Fungal oleogel compositions

Abstract

Oleogel compositions comprising filamentous fungus are disclosed, as well as methods of making such oleogel compositions. The fungal oleogels compositions are suitable for use in food compositions, cosmetics, and nutraceutical delivery systems, as well as uses associated therewith.

Description

[0001] Aty Docket No. 10087-82-PCT

[0002] FUNGAL OLEOGEL COMPOSITIONS

[0003] CROSS REFERENCE TO RELATED APPLICATION

[0004] This application claims the benefit of priority of U.S. provisional patent application 63 / 687,980, filed 28 August 2024, the entirety of which is incorporated herein by reference.

[0005] FIELD

[0006] The present disclosure relates to edible filamentous fungi and provides methods of preparing oleogel compositions of edible filamentous fungi, particularly fungal oleogel compositions for use in food compositions, cosmetics, and nutraceutical delivery systems, as well as uses and methods associated therewith.

[0007] BACKGROUND

[0008] Solid fats (i.e., fats that are solid at room temperature) play an important role in food products by serving as structure modifiers influencing food texture and sensory attributes (e.g., creaminess, fluffmess, or hardness), as well as providing flavor. Solid fats, such as butter and shortening, are particularly important in baked goods where they contribute to the tenderness and flakiness of pie crusts and provide structure and softness to cookies. Meat products also contain high levels of animal fat that contribute to the meat’s flavor profile and cooking characteristics. Solid fats used in cooking are mainly animal derived products.

[0009] Structure in solid fat-containing food products is provided by the network of crystalline triacylglycerols. However, these triacylglycerols contain high levels of saturated fatty acids. Instead of using naturally highly saturated solid fats, oils comprising triacylglycerols having high levels of unsaturated fatty acids are also transformed to more solid products by hydrogenation or partial hydrogenation of the oil. Trans fatty acids unavoidably emerge from partial hydrogenation of unsaturated oils. Research into the role fats and oils play in human health has indicated that the intake of trans fatty acids and saturated fatty acids can raise LDL cholesterol levels in the blood and have been associated with major health-related concerns, such as cardiovascular diseases and other non- communicative pathologies such as cancers and diabetes. Thus, the World Health Organization recommends the amount of total, saturated, and trans fats to be less than 30%, 10%, and 1% of total energy intake respectively.

[0010] In addition to health concerns, animal fat is also associated with environmental concerns relating to its production. Animal production for meat, fat, and dairy food Aty Docket No. 10087-82-PCT consumption is linked to a host of environmental impacts that threaten sustainability including increasing greenhouse gas emissions (GHGEs) and burdening water supplies.

[0011] Oleogels are solid or semisolid materials that can mimic solid fats in cooking while providing lower saturated fatty acids and trans fatty acids than common solid cooking fats (e.g., butter, lard, and shortening). Oleogels result from the gelation of an oil by structuring agents whereby the liquid oil is physically entrapped into a gel network formed by the oleogel structurant, also referred to as an oleogelator. Oleogels contain oil (an organic solvent) and an oleogelator which engulfs and immobilizes the oil. Examples of oleogelators include waxes, ethylcellulose, alcohols or fatty acid esters, phospholipids and phytosterols.

[0012] Protein based oleogels use proteins as an oleogel structurant. Protein based oleogels can be made by forming an emulsion that contains protein acting as an emulsifier and subsequently removing the water phase and shearing the resulting dried, concentrated emulsion. Emulsion stabilizers such as polysaccharides may be included in the emulsion to increase emulsion stability.

[0013] In addition to their use in the food industry, oleogels also have applications in a diverse range of industries including nutraceuticals and cosmetics.

[0014] Challenges with the use of oleogels in foods include identifying suitable food grade oleogelators that are capable of producing oleogels with desired functionality and sensory characteristics and improved nutritional profde as compared to common solid cooking fats.

[0015] SUMMARY

[0016] The present disclosure provides methods of making oleogel compositions from fdamentous fungal biomass to be utilized in food products, cosmetics, and nutraceutical delivery systems.

[0017] In one aspect of the present disclosure, a composition comprises an oleogel comprising an oil; and an oleogel structurant comprising fdamentous fungal biomass.

[0018] In embodiments, the oleogel may comprise about 75% to about 95% oil on a dry weight basis.

[0019] In embodiments, the oleogel may comprise about 5% to about 25% oleogel structurant on a dry weight basis.

[0020] In embodiments, the composition may comprise about 5% to about 25% fdamentous fungal biomass on a dry weight basis. Aty Docket No. 10087-82-PCT

[0021] In embodiments, the oleogel structurant is an aqueous dispersion comprising about 5% to about 99% fdamentous fungal biomass on a dry weight basis.

[0022] In embodiments, the composition may comprise less than about 1 wt. % water.

[0023] In embodiments, the composition may comprise at least about 1% protein on a dry weight basis.

[0024] In embodiments, the composition may comprise about 1% to about 10% protein on a dry weight basis.

[0025] In embodiments, the composition may comprise at least about 1% fiber on a dry weight basis.

[0026] In embodiments, the composition may comprise about 1% to about 10% fiber on a dry weight basis.

[0027] In embodiments, the oleogel structurant may further comprise a plant protein source, a polysaccharide source, or combinations thereof. In embodiments, the plant protein may, but need not, comprise hemp protein, soy protein, pea protein, chickpea protein, rice protein, or combinations thereof.

[0028] In embodiments, the composition may be solid at 20 °C.

[0029] In embodiments, the composition may be stable at 20 °C for at least about 24 hours.

[0030] In embodiments, the composition may have a turbiscan stability index of about 0 to about 3.

[0031] In embodiments, the oleogel may have a melting temperature of about 27 °C to about 37 °C.

[0032] In embodiments, the oleogel may have a crystallization temperature of about 5 °C to about 20 °C.

[0033] In embodiments, the oil may comprise acai oil, almond oil, avocado oil, blackcurrant seed oil, borage seed oil, canola oil, cashew oil, coconut oil, corn oil, cottonseed oil, evening primrose oil, grapeseed oil, hazelnut oil, hemp oil, macadamia oil, olive oil, palm oil, peanut oil, pecan oil, pine seed oil, pistachio oil, pumpkin seed oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, walnut oil, or combinations thereof.

[0034] In embodiments, the filamentous fungus may be selected from the group consisting of Rhizopus oligosporus, Ustilago esculenta, Hericulum erinaceus, Polyporous squamosus, Grifola fondosa, Hypsizygus marmoreus, Hypsizygus ulmarius, Calocybe gambosa, Pholiota nameko, Calvatia gigantea, Agaricus bisporus, Stropharia rugosoannulata, Hypholoma lateritium, Pleurotus eryngii, Pleurotus ostreatus, Pleurotus ostreatus var. Aty Docket No. 10087-82-PCT columbinus, Tuber borchii, Morchella esculenta, Morchella conica, Morchella importuna, Sparassis crispa, Fusarium venenatum, Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698), Disciotis venosa, Clonostachys rosea, Cordyceps militaris, Trametes versicolor, Ganoderma lucidum, Flammulina velutipes, Lentinula edodes, Pleurotus djamor, Pleurotus ostreatus, and Leucoagaricus spp.

[0035] In embodiments, the fdamentous fungus may be a Fusarium species.

[0036] In embodiments, the fdamentous fungus may be Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698).

[0037] In embodiments, the fdamentous fungus may be selected from Fusarium venenatum and Fusarium venenatum strain A3 / 5.

[0038] In embodiments, the fdamentous fungal biomass may be produced by submerged fermentation.

[0039] In embodiments, the fdamentous fungal biomass may be produced by nonsubmerged fermentation.

[0040] In embodiments, the fdamentous fungal biomass may be a cohesive fungal biomass.

[0041] In embodiments, the composition may be a flour.

[0042] In embodiments, the composition may be a dough.

[0043] In another aspect of the present disclosure, a food product may comprise a composition as disclosed herein, wherein the food product is selected from the group consisting of a butter analog, a shortening analog, a ghee analog, and a margarine analog. In embodiments, the food product may, but need not, be a savory food product. In embodiments, food product may, but need not, be selected from the group consisting of a processed meat product, a sausage, a meat patty, frankfurters, a pork analog, a beef analog, a chicken analog, and bread. In embodiments, the food product may, but need not, be sweet food product. In embodiments, the food product may, but need not, be selected from the group consisting of chocolate, chocolate compositions, cake, muffins, pastries, cookies, brownies, pie crust, and ice cream.

[0044] In another aspect of the present invention, a method of making an oleogel, comprises drying an oil-in-water emulsion, wherein the oil-in-water emulsion comprises filamentous fungal biomass and a continuous gelled water phase, to form a dried emulsion, and shearing the dried emulsion to form an oleogel. Aty Docket No. 10087-82-PCT

[0045] In embodiments, the oil-in-water emulsion may have an oil fraction volume of at least about 30 wt.%, at least about 40 wt.%, at least about 50 wt.%, at least about 60 wt.%, or at least about 70 wt.%.

[0046] In embodiments, the drying step may comprise lyophilization.

[0047] In embodiments, the dried emulsion may comprise less than about 1 wt. % water.

[0048] In embodiments, the shearing step may be conducted for about 5 seconds to about 30 seconds.

[0049] In embodiments, the method may further comprise forming the oil-in-water emulsion. In embodiments, the forming step may, but need not, comprise combining oil and an aqueous liquid dispersion of fdamentous fungal biomass and homogenizing the combined oil and aqueous liquid dispersion. In embodiments, the pH of the aqueous liquid dispersion may, but need not, be less than about 6.0, less than about 5.5, less than about 5.0, less than about 4.5, less than about 4.0, less than about 3.5, less than about 3.0, less than about 2.5, less than about 2.0, less than about 1.5, less than about 1.0, or less than about 0.5.

[0050] In embodiments, the homogenizing step may, but need not, be conducted at a temperature of at least 20°C. In embodiments, the homogenizing step may, but need not, be conducted at a temperature of at least 38°C.

[0051] In embodiments, the oil-in-water emulsion may be a stable emulsion.

[0052] In embodiments, the oleogel may comprise about 75% to about 95% oil on a dry weight basis.

[0053] In embodiments, the oleogel may comprise about 5% to about 25% on a dry weight basis of an oleogel structurant comprising the fdamentous fungal biomass. In embodiments, the oleogel structurant may, but need not, further comprise a plant protein source, a polysaccharide source, or a combination thereof.

[0054] In embodiments, the oleogel may comprise about 5% to about 25% fdamentous fungal biomass on a dry weight basis.

[0055] In embodiments, the oleogel may comprise at least about 1% protein on a dry weight basis.

[0056] In embodiments, the oleogel may comprise about 1% to about 10% protein on a dry weight basis.

[0057] In embodiments, the oleogel may comprise at least about 1% fiber on a dry weight basis. Aty Docket No. 10087-82-PCT

[0058] In embodiments, the oleogel may comprise about 1% to about 10% fiber on a dry weight basis.

