A method for cultivating fungal mycelium and then forming edible products from it.

Abstract

A method for growing fungal mycelia and forming an edible food product includes growing fungal cells in a growth medium such that the fungal cells produce mycelium. The growth medium includes sugars, nitrogen-containing compounds, and phosphate-containing compounds. The mycelium is separated from the growth medium. The mycelium is concentrated to obtain a fibrous mycelial mass having a protein content of greater than 40% by weight of the dry weight of the mycelium. A food additive can be added to the fibrous mycelial mass to produce biomass. The biomass can be formed into an edible food product. The biomass can be formed into an edible meat substitute product.

Description

Technical Field

[0001] Cross-Reference to Related Applications The present application claims priority and benefit from U.S. Provisional Application No. 62 / 682,301, filed on June 8, 2018, the entire disclosure of which is incorporated herein by reference.

[0002] The present disclosure generally relates to the field of fungal mycelium, and more specifically, to systems and methods for growing fungal mycelium and forming edible products therefrom.

Background Art

[0003] Food and beverage production and processing facilities are major consumers of water and also major producers of wastewater globally. Examples of such facilities include, inter alia, fruit and vegetable processing, meat processing, dairy processing, snack food processing, and beverage processing facilities. Water is typically used in these facilities for essential processes such as raw material treatment, cleaning of materials, conveyance of materials, and cleaning of equipment. Accordingly, residual water that is not consumed in food and beverage products arises as an effluent stream known as wastewater. Wastewater from food and beverage facilities has extremely high chemical oxygen demand (COD) and total suspended solids (TSS), which makes it difficult and costly to treat. These residual water streams are often considered a nuisance for facilities. The only widely implemented solutions are aerobic treatment, anaerobic digestion, and fixed films, and the only useful by-product is the output obtained from anaerobic digestion. Given the high capital cost of digesters and other external treatment technologies, wastewater is often discharged directly to municipal sewers for treatment. Furthermore, there is a growing demand for edible products that can provide high protein content derived from non-animal sources. Driven by increasing awareness of personal health, edible products containing components from non-animal sources, such as protein and fiber, are seen as healthier alternatives to animal protein-based products. In particular, there is a growing demand for edible meat substitutes that mimic the composition and texture of meat but are composed of non-animal components, which can reduce reliance on animals such as cattle and lower carbon emissions caused by such animals. Therefore, there is a need for non-animal protein sources that can facilitate large-scale production and the adoption of non-animal-based edible products. [Overview of the project]

[0004] The embodiments described herein generally relate to methods for cultivating fungal mycelium and forming edible food from it. In some embodiments, a method includes the step of growing fungal cells in a growth medium so that the fungal cells produce mycelium. The growth medium contains sugars, nitrogen-containing compounds, and phosphate-containing compounds. The method includes the steps of separating the mycelium from the growth medium and concentrating the mycelium to obtain fibrous mycelial masses having a protein content of more than 40% by mass of the dry mass of the mycelium. In some embodiments, a method for forming edible food includes the step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium. The method includes the step of adding a food additive to the fibrous mycelial mass to generate biomass, and the step of forming the biomass into edible food. In some embodiments, a method for forming an edible meat substitute product includes the step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium. The method includes the steps of adding a food additive to the fibrous mycelial mass to generate biomass, and forming the biomass into an edible meat substitute product. In some embodiments, the meat substitute product contains fibrous mycelial masses ranging from 10% to 100% by mass. The fibrous mycelial masses have a protein content exceeding 40% by mass of their dry mass. The meat substitute product contains a water content ranging from 0% to 90% by mass. In some embodiments, the edible chips contain edible substance. The edible substance contains fibrous mycelial masses ranging from 0.1% to 90% by mass. The fibrous mycelial masses have a protein content exceeding 40% by mass of their dry mass. The edible chips contain a water content of less than 20% by mass. The edible chips contain a carbohydrate content ranging from 0% to 90% by mass. The fibrous mycelial masses are distributed within the edible substance. In some embodiments, the powdered edible product includes powdered mycelial mass. The powdered mycelial mass has a protein content exceeding 40% by mass. Any combination of the above concepts and any further concepts discussed in more detail below (provided that such concepts are not contradictory) is intended to be part of the subject matter of the invention disclosed herein. In particular, any combination of the claimed subject matter found at the end of this disclosure is intended to be part of the subject matter of the invention disclosed herein. The features of this disclosure described above and other features are expected to become more fully apparent by combining the following description and the attached claims with the attached figures. These figures are intended to illustrate several implementations in accordance with this disclosure and are therefore not intended to limit its scope. This disclosure is described more specifically and in detail by using the attached figures. [Brief explanation of the drawing]

[0005] [Figure 1] A block diagram of an example method for cultivating fungal mycelium according to one embodiment is shown. [Figure 2] A process flow diagram of a brewery according to one embodiment is shown. [Figure 3]This shows a high-performance liquid chromatography (HPLC) scan of Vogel solution and residual water from a brewery according to one embodiment. [Figure 4] This shows an HPLC scan of N. crassa growth in residual water from a brewery according to one embodiment. [Figure 5] A block diagram of an example method for forming an edible product from fungal mycelium, according to one embodiment, is shown. [Figures 6A1-6A2] The top view and side view of a food product configured as chips according to one embodiment are shown, respectively. [Figures 6B1-6B2] Further, a top view and a side view of a food product configured as triangular chips according to another embodiment are shown. [Figures 6C1-6C2] The images show a top view and a side view of a food product configured as oval-shaped chips according to another embodiment. [Figure 7] A block diagram of an example method for forming an edible meat substitute product from fungal mycelium, according to one embodiment, is shown. [Figure 8A] A perspective view of an edible meat substitute food product configured as a patty, according to one embodiment, is shown. [Figure 8B] A perspective view of an edible meat substitute product configured as tenderizer according to one embodiment is shown. [Figure 8C] A perspective view of a ground meat substitute product according to one embodiment is shown.

[0006] The attached figures are referenced throughout the following detailed description. In the figures, unless otherwise indicated by context, similar symbols identify typically similar components. The exemplary implementations described in the detailed description, figures, and claims are not intended to be limiting. Other implementations may be used, and other modifications may be made without departing from the spirit or scope of the subject matter presented herein. As generally described herein and illustrated in the figures, aspects of this disclosure can be arranged, substituted, combined, and designed in a wide variety of different structures, all of which are clearly intended to form part of this disclosure. [Modes for carrying out the invention]

[0007] The embodiments described herein generally relate to methods for cultivating fungal mycelium and forming edible products therefrom. In particular, various embodiments described herein provide methods for cultivating fungal cells, separating mycelium, and concentrating mycelium to obtain fibrous masses. Various embodiments also relate to generating biomass by adding food additives and forming that biomass into edible foods or edible meat substitute products. The edible meat substitute products may contain fibrous mycelial masses having a protein content exceeding 40% by mass of the dry mass of the fibrous mycelium. Various embodiments of a method for cultivating fungal mycelium and forming edible products therefrom may provide one or more benefits, including, for example, (1) providing a batch or continuous process culture of fungal mycelium; (2) cultivating fungal mycelium in brewery wastewater or other wastewater streams to enable recycling and wastewater purification; (3) providing edible products containing proteins from a non-animal source, i.e., fungal mycelium, thereby reducing reliance on animal source proteins and reducing their carbon dioxide emissions; and (4) providing edible meat substitute products that have the texture and taste of real meat while simultaneously providing a high protein content. Fungal mycelia can include fungi from the Ascomycota and Zygomycota phyla, including the genera Aspergillus, Fusarium, Neurospora, and Monascus. Other species include edible species from the Basidiomycota and Lentinula genera. One genus, Neurospora, is used in solid-state fermentation food production. Neurospora species are known for their high biomass production efficiency and ability to break down complex carbohydrates. No known allergens have been detected for certain Neurospora species, and no mycotoxin levels are produced. In addition to single cultures of filamentous fungi, multiple strains can be cultured simultaneously to adjust the protein, amino acid, mineral, texture, and flavor profiles of the final biomass.

[0008] Figure 1 shows a block diagram of an example method for cultivating fungal mycelium according to one embodiment. Briefly, method 100 may include the step of cultivating fungal cells in a growth medium in step 102. Method 100 may include the step of separating the mycelium from the growth medium in step 104. Method 100 may include the step of concentrating the mycelium in step 106. Method 100 may include the step of obtaining fibrous hyphae in step 108. More specifically, method 100 may include the step of growing fungal cells in a growth medium in 102. For example, the growth medium may be contained in a container such as a large vat capable of growing several kilograms of fungal mycelium. The growth medium may be called the original growth medium. Method 100 may include the step of growing fungal cells in a growth medium so that the fungal cells produce mycelium. The growth medium may contain nutrients (e.g., sugars, nitrogen-containing compounds, or phosphate-containing compounds). The growth medium may contain sugars, nitrogen-containing compounds, and phosphate-containing compounds. The sugar may be in the range of 5 to 50 g / L. For example, the sugar may be 5 g / L, 10 g / L, 20 g / L, 30 g / L, 40 g / L, or 50 g / L, including both ends. The sugar may include sucrose, glucose, fructose, molasses, or a mixture of sugars. The nitrogen-containing compound may be in the range of 0.5 to 10 g / L. For example, the nitrogen-containing compound may be 0.5 g / L, 1 g / L, 2 g / L, 3 g / L, 4 g / L, 5 g / L, or 10 g / L, including both ends. The nitrogen-containing compound may include ammonium hydroxide, ammonium nitrate, ammonium sulfate, ammonium chloride, urea, yeast extract, peptone, or a mixture of nitrogen-containing compounds. The phosphate-containing compound may be in the range of 0.1 to 5 g / L. For example, the phosphate-containing compound may be 0.1 g / L, 0.2 g / L, 0.3 g / L, 0.4 g / L, 0.5 g / L, 1 g / L, 2 g / L, 3 g / L, 4 g / L, or 5 g / L, including both ends. The phosphate-containing compound may be potassium phosphate, sodium phosphate, phosphate, or a mixture of phosphate-containing compounds. Fungal cells can be grown at temperatures between 25°C and 40°C, including both ends. The fungal cells can be grown for 12 to 48 hours, including both ends. The growth of fungal cells can produce a dry mass of fungal cells yielding 5 to 20 g / L. The mycelium can have a protein content exceeding 40% by mass (dry mass). In some embodiments, the mycelium can have a protein content of 50 to 65% (dry mass), including both ends. The mycelium can contain at least 25 mg of a methionine and cysteine ​​combination per gram of crude protein. In some embodiments, method 100 may include a step of removing a certain volume of broth. The broth may contain fungal cells and growth medium. The step of removing a certain volume of broth may include removing a certain volume of broth separately. For example, a certain volume of broth may be removed from the container containing the broth by siphoning in a batch process, or continuously from the broth. For example, a certain volume of broth may be drained from the container containing the broth in a continuous process.

