. The temperature (e.g., about 100°C) and/or time (e.g., about 10 minutes to about 20 minutes) of the hot pressing may be selected to provide the desired physical, mechanical, and/or aesthetic properties. The fungal fabric material may, but not necessarily, be then laminated to a fabric backing; in these embodiments, a portion of the solution from step four may, but not necessarily, be used as an adhesive for adhering the fungal fabric material to the fabric backing.

Typically, the methods shown in Figures 1 and 2 result in a network of fungal filaments being crosslinked together by a combination of a polymer (e.g., chitosan) and a crosslinking agent (e.g., citric acid). The polymer and crosslinking agent can form bonds via esterification (between the alcohol groups of the fungal filaments and/or the polymer and the carboxylic acid groups of the crosslinking agent and/or the fungal filaments) and/or amidation (between the amide groups of the fungal filaments and/or the polymer and the carboxylic acid groups of the crosslinking agent and/or the fungal filaments). These reactions can be catalyzed, for example, by acidic conditions and/or by heat (e.g., in a hot-pressing step). The use of plasticizers such as glycerol can impart flexibility to the finished fungal fabric material. The method of Figure 1 can be used with both whole and broken fungal biomass.

Referring now to Figure 3, another embodiment of a method 300 for manufacturing a fungal fabric material is shown. In the lime treatment step 310 of method 300 shown in Figure 3, inactivated fungal biomass is added to an aqueous mixture or solution of the components and agitated, for example, on a shaker table. The aqueous mixture or solution comprises: an aqueous solvent, typically in a mass approximately equal to the mass of the fungal biomass; and a lime treatment substance, most commonly calcium hydroxide (i.e., slaked lime), in an amount of about 0.01 wt% to about 6 wt% or any subrange between these values, most commonly about 3 wt%, relative to the weight of the fungal biomass. The aqueous mixture or solution may optionally further comprise a solubilizer or surfactant, such as polysorbate, in an amount of about 0.01 wt% to about 1 wt% or any subrange between these values, most commonly about 0.2 wt%, relative to the weight of the fungal biomass. The agitation may be carried out for any suitable time from about 1 minute to about 180 minutes or any subrange between these values, most commonly about 90 minutes.

Prior to step 310, the fungal biomass may be produced and processed by any of several suitable methods (including, but not limited to, those described in '050, '626, '421, and '474), and the fungal biomass may be boiled, rinsed, irradiated, and/or pressed to inactivate the organism and/or remove excess water and/or other liquids. The biomass may also be frozen prior to step 310, particularly when it is desired or necessary to preserve the biomass for a period of time before proceeding to subsequent steps and thus extend its usable "shelf life"; and subsequently thawed.

Step 310 of method 300 shown in Figure 3 can be performed on whole fungal biomass, such as cohesive fungal biomass produced by surface fermentation, or it can be performed on fungal biomass that has been pre-crushed by any suitable method. As a non-limiting example, the method can be carried out in a mixer, food processor, mill, ultrasonic generator, or similar crushing device. Any such crushing can be carried out in any suitable device (e.g., a mixer) for any suitable duration (e.g., two minutes). In some embodiments, the fungal biomass may be active prior to crushing and may be inactivated by the crushing, for example, by disrupting the cellular structure of the fungus. More generally, it should be explicitly understood that the fungal biomass may be inactivated during, in combination with, or as a result of any one or more other steps of method 300, such as lime treatment step 310 (whereby the pH of the fungal biomass is raised to at least about 7, or to a further pH high enough to kill the fungus) or any subsequent steps (particularly if carried out at elevated temperatures).

In the delime step 320 of method 300 shown in Figure 3, the inactivated fungal biomass is added to the aqueous mixture or solution of the components and agitated, for example, on a vibrating table. The aqueous mixture or solution comprises: an aqueous solvent, typically about half the mass of the starting (i.e., prior to step 310) fungal biomass; and a delimeing agent, most commonly ammonium sulfate, in an amount of about 0.01 wt% to about 6 wt% or any subrange between these values, most commonly about 3 wt%, relative to the weight of the starting (i.e., prior to step 310) fungal biomass. The aqueous mixture or solution may optionally further comprise a solubilizer or surfactant, such as polysorbate, in an amount of about 0.01 wt% to about 1 wt% or any subrange between these values, most commonly about 0.2 wt%, relative to the weight of the starting (i.e., prior to step 310) fungal biomass. The agitation may be carried out for any suitable time from about 1 minute to about 180 minutes or any subrange between these values, most commonly about 90 minutes.

In the impregnation step 330 of method 300 shown in Figure 3, the inactivated fungal biomass is mixed with a solution of the polymer in an aqueous solvent. The polymer may, but need not, be a biopolymer, i.e., any polymer-type molecule naturally produced by animals, plants, or fungi, including, as non-limiting examples, cellulose, chitin, chitosan, collagen, silk fibroin, hyaluronic acid, keratin, alginate, starch, and combinations thereof. In various embodiments, the solution (or another solution of the inactivated fungal biomass and its combination in the same, preceding, or subsequent steps) may, in addition to the biopolymer, contain or contain as a substitute for the biopolymer: synthetic polymers soluble in the solvent (e.g., polyvinyl alcohol, polyethylene glycol, polysiloxane, polyphosphazene, low- and/or high-density polyethylene, polypropylene, polyvinyl chloride, polystyrene, nylon, polytetrafluoroethylene, thermoplastic polyurethane, polychlorotrifluoroethylene, polycaprolactone, polyacrylic acid, and/or any one or more synthetic polymers sold under various brand names (e.g., Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora, Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.). The solution may contain one or more additional components, such as plasticizers (e.g., glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g., mannitol, sorbitol) and their esters, epoxidized triglycerides, vegetable oils (e.g., from soybean oil), castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations thereof), crosslinking agents (e.g., homo-bifunctional crosslinking agents, hetero-bifunctional crosslinking agents, photoreactive crosslinking agents, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations thereof), solubilizers (e.g., hydrochloric acid, acetic acid, formic acid, lactic acid, etc.), and/or pH adjusters (e.g., hydrochloric acid, acetic acid, formic acid, lactic acid, etc.). Alkali metal halides (e.g., sodium chloride) may be provided to prevent swelling of the inactivated fungal biomass.

The solution can be prepared by combining the polymer and the solvent, along with optional one or more additional components, in a container and stirring or agitating the combination, optionally while simultaneously heating the combination. In various embodiments where the solution includes a solubilizer and/or a pH adjuster, either or both of these can be added to the solution after heating and stirring the other components. Preferably, the polymer (biopolymer, synthetic polymer, or a combination thereof) is completely dissolved in the solvent, and then the solution is mixed with an optionally degassed fungal paste. The mixture can be heated (e.g., to about 90°C and/or to boiling) and/or stirred sufficiently to ensure that the mixture is substantially homogeneous, for example, from about 1 minute to about 240 minutes or any subrange between these values, and most typically from about 30 minutes to about 45 minutes or about 120 minutes.

In step 330 of method 300, the polymer solution to which the inactivated fungal biomass is added typically comprises: an aqueous solvent, generally about equal to the mass of the starting (i.e., prior to step 310) fungal biomass; and a polymer in an amount of about 0.01 wt% to about 10 wt% or any subrange of these values, most commonly about 1 wt%, relative to the starting (i.e., prior to step 310) fungal biomass. Other components, if present during step 330, may be provided in any suitable amount; as a non-limiting example, a solubilizer or pH adjuster may be provided in an amount of about 0.01 wt% to about 10 wt% or any subrange of these values, most commonly about 0.5 wt% to about 2.5 wt%, relative to the starting (i.e., prior to step 310) fungal biomass, and the alkali metal halide may be provided in an amount of about 0.01 wt% to about 14 wt% or any subrange of these values, most commonly about 7 wt%, relative to the starting (i.e., prior to step 310) fungal biomass.

In the tanning step 340 of method 300 shown in Figure 3, the inactivated fungal biomass from impregnation step 330 is added to an aqueous solution containing a crosslinking or tanning agent and agitated, for example, on a vibrating table. The aqueous solution contains: an aqueous solvent, typically in a mass approximately equal to the mass of the starting (i.e., prior to step 310) fungal biomass; and a crosslinking or tanning agent, such as citric acid and/or tannic acid, in an amount of about 0.01% to about 12% by weight, most commonly about 5% by weight, relative to the weight of the starting (i.e., prior to step 310) fungal biomass. The agitation can be carried out for about 1 minute to about 360 minutes, or for any suitable time within any subrange of these values, most commonly about 180 minutes.

Although not shown in Figure 3, method 300 may optionally include one or more rinsing steps after any one or more of the lime treatment step 310, delime treatment step 320, impregnation step 330, and tanning step 340, wherein the inactivated fungal biomass is rinsed with water to remove excess aqueous solution. The rinsing step may include draining excess aqueous solution from a container (e.g., a shake flask) containing the inactivated fungal biomass, refilling the container with water, agitating the container, and draining water from the container.

In the plasticizing step 350 of method 300 shown in Figure 3, the inactivated fungal biomass is added to an aqueous solution containing a plasticizer and agitated, for example, on a vibrating table. The aqueous solution contains: an aqueous solvent, typically in a mass approximately equal to the mass of the starting (i.e., prior to step 310) fungal biomass; and a plasticizer, such as glycerol, in an amount of about 0.01% by weight to about 50% by weight, or any subrange between these values, most commonly about 25% by weight, relative to the weight of the starting (i.e., prior to step 310) fungal biomass. The agitation may be carried out for about 1 minute to about 180 minutes, or any suitable time within any subrange between these values, most commonly about 90 minutes. In some embodiments, the plasticizing step 350 may be a fatliquoring step, i.e., the plasticizer may be a fatliquoring oil such as sulfated castor oil, beeswax, coconut oil, vegetable oil, olive oil, linseed oil, oleic acid, sulfated fish oil, sulfated canola oil, soybean oil, palm oil, fatty acids, or combinations thereof.

In the drying step 360 of method 300 shown in Figure 3, the inactivated fungal biomass is dried, optionally after being cast into a desired shape (e.g., a flat or textured mold) if produced from crushed fungal biomass. This drying step may or may not involve the initiation of a chemical reaction, but generally results in at least a majority of any residual water, solvent, and other liquids being expelled from the inactivated fungal biomass. The drying can be passive (i.e., at room temperature without the use of blowers, fans, etc.) or active (i.e., under heating and/or with the use of forced ventilation, dry milling, etc.); when the drying is active, the temperature can be raised to a desired temperature above room temperature, most commonly about 80°F, and/or any suitable forced ventilation means (e.g., blowers, fans, forced ventilation dehydrators, etc.) can be used. In some embodiments, at least a portion of the fungal material can be clamped or otherwise pressed to reduce shrinkage. The curing can be allowed to continue for a time sufficient to provide the desired quality (e.g., about 20% of the quality before drying/curing) and/or moisture content of the cured material. In various embodiments, this time can be from about 1 minute to about 2 days or any subrange between these values, most commonly about 1 day.

In the hot-pressing step 370 of method 300 shown in Figure 3, the inactivated fungal biomass is hot-pressed to form a desired fungal fabric material. In various embodiments, the fungal fabric material may have at least one physical, mechanical, and/or aesthetic property that mimics or closely resembles the physical, mechanical, and/or aesthetic properties of conventional fabric materials such as leather; in particular, the hot-pressing step may be configured to impart a leather-like texture to the fungal fabric material. The temperature (e.g., about 100°C) and/or time (e.g., about 1 minute to about 20 minutes, most commonly about 10 minutes) of the hot pressing may be selected to provide the desired physical, mechanical, and/or aesthetic properties. The fungal fabric material may, but not necessarily, be then laminated to a non-fungal fabric backing.

Referring now to FIG4, another embodiment of the method 400 for manufacturing fungal fabric material is shown. In the inactivation step 405, the fungal biomass is inactivated to prevent the active growth and metabolism of the fungus. This inactivation can typically be achieved by boiling the fungal biomass in water of sufficient volume to completely submerge or surround it; the boiling typically lasts for a period of about 1 minute to about 60 minutes, or any subrange between these values, most commonly about 30 minutes. Of course, the inactivation step 405 can also be carried out by any other suitable means, such as by irradiation, freezing, crushing, or a combination thereof, in boiling or without boiling.

Prior to step 405, the fungal biomass may be produced and processed by any of several suitable methods, including but not limited to those described in applications '050, '626, '421, and '474. The biomass may also be frozen prior to step 405, particularly when it is desirable or necessary to preserve the biomass for a period of time before proceeding to subsequent steps and thus extend its usable "shelf life"; and subsequently thawed.

Step 405 of method 400 shown in Figure 4 can be performed on whole fungal biomass, such as cohesive fungal biomass produced by surface fermentation, or it can be performed on fungal biomass that has been pre-crushed by any suitable method. As a non-limiting example, the method can be carried out in a mixer, food processor, mill, ultrasonic generator, or similar crushing device. Any such crushing can be carried out in any suitable device (e.g., a mixer) for any suitable duration (e.g., two minutes). In some embodiments, the fungal biomass may be active prior to crushing and may be inactivated by the crushing, for example, by disrupting the cellular structure of the fungus.

Step 405 typically also includes dissolving, mixing, or suspending the inactivated fungal biomass in an aqueous solvent and may further include adding a solubilizer or surfactant, such as polysorbate, to the inactivated fungal biomass and combining the solubilizer or surfactant with the inactivated fungal biomass, for example, by agitation. The mass of the aqueous solvent may typically be about half to about six times the mass of the inactivated fungal biomass, most commonly about three times. The solubilizer or surfactant may be provided in an amount of about 0.01% by weight to about 1% by weight, or any subrange between these values, most commonly about 0.2% by weight, relative to the weight of the fungal biomass. The agitation or other mechanical operation of the inactivated fungal biomass with the aqueous solvent and optionally the solubilizer or surfactant may be carried out for a period of about 1 minute to about 60 minutes, or any subrange between these values, most commonly about 30 minutes.

In the lime treatment step 415 of method 400 shown in Figure 4, the inactivated fungal biomass is added to the aqueous mixture or solution of the components and agitated, for example, on a vibrating table. The aqueous mixture or solution comprises: an aqueous solvent, typically in a mass approximately equal to the mass of the fungal biomass; and a lime treatment substance, most commonly calcium hydroxide (i.e., slaked lime), in an amount of about 0.01 wt% to about 10 wt% or any subrange between these values, most commonly about 3 wt%, relative to the weight of the fungal biomass. The aqueous mixture or solution may optionally further comprise a solubilizer or surfactant, such as polysorbate, in an amount of about 0.01 wt% to about 1 wt% or any subrange between these values, most commonly about 0.2 wt%, relative to the weight of the fungal biomass. The agitation may be carried out for any suitable time from about 1 minute to about 300 minutes or any subrange between these values, most commonly about 150 minutes.

