Fungal ingredients and derived products

Abstract

The present invention relates to mycelial components obtained from submerged fermentation of at least one fungal strain in three different media: a defined medium to yield component A, a synthetic medium to yield component B, or a complex natural medium containing a sidestream extract from an agricultural food sidestream to yield component C. Such unique mycelial compositions are characterized chemically, biologically, physically, morphologically, nutritionally, and organoleptically. Three different filamentous mycelial masses of edible fungi obtained from the at least one fungal strain are further used to produce characterized food products, including meat substitutes, fish substitutes, dairy substitutes, beverages, or other food products. These three novel mycelial components can be used in the production of food, foodstuffs, beverages, pharmaceuticals, cosmetics, dietary supplements, biomaterials, and feed, as well as in industrial applications.

Description

[Technical Field]

[0001] The present invention relates to mycelial components obtained from submerged fermentation of at least one fungal strain in three different media, where the three different media are a defined medium yielding component A, a synthetic medium yielding component B, or a complex natural medium containing a sidestream extract selected from an agricultural food sidestream yielding component C. Such unique mycelial compositions are characterized chemically, biologically, physically, morphologically, nutritionally, and organoleptically. Three different filamentous mycelial masses of edible fungi obtained from the at least one fungal strain are further used to produce characterized food products, including meat substitutes, fish substitutes, dairy substitutes, beverages, or other food products. These three new mycelial components can be used in the production of food, foodstuffs, beverages, pharmaceuticals, cosmetics, dietary supplements, biomaterials, and feed, as well as in industrial applications. [Background technology]

[0002] Over the past decade, several food scandals have drawn unprecedented attention to our current food production systems and their lack of clarity and robustness when it comes to food safety. In 2011, long international supply chains were deemed the primary cause of an enteroaggregative E. coli outbreak in Germany, resulting in several deaths across Europe and the worldwide recall of related products. Then, in 2013, significant amounts of horse meat were found across Europe in products advertised as beef, with potential health implications linked to the contamination of phenylbutazone, a common painkiller for horses. That same year, Muslim and Jewish communities were affected when pork was found in beef products. In 2017, eggs contaminated with the common pesticide finopril were found in several European and Asian countries. In all these cases, determining the source of the contamination and then coordinating the removal of the spoiled food was hindered by the complexity of current global supply and distribution channels, thus posing long-term risks to consumers.

[0003] On a different level, the COVID-19 pandemic has also clearly highlighted that the global food system does not provide the necessary level of resilience with regard to food security. The first year of the pandemic resulted in an approximately 20% increase in world food prices. The World Food Programme (WFP) estimates that the number of people suffering from acute food insecurity has increased from 135 million to 272 million (worldbank.org / en / topic / agriculture / brief / food-security-and-covid-19 and csis.org / analysis / covid-19-and-global-food-security-one-year-later, both assessed on October 17, 2022). By analogy, food systems are vulnerable to disasters around the world.

[0004] At the same time, one-third of all food produced worldwide is lost before reaching the consumer or is subsequently wasted. This represents approximately 1.3 billion tons of food, much of which could potentially be recovered with optimal production and distribution logistics. Growing consumer awareness has led to the emergence of new purchasing trends, in which local, natural, healthy, and sustainable products are preferred over ultra-processed and unbalanced foods. Modern consumers expect to be able to trace the origins of what they buy, to be able to understand the ingredients on the packaging, and to understand the product's impact on their health and that of the planet.

[0005] Environmental awareness has increased in recent years as the unsustainability, and in some cases cruelty, of industrial production methods and practices for meat and fish has been highlighted by several published studies referenced here (https: / / doi.org / 10.3390 / foods9091227 and https: / / doi.org / 10.3390 / foods9091151). While plant-based alternatives can significantly reduce CO2 emissions and improve animal welfare compared to traditional meat production, they do not fully address the challenge of local production, as these products are mostly derived from three monocrops (soybeans, peas, and rice) that are grown in only a few countries and need to be exported worldwide. Furthermore, their cultivation requires large land areas, which unfortunately are often obtained by deforestation, and their efficient production still relies heavily on chemical agents, such as pesticides and fertilizers, which contribute to soil and water pollution and have long-term impacts on biodiversity. In addition, only concentrates and isolates from crops are used in the production of meat substitutes, and therefore a significant amount of waste is generated in the process. Finally, these plant proteins have a strong bitter taste and no inherent texture. Therefore, their use in food requires additional processing steps and a long list of ingredients. Therefore, plant-based alternatives cannot address all consumer concerns regarding food traceability and sustainability.

[0006] In parallel with the development of plant-based meat substitutes, interest in some traditional foods, such as mushrooms, is also increasing due to their potential as natural meat substitutes in terms of nutrition, texture, and / or taste. Mushrooms are of particular interest because they have a natural umami flavor and certain interspecies variations that allow for the production of products with a taste profile similar to that of meat or other flavorful foods without adding a long list of ingredients. Their fruiting bodies or caps also have a meat-like texture and can be further improved with minimal processing for specific uses. In terms of nutrition, mushrooms contain up to 40% complete protein and also contain prebiotic fiber, which is often lacking in the Western diet. Mushrooms also contain large amounts of important minerals, such as iron, zinc, calcium, potassium, or magnesium, and vitamins from the B group. Mushrooms can be considered one of the rare foods for providing a complete and balanced nutritional profile.

[0007] Mushroom cultivation is a highly environmentally friendly but lengthy process. In nature, mushrooms possess a wide range of unique enzymes that allow them to remove waste materials present on forest soils, such as fallen leaves or wood residues. Mushrooms can decompose complex plant compounds, providing nutrients that are otherwise generally inaccessible to other organisms. This characteristic makes mushrooms ideal candidates for upcycling waste from the agri-food industry, which is often highly unstable and, despite their residual nutrient content (e.g., wheat stalks or plant husks), is typically used as animal feed, burned, or simply discarded. To date, only approximately 144,000 fungal species have been described, and it is estimated that over 10 million species exist on Earth, including a wide range of unknown edible mushroom species. Many of these undiscovered species may offer new avenues for culinary experiences and upcycling opportunities for waste materials. Despite their attractive attributes as a food product, mushrooms are relatively slow-growing, with production cycles generally taking at least six weeks. Furthermore, traditional production methods are very basic and use techniques that are difficult to scale, such as growth on forest trees and / or in bags containing lignocellulosic material. In recent years, more modern techniques, including the use of incubation chambers where temperature and humidity are tightly controlled or hydroponic methods, have allowed for significant improvements in process standardization and production yields, but require large investments and do not fully address the scalability issue.

[0008] In this context, the use of fermentation to produce mushroom mycelium offers advantages in terms of sustainability, food safety, and traceability. Fermenters are sterile vessels that operate under controlled conditions. Therefore, the risk of spoilage is minimized. Fermenters can be vertically scaled, thus allowing for a smaller plant footprint and the potential for food to be produced locally using by-products from farms or food processors, which eliminates the need for long supply or distribution chains. Furthermore, the production of mushroom mycelium in fermenters is also more sustainable than the production of conventional plant-based alternatives. Water consumption, land area requirements, energy consumption, and CO2 emissions are estimated to be lower than those for conventional meat substitutes from soy.

[0009] Therefore, to avoid the above problems and limitations, three novel mushroom mycelium components or fungal biomass components were developed by liquid or submerged fermentation in a liquid medium. The taste, composition (carbohydrates, fats, proteins, nutrients, vitamins, fiber content, amino acids, etc.), texture, and structure are controlled during the fermentation process depending on the fermentation conditions (e.g., the medium, species, process configuration, and conditions used), which results in unique biological, physical, and chemical properties for each mycelium component. In addition, the biomass retains its typical umami flavor when cooked, which can also be controlled at the fermentation level. Therefore, the food products developed in the present invention, such as meat and milk substitutes, or other food products made with the mycelium pulp or components developed and disclosed in the present invention, require minimal processing and a very short list of ingredients that can be easily communicated to customers. Summary of the Invention

[0010] The present invention solves the above problems by introducing three new raw materials or ingredients, namely, edible mushroom mycelium, into the production of meat substitutes, fish substitutes, and milk substitutes, or other food products. These three new ingredients can also be used in the production of food, foodstuffs, beverages, pharmaceuticals, cosmetics, dietary supplements, biomaterials, and feed, as well as industrial applications. Due to their filamentous structure, mycelium or filamentous fungi have been widely applied in meat substitute products (GB Patent No. 2137226(A)), and in dairy drinks or yogurt as fat mimics (WO2002090527(A1)).