[0009] Method 100 may include the step of adding a new growth medium to a container containing broth. The broth may be a fermented broth. Nutrients (e.g., sugars, phosphate-containing compounds, or nitrogen-containing compounds) may be added in a batch growth configuration. For example, nutrients may be added after a predetermined time has elapsed (e.g., after 1 hour, 2 hours, 3 hours, 6 hours, or 12 hours). The concentration of one or at least one nutrient in the new growth medium may be the same as the concentration of the nutrient in the original growth medium as described in operation 102. The new growth medium may have a volume greater than, less than, or equal to the volume of growth medium lost from the original growth medium during the growth of fungal cells in the original growth medium. In one example, after 6 hours, the concentrations of sugars, phosphate-containing compounds, and nitrogen-containing compounds in the new growth medium are increased. Nutrients are added to bring the concentrations of sugars, phosphate-containing compounds, and nitrogen-containing compounds in the broth to the same concentrations as those in the original growth medium, respectively. In one example, 50–95% of the broth can be removed after at least 12 hours. A new medium containing nutrients (e.g., sugars, phosphate-containing compounds, or nitrogen-containing compounds) can be added. The nutrient concentration of the broth can be increased by the added new growth medium. Nutrients can be added in a continuous growth configuration. For example, a volume of broth (e.g., inclusive of the endpoints, 0.01 volume%, 1 volume%, 5 volume%, 10 volume%, 25 volume%, 50 volume%, or 95 volume%) can be removed from a container containing fungal cells and a growth medium. Fresh growth medium can be added to the container that contains the broth. The fresh growth medium can be provided as a continuous flow. The volume of broth in the container can be monitored to remain at a specified level. For example, the volume of broth in the container can remain at a constant volume. The volume of fresh growth medium added may be equal to the volume of broth lost from the container.

[0010] Method 100 comprises the step of separating mycelium from a growth medium at 104. Separation of mycelium from the growth medium can be performed using gravity pressure, centrifugation, a belt press, a filter press, a mechanical press, a drum dryer, or any other suitable process. During the separation process, the mycelium can be washed with water, ethanol, an acid, a base or other solvents. The recovered filtrate can be reused or discarded. The mycelium can have a water content of between 65% and 95%. The cell walls of the mycelium can be disrupted, for example by lysis. Lysis can be performed by adjusting the pH to less than 4 or more than 9, by adding a lytic enzyme, by increasing the temperature to between 40°C and 60°C for 1 to 24 hours, or by any other suitable lysis method. After separation, additives (e.g., food additives) can be mixed with the mycelium. For example, when the mycelium is formed into an edible product, the additives can include vegetable or animal proteins, fats, emulsifiers, thickeners, stabilizers, and flavors.

[0011] Method 100 may comprise the step of concentrating mycelium at 106. The step of concentrating mycelium may comprise increasing the proportion or level of fibrous mycelial mass obtained after separation from the growth medium. For example, the step of concentrating mycelium may comprise dehydrating the mycelium by removing water from the mycelium. For example, the mycelium may be heat-dried, for example, to a water content of less than 70%. The mycelium may be heat-dried at a specified temperature (for example, 30°C, 50°C, 75°C, or 90°C, inclusive of the endpoints). In some embodiments, the mycelium may be heat-dried using forced air. In some embodiments, removing water from the mycelium may comprise applying mechanical force (for example, a press, a sieve) to remove water from the mycelium. Dehydration of the mycelium (for example, via thermal or mechanical drying) can produce a partially dried fibrous mycelial mass. The partially dried fibrous mycelial mass can be pulverized to reduce particle size, for example, to produce a powder.

[0012] Method 100 comprises the step of obtaining a fibrous mycelial mass at 108. The fibrous mycelial mass may have a protein content exceeding 40% by mass of the dry mass of the mycelium. For example, the fibrous mycelial mass can have a protein content of 45% by mass, 50% by mass, 60% by mass, 70% by mass, 80% by mass, or 90% by mass, inclusive of the endpoints, based on the dry mass of the mycelium.

[0013] The following are several examples of growing fungi and obtaining a fibrous mycelial mass having a protein content exceeding 40% by mass therefrom. These examples are for illustrative purposes only, and should not be construed as limiting the present disclosure to any shape or form. In one example, Neurospora crassa (N. crassa) was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. Conidia or spores of N. crassa were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following culture medium: 20 g / L sucrose, 2 g / L ammonium nitrate, 2 g / L mononucleotide potassium phosphate, 1 g / L sodium nitrate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 24 hours, the mycelium was harvested using cheesecloth, water was removed using an apple press, and it was completely dried in a dehydrator set to 74°C. The dry mass of the total cells was 9.5 g / L. Protein analysis yielded a crude protein content of 57% by mass. Amino acid analysis yielded a PDCAAS score of 1.0 for the fibrous mycelial mass. The fibrous mycelial mass contained a combination of methionine and cysteine ​​of 26 mg per gram of crude protein.

[0014] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. Conidia or spores of N. crassa were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was aseptically transferred to a 10 L benchtop reactor containing the following medium: 20 g / L sucrose, 2 g / L ammonium nitrate, 1 g / L monopotassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 24 hours, the mycelium was harvested using cheesecloth, water was removed using an apple press, and it was completely dried in a dehydrator set to 74°C. The total dry mass of the cells was 9 g / L. Protein analysis revealed a crude protein content of 55% by mass. Amino acid analysis yielded a PDCAAS score of 1.0 for the fibrous mycelial mass. The fibrous mycelial mass contained a combination of methionine and cysteine ​​of 26 mg per gram of crude protein.

[0015] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. The conidia were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following medium: 30 g / L sucrose, 3 g / L ammonium nitrate, 1 g / L monopotassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 24 hours, the mycelium was harvested using cheesecloth, water was removed using an apple press, and it was completely dried in a dehydrator set to 74°C. The total dry mass of the cells was 11 g / L. Protein analysis revealed a crude protein content of 63% by mass. Amino acid analysis yielded a PDCAAS score of 1.0 for the fibrous mycelial mass. The fibrous mycelial mass contained a combination of methionine and cysteine ​​of 27 mg per gram of crude protein.

[0016] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. The conidia were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following medium: 20 g / L sucrose, 3.25 g / L urea, 1 g / L monobasic potassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 24 hours, the mycelium was harvested using cheesecloth, water was removed using an apple press, and it was completely dried in a dehydrator set to 74°C. The total dry mass of the cells was 8.5 g / L. Protein analysis revealed a crude protein content of 56% by mass. Amino acid analysis yielded a PDCAAS score of 1.0. The fibrous mycelial mass contained a combination of methionine and cysteine ​​of 25 mg per gram of crude protein.

[0017] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. The conidia were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following medium: 20 g / L sucrose, 2 g / L ammonium nitrate, 1 g / L monobasic potassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 15% ammonium hydroxide buffer and maintained at 5.8. After 24 hours, the mycelium was harvested using cheesecloth, water was removed using an apple press, and it was completely dried in a dehydrator set to 74°C. The total dry mass of the cells was 10 g / L. Protein analysis revealed a crude protein content of 60% by mass. Amino acid analysis yielded a PDCAAS score of 1.0. The fibrous mycelial mass contained a combined methionine and cysteine ​​content of 26 mg per gram of crude protein.

[0018] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. The conidia were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following medium: 20 g / L sucrose, 2 g / L ammonium nitrate, 1 g / L monopotassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 12 hours, 10 g / L of sucrose and 1 g / L of ammonium nitrate are added to the system. After a total of 24 hours, the mycelium is harvested using cheesecloth, water is removed using an apple press, and the mycelium is completely dried in a dehydrator set to 74°C. The dry mass of the total cells is 12 g / L. Protein analysis yields a crude protein content of 60% by mass. Amino acid analysis yields a PDCAAS score of 1.0 for the fibrous mycelium. The fibrous mycelium contains a combination of methionine and cysteine ​​at a rate of 26 mg per gram of crude protein.

[0019] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. The conidia were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following medium: 20 g / L sucrose, 2 g / L ammonium nitrate, 1 g / L monopotassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 24 hours, 90% of the culture medium was collected, and fresh medium was added at the aforementioned concentrations to return the entire system to 10 L. The time for the new batch series was shortened to 12 hours. 90% was collected every 12 hours, and the fed batch process was repeated. This process was carried out for 60 hours. The dry mass of the collected cells was 9.5 g / L. Protein analysis yielded a crude protein content of 60% by mass. Amino acid analysis yielded a PDCAAS score of 1.0 for the fibrous hyphal mass. The fibrous hyphal mass contained a combination of methionine and cysteine ​​of 26 mg per gram of crude protein.