[0059] In embodiments, the oleogel may be solid at 20 °C.

[0060] In embodiments, the oleogel may be stable at 20 °C for at least about 24 hours.

[0061] In embodiments, the oleogel may have a turbiscan stability index of about 0 to about 3.

[0062] In embodiments, the oleogel may have a melting temperature of about 27 °C to about 37 °C.

[0063] In embodiments, the oleogel may have a crystallization temperature of about 5 °C to about 20 °C.

[0064] In embodiments, the oil-in-water emulsion may comprise an oil selected from the group consisting of acai oil, almond oil, avocado oil, blackcurrant seed oil, borage seed oil, canola oil, cashew oil, coconut oil, com oil, cottonseed oil, evening primrose oil, grapeseed oil, hazelnut oil, hemp oil, macadamia oil, olive oil, palm oil, peanut oil, pecan oil, pine seed oil, pistachio oil, pumpkin seed oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, walnut oil, and combinations thereof.

[0065] In embodiments, the filamentous fungus may be selected from the group consisting of Rhizopus oligosporus, Ustilago esculenta, Hericulum erinaceus, Polyporous squamosus, Grifola fondosa, Hypsizygus marmoreus, Hypsizygus ulmarius, Calocybe gambosa, Pholiota nameko, Calvatia gigantea, Agaricus bisporus, Stropharia rugosoannulata, Hypholoma lateritium, Pleurotus eryngii, Pleurotus ostreatus, Pleurotus ostreatus var. columbinus, Tuber borchii, Morchella esculenta, Morchella conica, Morchella importuna, Sparassis crispa, Fusarium venenatum, Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698), Disciotis venosa, Clonostachys rosea, Cordyceps militaris, Trametes versicolor, Ganoderma lucidum, Flammulina velutipes, Lentinula edodes, Pleurotus djamor, Pleurotus ostreatus, and Leucoagaricus spp.

[0066] In embodiments, the filamentous fungus may be a Fusarium species.

[0067] In embodiments, the filamentous fungus may be Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698).

[0068] In embodiments, the filamentous fungus is selected from Fusarium venenatum and Fusarium venenatum strain A3 / 5.

[0069] In embodiments, the filamentous fungal biomass may be produced by submerged fermentation. Aty Docket No. 10087-82-PCT

[0070] In embodiments, the filamentous fungal biomass may be produced by nonsubmerged fermentation.

[0071] In embodiments, the filamentous fungal biomass may be a cohesive fungal biomass.

[0072] In embodiments, the oleogel may be a flour.

[0073] In embodiments, the oleogel may be a dough.

[0074] While specific embodiments and applications have been illustrated and described, the present disclosure is not limited to the precise configuration and components described herein. Various modifications, changes, and variations which will be apparent to those skilled in the art may be made in the arrangement, operation, and details of the methods and systems disclosed herein without departing from the spirit and scope of the overall disclosure.

[0075] As used herein, unless otherwise specified, the terms “about,” “approximately,” etc., when used in relation to numerical limitations or ranges, mean that the recited limitation or range may vary by up to 10%. By way of non-limiting example, “about 750” can mean as little as 675 or as much as 825, or any value therebetween. When used in relation to ratios or relationships between two or more numerical limitations or ranges, the terms “about,” “approximately,” etc. mean that each of the limitations or ranges may vary by up to 10%; by way of non-limiting example, a statement that two quantities are “approximately equal” can mean that a ratio between the two quantities is as little as 0.9: 1.1 or as much as 1.1:0.9 (or any value therebetween), and a statement that a four-way ratio is “about 5:3: 1:1” can mean that the first number in the ratio can be any value of at least 4.5 and no more than 5.5, the second number in the ratio can be any value of at least 2.7 and no more than 3.3, and so on.

[0076] The embodiments and configurations described herein are neither complete nor exhaustive. As will be appreciated, other embodiments are possible utilizing, alone or in combination, one or more of the features set forth above or described in detail below.

[0077] BRIEF DESCRIPTION OF THE DRAWINGS

[0078] Figure 1 A is a graph illustrating results of viscosity testing of a fungal oil-in-water emulsion prepared using fungal biomat, according to embodiments of the present invention.

[0079] Figure IB is a graph illustrating results of viscosity testing of a fungal oil-in-water emulsion prepared using fungal flour, according to embodiments of the present invention. Aty Docket No. 10087-82-PCT

[0080] Figure 2 A is a graph illustrating results of dynamic strain sweep testing of a fungal oil-in-water emulsion prepared using fungal biomat, according to embodiments of the present invention.

[0081] Figure 2B is a graph illustrating results of dynamic strain sweep testing of a fungal oil-in-water emulsion prepared using fungal flour, according to embodiments of the present invention.

[0082] Figure 3 A is a graph illustrating results of frequency sweep testing of a fungal oil- in-water emulsion prepared using fungal biomat, according to embodiments of the present invention.

[0083] Figure 3B is a graph illustrating results of frequency sweep testing of a fungal oil- in-water emulsion prepared using fungal flour, according to embodiments of the present invention.

[0084] Figure 4 A is a top perspective image of oleogels produced by methods according to the present disclosure.

[0085] Figure 4B is a side perspective image of oleogels produced by methods according to the present disclosure.

[0086] Figure 4C is an image of an oleogel produced by methods according to the present disclosure after shearing.

[0087] DETAILED DESCRIPTION

[0088] The present disclosure provides methods of making oleogel compositions from fdamentous fungal biomass. The fungal oleogel compositions are edible on their own, for example a shortening analog, or may be used as ingredients or components incorporated into other edible compositions, for example baked goods, chocolate, processed meats, meat analogs, and / or the like. Fungal oleogel compositions are also able to be used in a variety of cosmetic applications, as well as in nutraceutical delivery systems. In various aspects, the methods comprise drying an oil-in-water emulsion, the oil-in-water emulsion comprising an oleogel structurant made of fdamentous fungal biomass and a continuous gelled water phase. Thereafter, the resulting dried emulsion can be sheared to form an oleogel.

[0089] The general concept provided by the present disclosure, then, is the creation of a fungal-based oleogel via oleogelation. Oleogelation occurs by a process that takes advantage of certain features of the biomass disclosed herein, leading to a final product that effectively mimics soft spreadable structured fat products that are edible on their own, such as a margarine analog or shortening analog, or incorporated into other food products, such as Aty Docket No. 10087-82-PCT baked goods, chocolate, and meat analogs. Structure of the fungal oleogel and / or the food products, and their final flavor and texture, can be augmented via the addition of one or more enhancers, as may be desired and as disclosed. Suitable enhancers include, for example, polysaccharides, flavors, stabilizers, coloring agents, olfactory additions, preservatives, and the like. As will be appreciated, products produced by the present disclosure can provide vegan food products that accurately mimic fats in both taste and texture.

[0090] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. All patents, applications published applications, and other publications to which reference is made herein are incorporated by reference in their entirety. If there is a plurality of definitions for a term herein, the definition provided in the Summary prevails unless otherwise stated.

[0091] As used herein, unless otherwise specified, the term “analog” or “analog food product” refers to a food product comprising edible fungi that bears an aesthetic, culinary, nutritional, and / or sensory equivalence or resemblance to an identified non-fungal food product. By way of nonlimiting example, a “butter analog product,” as that term is used herein, refers to a food product comprising edible fungi that bears an aesthetic, culinary, nutritional, and / or sensory equivalence or resemblance to conventional butter made from animal milk, and a “shortening analog product,” as that term is used herein, refers to a food product comprising edible fungi that bears an aesthetic, culinary, nutritional, and / or sensory equivalence or resemblance to conventional shortening made using animal products.

[0092] As used herein, the term “biomass” refers to a mass of a living or formerly living organism. In some embodiments, this term specifically includes a mass of a living or formerly living filamentous fungi.

[0093] As used herein, the term “biomat” refers to a cohesive mass of filamentous fungal tissue comprising a network of interwoven hyphae filaments. Biomats may be produced by any one or more fungal fermentation methods, for example according to methods described in WO 2020 / 176758, WO 2019 / 099474, and / or WO 2018 / 014004.

[0094] As used herein, the phrase “mycelial biomass” refers to a biomass of filamentous fungus in which fungal mycelium makes up about 50 wt.% to about 100 wt.% of the biomass. In embodiments, fungal mycelium makes up at least about 50 wt.%, at least about 55 wt.%, at least about 60 wt.%, at least about 65 wt.%, at least about 70 wt.%, at least about 75 wt.%, at least about 80 wt.%, at least about 85 wt.%, at least about 90 wt.%, at least about Aty Docket No. 10087-82-PCT

[0095] 95 wt.%, at least about 96 wt.%, at least about 97 wt.%, at least about 98 wt.%, at least about 99 wt.%, or substantially all of the biomass. The remainder (i.e., the non-mycelial portion) of the mycelial biomass may contain other fungal tissues (conidia, fruiting bodies, etc.). “Mycelial biomass” specifically includes biomass produced by any fermentation method and thus encompasses biomass produced by submerged fermentation methods and nonsubmerged fermentation methods. In some embodiments, a mycelial biomass is produced by any one or more submerged fermentation methods known in the art, such as, for example, those described in U.S. Patent 7,635,492 to Finnigan et al. In other embodiments, a mycelial biomass is a “cohesive mycelial biomass” produced by a fermentation process other than a submerged fermentation process. A “cohesive mycelial biomass” has sufficient tensile strength and structural integrity to be picked up and moved by hand without disintegrating or tearing and is produced by any one or more fungal fermentation methods in which the filamentous fungus grows in such a way as to form a mass of interwoven mycelia, e.g., methods in which a fungal mycelium is grown in air or a controlled atmosphere out of a growth medium or feedstock (e.g., liquid surface fermentation, fermentation on the surface of a membrane or mesh scaffold, etc.). Examples of methods of producing a “cohesive mycelial biomass” are described in PCT Application Publications WO2020 / 176758, WO2019 / 099474, and WO2018 / 014004. Generally speaking, mycelial biomasses recovered from submerged fermentation processes are paste-like substances with poor tensile strength and structural integrity and thus are not cohesive mycelial biomasses.

[0096] As used herein, the term “nutraceutical” refers to substances that have health or medicinal benefits. In some instances, a nutraceutical not only supplements the diet but also aids in the prevention and / or treatment of disease and / or disorders. The term “nutraceutical” was coined from “nutrition” and “pharmaceutical” in 1989 by Stepen DeFelice, MD, founder and chairman of the Foundation for Innovation in Medicine (FIM).

[0097] As used herein, the term “oleogel” refers to a solidified or semi-solidified system, with a continuous phase made of a hydrophobic liquid (like a vegetable oil) where a selfassembled network, composed by a structurant, is responsible for the physical entrapment of the liquid.

[0098] As used here, the term “oleogelation” refers to the process whereby hydrophobic liquid oil, such as, for example, a plant based liquid oil is physically entrapped in a three- dimensional gel network formed by an oleogel structurant. Atty Docket No. 10087-82-PCT

[0099] As used herein, the phrase “oleogel structurant” or “structurant” refers to a molecule that is capable of physically entrapping a hydrophobic liquid (like a vegetable oil) by forming a three-dimensional network.