[0011] Chinese Patent No. 103184246(A) discloses a method for preparing ergothioneine using liquid culture of wild Pleurotus sapidus, Pleurotus pulmonarius, or Lepisa sordida to produce ergothioneine, with a low yield of 51 mg / L and a cultivation time of 10 days. For example, Chinese Patent Publication No. 110283856(A) discloses a method for producing ergothioneine at a yield of 300 mg / L by fermenting the fungal strain Pleurotus ostreatus 3210, but this process requires at least 25 days, including 15 days of growth of mycelium on PD followed by 10 days of fermentation. In the patent literature, it has been observed that a higher content of ergothioneine can be obtained by co-fermenting two or more fungal strains (CN Patent Publication No. 112195215 or CN Patent Publication No. 114214387). Finally, CN Patent No. 109939027(A) discloses a method for producing ergothioneine by fermenting hericium erinaceus with glucose and peptone, with a yield of 331 mg / L, but the production cost of the substrate is high and the process takes a long time (approximately 25 days).

[0012] Chinese Utility Model No. 212786880 reports that Pleurotus pulmonarius fruiting bodies have a low fiber content, which is supported by a review published in 2021 (Fungal Biotec 1(2):65-87(2021)). The review summarizes that the fiber content of Pleurotus species fruiting bodies ranges from 2.97% to at most 31% by weight, and that Pleurotus pulmonarius fruiting bodies in particular contain 4-9% fiber by weight on a dry basis. Chinese Patent No. 105054261 also discloses the finding that when Pleurotus pulmonarius is mixed with other strains, degradation of crude fiber occurs, particularly in oyster mushrooms (Pleurotus species), thus reducing the fiber content by 2.3-20.25% by weight. Another 2020 study (Int J Med Mushrooms. 2020;22(7):651-657. doi:10.1615 / IntJMedMushrooms.2020035449) also revealed that oyster mushroom (Pleurotus spp.) mycelium contains 22% insoluble fiber by weight. U.S. Patent Application Publication No. 2020 / 270559 discloses a specific method for producing an edible fungal biomat formulation and presents nutritional data from two Fusarium fungi with a total fiber content of up to 25% by weight and a fat content of 7% to 12% by weight.

[0013] In another respect, as disclosed in the developed methods in the publications US Pat. No. 4,041,189, US Pat. No. 4,501,765, and WO201802579, the RNA levels of currently available mycelial components are typically actively reduced by processes that include a final treatment step, which comprises a heating step at a certain temperature and / or adjusted pH for a certain time to actively reduce the RNA content to less than 4%, preferably less than 2%, on a dry basis, thus reducing undesired associated health risks and bitter taste and meeting regulatory requirements.

[0014] Jeng-Leun Mau (2015) summarized the equivalent umami concentration (EUC) values ​​of fruiting bodies and found that EUC values ​​varied widely, ranging from a high of 4465% (flat caps of Volvariella volvacea) to a low of 0.12% (Auricularia polytricha). EUC values ​​were grouped into four levels: >1000% (>10 g MSG / g dry matter), 100-1000% (1-10 g MSG / g), 10-100% (0.1-1 g MSG / g), and 10% (<0.1 g MSG / g), where MSG is the equivalent concentration of monosodium glutamate (MSG) (International Journal of Medicinal Mushrooms, Vol. 7, pp. 119-125 (2005)). Among the Pleurotus species, P. citrinopileatus had the highest EUC values ​​(511% at the second level), followed in descending order by P. eryngii small fruiting bodies (97.9%), P. cystidiosus (85.2%), and P. ostreatus (48.0%). Based on the morphology of fruiting bodies in culture jars or plastic bags (logs), the EUC values ​​of P. eryngii were 68.7, 97.9, and 32.1% for the large and small fruiting bodies and stalks, respectively. In a separate study, P. pulmonarius fruiting bodies were grown on three forestry waste materials (pine, poplar, and honeysuckle), which showed that the EUC values ​​of P. pulmonarius fruiting bodies ranged from 72.31% to 116.73% (Food Chemistry 397 (2022) 133714). It is understood that EUC is preferably expressed as g MSG / 100g dry matter, for example an EUC concentration of 1000% (or 1000% by weight) is equivalent to 1000g MSG / 100g dry matter.

[0015] In 1958, Eddy et al. reported several unsuccessful attempts to enhance the flavor of mycelium obtained from submerged fermentation based on patented inventions, e.g., U.S. Pat. No. 2,693,664 (J. Sci. Food Agric. 9 1958).

[0016] WO 2022 / 107388 discloses a method for providing an umami-enhancing composition of mushrooms of the genus Flammulina by enzymatic treatment, which results in a maximum reported equivalent umami concentration (EUC) value of 9.3 g / 100 g.

[0017] Chinese Patent No. 114027089 discloses a method for improving the flavor of edible mushrooms, in which the equivalent umami concentration value of Flammulina velutipes obtained by adding edible fungal root fermentation liquid increased the EUC from 4.48g / 100g to 10.8g MSG / 100g.

[0018] Chinese Patent No. 109156702(A) discloses a soaking method for Hericium erinaceus, which reduces the original EUC from 1131 g MSG / 100 g to the following values ​​by various treatments: steam treatment (959.82 g MSG / 100 g), water bath treatment (755.39 g MSG / 100 g), and ultrasonic treatment (189.84 g MSG / 100 g). A similar study was reported by Li-bin Sun et al. (Trends in Food Science & Technology 96 (2020) 176-187), in which mushroom fruiting bodies, rather than mycelium, were treated with various physical methods, which resulted in different EUC concentrations in the fruiting bodies.

[0019] Korean Patent No. 101535985 relates to a method comprising the step of initiating a Maillard reaction between powder of one or more mushroom varieties selected from the group comprising Yamabushitake (bearded tooth mushroom), Shiitake (Lentinula edodes), Oyster mushroom (Pleurotus ostreatus), and Enokitake (Flammulina velutipes) and other seasoning ingredients to reach a value of EUC of 176 mg MSG / 100 g. It should be noted that the measurements performed relate to the fruiting bodies of the fungi in question, not their mycelia.

[0020] Provided herein are three novel mycelial components A, B, and C that are chemically, biologically, physically, nutritionally, and organoleptically unique (i.e., characterized by improved, enhanced, or reduced taste, as applicable).

[0021] In one embodiment of the present invention, the mycelium provided is grown by submerged fermentation of at least one fungal strain in three different media, the three different media being a defined medium providing component A, a synthetic medium providing component B, or a natural medium comprising a sidestream extract selected from an agri-food sidestream providing component C.

[0022] In certain embodiments of the present invention, the mycelia provided have different carbon to nitrogen ratios, similar chitin content, but different portfolios of sugar content (sugars, polysaccharides, oligosaccharides).

[0023] In a further specific embodiment of the present invention, the mycelium provided has low indigenous RNA levels of at most 4% by weight, preferably at most 2% by weight, which avoids the need to have additional treatments to actively reduce RNA levels.

[0024] In a further specific embodiment of the present invention, the mycelium provided has an ergothioneine content of up to 800 mg / kg, which can be achieved within a shorter time compared to that reported in the prior art.

[0025] In further specific embodiments of the present invention, the mycelium provided has a specific / unique pore volume, pore size distribution, and / or characterized texture.

[0026] Further specific embodiments of the present invention relate to their adjustable insoluble fiber content of at least 20%, at least 30%, at least 40%, at least 50%, or at least 60% by weight, providing even higher health value of their high prebiotic insoluble fiber content, which is important for gut health.

[0027] In further specific embodiments of the present invention, with respect to their amino acid content, the amount of branched-chain amino acids (BCAAs) is at least about 19% by weight, at least about 20% by weight of the total amount of amino acids present (i.e., of the total protein).

[0028] In a further specific embodiment of the present invention, the protein content thereof is adjustable in the range of 10% to 65% by weight.

[0029] In a further specific embodiment of the invention, the umami amino acids in the mycelium component are at least about 20% by weight of the total amount of amino acids present.

[0030] In further specific embodiments, mycelium from at least one fungal strain is mixed with at least one protein-rich component, at least one lipid-rich component, and at least one compositional component to produce a meat or milk substitute.

[0031] In further specific embodiments, mycelia from at least one fungal strain have a very high equivalent umami concentration (EUC) ranging from up to at least 34%, more preferably at least 300%, and even more preferably at least 2800%, which is about 24-40 times higher than the EUC of the fruiting body of the same mushroom species and higher than the reported EUC values ​​of the mycelia contemplated in this invention.

[0032] Upon application of the mycelium component (any of A, B, and C) of the present invention, the flavor can also be significantly improved, since the typical natural umami flavor of milk, fish, or meat analogues can be brought about without the use of additional flavors, especially when the mycelium originates from a fungus that forms fruit bodies, such as a fungus selected from the Pleurotaceae family, for example, Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus citrinopileatus, Pleurotus florida, and Pleurotus salmoneostramineus, specifically when derived from Pleurotus pulmonarius, or when the mycelium is derived from a fungus selected from Morchella esculenta, Morchella angusticeps, Morchella deliciosa, and Morchella rufobrunnea, preferably Morchella rufobrunnea. This is due to the presence in such species of significant amounts of glutamate along with other amino acids (eg, aspartate) and / or 5'-nucleotides that are largely involved in the transduction of umami sensation.

[0033] In a further embodiment, the present invention relates to a method for producing fungal biomass by submerged fermentation of at least one fungal strain, wherein the at least one fungal strain is an edible fungus.