[0020] In another example, N. crassa was grown in batches in a 10 L benchtop reactor. N. crassa was first grown on an agar slant and incubated at 32°C for 3 days. The conidia were transferred to a 250 mL aerated Fernbach flask and grown on an orbital shaker table at 32°C for 48 hours. The resulting mycelium was then sterilely transferred to a 10 L benchtop reactor containing the following medium: 20 g / L sucrose, 2 g / L ammonium nitrate, 1 g / L monopotassium phosphate, 0.2 g / L magnesium sulfate, 0.1 g / L calcium chloride, and trace elements. Aeration was set to 0.75 vvm and stirring to 250 rpm. The pH was adjusted using 6N sodium hydroxide buffer and maintained at 5.8. After 24 hours, 90% of the medium is collected, and fresh medium is added at the above concentrations to return the entire system to 10 L. The time for the new batch series is shortened to 12 hours. 90% is collected every 12 hours, and the fed batch process is repeated. This process was carried out for 60 hours. After filtering using cheesecloth and a press, all the medium is collected, autoclaved, and reused with only the addition of 20 g / L sucrose, 2 g / L ammonium nitrate, and 1 g / L monopotassium phosphate. The repeated fed batch process is carried out for a total of 60 hours. The collected cell dry mass is 9.5 g / L. Protein analysis yields a crude protein content of 60% by mass. Amino acid analysis yields a PDCAAS score of 1.0 for the fibrous hyphal mass. The fibrous hyphal mass contains a combination of methionine and cysteine ​​of 26 mg per gram of crude protein.

[0021] Methods for cell maintenance and conidial isolation are described herein. A wild-type strain of Neurospora crassa (N. crassa) (FGSC number 4815) was purchased from the Fungal Genetic Stock Center. Cells used for inoculation were stored at -20°C on an agar slant composed of 2% Vogel 50x salt, 0.01% trace element solution, 0.005% biotin, 1.5% sucrose, and 1.5% agar. Growth experiments started with cells removed from the frozen agar slant, transferred to a fresh agar slant, and incubated at 30°C in complete darkness for 2-3 days. Conidia were isolated from the slant using standard methods for batch culture experiments and inoculated into 100 mL of fresh Vogel medium (50x salt of 2% Vogel, 0.01% trace element solution, 0.005% biotin, and 1.0% glucose). A suspension of conidia (1 mL in Vogel medium) with an optical density of approximately 0.7 was added to each culture.

[0022] A batch propagation method is described herein. The propagation experiment was carried out in 1 L of fresh residual water. The batch cultures were incubated at 30°C for 1 to 3 days under constant light (120 rpm). Biomass was collected using a vacuum filtration flask and then dried at 105°C.

[0023] The crude protein content of filamentous fungi can be increased by supplementing with additional nitrogen sources. Non-limiting examples include supplementing with gaseous ammonia, liquid ammonia, ammonium nitrate, ammonium sulfate, sodium nitrate, yeast extract, urea, peptone, or other organic nitrogen sources. Nitrogen sources can be added along with other pH buffering components. Non-limiting examples include acids, phosphates, borates, sulfates, and bases. In some embodiments, fungal mycelia can be grown using growth media derived from food-grade water streams produced in food and beverage processing facilities. For example, the methods and materials described herein can be used for water streams produced in breweries, which may include residual water streams from breweries, spent yeast, equipment cleaning, packaging, or mixtures thereof. Fungal mycelia can be grown from growth media of other industries where food-grade processes are performed and residual water streams are produced with significant levels of sugar, nutrients, and organic matter. As a non-limiting example, the following is a list of food and beverage facilities suitable for producing food-grade residual water streams: breweries, wineries, distilleries, nutritional drinks, sodas, fruit drinks, potato products, dairy products, meat processing, candies, baked goods, or mixtures thereof. Food-grade residual water can be collected for use as a growth medium for cultivating filamentous fungi, and the biomass of the filamentous fungi can be used for a number of products, including, but not limited to, edible proteins, chemical extracts, or materials. In other embodiments, fungal mycelia can be grown in a suitable culture medium containing controlled amounts of nutrients to enhance the mycelial culture efficiency.

[0024] Breweries are categorized into various types based on their annual production capacity. Beer is measured in barrels (bbl), where 1 bbl is equivalent to 31 gallons. Microbreweries generally produce less than 5,000 bbl per year, craft breweries produce between 5,000 and 1,000,000 bbl per year, and large national breweries are considered to produce more than 1,000,000 bbl per year. Breweries use significant amounts of water within their facilities, not only for product production but also for cleaning equipment. Residual water streams arise from the brewery, spent yeast, fermentation tank cleaning, filtration, and packaging processes. The resulting residual water is typically considered food grade, except for chemical rinsing of equipment and packaging. Residual streams also have extremely high chemical oxygen demand (COD) and total suspended solids (TSS), usually 20,000 mg L each. -1and 10,000 mg L -1 This exceeds [amount]. Depending on the size of the brewery, residual water can be disposed of in different ways. In most microbreweries and craft breweries, residual water can be discharged into the sewer system, and costs are levied by the local government based on the water's COD and TSS concentrations. In larger craft breweries and national giants, residual water is usually diverted to on-site wastewater treatment facilities. The residual water produced in these facilities often contains nutrients necessary for the growth of filamentous fungi, or requires further nutrient supplementation.

[0025] Figure 2 shows a brewery process 200 according to one embodiment. The brewery process diagram includes a schematic representation of how the fungal growth method described herein may be integrated into a commercial brewery. Certain residual water streams were identified as having a more optimized nutrient profile than other streams. The brewery process 200 may include a mash tun 202. The mash tun 202 may include a brewing vessel used to mix crushed malt with temperature-controlled water. The brewery process 200 may include grinding the malt within the mash tun 202. The brewery process 200 may include a lauter tun 204. The lauter tun 204 may include a vessel configured to perform lautering, i.e., to separate the mash into a clear liquid wort and residual grains. The residual water 203 may be removed from the lauter tun 204 to provide a growth medium for growing fungal cells. The brewery process 200 may include a kettle 206. The kettle 206 may include a vessel for boiling the wort with hops and other flavors. The kettle 206 may be made of copper. The brewery process 200 may include a whirlpool 208. The whirlpool 208 may include a vessel for separating solid particles from the hopped wort. The whirlpool 208 may also include a sedimentation tank. Residual water 203 may be removed from the whirlpool 208 to provide a growth medium for growing fungal cells. The brewery process 200 may include cooling of the wort 210. The brewery process 200 may include fermentation 212. Residual water 203 may be removed after fermentation 212 to provide a growth medium for growing fungal cells. The brewery process 200 may include maturation 214. The brewery process may include filtration 216. The brewery process 200 may include packaging 218.

[0026] As described herein, residual water 203 can be used as a growth medium for fungal cells. Fungal cells can undergo a growth process 220. The growth process 220 may include auxiliary agents as needed (e.g., sugars, phosphate-containing compounds, nitrogen-containing compounds). Fungal cells can be processed in 222 to produce biomass or products. Filamentous fungi are typically cultured in the laboratory in a synthetic solution known as Vogel solution. Table 1 shows the elemental analysis of Vogel solution and residual water from a brewery, as determined by inductively coupled plasma emission (optimal) spectroscopy (ICP-OES). All values ​​are expressed in mg / L. The elemental analysis provided in Table 1 represents residual water from a microbrewery. Brewery residual water contains the nutrients necessary for proper filamentous fungal growth. This data also reveals that the significant amounts of nutrients used in Vogel solution are not necessary for proper fungal growth.

[0027] [Table 1] The composition of the brewery water stream is very similar to that of Vogel, containing the necessary amounts of each essential organic matter and mineral for culturing filamentous fungi. The brewery water stream can be collected and stored in a fermentation vessel. Fungal spores can be added and incubated at 20-40°C for 6-72 hours, including both ends. The final filamentous fungi, being derived from food-grade stream, can be used for human consumption, can be broken down into individual chemicals such as chitin or chitosan, or used as raw materials for material production, such as activated carbon, but not limited to these. The water in which the fungal biomass is cultured can be sufficiently purified for both COD and TSS. Therefore, the water can be reused in food and beverage facilities for certain applications.

[0028] While residual water streams from breweries can be used as is, the efficiency of fungal growth can be maximized by adding further nutrients and supplements. Supplements include, but are not limited to, nitrogen sources, trace metal sources, phosphorus sources, potassium sources, magnesium sources, sulfate sources, vitamins, or mixtures thereof. Most large-scale food and beverage facilities produce only a single product, and therefore the composition of food-grade residual water streams is consistent. In smaller facilities (e.g., craft breweries), knowledge of the specific beer being brewed can indicate what elemental sources are present and what nutrients may need to be further supplemented. Supplements can serve a dual purpose: to enhance growth efficiency and to enrich the final mineral content of the biomass for food and material production purposes. Certain supplements can alter crude protein levels and amino acid profiles. Supplements can include pH buffers consisting of nitrogen sources. Spent yeast produced in the brewery can be used as a single auxiliary agent in Vogel solution, in other residual water streams, or on its own. Spent yeast may be inactivated, dissolved, or viable at the time of replenishment. Filamentous fungi can decompose the yeast for use as a source of carbon, nitrogen, and nutrients.

[0029] In addition to residual water sources from a single facility, water sources from multiple facilities can be blended to optimize the fermentation medium. Other residual solid nutrient sources from food and beverage facilities can also be added. For example, ground coffee can be added to the residual water in a brewery. Food-grade residual water streams can be sterilized before or after collection. Food-grade residual water streams can be sterilized using sterilization techniques such as ozone, heat, ultraviolet (UV), and hydrogen peroxide. In other applications, such as the production of activated carbon from fungal mycelium, iodine or chlorine dioxide can be used to sterilize the growth medium.

[0030] Figure 3 shows high-performance liquid chromatography (HPLC) scans of Vogel solution and residual water from the brewery. The HPLC scans show the intensity normalized to the sucrose / maltose peak for residual water from the brewery and Vogel solution. The maltotriose peak is stronger in residual water from the brewery than in Vogel solution. Furthermore, the glucose peak is stronger in residual water from the brewery than in Vogel solution, and the fructose peak is stronger in residual water from the brewery than in Vogel solution.