In the delime step 425 of method 400 shown in Figure 4, the inactivated fungal biomass is added to the aqueous mixture or solution of the components and agitated, for example, on a vibrating table. The aqueous mixture or solution comprises: an aqueous solvent, typically about half the mass of the starting (i.e., prior to step 405) fungal biomass; and a delimeing agent, most commonly ammonium sulfate or ammonium chloride, in an amount of about 0.01 wt% to about 10 wt% or any subrange between these values, most commonly about 3 wt%, relative to the weight of the starting (i.e., prior to step 405) fungal biomass. The aqueous mixture or solution may optionally further comprise a solubilizer or surfactant, such as polysorbate, in an amount of about 0.01 wt% to about 0.4 wt% or any subrange between these values, most commonly about 0.2 wt%, relative to the weight of the starting (i.e., prior to step 405) fungal biomass. The agitation may be carried out for any suitable time from about 1 minute to about 150 minutes or any subrange between these values, most commonly about 75 minutes.

In the impregnation step 435 of method 400 shown in Figure 4, the inactivated fungal biomass is mixed with an acid, most commonly hydrochloric acid, or other pH adjuster. Sufficient pH adjuster is added to achieve a target pH of no more than about 4.0, typically about 0.5 to about 3.5, more typically about 1.0 to about 3.0, even more typically about 1.5 to about 2.5, and most typically about 2.0. It is generally desirable to select a molar concentration and/or gravimetric molar concentration of the acid in an aqueous solvent, or a concentration of the pH adjuster, that allows the target to be achieved by adding a preselected mass or volume of acid or liquid solution. The aqueous solution of the acid or pH adjuster may further contain an alkali metal halide, such as sodium chloride, to prevent swelling of the fungal biomass; the alkali metal halide may be present in an amount of about 0.01% by weight to about 14% by weight, or any subrange between these values, most commonly about 7% by weight, relative to the starting (i.e., prior to step 405) fungal biomass. The inactivated fungal biomass can be stirred with the acid and/or pH adjuster and optionally the alkali metal halide for about 1 minute to about 180 minutes or any subrange between these values, most commonly about 90 minutes.

In the tanning step 445 of method 400 shown in Figure 4, a first crosslinking or tanning agent is added to the inactivated fungal biomass and the mixture is agitated, for example, in a drum or on a vibrating table. In various embodiments, the crosslinking or tanning agent may comprise an aldehyde, aluminum salt, chromium salt, or titanium salt, and may typically comprise an aluminosilicate. The crosslinking or tanning agent may be provided in an amount typically from about 0.01 wt% to about 15 wt%, or any subrange between these values, most commonly from about 1.5 wt% to about 7.5 wt%, relative to the weight of the starting (i.e., prior to step 405) fungal biomass. The agitation may be performed for any suitable time from about 1 minute to about 180 minutes, or any subrange between these values, most commonly from about 30 minutes to about 150 minutes. During this agitation, alkali or other pH adjusters, such as sodium hydroxide, may be added once or multiple times to achieve and/or maintain a target pH, which in various embodiments is typically about 2.0 to about 6.0, typically about 2.5 to about 5.5, more typically about 3.0 to about 5.0, even more typically about 3.5 to about 4.5, and most typically about 4.0.

Although not shown in Figure 4, method 400 may optionally include one or more rinsing steps after any one or more of the lime treatment step 415, the delime step 425, and the tanning step 445, wherein the inactivated fungal biomass is rinsed with water to remove excess aqueous solution. The rinsing step may include draining excess aqueous solution from a container (e.g., a shake flask) containing the inactivated fungal biomass, refilling the container with water, agitating the container, and draining water from the container.

In the retanning step 455 of method 400 shown in Figure 4, a second crosslinking or tanning agent is added to the inactivated fungal biomass and the mixture is agitated, for example, in a drum or on a vibrating table. In various embodiments, the second crosslinking or tanning agent may comprise, for example, citric acid and may be provided in an amount of about 0.01% to about 6% by weight, or any subrange of these values, most commonly about 3% by weight, relative to the weight of the starting (i.e., prior to step 410) fungal biomass. The agitation may be carried out for any suitable time, from about 1 minute to about 480 minutes, or any subrange of these values, most commonly about 60 minutes.

The retanning step 455 may optionally include an additional sub-step that imparts an additional substance or property to the inactivated fungal biomass and thus to the finished fungal fabric material. As a first non-limiting example, the inactivated fungal biomass may be mixed with an aqueous solution of any polymer disclosed herein and agitated, for example, in a drum or on a vibrating table. The polymer may be provided in an amount of about 0.01% by weight to about 30% by weight, or any subrange between these values, most commonly about 0.5% by weight to about 5% by weight, relative to the weight of the starting (i.e., prior to step 410) fungal biomass. The agitation may be carried out for any suitable time of about 1 minute to about 480 minutes, or any subrange between these values, most commonly about 60 minutes. As a second non-limiting example, a dye, such as anionic dye, may be added to the inactivated fungal biomass, and the combination may be agitated, for example in a drum or on a vibrating table, for a time sufficient to impart the desired color to the inactivated fungal biomass (typically about 1 minute to about 240 minutes, or any subrange between these values, and most typically about 120 minutes). The addition of this optional component (e.g., polymer, dye, etc.) may be performed before, after, or simultaneously with the addition of the second crosslinking or tanning agent.

Throughout the retanning step 455, acids, bases, and/or other pH adjusters may be added to maintain a target pH. As a first non-limiting example, in some embodiments, it may be desirable to begin the retanning step 455 at an initial pH of about 2.0 to about 6.0 (more typically about 2.5 to about 5.5, more typically about 3.0 to about 5.0, more typically about 3.5 to about 4.5, and most typically about 4.0) and gradually raise the pH to about 3.5 to about 7.5 (more typically about 4.0 to about 7.0, more typically about 4.5 to about 6.5, more typically about 5.0 to about 6.0, and most typically about 5.5) by adding bases or other pH raisers in one or more equal portions during agitation. As a second non-limiting example, when the retanning step 455 includes the addition of a polymer, in some embodiments, it is desirable to maintain a pH of about 3.5 to about 7.5 (more typically about 4.0 to about 7.0, more typically about 4.5 to about 6.5, more typically about 5.0 to about 6.0, and most typically about 5.5) during agitation of the inactivated fungal biomass with the polymer.

The polymer may, but need not, be a biopolymer, that is, any polymeric molecule naturally produced by animals, plants or fungi, including, as non-limiting examples, cellulose, chitin, chitosan, collagen, silk fibroin, hyaluronic acid, keratin, alginate, starch, and combinations thereof. In various embodiments, the solution (or another solution of the inactivated fungal biomass and its combination in the same, preceding, or subsequent steps) may contain, in addition to the biopolymer, the following or include the following as alternatives to the biopolymer: a synthetic polymer soluble in the solvent (e.g., polyvinyl alcohol, polyethylene glycol, polysiloxane, polyphosphazene, low and/or high density polyethylene, polypropylene, polyvinyl chloride, polystyrene, nylon, polytetrafluoroethylene, thermoplastic polyurethane, polychlorotrifluoroethylene, polycaprolactone, polyacrylic acid, and/or any one or more synthetic polymers sold under various brand names (e.g., Bakelite, Kevlar, Mylar, Neoprene, Nomex, Orlon, Rilsan, Technora, Teflon, Twaron, Ultem, Vectran, Viton, Zylon, etc.). In various further embodiments, the solution may contain one or more additional components, such as plasticizers (e.g., glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g., mannitol, sorbitol) and their esters, epoxidized triglyceride vegetable oils (e.g., from soybean oil), castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof), crosslinking agents (e.g., homo-bifunctional crosslinking agents, hetero-bifunctional crosslinking agents, photoreactive crosslinking reagents, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations thereof), solubilizers (e.g., hydrochloric acid, acetic acid, formic acid, lactic acid, etc.), and/or pH adjusters (e.g., hydrochloric acid, acetic acid, formic acid, lactic acid, etc.). Alkali metal halides (e.g., sodium chloride) may be provided to prevent swelling of the inactivated fungal biomass.

The solution can be prepared by combining the polymer and the solvent, along with optionally one or more additional components, in a container and stirring or agitating the combination, optionally while simultaneously heating the combination. In various embodiments where the solution includes a solubilizer and/or a pH adjuster, either or both of these can be added to the solution after heating and stirring the other components. Preferably, the polymer (biopolymer, synthetic polymer, or a combination thereof) is completely dissolved in the solvent, and then the solution is mixed with the inactivated fungal biomass. The mixture can be heated (e.g., to about 90°C and/or to boiling) and/or stirred sufficiently to ensure that the mixture is substantially homogeneous, for example, from about 1 minute to about 240 minutes, and most typically from about 30 minutes to about 45 minutes or about 120 minutes.

In the plasticizing step 465 of method 400 shown in Figure 4, a plasticizer is added to the inactivated fungal biomass and the mixture is agitated, for example, in a drum or on a vibrating table. In various embodiments, the plasticizing step may be a fatliquoring step, i.e., the plasticizer may comprise fatliquoring oils such as sulfated castor oil, beeswax, coconut oil, vegetable oils, olive oil, linseed oil, oleic acid, sulfated fish oil, sulfated canola oil, soybean oil, palm oil, fatty acids, or combinations thereof, and may be provided in any suitable amount. The agitation may be carried out for any suitable time from about 1 minute to about 120 minutes, or in any subrange of these values, most commonly about 60 minutes. The plasticizer may be provided as an emulsion, especially when the plasticizer is a conventional leather fatliquoring oil, and in some such embodiments, the plasticizing step 465 may be terminated by adding an acid, such as hydrochloric acid, to the emulsion to break it up and allow for easier drainage and removal of the plasticizer.

In the backing step 475 of method 400 shown in Figure 4, at least one backing layer of a non-fungal fabric material is applied to and adhered to the inactivated fungal biomass. In various embodiments, the non-fungal fabric material may include any one or more of the following: acrylic fabrics, alpaca wool fabrics, angora rabbit hair fabrics, cashmere fabrics, coconut fiber fabrics, cotton fabrics, iron yarn fabrics, hemp fabrics, jute fabrics, Kevlar fabrics, flax fabrics, microfiber fabrics, mohair fabrics, nylon fabrics, olefin fabrics, cashmere fabrics, polyester fabrics, pineapple fiber fabrics, ramie fabrics, rayon fabrics, seaweed fiber fabrics, silk fabrics, sisal fabrics, spandex fabrics, spider silk fabrics, and wool fabrics. The adhesive may be any suitable lamination adhesive used in the fabric, such as polyvinyl acetate, and in some embodiments may include any suitable amount of a crosslinking agent or plasticizer, such as citric acid.

In the hot-pressing step 485 of method 400 shown in Figure 4, the inactivated fungal biomass is hot-pressed together with the non-fungal fabric backing. In various embodiments, the fungal fabric material may have at least one physical, mechanical, and/or aesthetic property that mimics or closely resembles the physical, mechanical, and/or aesthetic properties of conventional fabric materials such as leather; in particular, the hot-pressing step may be configured to impart a leather-like texture to the fungal fabric material. The temperature (e.g., about 100°C) and/or time (e.g., about 1 minute to about 20 minutes, most commonly about 10 minutes) of the hot pressing may be selected to provide the desired physical, mechanical, and/or aesthetic properties.

In the drying step 495 of method 400 shown in Figure 4, the inactivated fungal biomass is dried, optionally after being cast into a desired shape (e.g., a flat or patterned mold), to form the fungal fabric material. This drying step may or may not involve the initiation of a chemical reaction, but generally results in at least a majority of any residual water, solvent, and other liquids being expelled from the inactivated fungal biomass. The drying can be passive (i.e., at room temperature without the use of blowers, fans, etc.) or active (i.e., under heating and/or with the use of forced ventilation, dry grinding, etc.); when the drying is active, the temperature can be raised to a desired temperature above room temperature, most commonly about 80°F, and/or any suitable forced ventilation means (e.g., blowers, fans, forced ventilation dehydrators, etc.). In some embodiments, at least a portion of the fungal material may be clamped or otherwise pressed to reduce shrinkage. The curing can be allowed to continue for a time sufficient to provide the desired quality (e.g., about 18% of the quality before drying/curing) and/or moisture content of the cured material. In various embodiments, this time can be from about 1 minute to about 2 days, most commonly about 1 day.

Although not shown in Figure 4, method 400 may include at least one additional post-processing or final disposal step. In particular, one or more conventional leather finishing waxes or oils (e.g., carnauba wax, candelilla wax) or nitrocellulose may be added to the fungal fabric material in any suitable amount and at any suitable time.

Referring now to FIG5, another embodiment of a method 500 for manufacturing a fungal fabric material is shown. In the inactivation step 510 of method 500 shown in FIG5, the fungal biomass is inactivated as described herein, for example, as described for inactivation step 405 of method 400 shown in FIG4. In the lime treatment step 520 of method 500 shown in FIG5, the inactivated fungal biomass is lime-treated as described herein, for example, as described for lime treatment step 310 of method 300 shown in FIG3 and/or lime treatment step 415 of method 400 shown in FIG4. In the delime step 530 of method 500 shown in FIG5, the inactivated fungal biomass is delime-treated as described herein, for example, as described for delime treatment step 320 of method 300 shown in FIG3 and/or delime treatment step 425 of method 400 shown in FIG4.

In the impregnation step 540 of method 500 shown in FIG. 5, the inactivated fungal biomass is impregnated as described herein, for example, as described in impregnation step 330 of method 300 shown in FIG. 3 and/or impregnation step 435 of method 400 shown in FIG. 4. However, one difference between the impregnation step 540 of method 500 shown in FIG. 5 and impregnation steps in other embodiments is that at least two equal portions of a crosslinking agent, such as tannic acid, are added to the combination of the inactivated fungal biomass and the polymer solution, or conversely, the inactivated fungal biomass may be contacted with the polymer solution before, or simultaneously with, or after contact with the crosslinking agent of the first equal portion but before, simultaneously with, or after contact with the crosslinking agent of the second equal portion. In this way, method 500 of Figure 5 can, in a sense, combine the immersion, tanning, and retanning steps, such as steps 330 and 340 of method 300 and/or steps 435, 445, and 455 of method 400, into a single step that includes the immersion, tanning, and retanning sub-steps.

In the neutralization step 550 of method 500 shown in Figure 5, the pH of the inactivated fungal biomass is neutralized by contacting it with a pH neutralizing agent, which in most embodiments is an alkaline pH neutralizer such as sodium bicarbonate, but in some embodiments may be an acidic pH neutralizer. The pH neutralizer may be provided as part of an aqueous solution and may (but not necessarily) be provided in an amount suitable for providing a pH of about 7. As with the other steps, neutralization step 550 may be carried out with agitation (e.g., in a shaking flask).