[0034] In still further embodiments, the present invention relates to methods for producing fungal biomass by submerged fermentation of at least one fungal strain, wherein the submerged fermentation is operated as a batch, fed-batch, or continuous process.

[0035] In a still further embodiment, the present invention relates to a method for producing fungal biomass by submerged fermentation of at least one fungal strain, wherein at least two fungal strains are co-fermented.

[0036] In a further embodiment, the present invention relates to a fungal biomass produced by the method of the present invention for producing a fungal biomass by submerged fermentation of at least one fungal strain, wherein the fungal strain is selected from the Pleurotaceae family, in particular the fungal strain is P. pulmonarius, P. ostreatus, P. citrinopileatus, or P. salmoneostramineus.

[0037] In a particular embodiment, the present invention relates to a fungal biomass produced by the method of the present invention for producing a fungal biomass by submerged fermentation of at least one fungal strain, wherein the fungal strain is selected from the Morchellaceae family, in particular the fungal strain is M. esculenta, M. angusticeps, or M. deliciosa.

[0038] In an alternative particular embodiment, the edible fibrous mycelial mass is obtained from at least one fungal strain, preferably at least one fungal strain capable of producing ergothioneine, preferably selected from Basidiomycota, Ascomycota, Hymenochaetaceae, Agaricomycetes, Sordariomycetes, Tremellomycetes, with preferred species herein being from at least one fungal species selected from Cordyceps spp., Inonotus spp., Grifola spp., Pleurotus spp., Ganoderma spp., Lentinula spp., Tremella spp., Trametes spp., Lepista spp., Tricholoma spp., Aspergillus spp., and / or Panus spp.

[0039] The present invention also relates to methods for producing the above-mentioned edible meat substitute, fish substitute, and dairy substitute products comprising the edible fibrous mycelial mass of the present invention, which methods are described in detail below.

[0040] The present invention also relates to the use of an edible fibrous mycelial mass for producing an edible meat substitute product, preferably selected from meatballs, sausages, tartare, minced meat, meat spreads, processed meat, met meat, foie gras, steak, beef jerky, burger patties, tenderloins, nuggets, salami, whole cuts, bacon, hot dogs, prosciutto, dried meat, and substitutes for extruded products.

[0041] The present invention also relates to the use of an edible fibrous mycelial mass for producing an edible dairy substitute, wherein the edible dairy substitute is selected from substitutes for milk, yogurt, fresh cheese, whey cheese, cream cheese, medium hard cheese, hard cheese, and soft mold cheese.

[0042] The present invention also relates to the use of edible fibrous mycelium mass for producing food substitute fish or seafood products, wherein the food substitute fish or seafood products are, for example, crab cakes, fish cakes, tuna, salmon, or shrimp.

[0043] In a further embodiment, the present invention relates to fungal-based food products prepared using the fungal biomass of the present invention.

[0044] In a further embodiment, the present invention also relates to the use of the supernatant produced during fermentation to develop specific health drinks containing antioxidants and specific aromas, tastes, and flavors generated during fermentation with edible mushrooms. Mushroom strains are known to produce pleasant volatiles and other compounds with, for example, apple or almond flavors. They also produce compounds known to regulate blood sugar levels.

[0045] The present invention also relates to the use of the further processed supernatant, for example to extract specific components thereof produced by the microorganism cultivated in the medium, such as proteins, in particular enzymes, polysaccharides, peptides, antioxidants, etc. [Brief explanation of the drawings]

[0046] [Figure 1] An overlay of pore size distribution curves (-dV / dlogD) of mycelial components A, B, and C calculated from normalized volume curves is shown. [Figure 2] 1 shows a thermogravimetric analysis curve for component A, illustrating the effect of temperature versus weight of the sample. Weight is expressed in terms of the percentage of sample remaining versus the weight at the start of the experiment at a given temperature / time. The second y-axis on the graph presents data for the first derivative of the TGA curve. This is known as the derivative thermogravimetric (DTG) curve and represents the rate of change of mass with respect to temperature (e.g., % mass loss per degree Celsius). [Figure 3] Figure 1 shows a thermogravimetric analysis curve for component B, illustrating the effect of sample temperature versus weight. Weight is expressed in terms of the percentage of sample remaining versus the weight at the start of the experiment at a given temperature / time. The second y-axis on the graph presents data for the first derivative of the TGA curve. This is known as the derivative thermogravimetric (DTG) curve and represents the rate of change of mass versus temperature (e.g., % mass loss per degree Celsius). [Figure 4] 1 shows a thermogravimetric analysis curve for component C, illustrating the effect of temperature versus weight of the sample. Weight is expressed in terms of the percentage of sample remaining versus the weight at the start of the experiment at a given temperature / time. The second y-axis on the graph presents data for the first derivative of the TGA curve. This is known as the derivative thermogravimetric (DTG) curve and represents the rate of change of mass with respect to temperature (e.g., % mass loss per degree Celsius). [Figure 5] The beef, chicken, green peas, and soybeans used in the comparative experiments are shown. DETAILED DESCRIPTION OF THE INVENTION

[0047] The present invention is described in detail below. It should be understood that all disclosed features can be combined with each other unless expressly stated to the contrary. In particular, features disclosed in different embodiments can be combined with each other unless expressly stated that they are not combinable.

[0048] The mycelial components A, B, and C of the present invention have unique organoleptic and biological / physical / chemical properties.

[0049] In particular, the present invention provides an edible mycelial ingredient comprising undifferentiated mycelial biomass, wherein the undifferentiated mycelial biomass has an elemental composition of the mycelium with a C:N ratio in the range of 2 to 12 (preferably 2 to 8, more preferably 2 to 6), and wherein the edible mycelial ingredient is characterized by an EUC of at least 500g MSG / 100g. As understood herein, undifferentiated mycelial biomass may be obtained, for example, in the course of submerged fermentation.

[0050] This particularly preferred edible mycelium component of the present invention may also be referred to as Component C.

[0051] EUC is

[0052]

number

[0053] In an alternative embodiment, the EUC comprises:

[0054]

number

[0055] Therefore, the present invention provides an edible mycelium ingredient characterized by a particularly very high EUC value, in other words, an ingredient with a very strong umami taste profile. The ingredient of the present invention has an EUC of at least 500g MSG / 100g, preferably at least 1000g MSG / 100g, more preferably at least 1500g MSG / 100g, and even more preferably at least 2000g MSG / 100g.

[0056] The composition of the present invention is further characterized by a 5'-AMP content of 3.5 to 10.0 g / kg and / or a 5'-GMP content of 3.5 to 10.0 g / kg. Preferably, the composition of the present invention is characterized by a 5'-AMP content of 4.5 to 6.5 g / kg and / or a 5'-GMP content of 4.5 to 6.5 g / kg. According to the present inventors, the composition of the present invention is preferably substantially free of 5'-IMP. In other words, the composition of the present invention is free of 5'-IMP.

[0057] As understood herein, preferably the sensory or morphological properties are selected from taste attributes, smell attributes, aroma attributes, mouthfeel attributes, texture attributes, consistency, edibility, and color. This list should be interpreted as exemplary and preferred, but not limiting. Those skilled in the art are in a position to extend the method by including one or more further sensory or morphological properties.

[0058] Preferably, as understood herein, taste and smell attributes can be determined by a taste panel composed of individuals who evaluate the taste and / or smell of provided samples. Preferably, the taste and smell panel is performed in parallel on several samples and includes a specific reference sample for normalization of the evaluation. Thus, in an exemplary manner of performing a taste panel, each trained panelist is blinded and receives samples sequentially. The panelists define the sensory attributes they recognize in the sample, consider the attributes together, select common attributes that all panelists can associate with the same taste and aroma of the sample and the reference sample, and compare them. Then, a second session begins, in which the panelists must evaluate the samples according to the selected attributes and assign a score, for example, a score of 0 to 5, for each attribute. The session can be repeated on different days to increase the statistical relevance of the data, and the average scores can be calculated and plotted in a spider web.

[0059] Preferably, the texture and mouthfeel attributes may also be determined by a specific panel.

[0060] The colors referred to herein are preferably determined at several locations on the sample, for example, 20 different locations, using an RGB system and a color analyzer. The average of these measurements is then used to compare the colors of the products. Preferably, a calibrated imaging device is used to determine the colors.

[0061] In the present invention, the nutritional properties are preferably selected from sugar content, amino acid composition, metabolite content, mineral content, vitamin content, carbohydrate content, fiber content, fatty acid content, lipid content and protein content, functional substance content, and / or C, H, N, O, S content. This list should be interpreted as exemplary and preferred, but not as limiting. The skilled person is in a position to extend the method by including one or more further nutritional properties.

[0062] As understood herein, in the present invention, sugar content or carbohydrate content preferably refers to the % w / w or weight % content of a biomass or product of the present invention, preferably expressed in terms of the dry mass of said biomass or said product. Information on sugar content may also include further details, such as the content of complex and simple carbohydrates, including the breakdown into pentoses or hexoses. The content of different types of sugars / carbohydrates referred to herein may also be expressed in % w / w or weight % of the total sugar / carbohydrate content of the product or biomass.