[0031] Figure 4 shows an HPLC scan of N. crassa growth in residual water from a brewery. The HPLC scan shows the growth of N. crassa in residual water over 48 hours. The sucrose / maltose peak is stronger at hour 0 than at 24 and 48 hours. The maltotriose peak is stronger at hour 0 than at 24 and 48 hours, the glucose peak is stronger at hour 0 than at 24 and 48 hours, and the fructose peak is stronger at hour 0 than at 24 and 48 hours. In one example, residual water from a brewery was collected from a microbrewery in Illinois. The water stream was the residue of the initial beer-making stream and therefore food-grade. A growth experiment was conducted over 48 hours. Figure 4 shows HPLC data over the growth process of N. crassa. Almost all sugars, including fructose, glucose, sucrose, maltose, maltotriose, and higher maltodextrins, were consumed. This is the first time that higher maltodextrins have been shown to be a suitable carbon source in a fermentation medium. Table 2 summarizes the yield characteristics, as well as the reductions in COD, TOC, and total nitrogen (TN). This example demonstrates that this particular food-grade water source can be used as a complete fermentation medium for growing fungal mycelia.

[0032] [Table 2] In another example, residual water and live yeast from a brewery in Illinois were collected. The water stream was the residue from the initial beer-making stream, and the yeast was the residue from the beer-making process; therefore, both were food-grade. The yeast was killed using an autoclave. The amount of yeast was increased and added to the residual water, which was used as a fermentation medium for N. crassa. Table 3 shows the yield data associated with this experiment. The final biomass yield with yeast addition exceeds the yield of live fungi plus the amount of yeast added at the start. This example demonstrates that yeast and other organic solids can be integrated into a medium, and that dead yeast can be used as a nutrient source for N. crassa.

[0033] [Table 3] In another example, residual water and spent grain flour were collected from a microbrewery in Illinois. The water stream was the residue from the initial beer-making stream, and the spent grain flour was the residue from the beer-making process; therefore, both were food-grade. The amount of spent grain flour was increased and added to the residual water, and used as a fermentation medium for N. crassa. Table 4 shows the yield data associated with this experiment. The final biomass yield with the addition of spent grain flour exceeded the yield of the live fungus plus the amount of flour added at the start. This example demonstrates that spent grain can be integrated into a culture medium and that spent grain can be used as a nutrient source for N. crassa.

[0034] [Table 4] In another example, residual water from a brewery was collected from a microbrewery in Illinois. The water stream was the residue of the initial beer-making stream and therefore food-grade. A growth experiment was conducted over 48 hours. As an adjuvant, the 50× salt portion of Vogel medium was added to the residual water stream at an appropriate concentration to act as a nutrient source. Table 5 shows the yield data associated with this experiment. The final biomass yield with the addition of 50× exceeded that without the addition. This example demonstrates that residual water from a brewery can be used as a carbon source in fermentation media for growing fungal mycelium.

[0035] [Table 5] In another example, residual water from a brewery was collected from a microbrewery in Illinois. The water stream was the residue of the initial beer-making stream and therefore food-grade. A growth experiment was conducted over 48 hours. The collected biomass was washed with deionized (DI) water, vacuum-filtered, dehydrated at 57°C for 8 hours, and used to produce edible chips. In another implementation, the vacuum-filtered biomass was brushed with sunflower oil before dehydration, dehydrated at 57°C for 8 hours, and used as edible chips. In yet another implementation, the vacuum-filtered biomass was deep-fried to produce edible chips. In yet another implementation, the dehydrated biomass was ground into a powder, mixed with water to form a dough, rolled into sheets, cut into circles, dried, and then deep-fried to form edible chips.

[0036] Figure 5 shows a block diagram of Method 500, which is an example of a method for forming an edible product from fungal mycelium according to one embodiment. Briefly, Method 500 includes the steps of providing a fibrous mycelial mass in 502, adding food additives to the fibrous mycelial mass to generate biomass in 504, generating biomass in 506, and forming the biomass into an edible food product. Further extension, Method 500 includes the step of providing a fibrous hyphal mass in 502. The fibrous hyphal mass has a protein content of more than 40% by mass of the dry mass of the mycelium. The fibrous hyphal mass can be produced by the operation of Method 100 or by any other method for producing fungal mycelium described herein. In 504, food additives are added to fibrous mycelial masses to produce biomass. Any suitable food additives can be used, such as plant or animal proteins, fats, emulsifiers, thickeners, stabilizers, flavors, pH adjusters, oils, spices, and salts. For example, the flavor of the biomass can be enhanced by adding various oils. Non-limiting examples of oils include nut-derived, vegetable-derived, plant-derived, and animal-derived oils. Oils can be added to food-grade residual water streams for their multipurpose role as defoamers, carbon sources for fungi, and extracellular / intracellular integration into the biomass. Alternatively, oils can be integrated into the biomass after harvesting or cooking. The formed and partially dried biomass may contain food additives introduced by various operations. For example, the biomass can be immersed in further ingredients. Further ingredients can be injected into the biomass. The biomass can be coated with further ingredients. Further ingredients can be dispersed into the biomass by applying further pressure or under vacuum conditions. Further ingredients may include plant or animal proteins, fats, emulsifiers, thickeners, stabilizers, flavors, pH adjusters, etc.

[0037] Method 500 includes a step of producing biomass in 506. The step of producing biomass may include adjusting the texture of the biomass. The texture of fungal biomass can be adjusted by chemically washing the biomass. Alternatively, the texture can be altered by controlling the water content of the biomass. The texture can also be altered by adding various nutrients that determine the growth and morphology of the biomass. The density of the final biomass can be controlled by changing the initial water content and drying conditions to produce a heavier or lighter final product. Raw biomass can be dried and ground into a powder or progressively finer powder. The powder can then be used as a protein component for human or animal feed. The powder can be formed into various shapes using other components, such as water and / or oil, and binders. Protein extraction can be performed to produce fungal protein isolates. These isolates can then be used as feed components for human or animal feed.

[0038] In 508, the biomass is formed into an edible food. The step of forming the biomass into an edible food may include dehydrating the biomass and / or grinding the biomass into a powder. For example, the powder can be added to smoothies, soups, stews, and puddings, among others. The edible food may include chips, protein bars, jerky, tortillas, bread, or crackers, which may be formed from the powdered mycelium mass, for example.

[0039] Edible foods may include powdered edible products. Powdered edible products may include powdered mycelial masses. Powdered mycelial masses may have a protein content exceeding 40% by mass of the powdered mycelial mass. Powdered edible products may include powder particles that have a particle size of less than 150 μm. For example, powder particles may have particle sizes of 149 μm, 135 μm, 120 μm, 100 μm, 50 μm, 25 μm, 10 μm, 5 μm, 2 μm, 1 μm, 0.5 μm, or 0.25 μm, including both ends. In some embodiments, method 500 may include a step of incorporating the powder into a baked food. For example, baked foods may include biscuits, bread, brownies, cakes, casseroles, cookies, crackers, custards, pastries, pies, puddings, roasts, and tarts. Baked foods may contain 0.1 to 80% by mass (e.g., 0.1%, 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80%) fungal mycelium, such as mycelium derived from N. crassa. Baked foods may contain 0 to 90% by mass (e.g., 0%, 1%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) carbohydrates, including both ends. Carbohydrates may include starch. The powdered mycelial mass of the powdered edible product may be in the range of 0.1% to 80% by mass of the total mass of the powdered edible product (e.g., 0.1%, 0.5%, 1%, 5%, 10%, 25%, 50%, 75%, or 80%, including both ends). The powdered edible product may contain carbohydrates in the range of 0% to 90% by mass (e.g., 0.1%, 0.5%, 1%, 5%, 10%, 25%, 50%, 75%, or 90%). The powdered mycelial mass may contain a combination of methionine and cysteine ​​of at least 20 mg per gram of crude protein (e.g., 20 mg / gram, 25 mg / gram, 30 mg / gram, etc.).

[0040] In one example, N. crassa was completely dehydrated at 74°C until its moisture content was less than 10% by mass, and then finely ground into a powder with particles of less than 150 μm. Then, conventional or similar recipes using wheat flour were used, substituting N. crassa powder for any amount between 1% and 100% of the total wheat mass. Products included chips, tortillas, bread, and crackers.

[0041] In some embodiments, Method 500 may include a step of shaping the biomass into a certain form. Raw biomass can be formed directly into edible food. Possible processes include extrusion, sheeting, and molding, among others, to form a variety of shapes, such as bars, discs, and cylinders. After drying and / or adding flavors, the biomass forms can be used for human or animal consumption, among other applications, such as chips, protein bars, and jerky. The formed biomass can be cooked at high temperatures, freeze-dried, dehydrated, or fried, among other things. Importantly, the final form factor can be produced without the use of a binder and becomes continuous biomass.

[0042] Biomass with added components can be cooked in a single method or a combination of methods. For example, biomass can be cooked in a dry or steam environment at temperatures below 100°C (e.g., 90°C, 80°C, 75°C, or 50°C including both ends) for 1 to 60 minutes. Biomass can be cooked in a dry or steam environment at temperatures between 100°C and 200°C (e.g., 100°C, 125°C, 150°C, or 200°C including both ends) for 1 to 60 minutes. Biomass can be cooked in a water bath below 100°C for 1 to 120 minutes.

[0043] In some embodiments, the biomass can be stored. The biomass may contain further components. The biomass can be cooked. The biomass can be frozen below 0°C under ambient or vacuum conditions, and / or refrigerated below 5°C under ambient or vacuum conditions. The biomass can be stored indefinitely in a sealed container.