In the plasticizing step 560 of method 500 shown in FIG. 5, the inactivated fungal biomass is plasticized as described herein, for example, as described for plasticizing step 350 of method 300 shown in FIG. 3 and/or plasticizing step 465 of method 400 shown in FIG. 4. In the hot-pressing step 570 of method 500 shown in FIG. 5, the inactivated fungal biomass is hot-pressed as described herein, for example, as described for hot-pressing step 370 of method 300 shown in FIG. 3 and/or hot-pressing step 485 of method 400 shown in FIG. 4.

Referring now to FIG6, another embodiment of a method 600 for manufacturing a fungal fabric material is shown. In the inactivation step 610 of method 600 shown in FIG6, the fungal biomass is inactivated as described herein, for example, as described in inactivation step 405 of method 400 shown in FIG4. Separately, in the polymer solution preparation step 615 of method 600 shown in FIG6, a polymer solution is prepared as described herein, for example, as described in impregnation step 330 of method 300 shown in FIG3. In the combination step 620 of method 600 shown in FIG6, the inactivated fungal biomass is combined with the polymer solution as described herein, for example, as described in impregnation step 330 of method 300 shown in FIG3 and/or impregnation step 435 of method 400 shown in FIG4. In the initial drying step 630 of method 600 shown in FIG6, the inactivated fungal biomass is dried as described herein, for example, as described in drying step 360 of method 300 shown in FIG3 and/or drying step 49 of method 400 shown in FIG4. In the lamination step 640 of method 600 shown in Figure 6, the inactivated fungal biomass is laminated together with one or more other inactivated fungal biomass and/or one or more non-fungal fabric material layers by any suitable method to form a composite fungal sheet. In the hot-pressing step 650 of method 600 shown in Figure 6, the composite fungal sheet is hot-pressed as described herein, for example, as described for hot-pressing step 370 of method 300 shown in Figure 3 and/or hot-pressing step 485 of method 400 shown in Figure 4. In the finishing step 660 of method 600 shown in Figure 6, one or more conventional leather finishing waxes or oils (e.g., carnauba wax, candelilla wax) or nitrocellulose may be added to the fungal fabric material in any suitable amount and at any suitable time. In the drying step 670 of method 600 shown in Figure 6, the composite fungal sheet is dried as described herein, for example, as described for drying step 360 of method 300 shown in Figure 3 and/or drying step 495 of method 400 shown in Figure 4, to form the fungal fabric material.

Typically, the methods shown in Figures 3-5 utilize a series of chemical washes, carried out under agitation, to enhance the diffusion of chemical species into the fungal structures and soften the hand feel of the finished fungal fabric material. The lime treatment step in these methods swells the matrix of the fungal structures and cleaves certain fungal proteins, thereby allowing for better diffusion of chemical species into the fungus and enabling chemically active sites to react in subsequent process steps. Vegetable tannins are thus effective in forming large hydrogen bond networks and thus crosslinking the fungal structures, thereby providing the strength, color, odor, and/or chemical stability properties of genuine leather. As in the methods shown in Figures 1 and 2, reinforcing polymers (e.g., chitosan) and additional non-tannin crosslinking agents (e.g., citric acid) can be used; in addition to having the effects described above for Figures 1 and 2, the reinforcing polymer and the non-tannin crosslinking agent can also form complexes with the tannin crosslinking agent. Similarly, plasticizers (e.g., glycerin) can be introduced into this method.

It should be clearly understood that any one or more filamentous fungi may be suitable for forming the fungal fabric material of the present invention, including but not limited to one or more filamentous fungi belonging to the following phyla: Ascomycota and Basidiomycota; one or more filamentous fungi belonging to the following orders: Ustilales, Russula, Agaricales, Discales, and Hypocreales; one or more filamentous fungi belonging to the following families: Ustilaginaceae, Hericaceae, Polyporaceae, Trichophyceae, Agaricales, Strophariaceae, Lycoperdonaceae, Agaricales, Pleurotaceae, Pleurotaceae, Gastrodiaceae, Trichaceae, Morchaceae, Hydrangeaceae, Agaricales, and Cordycepsaceae; and one or more filamentous fungi belonging to the following genera: Agaricales. Genus, including *Amanita*, *Lycoperdon*, *Cordyceps*, *Pleurotus*, *Flammulina*, *Fusarium*, *Ganoderma*, *Ganoderma*, *Hericium*, *Amanita*, *Mammillaria*, *Morchella*, *Morchella*, *Morchella*, *Leptochloa*, *Pleurotus*, *Polyporus*, *Hydrangea*, *Stropharia*, *Truffle*, *Ustilago*; and/or one or more filamentous fungi selected from the following species: *Ustilago maydis*, *Hericium*, *Polyporus floribunda*, *Sterculia*. The following fungi are listed: *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Tuber borscherita*, *Morchella*, *Morchella cuspidatum*, *Morchella esculenta*, *Morchella esculenta*, *Fusarium oxysporum*, *MK7 ATCC accession number PTA-10698*, *Pleurotus ribosus*, and *Cordyceps militaris*.

In practice, the inactivated fungal biomass is permitted to be immersed in and/or agitated together with the polymer, plasticizer, and/or crosslinking agent solution for a time sufficient to allow the pad to be permeated and/or saturated by the polymer, plasticizer, and/or crosslinking agent, typically at least about 1 hour. After immersion in and/or agitation together with the solution, the wet pad is removed from the solution (and excess solution can then be removed from one or more surfaces of the pad).

Plasticizers suitable for use in the fungal fabric materials of the present invention include, but are not limited to, glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g., mannitol, sorbitol) and their esters, epoxidized triglycerides, vegetable oils (e.g., from soybean oil), castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof. In practice of the invention, the plasticizer is typically present in the fungal fabric material in amounts ranging from about 0.5% by weight to about 50% by weight, or any subrange between these values, including, as non-limiting examples, about 50% by weight, about 37.5% by weight, about 25% by weight, or about 12.5% by weight.

Polymers suitable for use in the fungal fabric materials of the present invention include, but are not limited to, polyvinyl alcohol, chitosan, polyethylene glycol, polycaprolactone, polyacrylic acid, hyaluronic acid, alginate, and combinations and mixtures thereof. In various embodiments, two or more polymers may be included in any weight ratio of: about 99:1 to about 1:99; typically about 99:1, about 90:10, about 80:20, about 70:30, about 60:40, about 50:50, about 40:60, about 30:70, about 20:80, about 10:90, or about 1:99; and more typically about 50:50. The loading ratio of the fabric composition can take any of the following values: about 99:1 to about 1:99; typically about 99:1, about 95:5, about 90:10, about 85:15, about 80:20, about 75:25, about 70:30, about 65:35, about 60:40, about 55:45, about 50:50, about 45:55, about 40:60, about 35:65, about 30:70, about 25:75, about 20:80, about 15:85, about 10:90, about 5:95, and about 1:99; and more typically about 70:30.

Crosslinking agents suitable for use in the fungal fabric materials of the present invention include, but are not limited to, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations and mixtures thereof. In various embodiments, the fungal fabric material may contain proteins crosslinked with isopeptide bonds, and in some embodiments, its formation may be catalyzed by glutaminase.

The relative amounts of filamentous fungi, plasticizers, polymers, crosslinking agents, additives, etc., in the fungal fabric material of the present invention can be selected to provide a fungal fabric material having one or more desired physical, mechanical, sensory (e.g., olfactory, tactile, etc.) and/or aesthetic properties. In various embodiments, scented additives, such as leather fragrance, may be added to the fungal fabric material to provide the fungal fabric material with desired olfactory properties, such as a leathery aroma.

The filamentous fungus may constitute about 20% to 90% of the fungal fabric material, or any subrange between these values. In some embodiments, the filamentous fungus may constitute about 25-85%, about 30-80%, or about 35-75% of the fungal fabric material. For example, in a non-limiting example, in some embodiments, it may constitute about 40% to about 60% by weight of the fungal fabric material.

As a further non-limiting example, one or more polymers (e.g., chitosan) may constitute about 1% to about 40% by weight, or any subrange between these values, or about 5% to about 20% by weight, of the fungal fabric material. As a third non-limiting example, one or more crosslinking agents (e.g., citric acid) may constitute about 0.01% to about 8% by weight, or any subrange between these values, or about 0.05% to about 6% by weight, or about 0.1% to about 4% by weight, of the fungal fabric material. As a fourth non-limiting example, one or more plasticizers (e.g., glycerol) may constitute about 0.5% to about 80% by weight, or any subrange between these values, or about 9% to about 60% by weight, or about 17.5% to about 40% by weight, of the fungal fabric material.

Various embodiments of the present invention include fungal fabric materials having modified and/or adjusted thermal properties, and in particular fungal leather-like materials. As a first non-limiting example, according to the present invention, the heat storage coefficient of the fungal fabric material, i.e., the rate at which the fungal fabric material exchanges heat with its surroundings, can be modified or adjusted. As a second non-limiting example, according to the present invention, the thermal conductivity of the fungal fabric material, i.e., the heat transferred through the fungal fabric material, can be modified or adjusted. As a third non-limiting example, according to the present invention, the heat capacity, i.e., the amount of heat supplied to a given mass of the fungal fabric material to produce a unit change in its temperature, can be modified or adjusted. According to the present invention, the volumetric heat capacity of the fungal fabric material, i.e., the amount of heat that a volume of the fungal fabric material can store, can be modified or adjusted. The ability to thermally modify and/or adjust the fungal fabric material allows it to have the desired "warmth" and thus represents a significant improvement over prior art fungal or other non-animal fabric materials, which often suffer from the disadvantage of "feeling cold" (i.e., having poor thermal properties) and/or (e.g., providing insufficient insulation for the wearer of clothing made from the fungal fabric material). The present invention therefore allows for the production of fungal fabrics, for example, that retain greater heat and are therefore suitable for use in winter clothing. A further advantage and benefit of the invention lies in the ability to produce fabric materials that can have a combination of two or more of these or other thermal properties that are not achievable with conventional fabric materials, such as improving one thermal property while improving, maintaining, or reducing one or more other thermal properties, and/or maintaining one thermal property constant while improving, maintaining, or reducing one or more other thermal properties, and/or reducing one thermal property while improving, maintaining, or reducing one or more other thermal properties.

The thermal properties of the fungal fabric material of the present invention can be modified or adjusted by including a thermal dopant in the fungal fabric material. Suitable thermal dopants for use in the fungal fabric material of the present invention include one or more materials that alter the heat storage coefficient, thermal conductivity, and heat capacity of the fungal fabric material compared to the fungal fabric material without the thermal dopant. Such thermal dopants may include, but are not limited to, polymers, ceramics, and metallic materials having known thermal properties, and/or any other materials having desired thermal conductivity and/or thermal insulation properties. Other non-limiting examples of suitable thermal dopants for use in the present invention include activated carbon, alumina, bentonite, diatomaceous earth, ethylene vinyl acetate, lignin, nano-silica, polycaprolactone, polylactic acid, organosilicon, and yttrium oxide. In some embodiments, the thermal dopant may comprise a modified coating of thermally conductive and/or thermally insulating materials and/or a modified spatial distribution of thermally conductive and/or thermally insulating materials in the fungal fabric material to produce a preselected thermal profile.

In practice, the thermal dopant can be added to and/or introduced into the fungal fabric material at any suitable point in the manufacturing process. As a first non-limiting example, the thermal dopant can be provided in the polymer solution, i.e., in combination with the polymer and solvent, and subsequently in combination with the fungal biomass. As a second non-limiting example, the thermal dopant can be combined with the inactivated fungal biomass, water, and optionally pigments before or during the crushing and/or blending/homogenization steps of the manufacturing process. As a third non-limiting example, the thermal dopant can be added to a mixture of the fungal paste and the polymer solution while the paste/polymer solution mixture is stirred and/or heated. As a fourth non-limiting example, the thermal dopant, and in some embodiments, the thermal dopant with modified or designed spatial patterns or structures, can be integrated into the fungal fabric material. As a fifth non-limiting example, the thermal dopant can be provided before or during the casting step (e.g., by providing the thermal dopant in a tray or mold in which the cast sheet is placed, or by sprinkling or otherwise distributing dopant particles on the surface of the fungal material after casting). As a sixth non-limiting example, a thermal dopant may be added to the fungal fabric material after it has been cured.

The amount of the thermal dopant can be selected to provide the desired thermal properties to the resulting fungal fabric material without impairing other material properties of the fungal fabric material (e.g., flexibility, tensile strength, etc.). Typically, when provided, the thermal dopant may constitute about 0.1 wt% to about 25% of the fungal fabric material, or any subrange between these values. In some embodiments, the dopant may be present in about 0.1 to about 20 wt% of the fungal fabric material, or in about 0.1 to about 15 wt%. For example, in many embodiments, the dopant may constitute about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, about 15 wt%, or any tenths of a weight percentage between 0.1 and 25 wt%.

In various embodiments, the fungal composition formed after mixing with the polymer solution can be cast to at least partially cover a scaffold or substrate containing a thermal dopant. In various embodiments, a force can be applied to at least one of the fungal composition and the scaffold or substrate to provide a heterogeneous spatial distribution of the fungal composition and the scaffold or substrate in the cast sheet. In various embodiments, the fungal composition and the thermal dopant can be selectively applied to predetermined regions of the casting area to provide a heterogeneous spatial distribution of the blend composition and the thermal dopant in the cast sheet. In various embodiments, the cast sheet may comprise a multilayer structure having at least a first layer and a second layer, the first layer comprising at least a portion of the fungal composition and the second layer comprising at least a portion of the thermal dopant.

Various embodiments of the present invention include clothing articles constructed partially or entirely from the fungal fabric material of the present invention. Such clothing articles include, as non-limiting examples, protective clothing, shirts, trousers, shorts, jackets, coats, belts, hats, gloves, shoes, boots, sandals, flip-flops, watch straps, and aprons.

Various embodiments of the present invention include accessory articles constructed partially or entirely from the fungal fabric material of the present invention. Such accessory articles include, as non-limiting examples, wallets, handbags, cases, briefcases, luggage, backpacks, kiosks, and waist bags.

Various embodiments of the present invention include furniture articles constructed partially or entirely from the fungal fabric material of the present invention. Such furniture articles include, as non-limiting examples, chairs, recliners, daybeds, sofas, armchairs, and ottomans.

Various embodiments of the present invention include covers constructed partially or entirely from the fungal fabric material of the present invention. Such covers include, as non-limiting examples, covers for car seats, airplane seats, and train seats.