[0063] As understood herein, unless indicated to the contrary, the terms % w / w and % by weight are meant to be interchangeable.

[0064] As understood herein, in the present invention, amino acid composition preferably refers to the % w / w content of each amino acid relative to the total amino acid content in the biomass or product of the present invention. As known to those skilled in the art, in certain methods of amino acid analysis, it is not possible to distinguish between aspartate and asparagine, and between glutamate and glutamine, due to the hydrolysis conditions used in the process. Therefore, the Asp / Asn content and the Glu / Gln content are expressed as the total content of the two amino acids in each of these pairs.

[0065] As understood herein, umami amino acids are the aspartic acid (aspartate) and glutamic acid (glutamate) amino acids that contribute to flavor, and branched-chain amino acids (BCAAs) are a group of three essential amino acids, leucine, isoleucine, and valine, that are involved in muscle growth, athletic performance, weight loss, and reduced fatigue.

[0066] Umami or savory taste is also defined by 5'-ribonucleotides or 5'NMPs, which include the following 5'-nucleotides (5'-NMP), 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP), and 5'-adenosine monophosphate (AMP), preferably expressed in g / kg mycelial component.

[0067] 5'-GMP is known to impart a meaty flavor and is a much more potent flavor enhancer than monosodium glutamate (MSG) (JH Litchfield, "Morel Mushroom Mycelium as a Food Flavoring Material," Biotechnology and Bioengineering, 9 (1967), pp. 289-304). It has also been reported that a synergistic effect exists between umami 5'-nucleotides and umami amino acids, which can greatly enhance the umami flavor of mushrooms (Yamaguchi S, Yoshikawa T, Ikeda S, Ninomiya T. Measurement of the relative taste intensity of some α-amino acids and 5'-nucleotides. J. Food Sci. 1971;36:846-49). Yamaguchi (1967) then derived this synergistic effect of 5'-GMP or 5'-GMP versus MSG in a linear relationship, expressed as follows: EUC=A+1218(A)(N) where EUC (equivalent umami concentration) is the equivalent concentration of monosodium glutamate (MSG), a measure that takes into account the relative taste intensity of umami amino acids and 5'-nucleotides, since the intensities of umami amino acid taste and flavor nucleotides have been shown to be proportional to those of MSG versus IMP or GMP, respectively. EUC is expressed in g MSG per 100 g dry weight, and A and N are the concentrations of the amino acids and nucleotides, respectively, expressed in terms of the concentrations of MSG and GMP in solution. 1218 is a positive constant when IMP is used based on the concentration used in g / 100 g. This constant is 2800 when GMP is used for normalization; both yield the same results within acceptable standard deviations (Yamaguchi S, Yoshikawa T, Ikeda S, Ninomiya T. Measurement of the relative taste intensity of some α-amino acids and 5'-nucleotides. J Food Sci. 1971;36:846-49). In this case, both amino acids and nucleotides are considered to quantify the final taste or equivalent umami concentration relative to an equivalent concentration of MSG.

[0068] In further detail, the calculation is

[0069]

number

[0070] As understood herein, in the present invention, nucleic acid content refers to the total nucleic acid content (DNA and RNA) of a biomass or product as defined herein, preferably stated in % w / w or weight % of the dry mass of the biomass or product.

[0071] As understood herein, in the present invention, metabolite content refers to the amount of each metabolite in a biomass or product, expressed in mg per gram of biomass or product, respectively. For example, metabolite content can refer to any metabolite selected from those known to those skilled in the art. As referred to herein, metabolites typically refer to products of the metabolism of fungal species. Exemplary metabolites include alcohols, amino acids, nucleotides, antioxidants, organic acids (e.g., acetic acid, lactic acid), polyols (e.g., glycerol), and vitamins. However, this list is not meant to be construed as limiting.

[0072] As understood herein, in the present invention, mineral content refers to the content of any of the minerals considered essential in human nutrition, which content can be expressed in mg / kg of biomass or product for each mineral, mg / kg of biomass or product referring to the dry mass of the biomass or product. Preferably, the minerals referred to herein are selected from calcium, phosphorus, potassium, sodium, chloride, magnesium, iron, zinc, iodine, chromium, copper, fluoride, molybdenum, manganese and selenium.

[0073] As understood herein, in the present invention, vitamin content preferably refers to the content of a particular vitamin, expressed in μg / kg of dry biomass or dry product. Vitamins are known to those skilled in the art and include, inter alia, vitamin A, vitamin B 12 , Vitamin B1, Vitamin B3, Vitamin B6, Vitamin C, Vitamin D, Vitamin E, Vitamin K, and Vitamin O.

[0074] It is understood that the final fungal-derived product based on the disclosed mycelium component may include the supernatant obtainable in the production process of the mycelium component of the present invention. Thus, the final fungal-derived product based on the disclosed mycelium component may be the supernatant itself or the biomass (i.e., the mycelium component), or a combination thereof, or any related extract from each individual product or combination thereof. Thus, as will be apparent to those skilled in the art, the fungal component may be applicable to at least one form of these products.

[0075] As understood herein, in the present invention, fungal compositions preferably relate to those compositions of biomass or products derived from a species / strain, i.e., biomass or products derived from the mycelium component disclosed in the present invention.

[0076] As understood herein, in the present invention, biomass dry weight is the weight of the biomass obtained upon dehydration / water removal, preferably the weight of the biomass obtained after washing off the residual medium, preferably the weight of the biomass extrapolated to a minimum water content of 0% w / w.

[0077] As understood herein, whenever any content or any ratio is referred to, preferably determined "on a dry basis" or "on a dry weight basis", it refers to the material upon dehydration / water removal, preferably after washing away residual medium, preferably extrapolated to a minimum of 0% w / w water content.

[0078] As understood herein, in the present invention, fiber content preferably refers to the % w / w content of dietary fiber in the dry biomass or dry product.

[0079] As understood herein, in the present invention, protein content preferably refers to the % w / w content of protein in the dry biomass or dry product. Preferably, measured protein content refers to the protein content determined by the Kjeldahl method.

[0080] As understood herein, in the present invention, fatty acid content preferably refers to the % w / w content of fatty acids in the dry biomass or dry product.

[0081] As understood herein, in the present invention, lipid content preferably refers to the % w / w content of lipids in the dry biomass or dry product.

[0082] The terms mycelial ingredient or mushroom mycelial ingredient or mycelium ingredient or fungal ingredient or mycelium / mycelial biomass are understood to be synonymous.

[0083] As understood herein, in the method of the present invention, product titer preferably refers to the concentration of the product obtained, preferably expressed in g / L. In this specification, the term product preferably refers to fungal biomass, fungal metabolites, i.e. compounds obtainable from mycelium, colorants, dietary supplements, active compounds, enzymes, or cosmetics.

[0084] As understood herein, in the methods of the present invention, cultivation or fermentation conditions include data necessary to repeat a cultivation experiment, i.e., temperature, CO content / emission / production or other volatile gas emissions, agitation, humidity, pH, dissolved oxygen concentration, dissolved CO concentration, etc. This list is not meant to be limiting, as cultivation conditions will be apparent to those skilled in the art.

[0085] As understood herein, in the methods of the present invention, metabolic behavior preferably includes transcriptome information, metabolome information, proteome information, secretome information, and / or fluxome information. It is understood that the secretome preferably includes information regarding the structure and / or quantity of compounds produced by the fungal biomass and secreted outside the fungal cell, such as metabolites / proteins that may be secreted by the organism into the fermentation broth. As will be understood by those skilled in the art, the secretome can include valuable compounds that can be used for products, such as vitamins, enzymes, pigments, or mycelium-derived functional or active compounds. Therefore, information regarding the secretome can inform efforts to produce secreted compounds in processes involving fungal biomass.

[0086] Preferably, the molecular and metabolite information relates to metabolites present in or obtainable from a particular fungus. Metabolites should therefore preferably be understood as all nodes of a metabolic pathway map of a specific species under investigation. Information regarding the structure and abundance of specific metabolites is preferably included herein.

[0087] As understood herein, in the present invention, the fermentation medium preferably comprises at least one fungal strain, optionally other microorganisms co-cultured with the fungal strain (preferably comprising algae, bacteria, plant cells, archaeal cells, animal cells, adipocytes, or a combination thereof), and / or side streams from the agri-food industry.

[0088] As understood herein, chitin is a compound of (CH 13 O5N) n Chitin is a polysaccharide composed of connected N-acetylglucosamine subunits with the chemical formula: where n is the number of subunits. It is understood that n is a natural number. Chitin and its degradation products via the enzyme chitinase, present in humans and other mammals, are sensed in the skin, lungs, and digestive tract, eliciting an immune response potentially directed toward combating parasites. Additionally, chitin is commonly used as a food additive to improve flavor and as an emulsifier. Chitin also has anti-inflammatory properties, reduces cholesterol, and is beneficial for weight loss and blood pressure. Chitin can also be used to produce biomaterials or biodegradable packaging materials, and can be used in the food industry or other industries. Chitin can also be extracted from mushrooms or mycelium, making it a compound used as sausage casing.