[0100] Embodiments of the present invention include oleogel compositions of filamentous fungi, typically oleogel compositions of edible filamentous fungi, and most typically oleogel food compositions, i.e., edible oleogel compositions that are adapted for consumption by humans or domesticated, farmed (e.g., agriculture or aquaculture), or livestock animals, that include filamentous fungal particles. In some embodiments, the oleogel food composition may be a food product that is analogous to a conventional or known food product comprising a dairy or otherwise animal-derived ingredient (butter, shortening, etc.). In some embodiments, the oleogel food composition may be a non-dairy composition or food product and may be a vegan (i.e., no animal derived components) composition or food product. Embodiments of the oleogel food compositions include, without limitation, butter, ghee, shortening, margarine, and analogs thereof. Embodiments of the oleogel may be used as component of food compositions including, without limitation, chocolate, chocolate compositions, gummies, meat analogs (i.e. a chicken analog, a pork analog, a beef analog), processed meats (i.e. sausages, patties, frankfurters), bakery products (i.e. cakes, cookies, bread, pastries).

[0101] Fungal Taxonomy, Species, Strains

[0102] Filamentous fungi suitable for use in the disclosed methods can vary. Suitable strains that may be used in the disclosed methods include any filamentous fungal strains selected from the phyla or divisions of zygomycota, glomermycota, chytridiomycota, basidiomycota or ascomycota. In some embodiments, the fungus belongs to an order selected from the group consisting of Mucorales, Ustilaginales, Russulales, Agaricales, Pezizales, and Hypocreales.

[0103] In some embodiments, the fungus belongs to a family selected from the group consisting of Mucoraceae, Ustilaginaceae, Hericiaceae, Polyporaceae, Grifolaceae, Lyophyllaceae, Strophariaceae , Lycoperdaceae, Agaricaceae, Pleurolaceae, Physalacriaceae, Omphalotaceae, Tuberaceae, Morchellaceae, Sparassidaceae , Nectriaceae, Bionectriaceae, and Cordycipitaceae .

[0104] In some embodiments, the fungus belongs to a genus selected from the group consisting of Agaricus, Calocybe, Calvatia, Cordyceps, Disciotis, Pomes, Fusarium, Atty Docket No. 10087-82-PCT

[0105] Ganoderma, Grifola, Hericulum, Hypholoma, Hypsizygus, Morchella, Pholiota, Pleurotus, Poly porous. Sparassis, Stropharia, Tuber, and Ustilago.

[0106] In some embodiments, the fungus is selected from the group consisting of Rhizopus oligosporus, Ustilago esculenla, Hericulum erinaceus, Polyporous squamosus, Grifola fondosa, Hypsizygus marmoreus, Hypsizygus u Im arms, Calocybe gambosa, Pholiota nameko, Calvatia giganlea, Agaricus bisporus, Stropharia riigosoannulala, Hypholoma lalerilrum, Pleurotus eryngii, Pleurotus oslrealus, Pleurotus ostreatus var. coliimbinus. Tuber borchii, Morchella esculenla, Morchella conica, Morchella imporluna, Sparassis crispa. Fusarium venenatum, Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698), Disciotis venosa, Clonostachys rosea, Cordyceps militaris, Trametes versicolor, Ganoderma lucidum, Flammulina velutipes, Lentinula edodes, Pleurotus djamor, Pleurotus ostreatus, and Leucoagaricus spp.

[0107] In some embodiments, the fungus is a. Fusarium species.

[0108] In some embodiments the fungus is Fusarium strain flavolapis (ATCC Accession Deposit No. PTA-10698).

[0109] In some embodiments, the fungus is selected from Fusarium venenatum and Fusarium venenatum strain A3 / 5 (formerly classified as Fusarium graminearum). In some embodiments, the fungus is Fusarium venenatum A3 / 5.

[0110] In some embodiments, the fungus is Fusarium venenatum.

[0111] In some embodiments, the fungus is selected from Pleurotus ostreatus, Agrocybe brasiliensis, Flammulina velutipes, Hypholoma capnoides, Hypholoma sublaterium, Morchella angusticeps, Macrolepiota procera, Coprinus comatus, Agaricus arvensis, Ganoderma tsugae, and Inonotus obliquus. In some embodiments, the fungus is Pleurotus ostreatus. In some embodiments, the fungus is Ganoderma lucidum.

[0112] In some embodiments, the fungus is selected from a fungus belonging to the order Polyporales, a member of the family Ganodermataceae, Ganoderma lucidum, Ganoderma tsugae, Ganoderma applanatum, Ganoderma resinaceum, Ganoderma oregonense, Trametes versicolor, Trametes pubescens, Schizophyllum commune, and Polyporous squamosus.

[0113] In some embodiments, the fungus is selected from a fungus belonging to Ascomycota, Zygomycota, Aspergillus, Fusarium, Neurospora, Monascus, Basidiomycota, Lentinula, and Neurospora. In some embodiments, the fungus is Neurospora crassa. Atty Docket No. 10087-82-PCT

[0114] In some embodiments, the fungus is selected from Agaricus arvensis; Agrocybe brasiHensis Amylomyces rouxii; Amylomyces sp.; Armillaria mellea; Aspergillus nidulans; Aspergillus niger; Aspergillus oryzae; Ceriporia lacerata; Coprinus comatus; Fibroporia vaillantii; Fistulina hepatica; Flammulina velutipes; Fomitopsis officinalis; Ganoderma sessile; Ganoderma tsugae; Hericium erinaceus; Flypholoma capnoides; Flypholoma sublateritium; Inonotus obliguus; Lactarius chrysorrheus; Macrolepiota procera; Morchella angusticeps; Myceliophthora thermophila; Neurospora crassa; Penicillium camemberti; Penicillium chrysogenum; Penicillium rubens; Phycomyces blakesleeanus; Pleurotus djamor; Pleurotus ostreatus; Polyporus sguamosus; Psathyrella aguatica; Rhizopus microsporus; Rhizopus oryzae; Schizophyllum commune; Streptomyces venezuelae; Stropharia rugosoannulata; Thielavia terr e str is; Ustilago maydis; Fusarium sp. and Gibberella sp. In some embodiments, the fungus used includes Ganoderma sessile, Neurospora crassa, and / or Phycomyces blakesleeanus. In some embodiments, the fungus is a mutant strain of any of the above-identified fungal species. In some embodiments, the fungus is RMs2374, a Neurospora crassa mutant that can be obtained from the Fungal Genetics Stock Center (FGSC 5140). In some embodiments, the fungus is Ganoderma lucidum.

[0115] In some embodiments, the fungus is selected from Aspergillus oryzae, Rhizopus oryzae, Rhizopus oligosporus and Rhizopus microspores, Fusarium graminareum, Cordyceps militaris, Cordyceps sinensis, Tuber melanosporum, Tuber magnatum, Pennicillium camemberti, Neurospora intermedia, Neurospora sitophila, Xylaria hypoxion, and Neurospora crassa.

[0116] In some embodiments, the fungus is selected from brewer’s yeast (e.g., nutritional yeast, Saccharomyces cerevisiae, etc.), Brettanomyces bruxellensis, Brettanomyces anomalus, Brettanomyces custer sianus, Brettanomyces naardenensis, Brettanomyces nanus, Dekkera bruxellensis, Dekkera anomala, Candida stellata, Schizosaccharomyces pombe, Torulaspora delbrueckii, Zygosaccharomyces bailii, Pichia pastoris (also called, in some cases, Komagataella phaffii, K. pastoris, or K. pseudopastor is), Agaricus bisporus, Pleurotus ostreatus, Lentinula edodes, Auricularia auricula-judae, Volvariella volvacea, Flammulina velutipes, Tremella fuciformis, Hypsizygus tessellatus, Stropharia rugosoannulata, Cyclocybe aegerita, Hericium erinaceus, Boletus edulis, Calbovista subsculpta, Calvatia gigantean, Cantharellus cibarius, Craterellus tubaeformis, Clitocybe nuda, Cortinarius caperatus, Craterellus cornucopioides, Grifola frondosa, Gyromitra Atty Docket No. 10087-82-PCT esculenta, Hericium erinaceus, Hydnum repandum, Lactarius deliciosus, Morchella, Pleurotus ostreatus, Tricholoma matsutake, and Tuber sp.

[0117] In some embodiments, the fungus is selected from Rasamsonia composticola, Talaromyces emersonii, Rasamsonia emersonii, Thermomucor indicaeseudaticae, Rhizomucor miehei, Rhizomucor pusillus, Thielavia terricola var minor, aRhizopus sp., and Thermoascus thermophilus.

[0118] In some embodiments, the fungus is selected from Aspergillus oryzae, Rhizopus oryzae, Fusarium graminareum, Cordyceps militaris, Cordyceps sinensis, Tuber melanosporum, Tuber magnatum, Pennicillium camemberti, Neurospora intermedia, Neurospora sitophila, and Xylaria hypoxion. In some embodiments, the fungus is Neurospora.

[0119] In some embodiments, the fungus is selected from Candida etchellsii, Candida guilliermondii, Candida humilis, Candida utilis, Candida versatilis, Debaryomyces hansenii, Kluyveromyces lactis, Kluyveromyces marxianus, Kluyveromyces thermotolerans, Pichia pastoris, Rhodotorula sp., Saccharomyces bayanus, Saccharomyces beticus, Saccharomyces cerevisiae, Saccharomyces chevalieri, Saccharomyces diastaticus, Saccharomyces ellipsoideus, Saccharomyces exiguus, Saccharomyces florentinus, Saccharomyces pastorianus, Saccharomyces pombe, Saccharomyces sake, Saccharomyces uvarum, Sporidiobolus johnsonii, Sporidiobolus salmonicolor, Sporobolomyces roseus, Xanthophyllomyces dendrorhous, Yarrowia lipolytica, and Zygosaccharomyces rouxii.

[0120] Because they are edible, the filamentous fungi suitable for use in methods provided by the present disclosure have a low mycotoxin content. In some embodiments, the total amount of mycotoxins in a filamentous fungi utilized in the disclosed methods is less than about 10 ppm.