The fungal fabric materials according to the invention can be manufactured such that they are characterized by desired material, mechanical, and/or physical properties. As a first non-limiting example, the fungal fabric material can be manufactured to have a desired tensile strength, which in various embodiments may be at least about 15 MPa or about 4 MPa to about 15 MPa, or any subrange between these values. As a second non-limiting example, the fungal fabric material can be manufactured to have a desired breaking strain, which in various embodiments may be about 50% to about 60%, or about 10% to about 70%, or any subrange between these values. As a third non-limiting example, the fungal fabric material can be manufactured to have a desired degree of swelling, which in various embodiments may be about 50% to about 60%, or about 30% to about 120%, or any subrange between these values. As a fourth non-limiting example, the fungal fabric material can be manufactured to have a desired mass loss after immersion, which in various embodiments may be no more than about 5%. As a fifth non-limiting example, the fungal fabric material can be manufactured to have a desired average fungal particle size, which in various embodiments may be no greater than about 25 nanometers, no greater than about 50 nanometers, no greater than about 75 nanometers, no greater than about 100 nanometers, no greater than about 125 nanometers, no greater than about 150 nanometers, no greater than about 175 nanometers, no greater than about 200 nanometers, no greater than about 225 nanometers, no greater than about 250 nanometers, no greater than about 275 nanometers, no greater than about 300 nanometers, no greater than about 325 nanometers, no greater than about 350 nanometers, and no greater than about 375 nanometers. Not greater than approximately 400 nanometers, not greater than approximately 425 nanometers, not greater than approximately 2 micrometers, not greater than approximately 4 micrometers, not greater than approximately 6 micrometers, not greater than approximately 8 micrometers, not greater than approximately 10 micrometers, not greater than approximately 15 micrometers, not greater than approximately 20 micrometers, not greater than approximately 30 micrometers, not greater than approximately 40 micrometers, not greater than approximately 50 micrometers, not greater than approximately 75 micrometers, not greater than approximately 100 micrometers, not greater than approximately 150 micrometers, not greater than approximately 200 micrometers, not greater than approximately 250 micrometers, not greater than approximately 300 nanometers, not greater than approximately 400 micrometers, not greater than approximately 500 micrometers, and not greater than approximately 750 micrometers. In some embodiments, the fungal biomass may comprise fungal filaments of lengths of at least about 1 cm, at least about 2 cm, at least about 3 cm, at least about 4 cm, at least about 5 cm, at least about 6 cm, at least about 7 cm, at least about 8 cm, at least about 9 cm, at least about 10 cm, at least about 20 cm, at least about 30 cm, at least about 40 cm, at least about 50 cm, at least about 60 cm, at least about 70 cm, at least about 80 cm, or at least about 90 cm. As a sixth non-limiting example, the fungal fabric material may be manufactured to have a desired type of particle size distribution, which in various embodiments may be bimodal, approximately bimodal, trimodal, or approximately trimodal particle size distribution. As a seventh non-limiting example, the fungal fabric material may be manufactured to have a desired tear strength, which in various embodiments may be about 5 N/mm to about 25 N/mm, or any subrange between these values. As an eighth non-limiting example, the fungal fabric material can be manufactured to have a desired colorfastness to wet rubbing, dry rubbing, and/or xenon light with a grayscale of at least about 4. As a ninth non-limiting example, the fungal fabric material can be manufactured to have a desired flexural stiffness, which in various embodiments may be no greater than about 5 g-cm. One of the advantages and benefits of the invention is the ability to produce fabric materials that can have two or more combinations of these or other material, mechanical, and/or physical properties that are not achievable with conventional fabric materials, such as high tear strength (in some embodiments, at least about 1 N/mm, or at least about 2 N/mm, or at least about 3 N/mm, or at least about 4 N/mm, or at least about 5 N/mm, or at least about 6 N/mm, or at least about 7 N/mm, or at least about 8 N/mm, or at least about 9 N/mm, or at least about 10 N/mm, or at least about 11 N/mm, or at least about 12 N/mm, or at least about 13 N/mm). A combination of 1/mm or at least about 14 N/mm, or at least about 15 N/mm, or at least about 16 N/mm, or at least about 17 N/mm, or at least about 18 N/mm, or at least about 19 N/mm, or at least about 20 N/mm) and low flexural stiffness (in some embodiments, not greater than about 10 g-cm, or not greater than about 9 g-cm, or not greater than about 8 g-cm, or not greater than about 7 g-cm, or not greater than about 6 g-cm, or not greater than about 5 g-cm, or not greater than about 4 g-cm, or not greater than about 3 g-cm, or not greater than about 2 g-cm, or not greater than about 1 g-cm).

In some embodiments, a fungal leather analog material can be provided, made from pulverized, inactivated fungal biomass and free of any non-fungal fabric backing. Such a fungal leather analog material may have one or more of the following properties: a thickness of about 1 to about 2 mm or about 1.15 to about 1.6 mm; a tear strength of about 6 to about 12 N or about 7.4 to about 10.5 N; a tensile strength of about 3 to about 10 N/ ^mm² or about 4.7 to about 8.1 N/ ^mm² ; a flexural stiffness of about 1 to about 11 g·cm; a water stain grayscale grade of 4 to 5; a light color fastness to blue wool grade of at least 4; a rubbing fastness grayscale grade of 4 to 5 when dry; and a rubbing fastness grayscale grade of 4 to 5 when dry. Such a fungal leather analog material can readily exhibit various textures or embossments.

In some embodiments, a fungal leather analog material can be provided, made from pulverized, inactivated fungal biomass and with a non-fungal fabric backing adhered to one side. Such a fungal leather analog material may have any one or more of the following properties: a thickness of about 1 to about 3 mm or about 1.95 to about 2.09 mm; a tear strength of about 20 to about 50 N or about 33 to about 37 N; a tensile strength of about 3 to about 10 N/ ^mm² or about 5.8 to about 6.8 N/ ^mm² ; a flexural stiffness of about 1 to about 11 g·cm; a water stain grayscale grade of 4 to 5; a light color fastness to blue wool grade of at least 4; a rubbing fastness grayscale grade of 4 to 5 when dry; and a rubbing fastness grayscale grade of 4 to 5 when dry. Such a fungal leather analog material can readily produce various textures or embossed prints.

In some embodiments, a complex fungal leather analog material can be provided, made from pulverized, inactivated fungal biomass and in which a non-fungal fabric layer is adhered between two layers of fungal material (i.e., a material in which the non-fungal layer is "sandwiched" between fungal layers). Such a fungal leather analog material may have any one or more of the following properties: a thickness of about 1 to about 4 mm or about 2.2 to about 2.8 mm; a tear strength of about 25 to about 60 N or about 34 to about 52 N; a tensile strength of about 7 to about 14 N/ ^mm² or about 8.7 to about 11.4 N/ ^mm² ; a flexural stiffness of about 1 to about 11 g·cm; a water stain grayscale grade of 4 to 5; a light color fastness to blue wool grade of at least 4; a rubbing fastness grayscale grade of 4 to 5 when dry; and a rubbing fastness grayscale grade of 4 to 5 when dry. Such a fungal leather analog material can readily produce various textures or embossed prints.

In some embodiments, a fungal leather analog material can be provided, made from inactivated fungal biomass (e.g., biomass produced by surface fermentation and not subjected to breakage) in the form of one or more intact or whole biomasses, and free of any non-fungal fabric backing. Such a fungal leather analog material may have one or more of the following properties: a thickness of about 0.1 to about 1.5 mm per biomass or about 0.5 to about 0.9 mm per biomass, a tear strength of about 1 to about 3 N per biomass, a tensile strength of about 3 N/ ^mm² per biomass, a flexural stiffness of about 1 to about 11 g·cm, and a watermark grayscale grade of 4-5. Such a fungal leather analog material may have advantages that closely mimic the same quality of genuine leather, such as warmth, drape, softness, appearance, and odor.

In some embodiments, fungal leather analogs made from inactivated fungal biomass, and methods for manufacturing them, offer environmental advantages and benefits relative to genuine leather, in addition to the absence of animal products. Specifically, the manufacturing method of the present invention produces little or no of highly toxic or environmentally harmful materials used in conventional leather tanning processes, such as hexavalent chromium compounds. Furthermore, the leather analogs according to the present invention can be biodegradable, i.e., biodegrade more rapidly than genuine leather under the given set of conditions.

A feature of this invention is the ability to allow various chemical components (polymers, crosslinking agents, etc.) to infiltrate the mycelial matrix of inactivated fungal biomass. When the inactivated fungal biomass is fragmented fungal biomass, the infiltration can be a structure in which fungal particles with a high surface area come into contact with an infiltration fluid (one or more). When the inactivated fungal biomass is whole or cohesive fungal biomass (e.g., a biomass produced by surface fermentation), the infiltration can be achieved by any one or more of the following: prolonging the contact time between the fungal biomass and the fluid (one or more), agitating the fungal biomass and the fluid (one or more), applying pressure below or above atmospheric pressure to the fungal biomass and the fluid (one or more), etc.

One aspect of the present invention is to provide a method for preparing a durable sheet material comprising fungal biomass, comprising (a) combining inactivated fungal biomass with at least one component selected from plasticizers, polymers, crosslinking agents, and dyes to form a combined composition; (b) casting the combined composition to form a cast sheet; (c) removing solvent from the cast sheet; and (d) curing the cast sheet to form the durable sheet material. It should be clearly understood that this method can be used with whole cohesive biomass (e.g., biomats produced by surface fermentation) or broken fungal biomass.

In various embodiments, step (d) may include drying the cast sheet.

In various embodiments, step (d) may include initiating a chemical reaction within the cast sheet or on the surface of the cast sheet.

In various embodiments, the method may further include adding at least one of a natural fibrous material, a synthetic material, or a combination thereof to the blend composition. The natural fibrous material may, but does not necessarily, comprise a cellulose material. The natural fibrous material may, but does not necessarily, comprise cotton fibers. The at least one of the natural fibrous material and the synthetic material may, but does not necessarily, be in the form of multiple particles, sheets, or a combination thereof.

In various embodiments, the method may further include inactivating fungal biomass to form the inactivated fungal biomass.

In various embodiments, the method may further include breaking down fungal biomass.

In various embodiments, the method may further include adding a thermal dopant to at least one of the following: the inactivated fungal biomass, the blend composition, and the cast sheet.

Another aspect of the present invention is to provide a method for preparing a durable sheet material comprising fungal biomass, comprising (a) contacting inactivated fungal biomass with a solution comprising at least one component selected from plasticizers, polymers, crosslinking agents and dyes; (b) removing solvent from the biomass; and (c) curing the biomass to form the durable sheet material.

In various embodiments, the method may further include inactivating fungal biomass to form the inactivated fungal biomass.

Another aspect of the invention is to provide a fabric composition comprising inactivated fungal biomass and at least one component selected from plasticizers, polymers, crosslinking agents, and dyes.

In various embodiments, the fabric composition may include a plasticizer, a polymer, and a crosslinking agent.

In various embodiments, the fungal biomass may comprise fungi belonging to the phyla selected from Ascomycota and Basidiomycota.

In various embodiments, the fungal biomass may comprise fungi belonging to genera selected from the following: *Fusarium*, *Fomes*, and *Ganoderma*. The fungus may, but need not, belong to species selected from the following: *Fusarium veneriense*, *Fomes fomentarius*, *Ganoderma applanatum*, *Ganoderma curtisii*, *Ganoderma formosanum*, *Ganoderma nei-japonicum*, *Ganoderma resinaceum*, *Ganoderma sinense*, and *Ganoderma tsugae*.

In various embodiments, the fungal biomass may comprise fungi selected from Fusarium vesicae and MK7ATCC accession number PTA-10698.

In various embodiments, the plasticizer may comprise at least one selected from glycerol, polyethylene glycol, citric acid, and oleic acid. The plasticizer may, but does not necessarily, comprise glycerol. Glycerol may, but does not necessarily, be present in the fabric composition in an amount of about 0.5% by weight, about 50% by weight, or any subrange of these values. In various embodiments, glycerol may, but does not necessarily, be present in an amount of about 50% by weight, about 37.5% by weight, about 25% by weight, or about 12.5% by weight.

In various embodiments, the polymer may comprise at least one selected from: polyvinyl alcohol, chitosan, polyethylene glycol, and hyaluronic acid. The polymer may, but does not necessarily, comprise polyvinyl alcohol. The polymer may, but does not necessarily, comprise chitosan. The polymer may, but does not necessarily, comprise both polyvinyl alcohol and chitosan. The weight ratio of polyvinyl alcohol to chitosan may, but does not necessarily, be selected from about 99:1, about 90:10, about 80:20, about 70:30, about 60:40, about 50:50, about 40:60, about 30:70, about 20:80, about 10:90, and about 1:99, or any range formed by two of these ratios. The weight ratio of polyvinyl alcohol to chitosan may, but does not necessarily, be about 50:50.

In various embodiments, the fabric composition may comprise a polymer, wherein the loading ratio of the fabric composition is selected from about 99:1, about 95:5, about 90:10, about 85:15, about 80:20, about 75:25, about 70:30, about 65:35, about 60:40, about 55:45, about 50:50, about 45:55, about 40:60, about 35:65, about 30:70, about 25:75, about 20:80, about 15:85, about 10:90, about 5:95, and about 1:99, or any range formed by two of these ratios. The loading ratio may, but is not required, be about 70:30.

In various embodiments, the crosslinking agent may comprise at least one selected from: citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, glyoxal, and extracted vegetable tannins. The crosslinking agent may, but does not necessarily, comprise adipic acid.

Another aspect of the present invention is to provide clothing articles comprising the fabric composition of the present invention.

In various embodiments, the product can be protective clothing.

In various embodiments, the garment article may be selected from shirts, trousers, shorts, jackets, coats, belts, hats, gloves, shoes, boots, sandals, flip-flops, watch straps, and aprons.

Another aspect of the invention is to provide accessory articles comprising the fabric composition of the invention.

In various implementations, the accessory can be selected from wallets, handbags, cases, briefcases, luggage, backpacks, kiosks, and waist bags.

Another aspect of the invention is to provide furniture articles comprising the fabric composition of the invention.

In various implementations, the furniture item may be selected from chairs, recliners, daybeds, sofas, armchairs, footstools, and vehicle tables and chairs.

In various embodiments, the fabric composition may have a tensile strength of at least about 15 MPa.

In various embodiments, the fabric composition may have a breaking strain of about 30% to about 60%.

In various embodiments, the fabric composition may have a swelling degree of about 30% to about 60%.

In various embodiments, the fabric composition may have a post-immersion mass loss of no more than about 30%.