[0089] Preferably, the production of functional compounds includes information on compounds produced by mycelia. Preferably, the functional compound, which may also be referred to as an active compound, refers to any substance that has a beneficial (demonstrated or proven) effect on biological function, and the functional compound is preferably a mycelium-derived active compound selected from ergothioneine, lovastatin, ergosterol, resveratrol, glutathione, eritadenine, lentinan, and concanavalin A. However, this list is not intended to be construed as particularly limiting, and additional compounds produced in mycelia recognized by those skilled in the art may also be included. Exemplary compounds derived from Pleurotus ostreatus have recently been reviewed (Mishra et al., Int J Biol Macromol, 2021, 182, 1628-1637).

[0090] Ergothioneine is a sulfur-based amino acid found primarily in mushrooms and adzuki / black beans. Its antioxidant and anti-inflammatory properties are known to prevent chronic diseases of aging, such as heart or brain-related diseases, and can protect against cell and tissue damage in the body. Ergothioneine is often considered a longevity vitamin due to its beneficial effects. Preferably, ergothioneine has the formula:

[0091] [ka] or a salt thereof.

[0092] Oyster mushrooms are a major source of nutritional supplements and are known to have numerous therapeutic properties, such as antitumor, antiangiogenic, immunomodulatory, antioxidant, and antidiabetic mediation roles. Such macromolecules include β-glucan, α-glucan, ergosterol, linoleic acid, etc. It has also been shown that ergosterol intake can increase serum and liver vitamin D levels.

[0093] Polyphenols are the most abundant antioxidants consumed in the human diet. Total phenolic content (TPC) refers to phenolic compounds that have redox properties that contribute to antioxidant activity and are known to have potential beneficial effects on human health. Similarly, phytonutrients, such as flavonoids, also have anti-inflammatory properties and can act as antioxidants, protecting cells from oxidative damage that leads to disease.

[0094] The total phenolic content was measured by the Folin-Ciocalteu assay, preferably expressed in mg GAE per gram of biomass, where GAE is gallic acid equivalent. The total flavonoid content (TFC) was measured by an aluminum chloride colorimetric assay, preferably expressed in mg QE per gram of biomass, where QE is quercetin equivalent. Further polyphenol profiling was performed by high-performance liquid chromatography with diode-array detection (HPLC-DAD) to examine the content of potential polyphenols present in the biomass, such as catechin, vanillin, quercetin, chlorogenic acid, 3,4-dihydroxybenzoic acid (protocatechuic acid), salicylic acid, p-coumaric acid, syringic acid, vanillic acid, caffeic acid, ferulic acid, and gallic acid. The concentrations of the following substances are preferably expressed in mg / g: These methods are well known to those skilled in the art.

[0095] Catechin, in particular, is a natural antioxidant that helps prevent cell damage and provides other benefits. Quercetin is a plant pigment (flavonoid) that can reduce swelling, promote cancer cell death, and help prevent heart disease. Chlorogenic acid, found primarily in coffee and other foods, has been extensively investigated in neurodegenerative diseases due to its anti-inflammatory activity. 3,4-Dihydroxybenzoic acid (protocatechuic acid), commonly found in green tea, is known to have mixed effects on normal and cancer cells. Protocatechuic acid is considered an active component of several traditional Chinese herbal medicines, such as Cibotium barometz (L.) (Functional Foods in Health and Disease, vol. 7, pp. 232-244, 2011). For example, acai oil sourced from acai palm fruit (Euterpe oleracea) is rich in protocatechuic acid (630 mg / kg) (Journal of Agricultural and Food Chemistry, vol. 56, no. 12, pp. 4631-4636, 2008).

[0096] As understood herein, the C:N ratio (mycelium) preferably relates to the ratio between the carbon content and the nitrogen content in the mycelium component, and the C:N ratio (medium) preferably relates to the ratio between the carbon content and the nitrogen content in the fermentation medium, i.e., in the fermentation broth, which ratio is preferably understood herein as w / w ratio.

[0097] As understood herein, the degree of complexity of fermentation media can be divided into three categories: synthetic media, defined media, and complex media.

[0098] A synthetic medium is preferably a medium that is free of complex compounds, i.e., the concentrations of all components are known. For example, a medium that does not contain a complex nitrogen source is a synthetic medium.

[0099] A defined medium is preferably a medium that contains at most one complex compound (eg, yeast extract) and potential compounds with unknown concentrations.

[0100] Complex media are completely undefined compositions, preferably containing two or more complex compounds and unknown substances or natural extracts with unknown concentrations (e.g., compounds extracted from agricultural by-streams or waste streams from the food industry). Preferably, complex media are defined through their production process.

[0101] The Brunauer-Emmett-Teller (BET) surface area is a method used to characterize porous and finely dispersed solids via gas adsorption. During gas adsorption (DIN-ISO 9277, respectively, according to DIN 66131), the specific surface of a solid material is determined using the BET method, by default with nitrogen adsorption at 77.4 K. The evaluation is carried out in the respective relative pressure ranges described, in the general region of validity of the BET method, where p / p = 0.05 to 0.3. For the determination of very small surfaces, krypton adsorption (at 77.4 K) is used. Since the expected surface areas are very low, krypton is used because it is a suitable adsorbate for measuring low surface areas. The specific surface area is m 2 It is expressed as / g.

[0102] By median pore diameter, it is meant that 50% of the pore volume comes from pores larger than the median pore diameter, and the other 50% of the pore volume comes from pores smaller than the median pore diameter.

[0103] The specific pore volume can be determined from nitrogen adsorption measurements if the adsorbent is meso- or microporous. For macroporous adsorbents with pore diameters greater than 1000 Å, the pore volume can be determined by mercury porosimetry measurements by integrating the pressure-volume curve. The method is based on the Washburn equation, which describes the relationship between pore diameter and the pressure applied to a non-wetting liquid, such as mercury.

[0104] Fermentation processes, preferably biochemical processes, are not meant to be particularly limiting, and as will be appreciated by those skilled in the art, any process involving microorganisms can be encompassed by the present invention. Cultivation processes (e.g., production of biomass, in particular production of fungal biomass) and bioproduction processes (e.g., expression of enzymes in microbial cultures, bioproduction of ethanol, production of intracellular or extracellular compounds, such as colorants, flavor compounds, antioxidants, enzymes, moisturizing compounds, and similar processes) are particularly preferred.

[0105] Thus, the submerged fermentation in the method of the present invention can be operated as a batch process, a fed-batch process or a continuous process, these three main fermentation methods being known to those skilled in the art and differing in the flow of materials out of and into the fermentation vessel.

[0106] A batch process is characterized by the absence of any input of materials into the fermentation vessel. In a batch process, all nutrients are provided at the beginning of the culture, without further addition in subsequent bioprocesses. No additional nutrients are added during the entire bioprocess, except for gas, acid, and base. The bioprocess then continues until the nutrients are consumed. This strategy is suitable for rapid experiments such as strain characterization or nutrient medium optimization. A drawback of this convenient method is the limited yield of biomass and product. Typically, the microorganisms are not in logarithmic growth phase for long periods of time because the carbon source and / or oxygen transfer are limiting factors. After the end of a bioprocess run in batch mode, only the biomass or medium is recovered and appropriately processed to obtain the desired product. From the bioreactor's perspective, the process is repeatedly interrupted by cleaning and sterilization steps, and biomass is simply produced in stages.

[0107] In a fed-batch process, substrates, nutrients, and other substances can be added to a fermentation vessel to, among other things, extend the possible culture time or increase yield. The advantage of feeding during cultivation is that it allows for a higher overall production yield to be achieved. Under certain growth conditions, microorganisms and / or cells constantly double and thus follow a logarithmic growth curve. Therefore, in certain embodiments, the feed rate can be increased logarithmically as well. Typically, substrate is pumped from a feed bottle into the culture vessel, for example, through silicone tubing. The user can manually set the feed at any time (linear, logarithmic, or pulsed) or add nutrients when certain conditions are met, such as when a certain biomass concentration is reached or when nutrients are depleted. Fed-batch processes offer a wide range of control strategies and are also suitable for highly specialized applications. However, they can increase processing time and potentially lead to inhibition due to the accumulation of toxic by-products.