[0121] The amount of edible filamentous fungi used in the disclosed methods can vary based on the desired rheological properties of the final oleogel product. In various aspects, an amount of 0.25 wt.% - 45 wt.% of fungi, or any subrange included therein, can be used to prepare the disclosed aqueous dispersions. As used herein, wt.% is expressed as the weight of a component per unit volume. For example, if 1g of filamentous fungi is used to make 100ml of aqueous dispersion, it would be represented as a 1 wt.% fungal dispersion. In some embodiments, the amount of edible filamentous fungi used to prepare the disclosed aqueous dispersions is 1.0 wt.% - 5.0 wt.%. In some embodiments, the amount of edible filamentous fungi used to prepare the disclosed aqueous dispersions is 5.0 wt.% - 10.0 wt.%. Atty Docket No. 10087-82-PCT

[0122] In some embodiments, the amount of edible filamentous fungi used to prepare the disclosed aqueous dispersions is 20.0 wt.% - 28.0 wt.%. In some embodiments, the amount of edible filamentous fungi used to prepare the disclosed aqueous dispersions is selected from 2.0 wt.%, 3.0 wt.%, 4.0 wt.%, 5.0 wt.%, 6.0 wt.%, 7.0 wt.%, 8.0 wt.%, 9.0 wt.%, 10.0 wt.%, 11.0 wt.%, 12.0 wt.%, 13.0 wt.%, 14.0 wt.%, 15.0 wt.%, 16.0 wt.%, 17.0 wt.%, 18.0 wt.%,

[0123] 19.0 wt.%, 20.0 wt.%, 21.0 wt.%, 22.0 wt.%, 23.0 wt.%, 24.0 wt.%, 25.0 wt.%, 26.0 wt.%,

[0124] 27.0 wt.%, 28.0 wt.%, 29.0 wt.%, 30.0 wt.%, 31.0 wt.%, 32.0 wt.%, 33.0 wt.%, 34.0 wt.%,

[0125] 35.0 wt.%, 36.0 wt.%, 37.0 wt.%, 38.0 wt.%, 39.0 wt.%, 40.0 wt.%, 41.0 wt.%, 42.0 wt.%,

[0126] 43.0 wt.%, 44.0 wt.%, and 45.0 wt.%. As a result, the fungal oleogel compositions may have an enriched protein content, which may in embodiments be about at least 4.0 wt.%, at least about 4.5 wt.%, at least about 5.0 wt.%, at least about 5.5 wt.%, at least about 6.0 wt.%, at least about 6.5 wt.%, at least about 7.0 wt.%, at least about 7.5 wt.%, at least about 8.0 wt.%, at least about 8.5 wt.%, at least about 9.0 wt.%, at least about 9.5 wt.%, at least about 10.0 wt.%, at least about 10.5 wt.%, at least about 11.0 wt.%, 5 at least about 11.5 wt.%, at least about 12.0 wt.%, 12.5 wt.%, 13.0 wt.%, 13.5 wt.%, 14.0 wt.%, 14.5 wt.%, 15.0 wt.%, 15.5 wt.%, 16.0 wt.%, 16.5 wt.%, 17.0 wt.%, 17.5 wt.%, 18.0 wt.%, 18.5 wt.%, 19.0 wt.%, 19.5 wt.%, 20.0 wt.%, 20.5 wt.%, 21.0 wt.%, 21.5 wt.%, 22.0 wt.%, 22.5 wt.%, 23.0 wt.%, 23.5 wt.%, 24.0 wt.%, 24.5 wt.%, or at least about 25.5 wt.% on a dry weight basis.

[0127] In addition to having a high overall protein content, filamentous fungal particles in oleogel compositions of the present invention may provide advantageous protein compositions or chemistries. By way of first non-limiting example, the filamentous fungal particles may represent a “complete” protein source by providing all nine essential amino acids and / or all 20 proteinogenic amino acids. By way of second non-limiting example, the filamentous fungal particles may comprise at least one branched-chain amino acid (e.g., leucine, isoleucine, valine), and may in some embodiments contain such amino acids in amounts of at least about 10 wt.%, at least about 15 wt.%, at least about 20 wt.%, at least about 25 wt.%, or at least about 30 wt.%.

[0128] Filamentous fungal particles may provide various other nutritional or compositional advantages to the oleogel compositions of the present invention as well. By way of first nonlimiting example, the filamentous fungal particles may have an advantageously high content of dietary fiber to allow for the creation of high-fiber food products (and in particular high- fiber alternatives to or analogs of conventional food products that may have lower fiber contents); in some embodiments, the filamentous fungal particles may comprise at least Atty Docket No. 10087-82-PCT about 27 wt.%, at least about 28 wt.%, at least about 29 wt.%, at least about 30 wt.%, at least about 31 wt.%, at least about 32 wt.%, at least about 33 wt.%, at least about 34 wt.%, at least about 35 wt.%, or at least about 36 wt.% dietary fiber. A high fiber content may be advantageous for any one or more additional reasons not directly related to nutritional composition, e.g. improved hydration properties (such as decreased water activity to allow for easier preparation / storage and longer shelf life), increased satiation or “fullness” upon eating (which may encourage consumers to eat more moderate portions of “indulgence” products such as butter analog food products or shortening analog food products and thereby aid in preventing or mitigating adverse health effects such as high cholesterol), improved digestibility, etc.

[0129] As a result, the fungal oleogel compositions may have an enriched dietary fiber content, which may in embodiments be about at least 1.0 wt.%, at least about 1.5 wt.%, at least about 2.0 wt.%, at least about 2.5 wt.%, at least about 3.0 wt.%, at least about 3.5 wt.%, 4.0 wt.%, at least about 4.5 wt.%, at least about 5.0 wt.%, at least about 5.5 wt.%, at least about 6.0 wt.%, at least about 6.5 wt.%, at least about 7.0 wt.%, at least about 7.5 wt.%, at least about 8.0 wt.%, at least about 8.5 wt.%, at least about 9.0 wt.%, at least about 9.5 wt.%, or at least about 10.0 wt.% on a dry weight basis.

[0130] Methods

[0131] Oleogelation

[0132] An oleogel system is a solidified or semi-solidified system, with a continuous phase made of a hydrophobic liquid (e.g., a vegetable oil), in which a self-assembled network of a material referred to herein as a “structurant” physically entraps the liquid. Thus, oleogelation, as that term is used herein, is the process whereby the hydrophobic liquid (e.g. , vegetable oil) is physically entrapped in a three-dimensional gel network formed by the oleogel structurant. In various aspects, suitable oleogel structurants are a continuous gelled water phase of an oil-in-water emulsion. In many embodiments, the resulting oleogel may have the same functionality as a solid fat, but contains a reduced amount of trans fatty acids and saturated fatty acids.

[0133] Proteins can act as the oleogel structurant through emulsification by forming a three- dimensional network through hydrogen bonding and interfacial tension. The oleogel is formed by first preparing an oil-in-water emulsion by combining an oil with a proteincontaining aqueous liquid. In embodiments, a combination of proteins may be used. In embodiments, the emulsion contains at least 40%, 50%, 60%, 70%, 80% or 85% oil. In Atty Docket No. 10087-82-PCT some embodiments, polysaccharides can be added to the emulsion to reinforce the interface. Subsequently, the water is removed by drying the emulsion, which leads to the formation of a high internal phase emulsion consisting of a three-dimensional network of polymers filled with liquid oil. Drying may be done by, for example, freeze-drying, oven drying, and / or vacuum-oven drying. The emulsion is allowed to dry for at least about 12, at least about 24, at least about 30, at least about 36 hours, or at least about 48 hours or until the water content is less than approximately 1% . Then, the dried emulsion is sheared to obtain an oil- continuous material referred to as an oleogel. Shearing is performed for a duration of time necessary to break up the hard oleogel and blend it to a smooth consistency.

[0134] In embodiments, methods of making fungal oleogel compositions are provided, comprising combining an aqueous dispersion of filamentous fungal particles derived from a fungal biomass with an oil. In that regard, in various aspects an oleogel structurant is an aqueous dispersion of fungal particles. In various aspects, an oleogel structurant is an aqueous dispersion of fungal particles derived from a fungal biomass. Optionally, the pH of the resulting combination may be adjusted. Subsequently, the resulting combination is freeze-dried and sheared.

[0135] One non-limiting embodiment of such a method is as follows.

[0136] A dispersion of filamentous fungal particles in an aqueous liquid is prepared by combining water and fungal material (i.e., a fungal mycelial biomass) in a ratio of about 10: 1 to about 1 : 10 or any subrange in between. The water and fungal material are placed into an apparatus containing rotating blades, for example a blender or an impeller, and sheared until smooth. In this embodiment, shearing occurs by high speed blending or mixing, non-stop, for a period of at least 1 minute, or until a stable homogenous dispersion is achieved; shear mixing is desirable to achieve a satisfactory homogeneity of the fungal material in the water, and to fully disintegrate any possible agglomerates. The resulting dispersion is heat treated while mixing for a period of at least 30 minutes at 120 °F.

[0137] Then, optionally, the pH of the aqueous dispersion is adjusted to 4.5 by adding 1 M hydrochloric acid dropwise to the aqueous dispersion.

[0138] Next, the aqueous dispersion is combined with an oil or blend of two or more oils and homogenized at 120 °F and 12,000 rpm for 120 s using a high-speed homogenizer and then, cooled down to room temperature (e.g., 22-24 °C). In some embodiments, a single oil is added, in some embodiments, a blend of two or more oils is added. In one embodiment, sunflower oil is added in an amount of about 40 wt. %. In one embodiment, sunflower oil is Atty Docket No. 10087-82-PCT added in an amount of about 50 wt. %. In one embodiment, sunflower oil is added in an amount of about 60 wt.%. In one embodiment, sunflower oil is added in an amount of about 70 wt.%. In one embodiment, sunflower oil is added in an amount of about 80 wt.%. In some embodiments, the oil is heated at 120 °F for a period of at least 60 minutes prior to blending with the aqueous dispersion.

[0139] Then, the resulting emulsion is dried by freeze drying for a period of at least 36 hours. The dried emulsion is placed into an apparatus containing rotating blades, for example a blender or an impeller, and sheared for a period of 5 - 30 seconds.

[0140] The resulting oleogel composition contains less than about 1.0 wt.% water. The resulting oleogel composition is solid or semi-solid at 20 °C. The resulting oleogel composition is stable at 20 °C for at least about one day. Stability is determined by measuring whether there is any separation of the oil from other ingredients, known as “oiling-off,” in the composition after the drying process, after shearing, and / or after storage of the sheared oleogel. When oiling-off is not present during any of the three stages, the composition is stable. Stability may also be determined by measuring the turbiscan stability index (TSI). TSI is well-known for providing substantial information on the stability and dispersibility of any formulation. A TSI of less than 3 indicates a stable emulsion. In some embodiments, the resulting oleogel composition may have a TSI of about 0 to about 3.