In various embodiments, the fungal biomass may have an average particle size selected from the following: not greater than about 25 nanometers, not greater than about 50 nanometers, not greater than about 75 nanometers, not greater than about 100 nanometers, not greater than about 125 nanometers, not greater than about 150 nanometers, not greater than about 175 nanometers, not greater than about 200 nanometers, not greater than about 225 nanometers, not greater than about 250 nanometers, not greater than about 275 nanometers, not greater than about 300 nanometers, not greater than about 325 nanometers, not greater than about 350 nanometers, not greater than about 375 nanometers, not greater than about 400 nanometers. Not greater than approximately 425 nanometers, not greater than approximately 2 micrometers, not greater than approximately 4 micrometers, not greater than approximately 6 micrometers, not greater than approximately 8 micrometers, not greater than approximately 10 micrometers, not greater than approximately 15 micrometers, not greater than approximately 20 micrometers, not greater than approximately 30 micrometers, not greater than approximately 40 micrometers, not greater than approximately 50 micrometers, not greater than approximately 75 micrometers, not greater than approximately 100 micrometers, not greater than approximately 150 micrometers, not greater than approximately 200 micrometers, not greater than approximately 250 micrometers, not greater than approximately 300 nanometers, not greater than approximately 400 micrometers, not greater than approximately 500 micrometers, and not greater than approximately 750 micrometers.

In various embodiments, the fungal biomass may have a bimodal or near-bimodal particle size distribution.

In various embodiments, the fungal biomass may have a trimodal or near-trimodal particle size distribution.

In various embodiments, the fabric composition may contain glutamine transferase.

In various embodiments, the fabric composition may contain proteins cross-linked with isopeptide bonds. The formation of these isopeptide bonds may, but is not necessary, be catalyzed by glutamine transferase.

In various embodiments, the fabric composition may further comprise a thermal dopant. This thermal dopant may, but is not required, be selected from activated carbon, alumina, bentonite, diatomaceous earth, lignin, nano-silica, polycaprolactone, polylactic acid, organosilicon, and yttrium oxide.

Another aspect of the present invention is to provide a method for preparing a durable sheet material comprising fungal biomass, comprising (a) homogenizing inactivated fungal biomass with a fluid comprising water to form a fungal paste; (b) combining the fungal paste with an aqueous solution comprising a polymer to form a blend composition; (c) casting the blend composition to form a cast sheet; (d) removing the solvent from the cast sheet; and (e) curing the cast sheet to form the durable sheet material. It should be clearly understood that the fungal biomass suitable for use in this method can be produced by any of a number of methods known in the art and disclosed herein, including but not limited to surface fermentation methods, submerged fermentation methods, solid-substrate submerged fermentation (SSSF) methods, and methods as disclosed in '474 publication.

In various embodiments, the fluid in step (a) may further contain pigment.

In various embodiments, the method may further include inactivating fungal biomass to provide the inactivated fungal biomass.

In various embodiments, step (a) may further include simultaneously crushing the inactivated fungal biomass.

In various embodiments, the inactivated fungal biomass may be pulverized fungal biomass.

In various embodiments, the polymer may comprise chitosan.

In various embodiments, the aqueous solution of step (b) may further comprise at least one of a crosslinking agent, a plasticizer, a solubilizer, and a pH adjuster. The aqueous solution may, but does not necessarily, comprise a crosslinking agent, wherein the crosslinking agent comprises citric acid. The aqueous solution may, but does not necessarily, comprise a plasticizer, wherein the plasticizer comprises glycerol.

In various embodiments, the method may further include degassing the fungal paste between steps (a) and (b).

In various embodiments, the method may further include degassing the blend composition between steps (b) and (c).

In various embodiments, the method may further include adding at least one thermal dopant to at least one of the following: inactivated fungal biomass, the fluid of step (a), fungal paste, an aqueous solution of step (b), a blend composition, a cast sheet, and a tray, mold, or other container in which the blend composition is cast in step (c).

In various embodiments of any of the above methods, the fungal biomass can be produced by methods comprising: culturing a fungal inoculum by at least one of the following methods: surface fermentation, submerged fermentation, solid-substrate submerged fermentation, and fermentation as described in '474'. The fungal biomass may, but need not, be a biomass mat.

Any of the above methods may be further implemented by retaining or introducing at least one bubble.

Various embodiments of the above fabric composition may include at least one air bubble.

In various embodiments, the fungal biomass may comprise fungal filaments having the following lengths: at least about 1 cm, at least about 2 cm, at least about 3 cm, at least about 4 cm, at least about 5 cm, at least about 6 cm, at least about 7 cm, at least about 8 cm, at least about 9 cm, at least about 10 cm, at least about 20 cm, at least about 30 cm, at least about 40 cm, at least about 50 cm, at least about 60 cm, at least about 70 cm, at least about 80 cm, at least about 90 cm, at least about 100 cm, at least about 200 cm, at least about 300 cm, at least about 400 cm, at least about 500 cm, at least about 600 cm, at least about 700 cm, at least about 800 cm, or at least about 900 cm.

In various embodiments, the fungal biomass may comprise fungal filaments having the following lengths: not greater than about 1 cm, not greater than about 9 mm, not greater than about 8 mm, not greater than about 7 mm, not greater than about 6 mm, not greater than about 5 mm, not greater than about 4 mm, not greater than about 3 mm, not greater than about 2 mm, not greater than about 1 mm, not greater than about 900 μm, not greater than about 800 μm, not greater than about 700 μm, not greater than about 600 μm, not greater than about 500 μm, not greater than about 400 μm, not greater than about 30 μm. 0 micrometers, not greater than about 200 micrometers, not greater than about 100 micrometers, not greater than about 90 micrometers, not greater than about 80 micrometers, not greater than about 70 micrometers, not greater than about 60 micrometers, not greater than about 50 micrometers, not greater than about 40 micrometers, not greater than about 30 micrometers, not greater than about 20 micrometers, not greater than about 10 micrometers, not greater than about 9 micrometers, not greater than about 8 micrometers, not greater than about 7 micrometers, not greater than about 6 micrometers, not greater than about 5 micrometers, not greater than about 4 micrometers, not greater than about 3 micrometers, not greater than about 2 micrometers, or not greater than about 1 micrometer.

The present invention also relates to the following embodiments.

1. A method for preparing a durable sheet material comprising fungal biomass, comprising:

(a) To penetrate a solution into inactivated fungal biomass, the solution comprising a solvent and components selected from polymers, crosslinking agents, and combinations and mixtures thereof; and

(b) Solidify the biomass to remove the solvent from the biomass and form the durable sheet material.

2. The method of item 1, wherein the fungal biomass comprises fungal mycelium.

3. The method of claim 1, wherein the inactivated fungal biomass is crushed prior to step (a) and wherein step (a) comprises blending the crushed inactivated fungal biomass with the solution to form a blend composition.

4. The method of item 3, further comprising casting the blend composition to form a cast sheet, wherein in step (b), the solvent is removed from the cast sheet.

5. The method of item 3, wherein the ruptured inactivated fungal biomass has an average particle size of not more than about 125 micrometers.

6. The method of item 1, wherein the inactivated fungal biomass comprises cohesive fungal biomass and wherein step (a) comprises agitating the inactivated fungal biomass and the solution together for a period of time.

7. The method of item 6, wherein the cohesive fungal biomass is produced by a surface fermentation process or a deep solid surface fermentation process.

8. The method of item 6, wherein the period is selected from at least about 4 hours, at least about 5 hours, at least about 10 hours, at least about 15 hours, at least about 20 hours, or at least about 25 hours.

9. The method in item 6, wherein the period is approximately 10 hours to approximately 20 hours.

10. The method of item 6, wherein the agitation is carried out under a pressure different from atmospheric pressure.

11. The method of item 10, wherein the pressure is lower than atmospheric pressure.

12. The method of item 10, wherein the pressure is higher than atmospheric pressure.

13. The method of item 6, further comprising treating the inactivated fungal biomass with at least one chemical selected from calcium hydroxide and tannins.

14. The method of item 1, wherein the inactivated fungal biomass comprises a fungal paste produced by deep fermentation.

15. The method of claim 1, wherein the solution comprises a polymer selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations thereof.

16. The method of claim 1, wherein the solution comprises a polymer and the polymer is present in the durable sheet material in an amount selected from: no more than about 25% by weight of the durable sheet material, no more than about 20% by weight of the durable sheet material, no more than about 15% by weight of the durable sheet material, no more than about 10% by weight of the durable sheet material, and no more than about 5% by weight of the durable sheet material.

17. The method of claim 1, wherein the solution comprises a crosslinking agent selected from the group consisting of citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations thereof.

18. The method of item 1, wherein the solution further comprises a plasticizer.

19. The method of item 18, wherein the plasticizer is selected from glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols and their esters, epoxidized triglyceride vegetable oil, castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations thereof.

20. The method of item 1, wherein the fungal biomass comprises at least one filamentous fungus selected from the following orders: Ustilagoesales, Russulaesales, Agaricalesales, Discotylesales, and Hypocreales.

21. The method of claim 1, wherein the fungal biomass comprises at least one filamentous fungus belonging to the families selected from: Ustilagoeaceae, Hericaceae, Polyporaceae, Trichophyceae, Agaricaceae, Strophariaceae, Lycoperdonaceae, Agaricaceae, Pleurotaceae, Pleurotaceae, Gastromycetes, Gastromycetes, Tuberaceae, Morchellaceae, Hydrangeaceae, Agaricaceae, and Cordycepsaceae.

22. The method of claim 1, wherein the fungal biomass comprises at least one filamentous fungus belonging to the genera selected from the following genera: Agaricales, Lepidium, Lycoperdon, Cordyceps, Pleurotus, Fomitopsis, Fusarium, Ganoderma, Hericium, Hericium, Agaricus, Agaricus, Morel, Lepidium, Pleurotus, Polyporaceae, Pleurotus, Tricholoma, and Ustilago.

23. The method of item 1, wherein the fungal biomass comprises at least one filamentous fungus selected from the following: *Ustilago maydis*, *Heliotropium indicum*, *Polyporus floribunda*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus*, *Pleurotus ostreatus* var. *columbine*, *Tuberose*, *Morchella esculenta*, *Morchella esculenta*, *Morchella esculenta*, *Morchella esculenta*, *Morchella esculenta*, *Morchella esculenta*, *Morchella esculenta*, *Morchella esculenta*, *Fusarium oxysporum*, MK7 ATCC accession number PTA-10698, *Pleurotus ribosus*, and *Cordyceps militaris*.

24. The method of claim 1, wherein the solution further comprises at least one of a pigment, a solubilizer, and a pH adjuster.

25. The method of item 24, wherein the solution comprises a solubilizer selected from hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations and mixtures thereof.

26. The method of item 24, wherein the solution contains a pH adjuster selected from hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations and mixtures thereof.

27. The method of item 1, wherein the durable sheet material comprises a protein cross-linked with isopeptide bonds.

28. The method of item 1, further comprising at least one of the following: (i) adding a thermal dopant to the inactivated fungal biomass, and (ii) adding a thermal dopant to the durable sheet material after step (b).

29. The method of claim 28, wherein the amount of the thermal dopant is selected from at least about 2.5% by weight of the durable sheet material, at least about 5% by weight of the durable sheet material, at least about 7.5% by weight of the durable sheet material, at least about 10% by weight of the durable sheet material, at least about 12.5% by weight of the durable sheet material, at least about 15% by weight of the durable sheet material, and at least about 17.5% by weight of the durable sheet material.

30. The method of claim 28, wherein the amount of the thermal dopant is selected from no more than about 20% by weight of the durable sheet material, no more than about 17.5% by weight of the durable sheet material, no more than about 15% by weight of the durable sheet material, no more than about 12.5% by weight of the durable sheet material, no more than about 10% by weight of the durable sheet material, no more than about 7.5% by weight of the durable sheet material, and no more than about 5% by weight of the durable sheet material.

31. The method of item 28, wherein the thermal dopant is selected from ceramic materials, metallic materials, polymer materials, and combinations thereof.

32. The method of item 28, wherein the thermal dopant is selected from activated carbon, alumina, bentonite, diatomaceous earth, ethylene vinyl acetate, lignin, nano silica, polycaprolactone, polylactic acid, organosilicon, and yttrium oxide.

33. The method of item 1, wherein the inactivated fungal biomass is pulverized inactivated fungal biomass.

34. The method of item 1, wherein the inactivated fungal biomass comprises a biomat or a portion thereof produced by a surface fermentation process.

35. The method of item 1, wherein the inactivated fungal biomass is grown on a growth medium having a carbon to nitrogen molar ratio of about 5 to about 20, or about 7 to about 15.

36. A fabric composition comprising:

Inactivated fungal biomass; and

It is selected from at least one component of plasticizers, polymers, crosslinking agents, and dyes.

At least one of the components is distributed in the fungal mycelial biomass.

37. The fabric composition of item 36, having a thickness of at least about 1 mm.

38. The fabric composition of item 36, having a tear strength of at least about 30 N.

39. The fabric composition of item 36, having a tear strength of at least about 10 N/mm.

40. The fabric composition of item 36, having a flexural stiffness of not more than about 5 g-cm.

41. The fabric composition of item 36, having a tensile strength of at least about 10 MPa.

42. The fabric composition of item 36, having at least about 3 water spot gray level.

43. The fabric composition of item 36, having a light color fastness rating of at least about 4 for blue wool.

44. The fabric composition of item 36, having a gray scale grade of at least about 3 for color fastness to rubbing when dry.

45. The fabric composition of item 36, having a gray scale grade of at least about 2 for color fastness to rubbing when wet.

46. The fabric composition of item 36, further comprising at least one backing layer of non-fungal fabric material.

47. The fabric composition of item 46, wherein the non-fungal fabric material is selected from acrylic fabrics, alpaca wool fabrics, angora rabbit hair fabrics, cashmere fabrics, coconut fiber fabrics, cotton fabrics, iron yarn fabrics, hemp fabrics, jute fabrics, Kevlar fabrics, flax fabrics, microfiber fabrics, mohair fabrics, nylon fabrics, olefin fabrics, cashmere fabrics, polyester fabrics, pineapple fiber fabrics, ramie fabrics, rayon fabrics, seaweed fiber fabrics, silk fabrics, sisal fabrics, spandex fabrics, spider silk fabrics, wool fabrics, and combinations and mixtures thereof.

48. The fabric composition of item 36, further comprising a thermal dopant.

49. The fabric composition of item 48, wherein the thermal dopant is selected from ceramic materials, metallic materials, polymeric materials, and combinations and mixtures thereof.

50. The fabric composition of item 48, wherein the thermal dopant is selected from activated carbon, alumina, bentonite, diatomaceous earth, ethylene vinyl acetate, lignin, nano silica, polycaprolactone, polylactic acid, organosilicon, and yttrium oxide.

51. The fabric composition of item 48, wherein the thermal properties of the fabric composition are altered relative to the same thermal properties of the fabric composition without the thermal dopant, wherein the thermal properties are selected from heat storage coefficient, thermal conductivity, heat capacity, and combinations thereof.

52. An article of manufacture comprising the fabric composition of item 36, wherein the article of manufacture is selected from clothing articles, accessory articles, and furniture articles.