[0044] In one example, biomass is separated using cheesecloth and compressed in an apple press. The moisture content is 75%. The biomass is then dried at various temperatures using a commercially available dehydrator. At 40°C, drying for approximately 3 hours achieves a moisture content of less than 20%; at 60°C, drying for approximately 2 hours achieves a moisture content of less than 20%; and at 75°C, drying for approximately 1.5 hours achieves a moisture content of less than 20%. Lower temperatures take longer, but can result in a whiter appearance for the final biomass, and a lower density can be obtained as the individual hyphae remain separated. Higher temperatures promote the fusion of fungal hyphae together, resulting in higher density and increased resilience. In another example, biomass was separated using cheesecloth. The wet biomass was then added directly to a drum-type heating dryer and heated to 110°C using steam. The residual biomass collected from the drum dryer had a moisture content of less than 10% and had the quality of a thin film or flake. The dried biomass could be further processed into a powder. In another example, biomass is separated using cheesecloth. At this stage, food additives, including 10% bean protein, 10% potato starch, 5% canola oil, 5% flax meal, or other additives, can be added to the wet biomass. The concentration of the food additives depends on the dry biomass. Next, the wet biomass is compressed using an apple press. In another example, biomass is separated using cheesecloth. The wet biomass is then added to molds of various shapes. Pressure is applied to the top of the molds to collect the dried, formed biomass. The formed samples can then be partially dehydrated or used as is, placed in molds, or freely formed using a vacuum forming system. The biomass may be partially dried. Partially dried biomass can be formed using pressure forming. Partially dried biomass can be formed using vacuum forming. The formed samples can then be directly frozen and transported for continuous processing. In another example, a solution containing mainly water, 5% bean protein, and 0.25% carrageenan, as well as seasonings, was mixed. The formed and partially dried biomass was immersed in the solution. Alternatively, the solution was injected in precise amounts into the formed biomass. The sample was then compressed to disperse the solution in a mold. The sample was then cooked in an oven at 180°F for 30 minutes to denature the protein. Alternatively, the solution was dispersed using a vacuum mold, and the sample was then cooked in a sealed vacuum pouch in a water batch at 180°F for 30 minutes. The sample could then be cooked at a higher temperature and served, or frozen.

[0045] Figures 6A1 and 6A2 show a top view and a side view, respectively, of a food product configured as chips according to one embodiment. The chips may be corrugated chips, wavy chips, embossed chips, or have any other suitable size or shape. Figures 6B1 and 6B2 show a top view and a side view, respectively, of a food product configured as elliptical chips according to one embodiment. Figures 6C1 and 6C2 show a top view and a side view, respectively, of a food product configured as triangular chips according to another embodiment.

[0046] Edible food may include chips. Chips may include edible chips. Edible chips may include edible substance. Edible substance may include fibrous mycelial masses. The fibrous mycelial mass may be in the range of 0.1% to 90% by mass (e.g., 0.1%, 0.5%, 1%, 5%, 10%, 25%, 50%, 75%, or 90% by mass, including both ends). The fibrous mycelial mass may have a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial mass. Edible chips may contain a water content of less than 20% by mass (e.g., 15%, 10%, 5%, 2%, 1%, or 0% by mass, including both ends). The edible chips may contain a carbohydrate content ranging from 0% to 90% by mass (for example, 0%, 0.5%, 1%, 5%, 10%, 25%, 50%, 75%, or 90% by mass, including both ends). Fibrous mycelial masses may be distributed within the edible substance. For example, fibrous mycelial masses may be uniformly distributed within the edible substance. In other embodiments, fibrous mycelial masses may be located in concentrated areas within the edible substance. The edible substance may have a diameter greater than 10 times its thickness. For example, the thickness of the edible substance may be 1 mm and its diameter may be 40 mm. The thickness of the edible substance may be 1 mm and its diameter may be 60 mm. The thickness of the edible substance may be 2 mm and its diameter may be 50 mm. The thickness of the edible substance may be 2 mm and its diameter may be 75 mm. The thickness of the edible substance may be 1.5 mm and its diameter may be 45 mm. The thickness of the edible substance may be 1.5 mm and its diameter may be 50 mm. While this specification describes edible substances of specific dimensions, these are merely examples, and it should be understood that edible substances of other dimensions should also be considered within the scope of this disclosure. Edible chips may contain flavorings. Flavorings may include flavors or food additives. For example, flavorings may include oils, such as nut oils, vegetable oils, plant oils, and / or animal oils. In some embodiments, flavorings may include salt, pepper, or spices (e.g., black pepper, fennel, mustard, nutmeg, cinnamon, ginger, red pepper, clove, turmeric, etc.). In some embodiments, flavorings may include flavored powders (e.g., onion powder, garlic powder, BBQ powder, sour cream powder, lemon powder, lime powder, etc.). Fibrous mycelial masses may contain a methionine and cysteine ​​combination of at least 20 mg per gram of crude protein (e.g., 20 mg / gram, 25 mg / gram, or 30 mg / gram including both ends). Fibrous mycelial masses may have a Protein Digestive-Adjusted Amino Acid Score (PDCAAS) score of 1. PDCAAS may include a method for evaluating protein quality based on both human amino acid requirements and human protein digestibility.

[0047] Figure 7 shows a block diagram of Method 700, which is an example of a method for forming an edible meat substitute product from fungal mycelium according to one embodiment. Briefly, Method 700 includes the steps of providing a fibrous mycelial mass in 702, adding a food additive to the fibrous mycelial mass to generate biomass in 704, generating biomass in 706, and forming the biomass into an edible meat substitute product in 708. Further extension, method 700 includes the step of providing a fibrous hyphae in 702. The fibrous hyphae may have a protein content exceeding 40% by mass of the dry mass of the mycelium. The fibrous hyphae may be produced by the operation of method 100 or any other method described herein.

[0048] Method 700 may include the step of adding food additives to a fibrous mycelial mass in 704 to produce biomass. The step of adding food additives to a fibrous mycelial mass may include adding seasonings to the biomass. The flavor of the biomass can be enhanced by adding various oils. Non-limiting examples of oils include nut-derived, vegetable-derived, plant-derived, and animal-derived oils. Oils can be added to food-grade residual water streams because they have multipurpose functions, acting as defoaming agents, carbon sources for fungi, and for extracellular / intracellular integration into the biomass. Alternatively, oils can be integrated into the biomass after harvesting or cooking. The formed and partially dried biomass may have food additives introduced by various operations. For example, the biomass can be immersed in further ingredients. Further ingredients can be injected into the biomass. The biomass can be coated with further ingredients. Further ingredients can be dispersed into the biomass by applying pressure or under vacuum conditions. Further ingredients may include plant or animal proteins, fats, emulsifiers, thickeners, stabilizers, flavors, pH adjusters, spices, salt, etc.

[0049] Method 700 includes a step of producing biomass in 706. The step of producing biomass may include adjusting the texture of the biomass. The texture of fungal biomass can be adjusted by chemically washing the biomass. Alternatively, the texture can be altered by controlling the water content of the biomass. The texture can also be altered by adding various nutrients that determine the growth and morphology of the biomass. The density of the final biomass can be controlled by changing the initial water content and drying conditions to produce a heavier or lighter final product. Raw biomass can be dried and ground into a powder or progressively finer powder. The powder can then be used as a protein component for human or animal feed. The powder can be formed using other components, such as water and / or oil, and binders to create different forms. Protein extraction can be performed to produce fungal protein isolates. These isolates can then be used as feed components for human or animal feed.

[0050] Method 700 may include the step of forming biomass into an edible meat substitute product in 708. The step of forming biomass into an edible meat substitute product may include adding at least one soluble protein to the biomass. The step of forming biomass into an edible meat substitute product may include adding at least one thickener to the biomass. The step of forming biomass into an edible meat substitute product may include adding at least one fat source to the biomass. The step of forming biomass into an edible meat substitute product may include molding the biomass into an edible meat substitute product.

[0051] The meat substitute product may contain fibrous mycelial masses in the range of 10% to 100% by mass (for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 100%) including both ends. The meat substitute product may have a water content in the range of 0% to 100% by mass (for example, 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 100%) including both ends. In some embodiments, the fibrous mycelial masses are in the range of 10% to 50% by mass, and the water content is in the range of 50% to 90% by mass. In some embodiments, the meat substitute product contains soluble protein in the range of 1% to 20% by mass (e.g., 1% by mass, 2% by mass, 5% by mass, 10% by mass, 15% by mass, or 20% by mass, including both ends). The meat substitute product may contain a thickening agent in the range of 0.01% to 5% by mass (e.g., 0.01% by mass, 0.05% by mass, 0.1% by mass, 1% by mass, 2% by mass, or 5% by mass, including both ends). The meat substitute product may contain a fat source in the range of 0% to 10% by mass (e.g., 0% by mass, 0.5% by mass, 1% by mass, 2% by mass, 5% by mass, or 10% by mass, including both ends). Meat substitute products may contain flavorings. Flavorings may include flavors or food additives. For example, flavorings may include oils, such as nut oils, vegetable oils, plant oils, and animal oils. Flavorings may include spices (e.g., black pepper, fennel, mustard, nutmeg, cinnamon, ginger, red pepper, cloves, etc.). Flavorings may include flavored powders (e.g., onion powder, garlic powder, BBQ powder, sour cream powder, lemon powder, lime powder, etc.). The meat substitute product may contain a combined methionine and cysteine ​​content of at least 20 mg per gram of crude protein. In some embodiments, the combined methionine and cysteine ​​content in the meat substitute product is in the range of 20 mg / gram to 30 mg / gram (e.g., 20 mg / gram, 25 mg / gram, or 30 mg / gram including both ends). The meat substitute product may have a PDCAAS score of 1. The meat substitute product may have an internal pH in the range of 2 to 9 (e.g., 2, 3, 4, 5, 6, 7, 8, or 9 including both ends). The meat substitute product may have a dry protein mass in the range of 20% to 70% by mass (e.g., 20% by mass, 30% by mass, 40% by mass, 50% by mass, 60% by mass, or 70% by mass including both ends). The meat substitute product may have a fiber-free weight in the range of 5% to 30% by mass (e.g., 5% by mass, 10% by mass, 15% by mass, 20% by mass, 25% by mass, or 30% by mass, including both ends). The meat substitute product may have a dry fat weight in the range of 0% to 20% by mass (e.g., 0% by mass, 1% by mass, 5% by mass, 10% by mass, 15% by mass, or 20% by mass, including both ends). The meat substitute product may have a CIE L of more than 55. * The product may have a color represented by a value. The meat substitute product may have a Warner-Bratzler shear force exceeding 15N. The meat substitute product may have a hardness exceeding 50N. Meat substitutes may include chicken substitutes, beef substitutes, pork substitutes, veal substitutes, or fish substitutes. Meat substitutes may contain 10% to 90% by mass (for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% by mass, including both ends) of fibrous mycelial masses.