[0141] In some embodiments, the resulting oleogel composition may have a melting temperature from about 27 °C to about 37 °C. In some embodiments, the resulting oleogel compositions may have a crystallization temperature from about 5 °C to about 20 °C. Aqueous Dispersion

[0142] Methods provided by the present disclosure utilize a liquid dispersion made from one or more filamentous fungi. In various embodiments, the liquid dispersion is suitable for use as an oleogel structurant. The liquid dispersion comprises particles of filamentous fungi dispersed in an aqueous medium. In some embodiments, the fungal particles can be prepared from a submerged fermentation-derived fungal biomass, for example a flour-like material produced by drying (e.g., spray-drying), or a dewatered fungal biomass (e.g., a dough or paste-like material), and are dispersed in the medium to form the dispersion. In some embodiments, the fungal particles are size-reduced, which can occur by mechanical means such as cutting, chopping, dicing, mincing, grinding, milling, blending, etc. or via sonication. In other embodiments, the fungal particles are not size-reduced. Atty Docket No. 10087-82-PCT

[0143] In various embodiments, the liquid dispersion is prepared by combining and blending a filamentous fungal biomaterial with an aqueous phase. In various embodiments, the oleogel structurant is an aqueous dispersion prepared by combining and blending a filamentous fungal biomaterial with an aqueous phase. In embodiments, the filamentous fungal biomaterial is a dried, size reduced biomass product that is allowed to hydrate in water for a sufficient time to permit the size reduced particles to absorb water before a final blending step to produce a dispersion. The dispersion of filamentous fungal particles in an aqueous liquid utilized in the disclosed methods is preferably stable such that the particulates of filamentous fungus do not readily separate from the aqueous medium in which they are dispersed. For example, in various embodiments, upon forming the dispersion the resulting liquid appears to be homogeneous in appearance and does not visibly separate into distinct phases prior to further use in the process, or for at least about 1, 2, 3, 4, 5, 6, 12, 18, or 24 hours. No visibly discernable or significant sediment forms on the bottom of the container holding the dispersion.

[0144] The blending can occur at varying speeds and times. As can be appreciated, a typical blending apparatus is one that combines two or more components together via the use of moving or rotating blades. As such, longer blending times and / or higher blending speeds will often result in smaller fungal particles in the dispersion. In some embodiments, the fungal material is blended in an aqueous medium until it is reduced to fine particles. In some embodiments, the particle size of the fungal material is the same or similar to those seen in conventional flour-like materials, such as wheat flours, wherein the particle size is about 30 to about 400 microns. In embodiments the particle size of the fungal material is 500 microns or less. In embodiments the particle size of the fungal material is 400 microns or less. In embodiments the particle size of the fungal material is 300 microns or less. In embodiments the particle size of the fungal material is 200 microns or less. In embodiments the particle size of the fungal material is in the range of about 80 microns to about 150 microns. In embodiments the particle size of the fungal material is less than about 50, 40, 30, 20, or 10 microns.

[0145] Typically, the blending occurs at sufficient speed and for a sufficient period of time to produce filamentous fungal particles in the aqueous dispersions having a particle size of about 500 microns or less. In some embodiments, the blending is high shear blending, where the fungi and water are blended together for at least 2 minutes at a speed of 10,000 rpm or greater. In some embodiments, the blended mixture is heated gradually during blending, in Atty Docket No. 10087-82-PCT some embodiments to the boiling point of water, to facilitate production of the dispersion. Once blending is complete, the heated mixture is allowed to cool prior to further utilization in the disclosed methods.

[0146] The ratio of filamentous fungal biomaterial to water can be adjusted to produce an aqueous dispersion with a desired consistency and density. The ratio of the biomaterial to water is from about 1 :50 to about 50: 1 or any range of ratios in between. In some embodiments, the ratio of the fungal biomaterial to water is about 1 :30, about 1 :20, about 1 : 10, about 1 :9, about 1 :8, about 1 :7, about 1 :6, about 1 :5, about 1 :4, about 1 :3, about 1 :2, about 1 : 1, about 2: 1, about 3: 1, about 4:1, about 5: 1, about 6: 1, about 7: 1, about 8: 1, about 9: 1, or about 10: 1.

[0147] In some embodiments, the filamentous fungal particles in the dispersion comprise, and in some embodiments consist essentially of, fungal mycelia. In some embodiments, the filamentous fungal particles in the dispersion comprise 50 wt.% - 95 wt.% fungal mycelia. In some embodiments, the filamentous fungal particles in the dispersion comprise at least about 50 wt.% fungal mycelia, in some embodiments at least about 75 wt.% fungal mycelia, and in some embodiments at least about 95 wt.% fungal mycelia.

[0148] In some embodiments, the filamentous fungal particles in an oleogel structurant comprise, and in some embodiments consist essentially of, fungal mycelia. In some embodiments, the filamentous fungal particles in the oleogel structurant comprise 50 wt.% - 95 wt.% fungal mycelia. In some embodiments, the filamentous fungal particles in the oleogel structurant comprise at least about 50 wt.% fungal mycelia, in some embodiments at least about 75 wt.% fungal mycelia, and in some embodiments at least about 95 wt.% fungal mycelia.

[0149] In some embodiments, the oleogel structurant is an aqueous dispersion comprising about 5 wt% to about 99 wt%, about 5 wt% to about 95 wt%, about 5 wt% to about 90 wt%, about 5 wt% to about 85 wt%, about 5 wt% to about 80 wt%, about 5 wt% to about 75 wt%, about 5 wt% to about 70 wt%, about 5 wt% to about 65 wt%, about 5 wt% to about 60 wt%, about 5 wt% to about 55 wt%, about 5 wt% to about 50 wt%, about 5 wt% to about 45 wt%, about 5 wt% to about 40 wt%, about 5 wt% to about 35 wt%, about 5 wt% to about 30 wt%, about 5 wt% to about 25 wt%, about 5 wt% to about 20 wt%, about 5 wt% to about 15 wt%, about 5 wt% to about 10 wt%, or about 5 wt% filamentous fungal biomass. In some embodiments, the oleogel structurant is an aqueous dispersion comprising about 0.25 wt% to about 45 wt%, about 0.25 wt% to about 40 wt% about 0.25 wt% to about 35 wt%, about Atty Docket No. 10087-82-PCT

[0150] 0.25 wt% to about 30 wt%, about 0.25 wt% to about 25 wt%, about 0.25 wt% to about 20 wt%, about 0.25 wt% to about 15 wt%, about 0.25 wt% to about 10 wt%, about 0.25 wt% to about 5 wt%, about 0.25 wt% to about 4 wt%, about 0.25 wt% to about 3 wt%, about 0.25 wt% to about 2 wt%, about 0.25 wt% to about 1 wt%, about 0.25 wt% to about 0.5 wt%, or about 0.25 wt% filamentous fungal biomass. In some embodiments, the oleogel structurant is an aqueous dispersion comprising an amount of filamentous fungal biomass selected from about 10 wt%, about 8 wt%, or about 6.25 wt.%. In some embodiments, the oleogel structurant is an aqueous dispersion comprising about 10 wt% filamentous fungal biomass. In some embodiments, the oleogel structurant is an aqueous dispersion comprising about 8 wt% filamentous fungal biomass. In some embodiments, the oleogel structurant is an aqueous dispersion comprising about 6.25 wt% filamentous fungal biomass.

[0151] In some embodiments, the filamentous fungal particles comprise both fungal mycelia and one or more fruiting bodies. Some species of fungi produce multicellular fruit bodies for sexual reproduction and spore development. A fruiting body is the part of functional mushrooms that are typically thought of when considering the term “mushroom.” Fruiting bodies include mushroom caps, stems, gills, skirts, scales, volva, and the like. While a fruiting body grows above a growth substrate, mycelium typically grows within a substrate (e.g., soil, dead wood, etc.) from which it derives resources. Before fruiting, the mycelium typically must reach a certain biomass having a sufficient amount of storage mycelium to serve production of fruiting body production. For filamentous fungi that form fruiting bodies, the filamentous fungal particles can be completely or mostly formed of fruiting bodies. Further, the filamentous fungal particles can be derived from a fungal biomass that comprises conidia. In addition, the filamentous fungal particles can comprise a mixture of mycelium, conidia, and fruiting body material in any proportions. In embodiments, fungal particles are particles of mycelium from filamentous fungi, such as multicellular filamentous molds, that do not produce fruiting bodies. In embodiments, fungal particles comprise a blend of mycelia particles from more than one type of filamentous fungus. In embodiments, fungal particles comprise, or alternatively consist of, particles of mycelium from one or more multicellular filamentous molds that do not produce fruiting bodies wherein said particles comprise less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% conidia or particles from conidia. Atty Docket No. 10087-82-PCT

[0152] In some embodiments, the pH of the aqueous liquid dispersion is less than about 6.0, less than about 5.5, less than about 5.0, less than about 4.5, less than about 4.0, less than about 3.5, less than about 3.0, less than about 2.5, less than about 2.0, less than about 1.5, less than about 1.0, or less than about 0.5.

[0153] In some embodiments, a liquid dispersion is produced under nitrogen, which results in a creamier consistency of liquid dispersion with less fungal scent. Production under nitrogen can be accomplished by bubbling nitrogen gas into a closed vessel such that nitrogen replaces the available oxygen (and other non-nitrogen gases) in the vessel. This can occur during blending, while the nascent dispersion is created.

[0154] The amount of aqueous dispersion and / or the amount of oleogel structurant, that is the combined amount of edible filamentous fungi and aqueous liquid, used in the disclosed methods can vary based on the desired protein content, texture, and / or flavor of the final oleogel product. In various aspects, an amount of 10 wt.% - 60 wt.% of dispersion can be used in the disclosed methods. In some embodiments, the amount of aqueous dispersion used in the disclosed methods is 20 wt.% - 60 wt.%.

[0155] Non-fungal Proteins

[0156] Prior to the addition of an oil, or a combination of two or more oils, to the aqueous dispersion and / or oleogel structurant according to the disclosed methods, other components that impart beneficial qualities to the oleogel can be added to the dispersion. For example, additional, non-fungal proteins can be added to aid in the oleogelation process.

[0157] Non-fungal proteins can be added for any one or more of many reasons. For example, one or more non-fungal proteins can be added to aid the oleogelation process and to provide structural properties to the resulting oleogel. Additionally, certain non-fungal proteins can be added to boost the nutritional value of the resulting oleogel. For example, hemp protein is considered a “complete” protein because it is known to contain all of the essential amino acids humans are required to intake via food and hemp seeds contain more than 30g of protein per 100g seeds. Additionally, soy protein, like hemp, is considered a “complete” protein because it is known to contain all of the essential amino acids humans are required to intake via food and soy beans contain about 18g of protein per 100g. Other proteins contain similar benefits and can be mixed and matched in the disclosed methods to achieve desired nutritional profiles.

[0158] Suitable non-fungal proteins for use in the disclosed methods include plant proteins, such as, by way of non-limiting example, bean protein, broccoli protein, chickpea protein, Atty Docket No. 10087-82-PCT hemp protein, lentil protein, nut protein, pea protein, potato protein, quinoa protein, rice protein, seaweed protein, seed protein, spinach protein, and soy protein, among others. In some embodiments, the non-fungal protein is selected from hemp protein, soy protein, pea protein, chickpea protein, rice protein, and combinations of any of the foregoing. In some embodiments, the non-fungal protein is hemp protein. In some embodiments, the non-fungal protein is soy protein. In some embodiments, the non-fungal protein is a blend of hemp protein and soy protein.