53. A method for manufacturing a durable sheet material, comprising:

(a) Contacting inactivated fungal biomass with an aqueous solution containing calcium hydroxide to form lime-treated inactivated fungal biomass;

(b) Contact the lime-treated inactivated fungal biomass with an aqueous solution containing ammonium sulfate to form delime-treated inactivated fungal biomass;

(c) The deliquescent inactivated fungal biomass is contacted with an aqueous solution containing a polymer to form impregnated inactivated fungal biomass;

(d) Contact the impregnated inactivated fungal biomass with an aqueous solution containing a crosslinking agent to form tanned inactivated fungal biomass;

(e) Contact the tanned inactivated fungal biomass with an aqueous solution containing a plasticizer to form plasticized inactivated fungal biomass;

(f) Drying the plasticized inactivated fungal biomass to form dried inactivated fungal biomass; and

(g) The dried, inactivated fungal biomass is hot-pressed to form the durable sheet material.

54. The method of item 53, further comprising rinsing the inactivated fungal biomass with water to remove residual aqueous solution between any pair of steps selected from steps (a) and (b), steps (b) and (c), steps (c) and (d), and steps (d) and (e).

55. The method of claim 53, wherein at least one of steps (a) to (e) comprises agitating the inactivated fungal biomass together with the aqueous solution.

56. The method of item 53, wherein the aqueous solution of at least one of steps (a) to (c) further comprises a surfactant or a solubilizer.

57. The method of 56, wherein the surfactant or solubilizer is selected from polysorbate, hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations and mixtures thereof.

58. The method of 53, wherein the polymer is selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations and mixtures thereof.

59. The method of 53, wherein the aqueous solution of step (c) further comprises a plasticizer selected from: glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols and their esters, epoxidized triglyceride vegetable oil, castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof.

60. The method of item 53, wherein the aqueous solution of step (c) further comprises an alkali metal halide.

61. The method of item 60, wherein the alkali metal halide is sodium chloride.

62. The method of 53, wherein the crosslinking agent is selected from citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations and mixtures thereof.

63. The method of 53, wherein the plasticizer is selected from glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols and their esters, epoxidized triglyceride vegetable oils, castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebacic acid, maleate, bioplasticizers, and combinations and mixtures thereof.

64. A method for manufacturing a durable sheet material, comprising:

(a) The fungal biomass was inactivated by boiling it in water;

(b) Contact the inactivated fungal biomass with an aqueous solution containing calcium hydroxide to form lime-treated inactivated fungal biomass;

(c) The lime-treated inactivated fungal biomass is contacted with an aqueous solution containing ammonium sulfate to form delime-treated inactivated fungal biomass;

(d) The deliquescent inactivated fungal biomass is contacted with an aqueous solution containing alkali metal halides to form impregnated inactivated fungal biomass;

(e) The impregnated inactivated fungal biomass is brought into contact with a first cross-linking agent to form tanned inactivated fungal biomass;

(f) Contact the tanned inactivated fungal biomass with an aqueous solution containing at least one of a second crosslinking agent and a polymer to form a retanned inactivated fungal biomass;

(g) Contact the retanned inactivated fungal biomass with fat-added oil to form fat-added inactivated fungal biomass;

(h) Adhere a non-fungal fabric backing to the inactivated fungal biomass to form backed inactivated fungal biomass;

(i) The backed inactivated fungal biomass is hot-pressed to form hot-pressed inactivated fungal biomass;

(j) Drying the heat-pressed inactivated fungal biomass to form dried inactivated fungal biomass; and

(k) Apply at least one of a coating wax, a coating oil, and nitrocellulose to the dried, inactivated fungal biomass to form the durable sheet material.

65. The method of item 64 further comprises rinsing the inactivated fungal biomass with water to remove residual aqueous solution between any pair of steps selected from steps (b) and (c), steps (c) and (d), and steps (e) and (f).

66. The method of item 64, wherein at least one of steps (a) to (g) comprises agitating the inactivated fungal biomass together with the aqueous solution.

67. The method of item 64, wherein the aqueous solution of at least one of steps (b) and (c) further comprises a surfactant or solubilizer.

68. The method of 67, wherein the surfactant or solubilizer is selected from polysorbate, hydrochloric acid, acetic acid, formic acid, lactic acid, and combinations and mixtures thereof.

69. The method of 64, wherein the polymer is selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations and mixtures thereof.

70. The method of item 64, wherein the alkali metal halide is sodium chloride.

71. The method of item 64, wherein the aqueous solution of at least one of steps (d) to (f) contains a pH adjuster.

72. The method of 71, wherein the pH adjuster comprises hydrochloric acid, acetic acid, formic acid, lactic acid, or combinations or mixtures thereof, or a metal hydroxide.

73. The method of item 64, wherein the first crosslinking agent comprises an aluminum salt, a chromium salt, a titanium salt, an aldehyde, or a combination or mixture thereof.

74. The method of item 73, wherein the first crosslinking agent is an aluminosilicate.

75. The method of item 64, wherein the second crosslinking agent is selected from citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations and mixtures thereof.

76. The method of item 64, wherein the polymer is selected from polyvinyl alcohol, chitosan, polyethylene glycol, alginate, starch, polycaprolactone, polyacrylic acid, hyaluronic acid, and combinations and mixtures thereof.

77. The method of item 64, wherein the aqueous solution of step (f) further comprises anionic dye.

78. The method of item 64, wherein the fattening oil is selected from sulfated castor oil, beeswax, coconut oil, vegetable oil, olive oil, linseed oil, oleic acid, and combinations and mixtures thereof.

79. The method of item 64, wherein the fatliquoring oil comprises an emulsion, wherein the method further comprises, between steps (g) and (h), contacting the fatliquoring oil with an acid to dissociate the emulsion.

80. The method of 64, wherein the finishing wax is selected from carnauba wax, candelilla wax, and combinations and mixtures thereof.

The invention is further described by way of example through the following non-limiting examples.

Example 1

Fabric manufacturing process

In various embodiments, the fungal fabric material according to the invention can be manufactured according to the method described in this embodiment. Specifically, the method described in this embodiment can be used to manufacture leather-like fabric materials, i.e., fungal fabric materials that replicate, simulate (counterfeit), and/or replace genuine leather.

One or more of these methods for manufacturing fungal fabric materials typically involve obtaining a mat of fungal material comprising fungal mycelium from a suitable reactor, which in many embodiments may require the production of fungal biomats according to the methods described in '050, '626, '421, and/or '474 publications. These mats can then be inactivated, in some embodiments by steaming for at least 30 minutes, and the inactivated mats can then be cut to the desired size and geometry. In some embodiments, the mats can be partially or completely dried in a dehydrator at elevated temperatures (e.g., about 130°F to about 160°F).

The inactivated pad is then placed in a solution of one or more components selected to impart the desired properties to the final fungal fabric material. Typically, this solution contains one or more polymers, plasticizers, and crosslinking agents. Polymers suitable for use in the solutions according to the invention include, but are not limited to, polyvinyl alcohol, chitosan, polyethylene glycol, hyaluronic acid, polycaprolactone, polyacrylic acid, and combinations and mixtures thereof. Plasticizers suitable for use in the solutions according to the invention include, but are not limited to, glycerol and its esters, polyethylene glycol, citric acid, oleic acid, oleic acid polyols (e.g., mannitol, sorbitol) and their esters, epoxidized triglycerides, vegetable oils (e.g., from soybean oil), castor oil, pentaerythritol, fatty acid esters, carboxylic acid ester-based plasticizers, trimellitate, adipate, sebate, maleate, bioplasticizers, and combinations and mixtures thereof. Crosslinking agents suitable for use in the solutions according to the invention include, but are not limited to, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations and mixtures thereof.

The inactivated pad is allowed to be immersed in the polymer, plasticizer, and/or crosslinking agent solution for a time sufficient to allow the pad to be penetrated and/or saturated by the polymer, plasticizer, and/or crosslinking agent, typically at least about 2 hours and most typically about 24 hours. After immersion in the solution, the wet pad is removed from the solution (and excess solution can then be removed from one or more surfaces of the pad).

Optional steps (which may be preferred in some embodiments) in the method of the present invention include laminating two or more pads after soaking. In practice, multiple pads may be laminated by: stacking two or more pads vertically or arranging the pads in any desired spatial orientation (horizontal to vertical, parallel to vertical to inclined, etc.), which in some cases may include natural fibers in addition to fungal mycelium; and soaking the vertically stacked pads in a polymer solution, which may be the same as or different from the solution used in earlier soaking steps. Typically, laminating two or more pads according to the invention includes removing air bubbles trapped between the layers (e.g., by pressing the stacked pads, by rolling, by vacuum extraction, etc.).

The wet pad (or laminate of multiple pads) is then dried, typically in a dehydrator at elevated temperatures (e.g., about 130°F to about 160°F) for about 30 minutes to about 120 minutes, to remove substantially all the liquid from the outer surface of the pad (or laminate), but leaving at least some liquid inside the pad (or laminate). The pad is then removed from the dehydrator and, in some embodiments, hot-pressed, for example between textured silicone molds, at elevated temperatures (e.g., about 130°C); typically, the pad is hot-pressed at intervals of about 20 seconds to about 30 seconds for a total time of about 3 minutes to about 10 minutes.

Example 2

Fungal growth through fibers

In various embodiments, the fungal fabric material according to the invention can be manufactured according to the method described in this embodiment. Specifically, the method described in this embodiment can be used to manufacture fabric materials incorporating both filamentous fungi and other natural or synthetic fibers.

One or more of these methods for manufacturing fungal fabric materials typically include, in various embodiments, a growth medium for filamentous fungi, such as those described in applications '050, '626, '421, and/or '474, but may also include other types of growth media. In particular, the growth medium may be formulated with alternative carbon sources or different carbon contents, which in various embodiments promotes the consumption of natural fibers by the fungi to be cultured in the growth medium. As a non-limiting example, conventional growth media may be modified by replacing glycerol with hydrolyzed cellulose, crystalline cellulose, or other cellulose compounds to promote the production of cellulase by the filamentous fungi. As another non-limiting example, the total amount of cellulose material may be carefully controlled, for example, to about 10% by weight of the growth medium, to ensure the desired growth characteristics of the filamentous fungi. After the growth medium is prepared, it is typically boiled for at least 30 minutes to eliminate competing or pathogenic microorganisms, then sealed and allowed to cool. Typically, the cooled culture medium is pH-adjusted, for example using hydrochloric acid, and inoculated with an inoculum of filamentous fungi (e.g., MK7 ATCC accession number PTA-10698) at a ratio of about 5% by volume; the culture medium is usually stirred to provide a uniform dispersion of the fungal inoculum.

The reactor for producing filamentous fungal biomass is prepared as follows: a sanitary reactor, such as a Saran wrap reactor, is provided, and the interior of the reactor (e.g., walls, doors, racks, trays, etc.) is cleaned and/or sterilized (e.g., with ethanol). Specifically, natural fibers serving as the substrate and/or structural material of the fungal fabric are typically placed in one or more Pyrex trays at a ratio of about 0.5 g to about 5 g/tray, covered with aluminum foil, and dry-autoclaved to eliminate competing or pathogenic microorganisms, followed by cooling; the Pyrex tray is then placed in the cleaned reactor (typically on the reactor's trays).

The inoculated culture medium (typically at a ratio of approximately 200 mL/tray) is then poured into or otherwise introduced into the Pyrex tray in the reactor. It is generally desirable to introduce the inoculated medium into a corner of the Pyrex tray, rather than its center, to allow the growth medium to flow beneath the fibers within the Pyrex tray and thus allow the fibers to float on the surface of the liquid medium. After a typical incubation period of approximately 3 days to approximately 3 weeks, each Pyrex tray contains fungal biomass grown through the natural substrate and/or structural fibers, which can then be harvested for further processing.

Example 3

Introduction of oil (one or more)

In the production of genuine (i.e., non-fungal) leather, the leather material is typically subjected to an oiling process, whereby it is coated with one or more oils, or more commonly with a mixture of oils (one or more), emulsifiers, and penetrating agents. This oiling process lubricates the leather and improves its ability to flex without cracking (dry leather fibers are typically prone to cracking or breaking) and also imparts color and water resistance. In the practice of this invention, oil can be similarly introduced into fungal leather analog materials, or produced in situ by the filamentous fungus itself during the fermentation process, to provide similar advantages and benefits. This embodiment describes an implementation of such an oil introduction process for fungal leather analog materials.

In the oil introduction method of the "emulsion" according to the invention, one or more oils, fats, and/or waxes are provided. The oils, fats, and/or waxes may be selected based on their use as emulsifiers and/or surfactants (e.g., salts, soaps, and other amphiphilic molecules), and may include, as non-limiting examples, any one or more of the following: sulfated castor oil, beeswax, coconut oil, vegetable oils, olive oil, linseed oil and oleic acid, sulfated fish oil, sulfated canola oil, soybean oil, palm oil, and fatty acids. The emulsion formed by means of a surfactant can provide more stable conditions for penetration into leather; those skilled in the art can select anionic, cationic, or nonionic surfactants to improve the wetting effect of the emulsion on the fibers of the leather material. The oils, fats, and/or waxes are rapidly stirred in a container (e.g., via a magnetic stir bar), and in some embodiments, heat may be applied to melt one or more of the oils, fats, and/or waxes to ensure complete mixing, while water (preferably deionized water) is gradually added to the mixture until a milky white emulsion is formed; typically, water constitutes about 50% to about 70% by volume of the emulsion. The stirring rate is then reduced (e.g., via a magnetic stir bar or a track-type shaker), and the fungal leather analog material according to the invention is introduced into the container. The fungal leather analog material is typically allowed to remain in the stirred emulsion for about 20 minutes to about 4 hours, then removed from the emulsion and allowed to air dry for about 24 hours to about 48 hours. This oiling process can be performed before, after, and/or in lieu of hot pressing of the fungal leather analog material.

In the "stuffing" oil introduction method according to the invention, one or more liquefied oils or waxes (including, but not limited to, oils or waxes suitable for use in the above-described "emulsion" method) may be mechanically applied to the surface of the fungal leather analog material to allow the oil or wax to "work" into the structure of the fungal leather analog material. As in the "emulsion" method, the fungal leather analog material is then allowed to air dry for about 24 to about 48 hours, and the "stuffing" oiling process may be performed before, after, and/or in place of the hot pressing of the fungal leather analog material.

Example 4

Plant-based

In the practice of this invention, the use of dicarboxylic acids as crosslinking agents typically necessitates hot-pressing the fungal fabric material, as crosslinking of the chemical moieties found in the fungal fabric material by carboxylic acids usually only occurs at elevated temperatures (e.g., about 130°C). Alternatively, natural tannins, such as those extracted from plant (vegetable) materials or other plant (organic) materials, can bind to the fungal fabric material and/or induce chemical bonds (i.e., crosslinking) within it at lower temperatures compared to dicarboxylic acids, thus eliminating the need for hot pressing, which improves the water resistance of the fungal fabric material. Without wishing to be bound by any specific theory, it is believed that tannins interact with the fungal fabric material in largely the same way they interact with animal hides or skin, i.e., by binding to protein moieties to improve the material's strength and resistance to degradation.