[0052] Chicken substitute products may contain 50-90% by mass (e.g., 50%, 60%, 70%, 80%, or 90% by mass, including both ends) of water. Chicken substitute products may contain 10-50% by mass (e.g., 10%, 20%, 30%, 40%, or 50% by mass, including both ends) of fungal mycelium, such as mycelium derived from N. crassa. Chicken substitute products may contain 1-20% by mass (e.g., 1%, 2%, 5%, 10%, or 20% by mass, including both ends) of soluble protein. Soluble proteins may include beans, egg whites, and potatoes. Chicken substitute products may contain 0.01-5% by mass (e.g., 0.01%, 0.05%, 0.1%, 1%, 2%, or 5% by mass, including both ends) of thickeners. Thickeners may include pectin, carrageenan, and agar. Chicken substitutes may contain 0-10% by mass (0%, 1%, 2%, 3%, 4%, 5%, or 10%) of fat sources, including both ends. Fat sources may include vegetable oils and seed oils. Chicken substitutes may contain seasonings. Chicken substitutes can have a variety of physical properties. For example, chicken substitutes may have an internal pH between 2 and 9 (e.g., 2, 3, 4, 5, 6, 7, 8, or 9, including both ends). Chicken substitutes may have a dry protein mass of 40-70% by mass (e.g., 40%, 45%, 50%, 55%, 60%, 65%, or 70%). Chicken substitute products may have a fiber-free weight of 5-30% by mass (e.g., 5%, 10%, 15%, 20%, 25%, or 30% by mass, including both ends). Chicken substitute products may have a fat-free weight of 0-10% by mass (0%, 1%, 2%, 4%, 5%, or 10% by mass, including both ends). Chicken substitute products may have a CIE L of more than 55. * The chicken substitute can have a value. The chicken substitute can have a Warner-Bratzler shear force of more than 15N. The chicken substitute can have a hardness of more than 50N. In one example of producing a chicken substitute product, a solution containing mainly water, 5% bean protein, 0.25% carrageenan, and seasonings was mixed. The formed and partially dried biomass was immersed in the solution. Alternatively, the solution was injected into the formed biomass in precise amounts. The sample was then compressed to disperse the solution in a mold. The sample was then cooked in an oven at 180°F for 30 minutes to denature the protein. Alternatively, the solution was dispersed using a vacuum mold, and the sample was then cooked in a sealed vacuum pouch in a water batch at 180°F for 30 minutes. The sample could then be cooked at a higher temperature and served, or frozen.

[0053] Meat substitutes may contain 0-90% by mass (e.g., 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% by mass, including both ends) of water. Meat substitutes may contain 10-100% by mass (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% by mass, including both ends) of fungal mycelium, such as mycelium derived from N. crassa. Meat substitutes may contain 1-20% by mass (e.g., 1%, 2%, 5%, 10%, or 20% by mass, including both ends) of soluble protein. Soluble proteins may include those from beans, egg whites, and potatoes. Meat substitute products may contain 0-5% by mass (e.g., 0%, 0.01%, 0.05%, 0.1%, 1%, 2%, or 5%) of a thickening agent, including pectin, carrageenan, or agar. Meat substitute products may contain 0-50% by mass (e.g., 0%, 10%, 20%, 30%, 40%, or 50%) of a fat source, including vegetable oil or seed oil. Meat substitute products may contain seasonings. In one example of producing a meat substitute product, N. crassa was completely dehydrated at 74°C until the biomass had a water content of less than 10% by mass. The dried N. crassa was then rehydrated in a "marinade" consisting of sauce, Worcestershire sauce, vegetable oil, and seasonings. The N. crassa soaked in the marinade was then dehydrated at 74°C until the biomass had a water content of approximately 30% by mass.

[0054] The flavor of biomass can be enhanced by adding various oils. Non-limited examples of oils include those derived from nuts, vegetables, plants, and animals. Oils can be added to food-grade residual water streams because they have multipurpose uses, acting as defoamers, carbon sources for fungi, and for extracellular / intracellular integration into the biomass. Alternatively, oils can be integrated into the biomass after harvesting or cooking. The texture of fungal biomass can be adjusted by chemically washing the biomass. Alternatively, the texture can be altered by controlling the water content of the biomass. The texture can also be altered by adding various nutrients that determine the growth and morphology of the biomass. The final biomass density can be controlled by changing the initial water content and drying conditions to produce heavier or lighter final products.

[0055] In one example, N. crassa was propagated and harvested. The biomass was separated using cheesecloth. After draining by gravity, the water content was approximately 95%. The biomass was washed with various different solutions containing tap water, ethanol, 1M citric acid, or 0.1M calcium hydroxide. The biomass was compressed using an apple press. After compression, the water content was approximately 75% by mass. Samples washed with ethanol had a water content of 68%. Next, the wet biomass was tested for any residual sugar content derived from the culture medium. Unwashed biomass was recorded as having 3 g / L of sugar in the culture medium, biomass washed with water was recorded as having approximately 0.5 g / L of sugar, biomass washed with ethanol was recorded as having 0.4 g / L of sugar, and biomass washed with 1M citric acid and 0.1M calcium hydroxide was recorded as having less than 0.1 g / L of sugar.

[0056] Raw biomass can be directly formed into edible meat substitutes. Possible processes include extrusion, sheeting, and molding to form various shapes, among others, such as bars, discs, and cylinders. After drying and / or adding flavors to the biomass, it can be used to form edible products for human or animal consumption, among others, such as chips, protein bars, and jerky. The formed biomass can be cooked at high temperatures, freeze-dried, dehydrated, or fried, among others. Importantly, the final form factor can be produced without the use of a binder and becomes continuous biomass. Raw biomass can be dried and ground into a powder or progressively finer powder. The powder can then be used as a protein component for human or animal feed. The powder can be formed into various shapes using other components, such as water and / or oil, and binders. Protein extraction can be performed to produce fungal protein isolates. These isolates can then be used as feed components for human or animal feed.

[0057] Figure 8A shows a perspective view of a meat substitute food product configured as a patty according to one embodiment. The patty may be a patty for a burger or sandwich. Figure 8B shows a perspective view of a meat substitute product configured as a tender according to one embodiment. The tender may be a meat substitute tender, strip, fillet, or nugget. Figure 8C shows a perspective view of a ground meat substitute product according to one embodiment. In some embodiments, a method includes the step of growing fungal cells in a growth medium so that the fungal cells produce mycelium. The growth medium contains sugars, nitrogen-containing compounds, and phosphate-containing compounds. The method includes the steps of separating the mycelium from the growth medium and concentrating the mycelium to obtain fibrous mycelial masses having a protein content of more than 40% by mass of the dry mass of the mycelium. In some embodiments, the method includes the step of removing 0.01% to 95% by volume of the broth containing fungal cells and growth medium. The method includes the step of adding fresh growth medium to the container containing the broth. In some embodiments, the sugar is in the range of 5 to 50 g / L, the nitrogen-containing compound is in the range of 0.5 to 10 g / L, and the phosphate-containing compound is in the range of 0.1 to 5 g / L. In some embodiments, the sugar includes at least one of sucrose, glucose, fructose, or molasses. In some embodiments, the nitrogen-containing compound includes at least one of ammonium hydroxide, ammonium nitrate, ammonium sulfate, ammonium chloride, urea, yeast extract, or peptone. In some embodiments, the phosphate-containing compound includes at least one of potassium phosphate, sodium phosphate, or phosphate. In some embodiments, the method includes the step of lysing the cell walls of the mycelium. In some embodiments, the method includes the step of adding food additives to the mycelium. The food additives include at least one of plant protein, animal protein, fat, emulsifier, thickener, stabilizer, or flavor. In some embodiments, a method for forming edible food includes the step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium. The method includes the step of adding a food additive to the fibrous mycelial mass to generate biomass, and the step of forming the biomass into edible food. In some embodiments, the step of forming biomass into an edible food includes dehydrating the biomass. In some embodiments, the step of forming biomass into an edible food includes grinding the biomass into a powder. In some embodiments, the method includes incorporating the powder into a baked food. In some embodiments, the edible food is at least one of chips, protein bars, jerky, tortillas, bread, or crackers. In some embodiments, the method includes shaping the biomass into a certain form. In some embodiments, a method for forming an edible meat substitute product includes the step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium. The method includes the steps of adding a food additive to the fibrous mycelial mass to generate biomass, and forming the biomass into an edible meat substitute product. In some embodiments, the step of forming biomass into an edible meat substitute product includes adding at least one soluble protein to the biomass. In some embodiments, the step of forming biomass into an edible meat substitute product includes molding the biomass into an edible meat substitute product. In some embodiments, the step of forming biomass into an edible meat substitute product includes adding at least one thickener to the biomass. In some embodiments, the step of forming biomass into an edible meat substitute product includes molding the biomass into an edible meat substitute product. In some embodiments, the step of forming biomass into an edible meat substitute product includes adding at least one fat source to the biomass. In some embodiments, the step of forming biomass into an edible meat substitute product includes molding the biomass into an edible meat substitute product. In some embodiments, the method includes adding a seasoning to the biomass.