[0159] The amount of non-fungal protein used in the disclosed methods can vary based on the desired texture, and / or flavor of the final oleogel product. In various aspects, an amount of 1.0 wt.% - 5.0 wt.% of non-fungal protein can be used. This amount can reflect use of a single non-fungal protein source, or a combination of non-fungal protein sources, depending on the desired nutritional content of the resulting oleogel. In some embodiments, the amount of non-fungal protein used in the disclosed methods is selected from 0.5 wt.%, 1.0 wt.%, 1.5 wt.%, 2.0 wt.%, 2.5 wt.%, 3.0 wt.%, 3.5 wt.%, 4.0 wt.%, 4.5 wt.%, and 5.0 wt.%.

[0160] Oils

[0161] As provided herein, the present disclosure provides methods whereby an oil is, or one or more oils are, combined with a dispersion of filamentous fungal particles in an aqueous liquid, to produce an oil-in-water emulsion. Any suitable food grade oil, or blends of food grade oils, may be used in the disclosed methods. Oils may be selected for their effect on the thermal, textural, and rheological properties of the oleogel. For example, oils with a higher degree of saturation strengthen the structure of the oleogel. Oils may also be selected for their taste, texture, melting temperature, perceived moisture content, or any or all of the above. For example, some oils, such as olive oils, have a distinctive taste that can be imparted to the oleogel composition provided by the methods. Alternatively, a neutral flavored oil, such as sunflower or soybean oil, may be desired, to avoid flavoring the oleogel composition.

[0162] Oils suitable for use in the disclosed methods, either alone or in combinations of two or more, include acai oil, almond oil, amaranth oil, apricot oil, argan oil, artichoke oil, avocado oil, ben oil (extracted from the seeds of the Moringa oleifera), blackcurrant seed oil, borage seed oil, Borneo tallow nut oil, buffalo gourd oil, canola oil, carob pod oil, cashew oil, coconut oil, coriander seed oil, com oil, cottonseed oil, evening primrose oil, false flax oil, grapeseed oil, hazelnut oil, hemp oil, kapok seed oil, macadamia oil, meadowfoam seed oil, mustard oil, okra seed oil, olive oil, palm oil, peanut oil, pecan oil, Atty Docket No. 10087-82-PCT perilla seed oil, pequi oil, pine nut oil, pine seed oil, pistachio oil, poppyseed oil, prune kernel oil, pumpkin seed oil, quinoa oil, ramtil oil, rice bran oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, thistle oil, walnut oil, and wheat germ oil.

[0163] In some embodiments, the oil utilized in the disclosed methods is sunflower oil.

[0164] In some embodiments, the oil utilized in the disclosed methods is selected from acai oil, almond oil, avocado oil, blackcurrant seed oil, borage seed oil, canola oil, cashew oil, coconut oil, corn oil, cottonseed oil, evening primrose oil, grapeseed oil, hazelnut oil, hemp oil, macadamia oil, olive oil, palm oil, peanut oil, pecan oil, pine seed oil, pistachio oil, pumpkin seed oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, walnut oil, and combinations of any of the foregoing.

[0165] The amount of oil, or combinations of oils, used in the disclosed methods can vary based on the desired rheological properties of the final oleogel product. In various aspects, an amount of 30 wt.% - 85 wt.%, or any subrange in between, of oil can be used in the disclosed methods to form the oil-in-water emulsion. In some embodiments, the amount of oil used in the disclosed methods is selected from 30.0 wt.%, 31.0 wt.%, 32.0 wt.%, 33.0 wt.%, 34.0 wt.%, 35.0 wt.%, 36.0 wt.%, 37.0 wt.%, 38.0 wt.%, 39.0 wt.%, 40.0 wt.%, 41.0 wt.%, 42.0 wt.%, 43.0 wt.%, 44.0 wt.%, 45.0 wt.%, 46.0 wt.%, 47.0 wt.%, 48.0 wt.%, 49.0 wt.%, 50.0 wt.%, 51.0 wt.%, 52.0 wt.%, 53.0 wt.%, 54.0 wt.%, 55.0 wt.%, 56.0 wt.%, 57.0 wt.%, 58.0 wt.%, 59.0 wt.%, 60.0 wt.%, 61.0 wt.%, 62.0 wt.%, 63.0 wt.%, 64.0 wt.%., 65.0 wt.%, 66.0 wt.%, 67.0 wt.%, 68.0 wt.%, 69.0 wt.%, 70.0 wt.%, 71.0 wt.%, 72.0 wt.%, 73.0 wt.%, 74.0 wt.%, 75.0 wt.%, 76.0 wt.%, 77.0 wt.%, 78.0 wt.%, 79.0 wt.%, 80.0 wt.%, 81.0 wt.%, 82.0 wt.%, 83.0 wt.%, 84.0 wt.%., and 85.0 wt.%.

[0166] Combining the dispersion of filamentous fungal particles in an aqueous liquid with an oil is facilitated by the filamentous fungal particles having high water wettability (for highly stable oil-in-water emulsions), high oil wettability (for highly stable water-in-oil emulsions), and / or a balance between these two characteristics.

[0167] As a result, the fungal oleogel compositions may have an oil composition of about at least 75.0 wt.%, at least about 76.0 wt.%, at least about 77.0 wt.%, 78.0 wt.%, at least about 79.0 wt.%, at least about 80.0 wt.%, at least about 81.0 wt.%, at least about 82.0 wt.%, at least about 83.0 wt.%, at least about 84.0 wt.%, at least about 85.0 wt.%, at least about 86.0 wt.%, at least about 87.0 wt.%, at least about 88.0 wt.%, at least about 89.0 wt.%, at least about 90.0 wt.%, at least about 91.0 wt.%, at least about 92.0 wt.%, at least about 93.0 wt.%, at least about 94.0 wt.%, or at least about 95.0 wt.%, on a dry weight basis. Atty Docket No. 10087-82-PCT

[0168] Polysaccharides

[0169] In embodiments, polysaccharides can be added to the emulsion to strengthen the interfacial network and enhance the oil holding capacity of the system.

[0170] Polysaccharides suitable for use in the disclosed methods, either alone or in combinations of two or more, include, for example, carrageenan, xanthan gum, methylcellulose, carboxymethylcellulose (CMC), and hydroxyl -propyl methyl cellulose (HPMC).

[0171] Fungal Oleogel Food Compositions

[0172] The present disclosure provides methods of making oleogel compositions from filamentous fungal biomass. The fungal oleogel compositions are edible on their own, for example as a shortening analog or a butter analog, or may be used as ingredients or components incorporated into other edible compositions, for example as a fat replacer in processed meat products and chocolate.

[0173] The oleogels according to the present disclosure may be utilized as a wide range of products including, but not limited to a butter analog, a shortening analog, a ghee analog, or a margarine analog.

[0174] The oleogels according to the present disclosure may be incorporated into a wide range of savory food products including, but not limited to, processed meat products (such as sausages, patties, and frankfurters), meat analogs (such as a pork analog, a beef analog, a chicken analog), and baked products (such as bread).

[0175] The oleogels according to the present disclosure may be incorporated into a wide range of sweet food products including, but not limited to, chocolate, chocolate compositions, baked products (such as cake, muffins, pastries, cookies, brownies, pie crust), and ice cream.

[0176] Cosmetics

[0177] The present disclosure provides methods of making oleogel compositions from filamentous fungal biomass. The fungal oleogel compositions can be utilized in cosmetic compositions.

[0178] The oleogels according to the present disclosure may be incorporated into a wide range of cosmetics, including but not limited to, moisturizers, makeup, foundation, sun protection lotions, hair gels, mascara and eye shadows. Atty Docket No. 10087-82-PCT

[0179] Nutraceutical Delivery Systems

[0180] The present disclosure provides methods of making oleogel compositions from filamentous fungal biomass. The fungal oleogel compositions can be incorporated into nutraceutical delivery systems.

[0181] Edible fungal oleogels can be utilized to encapsulate and help in controlled and targeted release of lipid soluble nutraceuticals. Nutraceuticals are prone to crystallization and precipitation during storage, which significantly limits their effective loading content and bioavailability. Encapsulation mitigates common storage issues also increases solubility within the gut.

[0182] Compounds suitable for use in the disclosed methods, either alone or in combinations of two or more, include carotene, curcumin, lutein, lycopene, co-enzyme Q- 10, docosahexaenoic acid, eicosapentaenoic acid, and tannins.

[0183] Fungal Oleogel Compositions

[0184] In another aspect, the present disclosure provides fungal oleogel compositions, comprising an aqueous dispersion filamentous fungal biomass and an oil, optionally produced by the disclosed methods.

[0185] The oil present in the oleogel compositions is as disclosed hereinabove. That is, in some embodiments the oil is selected from a vegetable oil, acai oil, almond oil, avocado oil, blackcurrant seed oil, borage seed oil, canola oil, cashew oil, coconut oil, com oil, cottonseed oil, evening primrose oil, grapeseed oil, hazelnut oil, hemp oil, macadamia oil, olive oil, palm oil, peanut oil, pecan oil, pine seed oil, pistachio oil, pumpkin seed oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, walnut oil, and combinations of any of the foregoing.

[0186] In some embodiments, the fungal oleogel compositions comprise a non-fungal protein, for example hemp protein, soy protein, pea protein, chickpea protein, rice protein, or combinations of any of the foregoing. As noted above, such non-fungal proteins may aid in the oleogelation process.

[0187] In some embodiments, the fungal oleogel compositions contain less than about 1.0 wt. % water.

[0188] In embodiments, the fungal oleogel composition is solid or semi-solid at room 20 °C. In embodiments, the fungal oleogel composition is stable for at least about one day at 20 °C. Atty Docket No. 10087-82-PCT

[0189] In other embodiments, the filamentous fungal biomass used to make the oleogel comprises particles produced by size-reducing a cohesive filamentous fungal mycelial biomass. This biomass can be produced by a number of methods, including liquid surface fermentation and solid-state fermentation.

[0190] In one embodiment, the filamentous fungal particles consist essentially of fungal mycelia. In other embodiments, the filamentous fungal particles comprise at least about 50 wt. % fungal mycelia, at least about 75 wt. % fungal mycelia, or at least about 95 wt. % fungal mycelia.

[0191] The fungal oleogel compositions are edible on their own, but are also suitable for the production of food compositions, as described herein.

[0192] The following examples are provided for the purpose of illustration only. They depict specific embodiments and are not intended to limit the scope or breadth of the present disclosure.

[0193] EXAMPLES

[0194] Example 1

[0195] Fungal oleogel compositions were prepared from edible filamentous fungal biomass, derived the filamentous acidophilic fungus Fusarium strain flavolapis. Biomat growth and preparation of E flavolapis occur as described herein and / or as provided in PCT / US2020 / 020152 (WO 2020 / 176758 Al). Several batches suitable to produce the fungal oleogel composition are shown in Table 1. Samples were prepared utilizing different concentrations of filamentous fungal biomass and oil to test rheological properties of the resulting emulsions and fungal oleogels.