Eliminating the need for hot-pressing the fungal fabric material can offer further advantages and benefits for downstream processing. As a non-limiting example, oil introduction processes (such as those described in Example 3) typically require a relatively "open" structure of the fungal fabric material; hot pressing closes the structure of the fungal fabric material and thus makes it difficult for the oil to penetrate into the leather structure. While the oil introduction process can be performed before hot pressing, in some cases this can hinder the crosslinking reaction during hot pressing and/or cause the oil to leach out of the fungal fabric material. This example describes an implementation of a process using vegetable tannins to crosslink the fungal fabric material to avoid these and other disadvantages.

In the plant processing method according to the invention, a fungal biomass mat is produced by any suitable method (including, but not limited to, the methods disclosed herein and/or in '050, '626, and/or '421 applications) and steamed as described in Example 1. The steamed mat is washed once or several times with deionized water, brine, or a combination or mixture thereof, and then the washed mat is placed in a solution containing a tannin compound. The tannin compound may comprise any one or more commercially extracted tannins and/or pure tannic acid, and typically constitutes about 0.5% to about 20% by weight of the tanning solution. The fungal mat is typically allowed to remain in the tanning solution for 1 day to about 30 days, and in some embodiments, the fungal mat may be transferred between two or more tanning solutions, for example, tanning solutions with different compositions and/or concentrations of tannin compounds, during the tanning process.

Following tanning, the fungal mat may be oiled by any suitable method (e.g., one or both of the methods described in Example 3), and/or may undergo a plasticizing solution or process (e.g., using polyethylene glycol (PEG) and/or glycerin as plasticizers). Finally, the plasticized and/or oiled material is allowed to air-dry, typically for about 24 to about 72 hours. It should be clearly understood that in many embodiments, further cross-linking (e.g., using dicarboxylic acids as cross-linking agents) may be carried out after the plant-based processing described in this example.

Example 5

Effect of polymer-plasticizer ratio on fabric material properties

This embodiment describes the effect of the polymer-to-plasticizer ratio in the solution of the present invention on the material properties of fungal fabric materials, and particularly fungal leather analogues. The polymer (i.e., long-chain molecules of biological structures chemically bonded to the fungal fabric material) improves the tensile strength of the fungal fabric material, while the plasticizer (i.e., smaller molecules not chemically bonded to the biological structures or the polymer) improves the flexibility of the fungal fabric material and reduces its brittleness. Therefore, without being bound by any specific theory, it is believed that changing the polymer-to-plasticizer ratio (hereinafter “PP ratio”) allows those skilled in the art to precisely control, select, or adjust the physical properties of the fungal fabric materials produced according to the present invention.

Biomasses with ATCC accession number PTA-10698 (hereinafter "MK7") were grown and steamed or boiled for 30 minutes to inactivate the fungus. The inactivated biomasses were cut into approximately 4 cm x 6 cm rectangles, each soaked overnight in a solution containing both a polymer (polyvinyl alcohol (PVA) or chitosan) and a plasticizer (glycerol). After soaking, each rectangle was dried in a benchtop dehydrator for approximately 45 minutes and 1 hour, then hot-pressed at 275°F for a total of 4 minutes at 30-second intervals. The samples were then air-dried overnight at room temperature, and the degree of swelling (DOS), loss of mass (ML) after soaking, tensile strength (TS), and subjective flexibility (six evaluations, 0-10 scale) were subsequently tested. The results are presented in Table 1.

Table 1:

Material properties of MK7 leather analog samples with varying polymer-plasticizer ratios

Regardless of the type of polymer used (PVA versus chitosan), certain trends are evident: tensile strength increases with increasing PP ratio, swelling increases with increasing PP ratio, mass loss decreases with increasing PP ratio, and flexibility decreases with increasing PP ratio. Introducing the polymer into the MK7 biomat and subsequently hot-pressing results in the formation of covalent and non-covalent bonds between the fungal mycelium and the polymer molecules. These polymer molecules also bind to each other to create entanglements in the bonded structure. Plasticizers such as glycerol retain the "free-floating" molecules unbound to both the polymer and the MK7 structure and prevent the formation of chemical bonds between the polymer and biomass. When plasticizers are sparsely present, more chemical bonding occurs, resulting in materials with increased strength and brittleness. When plasticizers are abundant, they prevent the formation of chemical bonds and result in flexible but weak materials. This phenomenon is demonstrated by the wide range of tensile strengths (2.70 MPa–8.61 MPa) and flexibility (0.67–9.83 based on a subjective 0–10 scale) obtained by varying the concentration of the polymer relative to the plasticizer.

Samples using PVA as the polymer showed more consistent results across the middle range of the test parameters, while samples containing chitosan as the polymer showed less consistent results across the extremes of the parameter range. This result can be partly attributed to the difference between steamed and boiled biomat samples; boiled samples were able to introduce the polymer solution of the specified content more uniformly and therefore showed better performance, while steamed samples tended to be much more brittle. It appears that PVA is absorbed more readily and uniformly by the steamed biomat compared to chitosan, which could explain the more consistent data obtained for the PVA samples.

Example 6

Effect of glycerol content on the properties of fabric materials

The procedure of Example 5 was repeated, except that the polymer/plasticizer solution was free of polymer (i.e., free of PVA or chitosan) and the content of the plasticizer (i.e., glycerol) was varied to evaluate the effect of the glycerol content on the material properties of the fungal fabric material.

Within the range of glycerol concentrations tested on MK7 leather samples, distinct trends were observed in terms of TS, Strain at Break (SAB), DOS, and ML. As shown in Figure 6, the TS of MK7 leather decreased with increasing glycerol concentration; a maximum TS of 8.65 MPa was achieved for samples made from pre-boiled biomass without added glycerol, while a minimum TS of 1.55 MPa was recorded for raw biomass samples with 37.5% added glycerol. As shown in Figure 7, the SAB of MK7 leather increased with increasing glycerol concentration, although this trend was less representative of samples made from pre-boiled biomass, most likely due to incomplete drying of some samples prior to strain testing. Furthermore, as shown in Figure 8, the DOS of MK7 leather decreased with increasing glycerol concentration, while as shown in Figure 9, ML increased with increasing glycerol concentration.

Glycerin acts by disrupting polymer-polymer interactions, increasing free space, and thus enhancing the mobility of polymer molecules. In various embodiments of MK7 leather according to the invention, a mixture of PVA and/or chitosan polymers is present, along with natural MK7 cells and secreted biopolymers (EPS). In the absence of glycerin, the added polymers, cells, and biopolymers can form more hydrogen bonds, ionic bonds, and covalent bonds with each other; molecular mobility and free space are low, while bond concentration is high. In this state, the material is more rigid and requires more energy to stretch or bend. Therefore, when the glycerin concentration is low, the measured TS is higher and the strain is lower, and vice versa.

Example 7

Effect of loading ratio on fabric material properties

The procedure of Example 5 was repeated, except that the polymer/plasticizer solution contained no plasticizer (i.e., no glycerin was added) and the total polymer content (i.e., the total amount of PVA and/or chitosan) was varied to evaluate the effect of the loading ratio on the material properties of the fungal fabric material.

As shown in Figure 10, the tensile strength (TS) of MK7 leather was observed to increase with increasing polymer concentration; in other words, the highest tensile strength was observed at the lowest loading ratio, and vice versa. TS was observed to increase linearly with polymer concentration up to approximately 36.5%, and then decreased at approximately 47.5% polymer concentration. Above approximately 47.5% polymer concentration, TS increased linearly, reaching a maximum of 6.89 MPa at 73% polymer concentration. Without being bound by any specific theory, this effect is believed to be attributable to the numerous hydroxyl and amine groups present in PVA and chitosan molecules; these groups can form covalent and non-covalent bonds with the biological structure as well as with other polymer molecules. With increasing polymer concentration, the concentration of intermolecular bonds also increases. The high bond concentration then leads to improved strength of the material. Additionally, the unprocessed biomass used to produce the leather samples contained both residual glycerol from the culture medium and EPS molecules produced by the organism. Glycerol, and possibly some components of EPS, act as plasticizing agents for the leather structure. Based on the results of the glycerol concentration experiment, it is reasonable to infer that increasing the biomass concentration, and therefore the plasticizer concentration, may lead to a decrease in the TS of the sample.

As shown in Figure 11, the SAB of the MK7 leather sample was observed to increase linearly with polymer concentration, reaching a maximum of 182% at a polymer concentration of 36.5%. Beyond 36.5%, the SAB then decreased linearly. Without being bound by any specific theory, this effect is believed to be attributed to a competitive effect of intermolecular bonding and plasticization within the leather structure. At high loading ratios, the high concentrations of biomass and plasticizer, coupled with low bond concentrations and a lack of intermolecular bonds, result in a material with a low tensile limit; therefore, during tensile testing, the tensile limit is likely to be reached before significant material strain, leading to material failure at low strain. In contrast, at the median loading ratio (37.5% polymer concentration), significant intermolecular bonding is more likely to occur. Additionally, the sample is also significantly plasticized due to the significant introduction of biomass at the median loading ratio. These properties result in a material with both a moderately high tensile limit and a moderately high strain limit. During tensile testing, the material can significantly elongate before reaching its tensile or strain limit. At lower loading ratios, the sample is no longer significantly plasticized. They contain high concentrations of polymers that form intermolecular bonds and therefore have high tensile limits. However, the lack of plasticizing molecules results in low strain limits and maximum strain values at correspondingly lower SAB values.

A similar trend to that observed in SAB was observed for the DOS of the leather samples, as shown in Figure 12. The DOS of the samples increased linearly with increasing polymer concentration, reaching a maximum of 405% at a polymer concentration of 47.5%. A linear decrease in DOS was observed as the polymer concentration increased beyond 47.5%. PVA and chitosan are known to form hydrogels, which are materials containing a three-dimensional network or web of physically and chemically bonded polymer molecules. When not fully cross-linked, this hydrogel network is flexible and contains spaces between the polymer chains, and therefore the hydrogel is stretchable and retains a large amount of water in the spaces between the polymer chains. When fully cross-linked, the spaces between the polymer chains are bound, and the material is less able to bend as water is absorbed. In this cross-linked state, the hydrogel has less water retention capacity. Without being bound by any specific theory, it is believed that at high loading ratios, due to the smaller amount of polymer molecules, there are fewer bonding sites available for water molecules. Therefore, at high loading ratios, the ability to absorb water is smaller. Maximum DOS values were observed at median polymer concentrations; at these concentrations, there are relatively high polymer concentrations and relatively high biomass concentrations. Biomass contains absorbed glycerol, resulting in a plasticized polymer network with low cross-linking and high water retention capacity. As polymer concentration increases further, the plasticizing effect decreases as the absorbed glycerol decreases. This leads to more highly cross-linked materials that cannot retain as much water.

However, the ML values of MK7 leather did not show any local maxima. Instead, as shown in Figure 13, a linear decrease in ML values was observed with increasing polymer concentration. Without being bound by any specific theory, this effect is believed to be attributable to the reduced biomass associated with the decreased loading ratio. The biomass contains a large proportion of water-soluble compounds that diffuse into the aqueous phase when the biomass is soaked. Therefore, for samples with high loading ratios containing a significant amount of soluble compounds, the difference in mass before and after soaking is much greater.

Example 8

The effect of polyvinyl alcohol-chitosan ratio on the properties of fabric materials

The procedure of Example 5 was repeated, except that the polymer/plasticizer solution contained no plasticizer (i.e., no glycerin was added), the total polymer content (i.e., the total amount of PVA and/or chitosan) was kept constant, and the ratio of PVA to chitosan was varied to evaluate the effect of different polymer compositions on the material properties of the fungal fabric material.

As shown in Figure 14, local maxima were observed in the TS (transferase) of the MK7 leather samples at PVA:chitosan weight ratios of 0:100, 50:50, and 100:0. These points correspond to PVA concentrations of 0%, 11.7%, and 23.4%, and the TS values at these points are 3.55 MPa, 3.53 MPa, and 4.32 MPa, respectively. As shown in Figure 15, local maxima were observed in the SAB (saturated fatty acid) of the samples at the same PVA:chitosan ratios observed for TS. The SAB values at these points are 143%, 138%, and 132%, respectively. Without being bound by any specific theory, it is possible that local TS maxima are observed when one polymer is absent and when multiple polymers are present in equal amounts, because the chemical bonding of one polymer can be disrupted by including a small amount of another polymer; that is, a small amount of chitosan can form aggregates within a larger PVA matrix, and vice versa. The polymer aggregates will compete with the biomass for binding sites in the large polymer matrix, resulting in reduced strength and disrupting the ability of polymer molecules to move and elongate. This explains the low TS and SAB values observed at PVA:chitosan ratios of 80:20, 60:40, 40:60, and 20:80. When approximately equal amounts of each polymer are added, aggregates may not form and a homogeneous polymer matrix can be present. In the absence of aggregates, the degree of bonding between the polymer and biomass will increase. Furthermore, the lack of aggregates will allow for the movement and deflection of polymer molecules. This explains the increase in TS and SAB observed at a 50:50 PVA:chitosan ratio.

As shown in Figure 16, the DOS of the leather samples decreased with increasing PVA concentration. As shown in Figure 17, the ML of the samples showed the opposite trend. In samples using chitosan as the sole polymer, the DOS value was three times higher than that of samples using PVA as the sole polymer. Without being bound by any specific theory, it is believed that chitosan molecules have a higher affinity for water molecules than PVA molecules, possibly due to the positive charge on the amine groups of chitosan at low pH. These charged amine groups are also more likely to bind to other molecules in the system, such as biomass particles, glycerol, or EPS components. Because of the charged amine groups, chitosan molecules are more likely to remain bound (tethered) during immersion, which would explain the lower ML values observed at higher Ch concentrations and vice versa.

Example 9

Effect of blending time on fungal particle length

40 g of raw (unprocessed) fungal biomass and 40 mL of deionized water were placed in a small Oster mixer and blended for 10 seconds. 3 mL of the resulting mixture was removed from the mixer and combined with 27 mL of deionized water to form a 30 mL "10-second blend" test material. The remaining mixture in the mixer was blended for another 10 seconds, and another 3 mL of the sample was removed and combined with 27 mL of deionized water to form a 30 mL "20-second blend" test material. This process was repeated with another 20 seconds of blending to obtain a "40-second blend," and once again, after another 20 seconds of blending, a "60-second blend" was obtained. The test materials were then further diluted individually in deionized water at a 9:1 ratio to form four 300 mL samples, each containing 1% by volume of the blended mixture.