[0058] In some embodiments, the edible meat substitute product includes at least one of a chicken substitute product, a beef substitute product, a pork substitute product, a veal substitute product, or a fish substitute product. In some embodiments, the edible meat substitute product includes 10% to 90% by mass of fibrous mycelium. In some embodiments, the edible meat substitute product comprises fibrous mycelial mass in the range of 10% to 100% by mass, having a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial mass, and a water content in the range of 0% to 90% by mass. In some embodiments, the fibrous mycelial mass is in the range of 10% to 50% by mass, and the water content is in the range of 50% to 90% by mass. In some embodiments, the meat substitute product further comprises soluble protein in the range of 1% to 20% by mass and a thickening agent in the range of 0.01% to 5% by mass. In some embodiments, the meat substitute product further comprises a fat source in the range of 0% to 10% by mass. In some embodiments, the meat substitute product further comprises soluble protein in the range of 0.01% to 20% by mass, a thickening agent in the range of 0.01% to 5% by mass, and a fat source in the range of 0% to 50% by mass.

[0059] In some embodiments, the meat substitute product further comprises flavoring agents. In some embodiments, the fibrous mycelium contains a content of at least 20 mg of a methionine and cysteine ​​combination per gram of crude protein. In some embodiments, the fibrous mycelium has a protein digestibility-corrected amino acid (PDCAAS) score of 1. In some embodiments, the meat substitute product has an internal pH in the range of 2 to 9, a dry protein mass in the range of 20% to 70% by mass, a dry fiber mass in the range of 5% to 30% by mass, and a dry fat mass in the range of 0% to 20% by mass. In some embodiments, the meat substitute product has a CIE L greater than 55. * It has a color represented by a value, a Warner-Bratzler shear force exceeding 15N, and a hardness exceeding 50N. In some embodiments, the edible chips contain an edible substance comprising fibrous mycelial masses ranging from 0.1% to 90% by mass, with a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial masses, a water content of less than 20% by mass, and a carbohydrate content ranging from 0% to 90% by mass. The fibrous mycelial masses are distributed within the edible substance. In some embodiments, the edible substance has a diameter greater than 10 times the thickness of the edible substance. In some embodiments, the edible substance also contains flavoring agents. In some embodiments, the fibrous mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein. In some embodiments, the fibrous mycelial mass has a PDCAAS score of 1.

[0060] In some embodiments, the powdered edible product includes powdered mycelium having a protein content exceeding 40% by mass of the powdered mycelium. In some embodiments, the particles of the powdered mycelium have a particle size of less than 150 μm. In some embodiments, the powdered mycelium is in the range of 0.1% to 80% by mass of the total mass of the powdered edible product, and the powdered edible product further contains carbohydrates in the range of 0% to 90% by mass. In some embodiments, the powdered mycelium contains a content of at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein. In some embodiments, the powdered mycelium has a PDCAAS score of 1.

[0061] This specification contains many details of specific implementations, but these should not be construed as limiting the scope of either the invention or the claims, but rather as describing characteristics specific to a particular implementation of the invention. Certain characteristics described herein in the context of a separate implementation may also be implemented in combination in a single implementation. Conversely, various characteristics described in the context of a single implementation may also be implemented in multiple implementations, individually or in any suitable partial combination. Furthermore, characteristics may be described above as acting in a particular combination and may be initially claimed as such, but one or more characteristics resulting from a claimed combination may, in some cases, be excluded from that combination, and the claimed combination may be a partial combination or a variation of a partial combination. Similarly, while operations are illustrated in a specific order in the figures and tables, this should not be understood as requiring that such operations be performed in the order or sequence shown, or that all illustrated operations be performed, in order to achieve the desired result. In certain environments, concurrent parallel processing may be advantageous. Furthermore, the separation of various system components in the aforementioned implementation forms should not be understood as requiring such separation in all implementation forms, and it should be understood that the program components and systems described can generally be integrated into a single software product or packaged into multiple software products. Accordingly, a specific implementation of the present invention is described. Other implementations are also within the scope of the following claims. In some cases, the operations described in the claims can be performed in a different order to achieve the desired results. Furthermore, the processes illustrated in the accompanying figures do not necessarily require the specific order or sequence shown to achieve the desired results. In certain particular implementations, concurrent parallel processing may be advantageous.

[0062] As used herein, the singular forms “a,” “an,” and “the” refer to multiple objects unless otherwise specified by the context. Thus, for example, the term “a member” is intended to mean a single member or combination of members, and “a material” is intended to mean one or more materials or combination thereof.

[0063] As used herein, the terms “about” and “approximately” generally mean plus or minus 10% of the stated value. For example, about 0.5 may include 0.45 and 0.55, about 10 may include 9 to 11, and about 1000 may include 900 to 1100. When the term “exemplary” is used herein to describe various embodiments, it is intended to indicate that such embodiments are possible examples, representative, and / or illustrative of possible embodiments (it is not intended to imply that such embodiments necessarily represent special or best examples). As used herein, terms such as “joined” and “connected” mean that two members are joined to each other directly or indirectly. Such joining may be fixed (e.g., permanent) or movable (e.g., removable or detachable). Such joining may be achieved by two members or two members and any further intermediate members forming a single unit with each other, or by two members or two members and any further intermediate members joining to each other. It is important to note that the configurations and arrangements of the various exemplary embodiments are merely illustrative. Although only a few embodiments have been described in detail in this disclosure, those skilled in the art who reconsider this disclosure will readily understand that many modifications (e.g., changes in the size, dimensions, structure, shape and proportions, parameter values, mounting arrangements, material use, color, orientation, etc.) are possible without substantially departing from the novel teachings and advantages of the subject matter described herein. Other substitutions, modifications, changes, and omissions can also be made to the designs, operating conditions, and arrangements of the various exemplary embodiments without departing from the scope of the invention.

[0064] Throughout this specification, any reference to “one embodiment,” “an embodiment,” or similar terms means that a particular characteristic, structure, or feature described in relation to an embodiment is included in at least one embodiment of this disclosure. Throughout this specification, any occurrence of the phrases “in one embodiment,” “an embodiment,” and similar terms may or may not refer to the same embodiment. Similarly, the use of the term “implementation” means an implementation having a particular characteristic, structure, or feature described in relation to one or more embodiments of this disclosure, but unless there is a clear correlation indicating otherwise, one implementation may be related to one or more embodiments.

[0065] This specification contains many details of specific implementations, but these should not be construed as limiting the scope of either the invention or the claims, but rather as describing characteristics specific to a particular implementation of the invention. Certain characteristics described herein in the context of a separate implementation may also be implemented in combination in a single implementation. Conversely, various characteristics described in the context of a single implementation may also be implemented in multiple implementations, individually or in any suitable partial combination. Furthermore, characteristics may be described above as acting in a particular combination and may be initially claimed as such, but one or more characteristics resulting from a claimed combination may, in some cases, be excluded from that combination, and the claimed combination may be a partial combination or a variation of a partial combination. Similarly, while operations are illustrated in a specific order in the figures and tables, this should not be understood as requiring that such operations be performed in the order or sequence shown, or that all illustrated operations be performed, in order to achieve the desired result. In certain environments, concurrent parallel processing may be advantageous. Furthermore, the separation of various system components in the aforementioned implementation forms should not be understood as requiring such separation in all implementation forms, and it should be understood that the program components and systems described can generally be integrated into a single software product or packaged into multiple software products. Accordingly, a specific implementation of the present invention is described. Other implementations are also within the scope of the following claims. In some cases, the operations described in the claims can be performed in a different order to achieve the desired results. Furthermore, the processes illustrated in the accompanying figures do not necessarily require the specific order or sequence shown to achieve the desired results. In certain particular implementations, concurrent parallel processing may be advantageous. Another aspect of the present invention may be as follows: [1] The step of growing fungal cells in a growth medium containing sugar, nitrogen-containing compounds, and phosphate-containing compounds so that the fungal cells produce mycelium, The steps include: separating the mycelium from the growth medium, The steps include: concentrating the mycelium to obtain a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium; Methods that include... [2] A step of removing 0.01% to 95% by volume of the broth containing fungal cells and growth medium, The steps include adding new growth medium to a container containing broth and The method according to [1], further comprising: [3] The method according to [1], wherein the sugar is in the range of 5 to 50 g / L, the nitrogen-containing compound is in the range of 0.5 to 10 g / L, and the phosphoric acid-containing compound is in the range of 0.1 to 5 g / L. [4] The method according to [1], wherein the sugar comprises at least one of sucrose, glucose, fructose, or molasses. [5] The method according to [1], wherein the nitrogen-containing compound comprises at least one of ammonium hydroxide, ammonium nitrate, ammonium sulfate, ammonium chloride, urea, yeast extract, or peptone. [6] The method according to [1], wherein the phosphate-containing compound comprises at least one of potassium phosphate, sodium phosphate, or phosphate. [7] Step of lysing the cell wall of the mycelium The method according to [1], further comprising: [8] Adding a food additive to the mycelium, which includes at least one of plant protein, animal protein, fat, emulsifier, thickener, stabilizer, or flavor. The method according to [1], further comprising: [9] A method for forming edible food, A step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium, A step of adding food additives to fibrous mycelial masses to generate biomass, Steps for forming biomass into edible food and Methods that include...

[10] The step of forming biomass into edible food is Dehydrating biomass, and The process of finely grinding biomass into a powder. The method according to [9], further comprising:

[11] The method according to

[10] , further comprising the step of incorporating the powder into a baked food product.

[12] The method according to [9], wherein the edible food is at least one of chips, protein bars, jerky, tortillas, bread, or crackers.

[13] The method according to [9], further comprising the step of molding biomass into a certain shape.

[14] A method for forming an edible meat substitute product, A step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium, A step of adding food additives to fibrous mycelial masses to generate biomass, Steps for forming biomass into edible meat substitute products and Methods that include...