[0108] In the method of the present invention, submerged fermentation is preferably operated as a continuous process. After a batch growth phase, equilibrium is established with respect to certain components (also called steady state). Under these conditions, fresh culture medium is added in an amount equal to that removed (chemostat). These bioprocesses, called continuous cultures, are particularly suitable when excess nutrients cause inhibition, for example, due to acid or ethanol accumulation or excessive heating. After reaching steady state, continuous mode is understood to always operate in the logarithmic growth phase, where cells are maintained at a constant concentration. The transient state is the state before a certain steady-state condition is reached in continuous mode and is actually similar to the start of batch mode operation. Other advantages of this method include reduced product inhibition and improved space-time yield. Cells are recovered when medium is removed, which is why the inflow and outflow rates must be less than the doubling time of the microorganism. Alternatively, cells can be retained in a variety of ways (e.g., spin filters) called perfusion. In continuous processes, the space-time yield of the bioreactor can be further improved compared to that of fed-batch processes. However, long culture periods also increase the risk of contamination and long-term changes in the culture. The three most common types of continuous culture are chemostat (the rate of addition of a single growth-limiting substrate controls cell growth), turbidostat (an indirect measurement of cell number, i.e., turbidity or optical density, which requires an additional sensor, is driven by real-time feedback to control the addition and removal of fluid), and perfusion (this type of continuous bioprocessing mode is based on either retaining cells in the bioreactor or recycling cells back to the bioreactor, providing fresh medium and removing cell-free supernatant at the same rate).

[0109] In one embodiment of the invention, in the method of the invention, the submerged fermentation is not operated as a continuous process.

[0110] In a preferred embodiment, the edible fibrous mycelial mass is derived from submerged fermentation. Submerged fermentation allows mycelium to grow without the need for a substrate to structurally support the mycelium, as in solid-state fermentation. Furthermore, growth rates are higher because nutrients can be transported to all points of the mycelium and it is easier to maintain sterile conditions. Depending on the fermentation conditions, mycelium has the potential to grow into either pellets or threads in submerged fermentation. This structure is maintained at harvest and, when gently processed, imparts different textural characteristics to the resulting product, such as a cheese product.

[0111] Furthermore, submerged fermentation offers the additional advantage that washing of the mycelium grown in particular in submerged fermentation can be easily accomplished, resulting in a clean product that is not contaminated with residues of the solid substrate that may subsequently result in an unpleasant taste or allergies. Likewise, by-products from fermentation, such as acids or alcohols that may also alter the taste, can also be easily removed.

[0112] Also, in submerged fermentation, it is easier to better control fermentation conditions, such as pH and oxygen content, thus allowing for maximum mycelial growth compared to substrate limitation in solid-state fermentation, resulting in a more homogeneous product composition and reduced batch-to-batch variability.

[0113] Furthermore, the material obtained by submerged fermentation is more malleable and can be formed into any kind of shape. For example, the mycelium can be homogenized into a liquid that can be fermented, which is particularly advantageous if the mycelium is used to produce dairy substitutes. The mycelium obtained from solid-state fermentation is usually dried and therefore needs to be resuspended in water to obtain a milk-like substance.

[0114] Furthermore, in submerged fermentation, the fermentation medium can be agitated, which makes it easy and efficient to prevent sporulation or fruiting body formation, which can lead to toxin production. It is further apparent to those skilled in the art that when growth is carried out by submerged fermentation, differentiation of the mycelium into spores and / or fruiting bodies preferably does not occur. Therefore, it is apparent to those skilled in the art that the mycelial mass derived from submerged fermentation can also be referred to as undifferentiated mycelial biomass. Those skilled in the art will recognize further advantages of using submerged fermentation, such as better control over the texture and composition of the biomass compared to solid-state fermentation.

[0115] Furthermore, submerged fermentation methods are easier to scale up because a first culture can be inoculated into a second culture, since both cultures are in a liquid state. This is not easily possible in solid-state fermentation because the first culture cannot flow. This drawback can be partially ameliorated by using a rotating drum reactor, which rotates the reactor to mix the solid and mycelium. However, drum reactors are known to have a major problem: heat transfer during fermentation can result in different temperatures within the reactor. Because heat cannot be properly removed, some of the fermentation volume may be overheated at some point, or some of the fermentation volume may be underheated, especially at the beginning of the fermentation. In submerged fermentation, heat transfer is more efficient because liquid is a good medium for transferring heat, resulting in a more uniform heat distribution and therefore a more controllable and efficient fermentation process.

[0116] Therefore, preferably, as encompassed by the present invention, the process is carried out with at least one fungal species. The at least one fungal species may be combined with an edible fungus. The at least one fungal species may be combined with an edible fungus, algae, bacteria, plant cells, archaeal cells, animal cells, fat cells, or a combination thereof.

[0117] Preferably, in one embodiment, currently discovered or yet to be discovered edible fungi and the co-cultivation or co-fermentation of such edible fungi with each other are used. In another embodiment, the above embodiment is further combined with the use of algae or bacteria or plants or archaea or animal cells / adipocytes, or combinations thereof.

[0118] Preferably, the at least one fungal species is selected from the group consisting of Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Pezizomycetes, Agaricomycetes, Sordariomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricomyceae, Marasmiaceae, Canth arellaceae, Hydnaceae, Boletaceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizop selected from hyllaceae, Sclerodermataceae, Ganodermaceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinaceae.

[0119] In one embodiment, currently discovered or yet to be discovered edible fungi and the co-cultivation or co-fermentation of such edible fungi with each other are used. In another embodiment, the above embodiment is further combined with the use of algae or bacteria or plants or archaea or animal cells / adipocytes, or combinations thereof.

[0120] According to the present invention, the at least one fungal strain may be selected from the phylum Basidiomycota. Preferably, the at least one fungal strain selected from Basidiomycota may be a fungal strain selected from the subphylum Agaromycotina. As defined herein, a fungal strain selected from the subphylum Agaromycotina may be a fungal strain selected from the class Agaricomycetes. Preferably, a fungal strain selected from the Agaricomycetes may be a fungal strain selected from the orders Agaricales, Auriculariales, Boletales, Cantharellales, Polyporales, and Russulales.

[0121] If the fungal strain is selected from the order Agaricales, the fungal strain is preferably selected from the families Agariaceae, Fistulinaceae, Lyophyllaceae, Marasmiaceae, Omphalotaceae, Physalacriaceae, Pleurotaceae, Schizophyllaceae, Strophariaceae, and Tricholomataceae.

[0122] The fungal strain selected from the Agaricaceae family can be Agaricus bisporus or Agaricus blazei, more preferably Agaricus bisporus.

[0123] The fungal strain selected from the Fistulinaceae family is preferably Fistulina hepatica.

[0124] The fungal strain selected from the Lyophyllaceae family is preferably Calocybe indica.

[0125] The fungal strain selected from the Marasmiaceae family is preferably Lentinula edodes.

[0126] The fungal strain selected from the Omphalotaceae family is preferably Calvatia gigantea.

[0127] The fungal strain selected from the Physalacriaceae family is preferably Flammulina velutipes.

[0128] More preferably, the at least one fungal strain selected from Agaricales may be a fungal strain selected from the Pleurotaceae family. Even more preferably, the at least one fungal strain of the present invention is selected from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus citrinopileatus, Pleurotus florida, Pleurotus eunosmus, Pleurotus columbinus, Pleurotus ferulae, Pleurotus salmoneo-stramineus, and Pleurotus salmoneostramineus, even more preferably from Pleurotus pulmonarius or Pleurotus ostreatus, and most preferably from Pleurotus pulmonarius.

[0129] The fungal strain selected from the Schizophyllaceae family is preferably Schizophyllum commune.

[0130] The fungal strain selected from the family Strophariaceae is preferably a fungal strain selected from Agrocybe aegerita and Hypholoma capnoides.

[0131] The fungal strain selected from the Tricholomataceae family is preferably a fungal strain selected from Hypsizygus tesselatus and Clitocybe nuda.

[0132] Alternatively, the fungal strain selected from the Agaricomycetes may be a fungal strain selected from the order Auriculariales, more preferably a fungal strain selected from the family Auriculariaceae. Preferably, the fungal strain selected from the Auriculariaceae is Auricularia auricula-judae.

[0133] If the fungal strain is selected from the order Boletales, the fungal strain is preferably selected from the families Boletaceae and Sclerodermataceae. A fungal strain selected from the family Boletaceae is preferably Boletus edulis.

[0134] When the fungal strain is selected from the order Cantharellales, the fungal strain is preferably selected from the families Cantharellaceae and Hydnaceae. The fungal strain selected from Cantharellaceae can be Cantharellus cibarius. The fungal strain selected from Hydnaceae can be Hydnum repandum.

[0135] When the fungal strain is selected from the order Polyporales, the fungal strain is preferably selected from the families Ganodermataceae, Meripillaceae, Meruliaceae, Polyporaceae, and Sparassidaceae, more preferably Ganodermataceae, Meripillaceae, Polyporaceae, and Sparassidaceae.

[0136] The fungal strain selected from Meripilaceae is preferably Grifola frondosa. The fungal strain selected from Polyporaceae can be from Polyporus umbellatus and Laetiporus sulphureus (L. sulphureus). The fungal strain selected from Sparassidaceae can be Sparassis crispa. The fungal strain selected from Meruliaceae is preferably from B. adusta and B. fumosa.

[0137] When the fungal strain is selected from the order Russulales, the fungal strain may be selected from the families Bondarzewiaceae and Hericiaceae. Preferably, the fungal strain selected from Russulales is selected from the family Hericiaceae, preferably from Hericium erinaceus and Hericium coralloides. The fungal strain selected from Bondarzewiaceae may be Bondarzewia berkeleyi.