Each of the four 1% (v/v) samples, in 75 μL increments, was placed on a microscope slide, and photomicrographs of each sample were taken. In each photomicrograph, the apparent length of 30 fungal particles was measured, and these apparent lengths were converted to the true length of each particle based on the magnification used in the microscope. Histograms of the particle lengths of the blends are shown in Figures 19A, 19B, 19C, and 19D, respectively.

Example 10

Effect of loading ratio on foaming during manufacturing

Raw (unprocessed) fungal biomass was chopped into approximately 1 cm cubes and added in varying amounts to several 400 mL beakers along with water, PVA solution, chitosan, and adipic acid. The height of the mixture in each beaker was measured, and then the mixtures were blended for 1 minute using a Hamilton Beach HB08 manual mixer, with the height measured again. The mixtures were then stirred at 180°C (large stir bar, 60 rpm) for 30 minutes, with the height measured a third time, followed by a fourth measurement after the addition of a certain volume of acetic acid to the beakers; the mixtures were stirred for an additional 10 minutes while cooling. The mixtures were then poured into flat trays and allowed to dry at room temperature for 2 days. They were then removed from the trays and hot-pressed in silicone-textured molds at 275°F for 10 minutes at a time, in 5 batches. The density of each hot-pressed sample was then measured. Figure 19 illustrates the “blending excess,” “heating excess,” and “total excess” of each mixture as a function of the loading ratio—which are the volume changes relative to the starting mixture after blending, heating, and the addition of acetic acid, respectively—and density.

Example 11

Physical properties of fungal leather analogues—the effect of fragmentation on intact biomass

In addition to the notes, eight samples of fungal leather analogue materials were produced according to the method shown in Figure 3 and the associated description. Of these eight samples, three were made from inactivated fungal biomass that was broken down prior to step 310—the first sample had no non-fungal fabric backing, the second sample had a non-fungal fabric (cotton) backing on one side of the fungal layer, and the third sample had a non-fungal fabric (cotton) layer “sandwiched” between two fungal layers. The other five samples were made from intact (unbroken) biomass produced by a surface fermentation process—the fourth, fifth, and sixth samples had no non-fungal fabric backing, the seventh sample had a non-fungal fabric (cotton) backing on one side of the fungal layer, and the third sample had a non-fungal fabric (cotton) layer “sandwiched” between two fungal layers.

The pulverized fungal biomass was prepared as follows: Water and thawed (previously frozen) processed biomass were added to a Vitamix mixer at a 1:1 mass ratio. These were blended together for approximately 2 minutes to produce a homogeneous mixture of pulverized biomass in water. Separately, solutions of water, glycerol, chitosan, citric acid, and hydrochloric acid were prepared, with each component in a mass ratio of 200:17.5:6.3:1:13.5. The total mass of the solutions was equal to the total mass of the biomass-water blend. Once the chitosan was dissolved, the polymer aqueous solution and the biomass-water mixture were combined. The newly formed mixture was stirred under heat for approximately 30 minutes to form a homogeneous paste. The paste was then cast onto a flat, non-stick tray and allowed to dry under ambient conditions. Once dried, the newly formed sheet material was hot-pressed at 100°C for 10 minutes.

For samples with a non-fungal (cotton) layer, the cotton backing material is adhered to the sample using an aqueous solution of chitosan (1% w/v), citric acid (1% w/v), and hydrochloric acid (1% v/v). The chitosan solution is applied to a suitable side (one or more) of the fungal layer, and the cotton is applied to the wetted surface. The chitosan adhesive is allowed to dry for approximately 20 minutes, and then the sample is hot-pressed at 275°F for two minutes to bond the backing material.

Nine physical properties of the eight fungal leather analogue samples were tested—thickness, tensile strength, tensile force, elongation at break, tear resistance, density, flexural stiffness, degree of swelling, and loss of mass after immersion. The results of these tests are given in Table 2 below.

Table 2

Example 12

Effect of carbon-nitrogen ratio on the properties of fungal leather analogues

Four growth media with the same fructose content were prepared (as described in, for example, applications '050, '626, and '421) for surface fermentation of fungal biomass. The carbon-to-nitrogen molar ratio (“CN ratio”) of each medium was adjusted by increasing or decreasing the combined content of ammonium sulfate and urea (keeping the ratio of these two components relative to each other constant) until the media had CN ratios of 5, 8.875, 10, and 20, respectively. Each medium was inoculated with 5% v/v MK7 inoculum via shake flask inoculation.

250 mL of each of the inoculated culture medium was poured into each of four glass trays, resulting in a total of 16 inoculated trays. The trays were placed in a sealed reactor at 27°C and incubated for 120 hours, with photographs taken at 72, 96, and 120 hours. The biomass from each tray was then harvested and inactivated in deionized water at 70°C for 30 minutes. After inactivation, the wet yield of each sample was determined to evaluate relative growth performance.

The biomass samples were then converted into fungal leather analogues according to the method described in Example 11 above. After tanning, various physical parameters of each sample were measured. The results are given in Table 3 (values shown are averages for each CN ratio).

Table 3

Various qualitative differences were observed between samples before and after the tanning process. Biomass grown on a medium with a CN ratio of 5 was more "slippery" and significantly thinner in some areas, especially in the portion of the biomass grown near the center of the tray; these fungal samples were extremely flexible once inactivated. Biomass grown on mediums with CN ratios of 8.875 and 10 became very stiff after inactivation, possibly due to the thickness of the biomass. Biomass grown on a medium with a CN ratio of 20 was more flexible than those grown on mediums with CN ratios of 8.875 and 10, both before and after the inactivation step.

Following the tanning process, samples obtained from the medium with a CN ratio of 5 were observed to have uneven thickness and were flexible in the thickest areas; areas subjected to greater pressure during the hot-pressing step were shinier and had a smoother texture, likely due to the compression of the hot press causing the filaments of the filamentous fungus to align and compact. Samples obtained from the medium with a CN ratio of 8.875 were the thickest and shrank the most during the drying step, exhibiting an uneven surface texture and feeling stiffer compared to the other samples. Samples obtained from the medium with a CN ratio of 10 had a moderate thickness and were more flexible than those obtained from the medium with a CN ratio of 8.875, but also showed an uneven surface; like the sample obtained from the medium with a CN ratio of 5, the areas exposed to maximum compression during the hot-pressing step were significantly shinier. The sample obtained from the culture medium with a CN ratio of 20 was intermediate in thickness between the samples with CN ratios of 5 and 10, and was relatively flexible with a slightly more uniform surface; again, the areas that were most compressed during hot pressing were the shiniest.

Example 13

Thermal doping of fungal leather analogues

Each of the five experimental samples was prepared as follows: 75 g of glycerol, 27 g of chitosan, 4.3 g of citric acid, 880 mL of water, and 13.5 mL of concentrated hydrochloric acid were placed in a beaker and stirred until the chitosan dissolved. Separately, 80 g of wet filamentous fungal biomass produced by the surface fermentation method as described herein and in applications '050, '626, and '421, and 80 mL of water were placed in a kitchen blender and mixed until homogeneous. (Except in the case of the control sample) 7.2 g of thermal dopant was then added to the blender, and the mixture was mixed again until homogeneous. 160 g of the chitosan solution was then added to the blender, and the mixture was homogenized again; 300 g of the resulting mixture was poured into a small non-stick tray and dried at 90°F for 23 hours. The dried sample was hot-pressed at 100°C for 10 minutes to produce a medium-flexibility flat sheet approximately 2 mm thick.

The thermal properties of the samples were measured. The results of these measurements are given in Table 4; control samples of unadulterated raw hide leather, unadulterated blended fungal leather analogues, and unadulterated leather made from intact biomass (CN ratios of 8.875, 10, and 20, respectively, denoted as “CN8”, “CN10”, and “CN20”) were also tested for comparison.

Table 4

Example 14

The Influence of Polyvinyl Acetate on Material Properties

A large mixture of biomass, water, glycerol, and adipic acid was mixed in a mixer and divided into five equal portions. Five separate 6% polymer solutions containing polyvinyl alcohol (PVA) and chitosan in an 80:20 mass ratio were prepared, each containing a different type of Kuraray PVA. Each polymer solution was combined with a portion of the biomass mixture to prepare five separate leather precursor mixtures. These leather precursor mixtures were individually mixed using a hand-held immersion mixer and poured into small Pyrex trays, then dried at room temperature with a fan blowing on the trays. When each sample reached a moisture content of no more than 20%, it was hot-pressed at 100°C for 10 minutes, in two batches. The samples were then allowed to dry overnight at room temperature, and tensile and tear strengths were subsequently tested, and texture and water resistance were qualitatively assessed. Each leather sample had a 75:25 loading ratio and a plasticizer content of 22.5% by weight. The results of these tests are provided in Table 5.

Table 5

The viscosity of PVA (which is directly related to molecular weight) has a significant impact on tensile and tear strength, with low-viscosity PVA resulting in poor tensile and tear properties. For higher-viscosity PVA types, the degree of hydrolysis appears to be the determining factor for tensile and tear properties; samples with a low degree of hydrolysis exhibit better tensile and tear properties. Without being bound by any specific theory, the inventors hypothesize that a higher concentration of acetate groups acts as a plasticizer, allowing free movement of internal molecules and reducing cracking and brittleness at the microscale, thereby improving the overall strength and flexibility of the sample.

This disclosure, in various aspects, implementations, and configurations, includes components, methods, processes, systems, and/or apparatuses substantially as depicted and described herein, encompassing multiple aspects, implementations, configurations, sub-combinations, and subsets. Those skilled in the art, upon understanding this disclosure, will understand how to make and utilize the aforementioned aspects, implementations, and configurations. This disclosure, in various aspects, implementations, and configurations, includes providing apparatus and processes in the absence of items not depicted and/or described herein or in its various aspects, implementations, and configurations (including the absence of items such as those that could have already been used in previous apparatuses or processes, for example, to improve performance, ease of implementation, and/or reduce implementation costs).

The foregoing discussion of this disclosure is presented for illustrative and descriptive purposes. It is not intended to limit this disclosure to the one or more forms disclosed herein. In the detailed description above, for example, to simplify this disclosure, various features of this disclosure are grouped together in one or more aspects, embodiments, and configurations. Features of aspects, embodiments, and configurations of this disclosure may be combined in alternative aspects, embodiments, and configurations other than those discussed above. The methods of this disclosure should not be construed as reflecting an intention that the claimed disclosure requires more features than expressly recited in the claims. Rather, as reflected in the appended claims, aspects of the invention lie in fewer than all the features of a single foregoingly disclosed aspect, embodiment, and configuration. Therefore, the appended claims are hereby incorporated into this detailed description, wherein each claim independently represents a single embodiment of this disclosure.

Furthermore, while the description of this disclosure includes descriptions of one or more aspects, implementations, or configurations, and certain variations and modifications, other variations, combinations, and modifications within the scope of this disclosure, for example, are possible to those skilled in the art upon understanding this disclosure. It is intended to include, to the permissible extent, alternative aspects, implementations, and configurations, including alternative, interchangeable, and/or equivalent structures, functions, scopes, or steps, regardless of whether such alternative, interchangeable, and/or equivalent structures, functions, scopes, or steps are disclosed herein; and not to provide any patentable subject matter to the public.

Claims (30)

1. Fabric materials, which include: a. Inactivation of fungal biomass; and b. Polymer, The fungal biomass:polymer loading ratio is approximately 10:90 to approximately 40:60. and The fabric material has a tensile strength of at least about 3 MPa. 2. The fabric material of claim 1, wherein the fungal biomass:polymer loading ratio is about 10:90 to about 30:70. 3. The fabric material of claim 1, wherein the fungal biomass:polymer loading ratio is about 15:85 to about 25:75. 4. The fabric material of claim 1, wherein the polymer comprises two or more polymers. 5. The fabric material of claim 1, wherein the polymer is selected from synthetic polymers, biopolymers, and combinations thereof. 6. The fabric material of claim 1, wherein the polymer comprises a synthetic polymer. 7. The fabric material of claim 1, comprising about 20% to about 40% by weight of inactivated fungal biomass. 8. The fabric material of claim 1, comprising about 20% to about 30% by weight of inactivated fungal biomass. 9. The fabric material of claim 1, comprising about 20% to about 25% by weight of inactivated fungal biomass. 10. The fabric material of claim 1, wherein the inactivated fungal biomass has an average particle size of not more than about 75 micrometers. 11. The fabric material of claim 1, wherein the inactivated fungal biomass has an average particle size of not more than about 30 micrometers. 12. The fabric material of claim 1, wherein the inactivated fungal biomass has an average particle size of not more than about 20 micrometers. 13. The fabric material of claim 1, having a thickness of at least about 1 mm. 14. The fabric material of claim 1, having a tensile strength of at least about 10 MPa. 15. The fabric material of claim 1, having a tensile strength of at least about 15 MPa. 16. The fabric material of claim 1, having a flexural stiffness of not more than about 10 g-cm. 17. The fabric material of claim 1, having a flexural stiffness of not more than about 5 g-cm. 18. The fabric material of claim 1, having a flexural stiffness of not more than about 2 g-cm. 19. The fabric material of claim 1, having a tear strength of at least about 30 N. 20. The fabric material of claim 1, having a tear strength of at least about 10 N/mm. 21. The fabric material of claim 1, having a fracture strain between about 75% and about 275%. 22. The fabric material of claim 1, having a swelling degree of about 30% to about 120%. 23. The fabric material of claim 1, having a mass loss of about 20% to about 30% after immersion. 24. The fabric material of claim 1, wherein the polymer is crosslinked by a crosslinking agent. 25. The fabric material of claim 24, wherein the crosslinking agent is selected from homo-bifunctional crosslinking agents, hetero-bifunctional crosslinking agents, photoreactive crosslinking reagents, citric acid, tannic acid, octanoic acid, adipic acid, succinic acid, extracted vegetable tannins, glyoxal, and combinations thereof. 26. The fabric material of claim 1, further comprising a non-fungal fabric material backing layer. 27. The fabric material of claim 26, wherein the non-fungal fabric material is selected from acrylic fabrics, alpaca wool fabrics, angora rabbit hair fabrics, cashmere fabrics, coconut fiber fabrics, cotton fabrics, iron yarn fabrics, hemp fabrics, jute fabrics, Kevlar fabrics, flax fabrics, microfiber fabrics, mohair fabrics, nylon fabrics, olefin fabrics, cashmere fabrics, polyester fabrics, pineapple fiber fabrics, ramie fabrics, rayon fabrics, seaweed fiber fabrics, silk fabrics, sisal fabrics, spandex fabrics, spider silk fabrics, wool fabrics, and combinations thereof. 28. The fabric material of claim 26, further comprising an adhesive for adhering a fabric backing. 29. The fabric material of claim 1, wherein the fabric material is embossed. 30. The fabric material of claim 1, further comprising a dye.