[15] The step of forming biomass into an edible meat substitute product is Adding at least one soluble protein to the biomass, and Molding biomass into edible meat substitute products The method according to

[14] , including the method described above.

[16] The step of forming biomass into an edible meat substitute product is Adding at least one thickening agent to the biomass, Molding biomass into edible meat substitute products The method according to

[14] , including the method described above.

[17] The step of forming biomass into an edible meat substitute product is Adding at least one fat source to the biomass, and Molding biomass into edible meat substitute products The method according to

[14] , including the method described above.

[18] The method according to

[14] , further comprising the step of adding a seasoning to biomass.

[19] The method according to

[14] , wherein the edible meat substitute product comprises at least one of a chicken substitute product, a beef substitute product, a pork substitute product, a veal substitute product, or a fish substitute product.

[20] The method according to

[14] , wherein the edible meat substitute product contains 10% to 90% by mass of fibrous mycelial masses.

[21] Fibrous mycelial masses having a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial mass, and having a protein content in the range of 10% to 100% by mass, Water content in the range of 0% to 90% by mass Meat substitute products that include edible meat.

[22] The edible meat substitute product according to

[21] , wherein the fibrous mycelial mass is in the range of 10% to 50% by mass and the water content is in the range of 50% to 90% by mass.

[23] Soluble protein in the range of 1% to 20% by mass, and Thickening agent content in the range of 0.01% to 5% by mass The edible meat substitute product described in

[22] further includes the following:

[24] The edible meat substitute product according to

[23] , further comprising a fat source in the range of 0% to 10% by mass.

[25] Soluble protein in the range of 0.01% by mass to 20% by mass, Thickening agent content in the range of 0.01% by mass to 5% by mass, Fat sources in the range of 0% to 50% by mass The edible meat substitute product described in

[21] further includes the following:

[26] The edible meat substitute product according to

[21] , further comprising a flavoring agent.

[27] The edible meat substitute product according to

[21] , wherein the fibrous mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein.

[28] The edible meat substitute product according to

[21] , wherein the fibrous mycelial mass has a protein digestibility-corrected amino acid (PDCAAS) score of 1.

[29] The edible meat substitute product according to

[21] , having an internal pH in the range of 2 to 9, a dry protein mass in the range of 20% to 70% by mass, a dry fiber mass in the range of 5% to 30% by mass, and a dry fat mass in the range of 0% to 20% by mass.

[30] CIE L exceeding 55 * The edible meat substitute product described in

[29] , having a color represented by a value, a Warner-Bratzler shear force greater than 15 N, and a hardness greater than 50 N.

[31] Edible chips, Edible substance containing fibrous mycelial masses ranging from 0.1% to 90% by mass, with a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial mass. Water content of less than 20% by mass, and Carbohydrate content in the range of 0% to 90% by mass Includes, Edible chips in which fibrous mycelial masses are distributed within the edible substance.

[32] The edible chips according to

[31] , wherein the edible portion has a diameter that is more than 10 times the thickness of the edible portion.

[33] Edible chips as described in

[31] , further comprising a flavoring agent.

[34] The edible chips according to

[31] , wherein the fibrous mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein.

[35] Edible chips according to

[31] , wherein the fibrous mycelial mass has a PDCAAS score of 1.

[36] Powdered edible product, Powdered mycelial mass having a protein content exceeding 40% by mass A powdered edible product containing [the specified ingredient].

[37] The powdered edible product according to

[36] , wherein the powder particles have a particle size of less than 150 μm.

[38] The powdered edible product according to

[36] , wherein the powdered mycelial mass is in the range of 0.1% to 80% by mass of the total mass of the powdered edible product, and the powdered edible product further contains carbohydrates in the range of 0% to 90% by mass.

[39] The powdered edible product according to

[36] , wherein the powdered mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein.

[40] The powdered edible product according to

[36] , wherein the powdered mycelial mass has a PDCAAS score of 1.

Claims

1. The steps include growing fungal cells in a growth medium containing sugar, nitrogen-containing compounds, and phosphate-containing compounds so that the fungal cells produce mycelium, The steps include: separating the mycelium from the growth medium, The steps include: concentrating the mycelium to obtain a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium; Includes, A method in which mycelium is a mass of fungal cells of the species Neurospora crassae.

2. A step of removing 0.01% to 95% by volume of the broth containing fungal cells and growth medium, The steps include adding new growth medium to a container containing broth and The method according to claim 1, further comprising:

3. The method according to claim 1, wherein the sugar is in the range of 5 to 50 g / L, the nitrogen-containing compound is in the range of 0.5 to 10 g / L, and the phosphoric acid-containing compound is in the range of 0.1 to 5 g / L.

4. The method according to claim 1, wherein the sugar comprises at least one of sucrose, glucose, fructose, or molasses.

5. The method according to claim 1, wherein the nitrogen-containing compound comprises at least one of ammonium hydroxide, ammonium nitrate, ammonium sulfate, ammonium chloride, urea, yeast extract, or peptone.

6. The method according to claim 1, wherein the phosphoric acid-containing compound comprises at least one of potassium phosphate, sodium phosphate, or phosphoric acid.

7. The step of adding a food additive to the mycelium, which includes at least one of plant protein, animal protein, fat, emulsifier, thickener, stabilizer, or flavor. The method according to claim 1, further comprising:

8. A method for forming an edible meat substitute product, A step of providing a fibrous mycelial mass having a protein content exceeding 40% by mass of the dry mass of the mycelium, A step of adding food additives to fibrous mycelial masses to generate biomass, Steps for forming biomass into edible meat substitute products and Includes, A method for determining whether a fibrous hyphae is a mass of fungal cells of the species Neurospora crassae.

9. The step of forming biomass into edible meat substitute products is Adding at least one soluble protein to the biomass, Molding biomass into edible meat substitute products The method according to claim 8, including the method described in claim 8.

10. The step of forming biomass into edible meat substitute products is Adding at least one thickening agent to the biomass, Molding biomass into edible meat substitute products The method according to claim 8, including the method described in claim 8.

11. The step of forming biomass into edible meat substitute products is Adding at least one fat source to the biomass, and Molding biomass into edible meat substitute products The method according to claim 8, including the method described in claim 8.

12. The method according to claim 8, further comprising the step of adding a seasoning to biomass.

13. The method according to claim 8, wherein the edible meat substitute product comprises at least one of a chicken substitute product, a beef substitute product, a pork substitute product, a veal substitute product, or a fish substitute product.

14. The method according to claim 8, wherein the edible meat substitute product contains 10% to 90% by mass of fibrous mycelial masses.

15. Fibrous mycelial masses having a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial mass, and fibrous mycelial masses in the range of 10% to 100% by mass, and Water content in the range of 0% to 90% by mass Includes, A meat substitute product in which fibrous mycelial masses are clumps of fungal cells of the Neurospora crassae species.

16. The edible meat substitute product according to claim 15, wherein the fibrous mycelial mass is in the range of 10% to 50% by mass, and the water content is in the range of 50% to 90% by mass.

17. Soluble protein in the range of 1% to 20% by mass, and Thickening agent content in the range of 0.01% to 5% by mass The edible meat substitute product according to claim 16, further comprising:

18. The edible meat substitute product according to claim 17, further comprising a fat source in the range of 0% to 10% by mass.

19. Soluble protein in the range of 0.01% to 20% by mass, A thickening agent content in the range of 0.01% by mass to 5% by mass, and Fat sources in the range of 0% to 50% by mass The edible meat substitute product according to claim 15, further comprising:

20. The edible meat substitute product according to claim 15, further comprising a flavoring agent.

21. The edible meat substitute product according to claim 15, wherein the fibrous mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein.

22. The edible meat substitute product according to claim 15, wherein the fibrous mycelial mass has a protein digestibility correction amino acid (PDCAAS) score of 1.

23. The edible meat substitute product according to claim 15, having an internal pH in the range of 2 to 9, a dry protein mass in the range of 20% to 70% by mass, a dry fiber mass in the range of 5% to 30% by mass, and a dry fat mass in the range of 0% to 20% by mass.

24. Over 55 CIE L * The edible meat substitute product according to claim 23, having a color represented by a value, a Warner-Bratsler shear force greater than 15 N, and a hardness greater than 50 N.

25. Edible chips, Edible substance containing fibrous mycelial masses ranging from 0.1% to 90% by mass, with a protein content exceeding 40% by mass of the dry mass of the fibrous mycelial masses. Water content of less than 20% by mass, and Carbohydrate content in the range of 0% to 90% by mass Includes, Edible chips in which fibrous mycelial masses are distributed within the edible substance, and these fibrous mycelial masses are clumps of fungal cells of the Neurospora crassae species.

26. The edible chips according to claim 25, wherein the edible portion has a diameter that is more than 10 times the thickness of the edible portion.

27. Edible chips according to claim 25, further comprising a flavoring agent.

28. The edible chips according to claim 25, wherein the fibrous mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein.

29. The edible chips according to claim 25, wherein the fibrous mycelial mass has a PDCAAS score of 1.

30. It is a powdered edible product, Powdered mycelial mass having a protein content exceeding 40% by mass Includes, Powdered mycelial mass is a powdered edible product consisting of clumps of fungal cells from the Neurospora crassae species.

31. The powdered edible product according to claim 30, wherein the powder particles have a particle size of less than 150 μm.

32. The powdered edible product according to claim 30, wherein the powdered mycelial mass is in the range of 0.1% to 80% by mass of the total mass of the powdered edible product, and the powdered edible product further contains carbohydrates in the range of 0% to 90% by mass.

33. The powdered edible product according to claim 30, wherein the powdered mycelial mass contains at least 20 mg of a combination of methionine and cysteine ​​per gram of crude protein.

34. The powdered edible product according to claim 30, wherein the powdered mycelial mass has a PDCAAS score of 1.

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