[0138] According to the present invention, the at least one fungal strain may be selected from the phylum Ascomycota. Preferably, the at least one fungal strain selected from Ascomycota may be a fungal strain selected from the subphylum Pezizomycotina.

[0139] The fungal strain selected from Pezizomycotina may be selected from the class Pezizomycetes. Preferably, the fungal strain selected from Pezizomycetes may be selected from the order Pezizales. Preferably, the fungal strain selected from Pezizales may be selected from the families Morchellaceae and Tuberaceae.

[0140] In a preferred embodiment, the mycelium is not from the Sordoriomycetes, and in particular not from the genus Neurospora, such as Neurospora crassa, or the genus Fusarium, such as Fusarium venenatum.

[0141] Preferably, the fungal strain selected from the Morchellaceae family is Morchella esculenta, Morchella angusticeps, Morchella deliciosa, Morchella sceptrifomtis, Morchella steppicola, Morchella puncripes, Morchella rufobrunnea, Morchella importuna, Morchella Jaurentinaa, or Morchella purpumscens, preferably Morchella esculenta, Morchella angusticeps, or Morchella deliciosa.

[0142] Preferably, the fungal strain selected from the Tuberaceae is Tuber magnatum, T. estivum, T. uncinatum, T. indicum, T. rufum, or T. melanosporum, more preferably T. melanosporum and T. magnatum.

[0143] Alternatively, the at least one fungal strain selected from Ascomycota may be a fungal strain selected from the class Sordariomycetes.

[0144] Preferably, the fungal strain selected from the Sordariomycetes may be a fungal strain selected from the order Hypocreales.

[0145] The fungal strain selected from the Hypocreales may be a fungal strain selected from the family Cordycipitaceae. The fungal strain selected from the Cordycipitaceae may be a fungal strain selected from Cordyceps militaris and Cordyceps sinensis.

[0146] Alternatively, the fungal strain selected from Hypocreales may be a fungal strain preferably selected from the family Nectriaceae. The fungal strain selected from Nectriaceae may be a Fusarium strain, for example Fusarium venenatum.

[0147] In another embodiment, the fungal strain selected from the Sordariomycetes can be a fungal strain selected from the family Sordariaceae. The fungal strain selected from the Sordariaceae can be a Neurospora strain, for example, Neurospora crassa.

[0148] Preferably, the edible fibrous mycelial mass is selected from the group consisting of Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Pezizomycetes, Agaricomycetes, Sordariomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricomyceae, Marasmiaceae, Cantharellaceae ae, Hydnaceae, Boletaceae, Meripillaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinacea.

[0149] More preferably, the edible fibrous mycelial mass is selected from the group consisting of Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Agaricomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, B The fungal extract is obtained from at least one fungal strain selected from the group consisting of: Oletaceae, Meripillaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinacea.

[0150] Even more preferably, the mycelial mass is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus florida, Pleurotus citrinopileatus, Pleurotus salmoneostramineus, Morchella esculenta, Morchella angusticeps, or Morchella deliciosa.

[0151] Even more preferably, the mycelial mass is obtained from Pleurotus pulmonarius, Pleurotus florida, Pleurotus citrinopileatus, Pleurotus salmoneostramineus, Morchella esculenta, Morchella angusticeps, or Morchella deliciosa, or Morchella rufobrunnea.

[0152] Most preferably, the mycelial mass is obtained from Pleurotus pulmonarius or Morchella rufobrunnea.

[0153] In a preferred embodiment, the mycelial mass is obtained from Pleurotus pulmonarius. In another preferred embodiment, the mycelial mass is obtained from Morchella rufobrunnea. In a preferred embodiment, the mycelial biomass comprises Pleurotus pulmonarius and / or Morchella rufobrunnea. In a further preferred embodiment, the mycelial biomass comprises Pleurotus pulmonarius and Morchella rufobrunnea.

[0154] In one embodiment, the mycelial biomass is obtained from L. sulphureus or B. adusta.

[0155] In a separate embodiment, the edible fibrous mycelial mass is obtained from at least one fungal strain selected from Basidiomycota, Ascomycota, Hymenochaetaceae, Agaricomycetes, Sordariomycetes, Tremellomycetes, with preferred species herein being from at least one fungal species selected from Cordyceps, Inonotus, Grifola, Pleurotus, Ganoderma, Lentinula, Tremella, Trametes, Lepista, Tricholoma, Aspergillus, and / or Panus. Thus, preferably, the at least one fungal strain is an ergothioneine-producing strain.

[0156] Typically, a constant temperature is maintained throughout the process, which can be selected for optimal growth of a particular fungal strain, as known to those skilled in the art. For example, for P. ostreatus, cultivation is preferably carried out at a temperature of 25-30°C. More preferably, cultivation is carried out at a pH of 3.0-8.5. As will be appreciated by those skilled in the art, the choice of pH may depend on the fungal strain being cultivated or on potential contaminating strains to be excluded from growth. More preferably, the cultivation step is carried out for a period of 12-240 hours.

[0157] Therefore, the present invention further relates to a method for producing mycelial components by submerged fermentation, wherein the fermentation medium comprises 5-60 g / L of carbon source, 0.1-60 g / L of nitrogen source, 0.01-15 g / L of minerals, and 0.01-50 mg / L of vitamins. Those skilled in the art can extend the method by adjusting the carbon source (i.e., total sugars added) and / or other parameters to meet the requirements of the reactor configuration used (batch mode vs. fed-batch mode vs. continuous mode).

[0158] In the method for producing a mycelium component by submerged fermentation, the fermentation medium preferably contains 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.5 to 15 g / L of minerals, and 0.1 to 10 mg / L of vitamins. Preferably, the carbon source is selected from potato dextrose agar, starch, cellulose, malt extract, sugar beet molasses, corn molasses, sucrose, glycerol, glucose, fructose, lactose, galactose, xylose, arabinose, and / or maltose. Preferably, the nitrogen source is selected from corn steep liquor (CSL), yeast extract, peptone, ammonia, urea, ammonium sulfate, ammonium chloride, and / or ammonium carbonate. Preferably, the minerals are selected from sodium selenate, magnesium sulfate, magnesium chloride, iron sulfate, iron chloride, manganese chloride, manganese sulfate, zinc sulfate, calcium sulfate, calcium chloride, calcium carbonate, copper chloride, copper sulfate, diammonium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium phosphate monobasic hydrate, and / or sodium chloride. Preferably, the vitamins are selected from biotin, choline chloride, folic acid, myo-inositol, niacinamide, pantothenic acid, pyridoxal, riboflavin, thiamine, cobalamin, and / or ascorbic acid. pH control is preferably controlled via a substance selected from sodium chloride, sodium hydroxide, potassium hydroxide, sulfuric acid, phosphoric acid, hydrochloric acid, citric acid, acetic acid, and hypochlorous acid. Note that other components, such as agar, or food-grade antifoaming agents, such as edible oils, or others, can be used. Those skilled in the art can expand the method by including additional substances, metabolites, and additional types of carbon, nitrogen, mineral, and vitamin sources.

[0159] In one embodiment, the medium is enriched with vitamin B12 (cobalamin) by adding it in its pure chemical form to replenish the mycelial biomass. In a further embodiment, the medium is enriched with vitamin B12 in the range of 1 μg / L to 200 μg / L, preferably in the range of 10 μg / L to 100 μg / L. In certain embodiments, the accumulated vitamin B12 in the biomass is at least 10%, preferably at least 5%, preferably at least 2.5%, preferably at least 1%, preferably at least 0.5% of the initial amount added. In a preferred embodiment, the accumulation of vitamin B12 in the biomass is at most 10%, preferably at most 7.5%, more preferably at most 5%, more preferably at most 2.5%, preferably at most 1%, more preferably at most 0.5%.

[0160] In one embodiment, the fungal strain is capable of growing in a defined medium. The mycelial component obtainable when grown in a defined medium as defined herein may also be referred to as mycelial component A.

[0161] Thus, the present invention relates to an edible mycelial component (also referred to as mycelial component A) preferably comprising (undifferentiated) mycelial biomass, wherein the mycelial biomass has an elemental composition with a C:N ratio in the range of 6-8 of the mycelium, and the edible mycelial component is characterized by an EUC of 200-500g MSG / 100g. Preferably, the mycelium is Pleurotus pulmonarius mycelium. EUC is as defined herein.

[0162] In another embodiment, the fungal strain may be grown in a synthetic medium. The mycelial component obtainable when grown in a synthetic medium as defined herein may also be referred to as mycelial component B.

[0163] Accordingly, the present invention relates to an edible mycelial component (also referred to as mycelial component B) comprising undifferentiated mycelial biomass, wherein the undifferentiated mycelial biomass has an elemental composition with a C:N ratio in the range of 8-12 of the mycelium, and wherein the edible mycelial component is characterized by an EUC of less than 200g MSG / 100g, EUC being as defined herein.