[0196] T1 Atty Docket No. 10087-82-PCT

[0197] Table 1

[0198] Aqueous Dispersion of Filamentous Fungal Biomat: A biomat of Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698) was produced by a surface fermentation process. The biomat was pureed using a grinder. The moisture content of the pureed material was measured and determined to be 78-80 wt.%. Water was added to the pureed biomass at 2: 1 and 3 : 1 w / w ratios of water to biomass to create aqueous dispersions with a concentration of filamentous fungal particles of 8.33 wt.% and 6.25 wt.%. The water / biomass mixture was placed in a blender (Vitamix) and blended at the maximum setting for 1 minute. The blended composition was transferred to a multicooker (Thermomix) and mixed for 30 minutes at 120 °F and a speed of 3.0.

[0199] Oleogel preparation: Emulsions were prepared using the above described aqueous dispersions at two different pHs. The first pH was “as is” (approximately 5.2 - 6.2) and the second pH was adjusted to 4.5 by adding 1 M HC1 dropwise. Sunflower oil was used to create the emulsions. The oil was heated to 120 °F for at least 1 hour before emulsion preparation. The emulsions were made by adding the aqueous phase to the oil phase, where the oil phase volume fraction was varied from 40.00% to 70.00% (Table 1). The emulsions were produced by homogenizing the water and oil mixtures at 120 °F and 12,000 rpm for 120 seconds using a high-speed homogenizer and then, cooling them at room temperature (22 - 24 °C) for 2 hours. Then, the emulsion samples were dried by freeze drying for 36 hours. To prepare oleogel samples, the dried emulsions were uniformly sheared using a coffee grinder (Secura) for 10-15 seconds. Atty Docket No. 10087-82-PCT

[0200] Rheological characterization: All rheological properties of emulsions were characterized using a rheometer equipped with a cross-hatched parallel plate geometry and a solvent trap. First, the steady shear behavior of the emulsions was examined by measuring the apparent viscosity (r|a, Pa s) of each sample as a function of shear rate from 0.01 to 200 1 / s (Figure 1 A). All emulsion viscosities decreased with increased shear rate, representing a typical shear-thinning effect for pseudoplastic materials. For samples 1 - 4, pH and oil volume fraction did not have significant impact on flowing behavior and viscosity values.

[0201] Next, strain sweeps were performed (0.01-200%, 1 rad / s) to determine the linear viscoelastic region (LVR) (Figure 2A). For samples 1 - 4, the oil phase volume fraction and pH did not have a significant impact on structural strength and mechanical deformation of the samples.

[0202] Lastly, the small-amplitude oscillatory shear behavior of the emulsions was determined by frequency sweeps, where the elastic modulus (G', Pa) and viscous modulus (G", Pa) of each emulsion were measured as a function of frequency (co, rad / s) in the range of 0.1-100 rad / s (Figure 3 A). Dynamic oscillatory measurements were performed to establish relationships between internal structure and emulsion viscoelastic behaviors since the viscosity measurements alone were not capable of giving a full understanding of the relationships between emulsion microstructures and macroscopic properties. In all of the studied emulsions, the values of G' were significantly higher than those of G" at all frequencies, and no crossover of G' and G" was observed. These results indicated that the emulsions could be classified as weak physical gels with predominantly elastic gel-like behaviors.

[0203] All measurements were performed at 50% of the smallest critical strain determined to ensure that testing was done within the LVR.

[0204] Example 2

[0205] Fungal oleogel compositions were prepared from a submerged dough (SMD) of edible filamentous fungal biomass, z.e., a fungal dough obtained by a submerged fermentation process, derived from the filamentous acidophilic fungus Fusarium strain flavolapis. Fungal biomass was grown via a stirred-tank submerged fermentation process followed by steam injection into the fermentation vessel until the temperature inside is sufficient to inactivate growth of the biomass. The mycelium was then washed with deionized water and the washed biomass having a water content of about 75-85% collected (“dough”). Several emulsion batches suitable to produce the fungal oleogel composition Atty Docket No. 10087-82-PCT are shown in Table 2. Samples were prepared utilizing different concentrations of filamentous fungal biomass and oil to test rheological properties of the resulting emulsions and fungal oleogels.

[0206] Table 2

[0207] Aqueous Dispersion of Fungal Submerged Dough: The submerged dough (SMD) biomass was pureed using a grinder, and the moisture content of the pureed material was measured and determined to be 83-86 wt.%. Water was added to the pureed biomass to create dispersions with a concentration of filamentous fungal particles of 6.25 wt.% and 10 wt.%. The water / biomass mixtures were blended in a blender (Vitamix) at the maximum setting for 1 minute. The obtained mixtures were transferred to a multicooker (Thermomix) and heat-treated for 30 minutes at 120 °F and a speed of 3.0.

[0208] Oleogel preparation: Emulsions were prepared using the resulting aqueous dispersions at two different pHs. The first pH was “as is” (approximately 5.2 - 6.2) and the second pH was adjusted to 4.5 by adding 1 M HC1 dropwise. Sunflower oil was used to create the emulsions. The oil was heated to 120 °F for at least 1 hour before emulsion preparation. The emulsions were made by adding the aqueous phase to the oil phase, where the oil phase volume fraction was varied from 40.00% to 70.00% (Table 2). The emulsions were obtained by homogenizing the water and oil mixtures at 120 °F and 12,000 rpm for 120 seconds using a high-speed homogenizer and then cooling at room temperature (22 - 24 °C) for 2 hours. Then, the emulsion samples were dried by freeze drying for 36 hours (Figures 4A and 4B). To prepare oleogel samples, the dried emulsions were uniformly sheared using a coffee grinder (Secura) for 10-15 seconds (Figure 4C). Atty Docket No. 10087-82-PCT

[0209] Batches 1-4 produced oleogels that remained stable even after shearing. However, oiling-off was observed in batches 5-8, indicating that these batches did not produce stable oleogels.

[0210] Example 3

[0211] Fungal oleogel compositions were prepared from a submerged flour (SMF) of edible filamentous fungal biomass, i.e., a fungal flour obtained by a submerged fermentation process, derived from the filamentous acidophilic fungus Fusarium strain flavolapis. Briefly, Fusarium strain flavolapis biomass was grown via a stirred-tank submerged fermentation process. Steam was injected into the fermentation vessel until the temperature inside the was sufficient to inactivate growth of the biomass. The mycelium was then washed with deionized water and the washed biomass was collected and spray-dried from a moisture content of about 75 wt.% to a moisture content of about 2 wt.%. Several batches suitable to produce the fungal oleogel composition are shown in Table 3. Samples were prepared utilizing different concentrations of filamentous fungal biomass and oil to test rheological properties of the resulting emulsions and fungal oleogels.

[0212] Table 3

[0213] Aqueous Dispersion of Fungal Submerged Flour Milk: Submerged flour (SMF) milk was prepared by adding fungal flour to water to create aqueous dispersions with a concentration of filamentous fungal particles of 6.25 wt.% and 10 wt.%. The obtained mixture was transferred to a multicooker (Thermomix) and heat-treated for 30 minutes at 120 °F and a speed of 3.0. Atty Docket No. 10087-82-PCT

[0214] Oleogel preparation: Emulsions were prepared using the resulting aqueous dispersion at two different pHs. The first pH is “as is” (approximately 5.2 - 6.2) and the second pH was adjusted to 4.5 by adding 1 M HC1 dropwise. Sunflower oil was used and to create the emulsion. The oil was heated 120 °F for at least 1 hour before emulsion preparation. The emulsions were made by adding the aqueous phase to the oil phase, where the oil phase volume fraction was varied from 40.00% to 70.00% (Table 1). The emulsions were obtained by homogenization of the water and oil mixtures at 120 °F and 12,000 rpm for 120 s using a high-speed homogenizer and then, cooled down to room temperature (22 - 24 °C) by store them at room temperature for 2 hours. Then, the emulsion samples were dried by freeze drying for 36 hours.

[0215] Rheological characterization: All rheological properties of emulsions were characterized using a rheometer equipped with a cross-hatched parallel plate geometry and a solvent trap. First, the steady shear behavior of the emulsions was examined by measuring the apparent viscosity (qa, Pa s) of each sample as a function of shear rate from 0.01 to 200 1 / s (Figure IB). All emulsion viscosities decreased with increased shear rate, representing a typical shear-thinning effect for pseudoplastic materials. For samples 1 - 4 with “as is” pH, oil volume fraction did not have significant impact on flowing behavior and viscosity values.

[0216] Next, strain sweeps were performed (0.01-200%, 1 rad / s) to determine the linear viscoelastic region (LVR) (Figure 2B). For samples 1- 4 with “as is” pH, complex modulus (G*) decreased by increasing the oil volume fraction, indicating decreased rigidity of the system. Decreasing the pH had a significant impact on mechanical behavior of the emulsions.

[0217] Lastly, the small-amplitude oscillatory shear behavior of the emulsions was determined by frequency sweeps, where the elastic modulus (G', Pa) and viscous modulus (G", Pa) of each emulsion were measured as a function of frequency (co, rad / s) in the range of 0.1-100 rad / s (Figure 3B). Dynamic oscillatory measurements were performed to establish relationships between internal structure and emulsion viscoelastic behaviors since the viscosity measurements alone were not capable of giving a full understanding of the relationships between emulsion microstructures and macroscopic properties. In all the studied emulsions, the values of G' were significantly higher than those of G" at all frequencies, and no crossover of G' and G" was observed. These results indicated that the emulsions can be classified as weak physical gels with predominantly elastic gel-like behaviors. Atty Docket No. 10087-82-PCT

[0218] All measurements were performed at 50% of the smallest critical strain determined to ensure that testing was done within the LVR.

[0219] The aqueous dispersions having 6.25 wt.% and 10 wt.% submerged flour particles as described above were unable to form oleogels in this experiment.

[0220] The concepts illustratively disclosed herein suitably may be practiced in the absence of any element which is not specifically disclosed herein. It is apparent to those skilled in the art, however, that many changes, variations, modifications, other uses, and applications of the disclosure are possible, and changes, variations, modifications, other uses, and applications which do not depart from the spirit and scope of the disclosure are deemed to be covered by the disclosure.

[0221] The foregoing discussion has been presented for purposes of illustration and description. The foregoing is not intended to limit the disclosure to the form or forms disclosed herein. In the foregoing Detailed Description, for example, various features are grouped together in one or more embodiments for the purpose of streamlining the disclosure. The features of the embodiments may be combined in alternate embodiments other than those discussed above. This method of disclosure is not to be interpreted as reflecting an intention that the claims require more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive aspects lie in less than all features of a single foregoing disclosed embodiment. Thus, the following claims are hereby incorporated into this Detailed Description, with each claim standing on its own as a separate embodiment.

[0222] Moreover, though the present disclosure has included description of one or more embodiments and certain variations and modifications, other variations, combinations, and modifications are within the scope of the disclosure, e.g., as may be within the skill and knowledge of those in the art, after understanding the present disclosure. It is intended to obtain rights which include alternative embodiments to the extent permitted, including alternate, interchangeable, and / or equivalent structures, functions, ranges, or steps to those claimed, regardless of whether such alternate, interchangeable, and / or equivalent structures, functions, ranges, or steps are disclosed herein, and without intending to publicly dedicate any patentable subject matter.

Claims

Atty Docket No. 10087-82-PCTCLAIMS1. A composition, comprising: an oleogel comprising: an oil; and an oleogel structurant comprising filamentous fungal biomass.

2. The composition of claim 1, wherein the oleogel comprises about 75% to about 95% oil on a dry weight basis.

3. The composition of claim 1 or claim 2, wherein the oleogel comprises about 5% to about 25% oleogel structurant on a dry weight basis.

4. The composition of any one of claims 1-3, wherein the composition comprises about 5% to about 25% filamentous fungal biomass on a dry weight basis.

5. The composition of any one of claims 1-4, wherein the oleogel structurant is an aqueous dispersion comprising about 5 wt% to about 99 wt% filamentous fungal biomass.

6. The composition of any one of claims 1-5, wherein the composition comprises less than about 1 wt. % water.

7. The composition of any one of claims 1-6, wherein the composition comprises at least about 1% protein on a dry weight basis.

8. The composition of any one of claims 1-7, wherein the composition comprises about 1% to about 10% protein on a dry weight basis.

9. The composition of any one of claims 1-8, wherein the composition comprises at least about 1% fiber on a dry weight basis.

10. The composition of any one of claims 1-9, wherein the composition comprises about 1% to about 10% fiber on a dry weight basis.

11. The composition of any one of claims 1-10, wherein the oleogel structurant further comprises a plant protein source, a polysaccharide source, or combinations thereof.

12. The composition of claim 11, wherein the plant protein comprises hemp protein, soy protein, pea protein, chickpea protein, rice protein, or combinations thereof.

13. The composition of any one of claims 1-12, wherein the composition is solid at 20 °C.

14. The composition of any one of claims 1-13, wherein the composition is stable at 20 °C for at least about 24 hours.Atty Docket No. 10087-82-PCT15. The composition of any one of claims 1-14, wherein the composition has a turbiscan stability index of about 0 to about 3.

16. The composition of any one of claims 1-15, wherein a melting temperature of the oleogel is about 27 °C to about 37 °C.

17. The composition of any one of claims 1-16, wherein a crystallization temperature of the oleogel is about 5 °C to about 20 °C.

18. The composition of any one of claims 1 -17, wherein the oil comprises acai oil, almond oil, avocado oil, blackcurrant seed oil, borage seed oil, canola oil, cashew oil, coconut oil, corn oil, cottonseed oil, evening primrose oil, grapeseed oil, hazelnut oil, hemp oil, macadamia oil, olive oil, palm oil, peanut oil, pecan oil, pine seed oil, pistachio oil, pumpkin seed oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, walnut oil, or combinations thereof.

19. The composition of any one of claims 1-18, wherein the filamentous fungus is selected from the group consisting of Rhizopus oligosporus, Ustilago esculenla, Hericulum erinaceus. Polyporous squamosiis. Grifola fondosa. Hypsizygus marmoreus. Hypsizygus ulmarius. Calocybe gambosa. Pholiota nameko. Calvatia giganlea. Agaricus bisporus. Stropharia riigosoanmilala. Hypholoma lalerilnim, Pleurotus eryngii. Pleurotus oslrealiis. Pleurotus ostreatus var. columbinus. Tuber borchii. Morchella esculenla. Morchella conica. Morchella imporluna. Sparassis crispa. Fusarium venenatum, Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698), Disciotis venosa, Clonostachys rosea, Cordyceps militaris, Trametes versicolor, Ganoderma lucidum, Flammulina velutipes, Lentinula edodes, Pleurotus djamor, Pleurotus ostreatus, and Leucoagaricus spp.

20. The composition of any one of claims 1-18, wherein the filamentous fungus is Fusarium species.

21. The composition of any one of claims 1-18, wherein the filamentous fungus is Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698).

22. The composition of any one of claims 1-18, wherein the filamentous fungus is selected from Fusarium venenatum and Fusarium venenatum strain A3 / 5.

23. The composition of any one of claims 1-22, wherein the filamentous fungal biomass is produced by submerged fermentation.

24. The composition of any one of claims 1-22, wherein the filamentous fungal biomass is produced by non-submerged fermentation.Atty Docket No. 10087-82-PCT25. The composition of any one of claims 1-22, wherein the filamentous fungal biomass is a cohesive fungal biomass.

26. The composition of any one of claims 1-25, wherein the composition is a flour.

27. The composition of any one of claims 1-25, wherein the composition is a dough.

28. A food product comprising the composition of any one of claims 1-25, wherein the food product is selected from the group consisting of a butter analog, a shortening analog, a ghee analog, and a margarine analog.

29. A food product comprising the composition of any one of claims 1-25, wherein the food product is a savory food product.

30. The food product of claim 29, wherein the food product is selected from the group consisting of a processed meat product, a sausage, a meat patty, frankfurters, a pork analog, a beef analog, a chicken analog, and bread.

31. A food product comprising the composition of any one of claims 1-25, wherein the food product is a sweet food product.

32. The food product of claim 31, wherein the food product is selected from the group consisting of chocolate, chocolate compositions, cake, muffins, pastries, cookies, brownies, pie crust, and ice cream.

33. A method of making an oleogel, comprising: drying an oil-in-water emulsion, wherein the oil-in-water emulsion comprises filamentous fungal biomass and a continuous gelled water phase, to form a dried emulsion; shearing the dried emulsion to form an oleogel.

34. The method of claim 33, wherein the oil-in-water emulsion has an oil fraction volume of at least about 30 wt.%, at least about 40 wt.%, at least about 50 wt.%, at least about 60 wt.%, or at least about 70 wt.%.

35. The method of claim 33 or claim 34, wherein the drying step comprises lyophilization.

36. The method of any one of claims 33-35, wherein the dried emulsion comprises less than about 1 wt. % water.

37. The method of any one of claims 33-36, wherein the shearing step is conducted for about 5 to about 30 seconds.Atty Docket No. 10087-82-PCT38. The method of any one of claims 33-37, further comprising forming the oil- in-water emulsion.

39. The method of claim 38, wherein the forming step comprises: combining oil and an aqueous liquid dispersion of filamentous fungal biomass; and homogenizing the combined oil and aqueous liquid dispersion.

40. The method of claim 39, wherein the pH of the aqueous liquid dispersion is less than about 6.0, less than about 5.5, less than about 5.0, less than about 4.5, less than about 4.0, less than about 3.5, less than about 3.0, less than about 2.5, less than about 2.0, less than about 1.5, less than about 1.0, or less than about 0.5.

41. The method of claim 39 or claim 40, wherein the homogenizing step is conducted at a temperature of at least 20°C.

42. The method of claim 39 or claim 40, wherein the homogenizing step is conducted at a temperature of at least 38°C.

43. The method of any one of claims 33-42, wherein the oil-in-water emulsion is a stable emulsion.

44. The method of any one of claims 33-43, wherein the oleogel comprises about 75% to about 95% oil on a dry weight basis.

45. The method of any one of claims 33-44, wherein the oleogel comprises about 5% to about 25% on a dry weight basis of an oleogel structurant comprising the filamentous fungal biomass.

46. The method of claim 45, wherein the oleogel structurant further comprises a plant protein source, a polysaccharide source, or a combination thereof.

47. The method of any one of claims 33-46, wherein the oleogel comprises about 5% to about 25% filamentous fungal biomass on a dry weight basis.

48. The method of any one of claims 33-47, wherein the oleogel comprises at least about 1% protein on a dry weight basis.

49. The method of any one of claims 33-48, wherein the oleogel comprises about 1% to about 10% protein on a dry weight basis.

50. The method of any one of claims 33-49, wherein the oleogel comprises at least about 1% fiber on a dry weight basis.

51. The method of any one of claims 33-50, wherein the oleogel comprises about 1% to about 10% fiber on a dry weight basis.Atty Docket No. 10087-82-PCT52. The method of any one of claims 33-51, wherein the oleogel is solid at 20°C.

53. The method of any one of claims 33-52, wherein the oleogel is stable at 20 °C for at least about 24 hours.

54. The method of any one of claims 33-53, wherein the oleogel has a turbiscan stability index of about 0 to about 3.

55. The method of any one of claims 33-54, wherein the oleogel has a melting temperature of about 27 °C to about 37 °C.

56. The method of any one of claims 33-55, wherein the oleogel has a crystallization temperature of about 5 °C to about 20 °C.

57. The method of any one of claims 33-56, wherein the oil-in-water emulsion comprises an oil selected from the group consisting of acai oil, almond oil, avocado oil, blackcurrant seed oil, borage seed oil, canola oil, cashew oil, coconut oil, com oil, cottonseed oil, evening primrose oil, grapeseed oil, hazelnut oil, hemp oil, macadamia oil, olive oil, palm oil, peanut oil, pecan oil, pine seed oil, pistachio oil, pumpkin seed oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea oil, walnut oil, and combinations thereof.

58. The method of any one of claims 33-57, wherein the filamentous fungus is selected from the group consisting of Rhizopus oligosporus, Ustilago esculenla, Hericulum erinaceus. Polyporous squamosiis. Grifola fondosa, Hypsizygus marmoreus. Hypsizygus ulmarius, Calocybe gambosa. Pholiota nameko. Calvatia giganlea. Agaricus bisporus. Stropharia riigosoanmilala. Hypholoma lalerilnim, Pleurotus eryngii. Pleurotus oslrealus, Pleurotus ostreatus var. coliimbinus. Tuber borchii. Morchella esculenla. Morchella conica. Morchella imporluna. Sparassis crispa. Fusarium venenatum, Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698), Disciotis venosa, Clonostachys rosea, Cordyceps militaris, Trametes versicolor, Ganoderma lucidum, Flammulina velutipes, Lentinula edodes, Pleurotus djamor, Pleurotus ostreatus, and Leucoagaricus spp.

59. The method of any one of claims 33-57, wherein the filamentous fungus is a Fusarium species.

60. The method of any one of claims 33-57, wherein the filamentous fungus is Fusarium strain flavolapis (ATCC Accession Deposit No. PTA- 10698).

61. The method of any one of claims 33-57, wherein the filamentous fungus is selected from Fusarium venenatum and Fusarium venenatum strain A3 / 5.Aty Docket No. 10087-82-PCT62. The method of any one of claims 33-61, wherein the filamentous fungal biomass is produced by submerged fermentation.

63. The method of any one of claims 33-61, wherein the filamentous fungal biomass is produced by non-submerged fermentation.

64. The method of any one of claims 33-61, wherein the filamentous fungal biomass is a cohesive fungal biomass.

65. The method of any one of claims 33-64, wherein the oleogel is a flour.

66. The method of any one of claims 33-64, wherein the oleogel is a dough.

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