[0164] In one preferred embodiment, the present invention provides an edible mycelial ingredient comprising Pleurotus pulmonarius mycelial biomass (preferably undifferentiated biomass), wherein the Pleurotus pulmonarius mycelial biomass has an elemental composition of the mycelium with a C:N ratio in the range of 2 to 12, and wherein the edible mycelial ingredient is characterized by an equivalent umami concentration (EUC) of at least 30 g monosodium glutamate / 100 g MSG, where EUC is as defined herein. The edible mycelial component described herein can be obtained by a method for producing the edible mycelial component described herein by submerged fermentation, the method comprising culturing Pleurotus pulmonarius in a fermentation medium, the fermentation medium provided at the start of the fermentation characterized by a C:N ratio ranging from 1 to 50, preferably 5 to 50, the fermentation medium provided at the start of the fermentation comprising 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins, the medium comprising arginine and glutamate as the only amino acids. In one embodiment, the w / w ratio of arginine to glutamate is preferably 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, or 90:10. This is preferably determined by the specific EUC required for a particular mycelial component or food product comprising said mycelial component. In a preferred embodiment, the total arginine to glutamate w / w ratio is preferably 20:80, more preferably 50:50, even more preferably 40:60, and most preferably 30:70. In a preferred embodiment, the medium comprises glutamic acid as the only amino acid. It is understood that the present invention further provides an edible product, preferably a meat substitute product or a dairy substitute product, comprising an edible mycelial component described herein in the range of 1-99% by weight.

[0165] Preferably, the defined medium contains two or more amino acids and two or more vitamins, each preferably at different concentrations. In this case, the biomass may have the potential to grow on different sources. Preferably, the amino acids and vitamins are supplied from at least one complex nitrogen source, which is preferably selected from yeast extract, CSL, and peptone, more preferably yeast extract. The carbon source is preferably supplied from dextrose, glucose, maltose, galactose, or malt extract, or cellulose. The carbon-to-nitrogen ratio (medium), i.e., the C:N ratio in this defined medium, is in the range of 5 to 50, preferably 10 to 25, preferably 14 to 19. In one embodiment, the carbon-to-nitrogen ratio (medium), i.e., the C:N ratio in this defined medium, is in the range of 10 to 25, preferably 16 to 18, more preferably 16.5 to 17.5.

[0166] Preferably, the synthetic medium comprises at least one amino acid and at least one type of vitamin, (1) the at least one vitamin used has an amount characterized by a ratio of the at least one vitamin used to the total amount of all vitamins present in the defined medium (with reference to the preferred nitrogen complexes, the ratio being expressed as w / w) ranging from 0.01 to 10, preferably from 0.1 to 10, more preferably from 0.5 to 10, and (2) the at least one amino acid used has an amount characterized by a ratio of the at least one amino acid used to the total amount of all amino acids present in the defined medium (the ratio being expressed as w / w ratio) ranging from 0.01 to 10, preferably from 0.1 to 10, more preferably from 0.5 to 10.

[0167] Preferably, the nitrogen source concentration is in the range of 0.1 to 60 g / L, more preferably 1 to 40 g / L, even more preferably 1 to 30 g / L, and most preferably 1 to 20 g / L. In another preferred embodiment, the nitrogen source concentration is in the range of 0.1 to 10 g / L, preferably about 6 g / L.

[0168] The carbon to nitrogen ratio (medium), i.e., the C:N ratio in this synthetic medium, is in the range of 1 to 50, preferably 5 to 50, more preferably 10 to 25, and even more preferably 16 to 23. In one embodiment, the C:N ratio in this synthetic medium is 2 to 22, preferably 15 to 22, more preferably 20 to 25, and even more preferably 20 to 22. In an alternative embodiment, the C:N ratio in this synthetic medium is in the range of 2 to 22, preferably 2 to 16, and more preferably 2 to 14. In one embodiment, the C:N ratio in this synthetic medium is in the range of 1 to 5.

[0169] The at least one amino acid is preferably selected from alanine, asparagine, aspartate, arginine, tryptophan, glycine, glutamic acid, glutamine, methionine, phenylalanine, serine, valine, cystine, proline, leucine, tyrosine, threonine, isoleucine, histidine, lysine, and selenocysteine, or a combination thereof.

[0170] The at least one amino acid is preferably selected from essential amino acids selected from phenylalanine, valine, tryptophan, threonine, isoleucine, methionine, histidine, leucine, lysine, or combinations thereof.

[0171] Alternatively, the at least one amino acid is preferably selected from a non-essential amino acids selected from alanine, arginine, asparagine, aspartic acid (or aspartate), cysteine, glutamic acid (or glutamate), glutamine, glycine, proline, serine, tyrosine, selenocysteine, or a combination thereof. More preferably, the at least one amino acid is preferably selected from a non-essential amino acids selected from alanine, arginine, aspartic acid (or aspartate), cysteine, glutamic acid (or glutamate), glutamine, glycine, proline, serine, tyrosine, selenocysteine, or a combination thereof.

[0172] The at least one amino acid is preferably selected from non-essential amino acids selected from alanine, arginine, cysteine, glycine, proline, serine, tyrosine, selenocysteine, or a combination thereof. This list does not include umami amino acids. Five or fewer, preferably three or fewer, and even more preferably two or fewer types of amino acids are preferably added to the culture medium. The at least one amino acid, preferably five or fewer types of amino acids, more preferably three or fewer types of amino acids, and even more preferably two or fewer types of amino acids, is most preferably selected from alanine, arginine, cysteine, glycine, proline, serine, tyrosine, and selenocysteine.

[0173] Therefore, a defined medium in the present invention can also be defined as a medium characterized by a C:N ratio ranging from 1 to 50, comprising at least one amino acid selected from alanine, arginine, cysteine, glycine, proline, serine, tyrosine, and selenocysteine. Preferably, the at least one amino acid defined herein comprises arginine. It is clear to those skilled in the art that certain amino acids are considered essential for mycelial growth and, therefore, it may not be possible to grow mycelia without adding them to the medium. The inventors surprisingly found that it is possible to grow mycelia in the presence of only one non-essential amino acid, arginine, which allows obtaining mycelia with a low umami content (defined below and represented by the EUC parameter). The minimum amount of amino acid, preferably arginine, to be added is determined by the desired C:N ratio of the medium. In other words, the minimum amount of amino acid added preferably correlates with the minimum amount of carbon source required for growth, with the carbon source required for growth being at least 5 g / L, more preferably at least 10 g / L.

[0174] Preferably, the at least one non-essential amino acid added is no more than five (preferably no more than four, even more preferably no more than three, and even more preferably no more than two) amino acids, including arginine and (i) a non-essential amino acid selected from alanine, cysteine, glycine, proline, serine, tyrosine, and selenocysteine, and / or (ii) at most four further amino acids selected from essential amino acids selected from phenylalanine, valine, tryptophan, threonine, isoleucine, methionine, histidine, leucine, lysine, more preferably selected from alanine, cysteine, glycine, proline, serine, tyrosine, and selenocysteine.

[0175] Furthermore, in one embodiment, the medium may optionally further comprise at least one umami amino acid, as defined below.

[0176] In one embodiment, the medium contains only one amino acid, arginine, and it is understood that the mycelium component obtainable using this medium is characterized by an EUC of 30-100 g MSG / 100 g, more preferably 30-60 g MSG / 100 g.

[0177] Preferably, the C:N ratio is in the range of 10 to 25, more preferably 16 to 23. In one embodiment, the C:N ratio in this synthetic medium is 2 to 22, preferably 15 to 22, more preferably 20 to 25, and even more preferably 20 to 22. In one embodiment, the C:N ratio is in the range of 5 to 50. In one embodiment, the C:N ratio is in the range of 1 to 5.

[0178] The inventors have surprisingly found that culturing mycelia using a medium disclosed herein, where the medium lacks any of the umami amino acids described herein, can result in an edible mycelial ingredient characterized by an EUC of less than 200 g MSG / 100 g, preferably less than 100 g MSG / 100 g, and even more preferably less than 50 g MSG / 100 g. Optionally, at least one umami amino acid is used at a low concentration, preferably at most 1 g / L, more preferably at most 0.2 g / L, and even more preferably at most 0.1 g / L, to obtain the desired EUC of mycelia needed for specific food applications: less than 200 g MSG / 100 g, preferably 30-200 g MSG / 100 g.

[0179] The present inventors have surprisingly found that by culturing mycelia using a defined medium under the fermentation conditions of the present invention, an edible mycelial ingredient can be obtained that is preferably characterized by an EUC in the range of 200-500 g MSG / 100 g, and preferably characterized by an EUC in the range of 250-450 g MSG / 100 g.

[0180] After one of the nutrients becomes limiting (e.g., carbon source, nitrogen source, oxygen) and is completely depleted, the flask is used to inoculate either another flask with a 10-100 times larger volume or a similarly sized pre-fermentor. The fermentation seed train then consists of several pre-fermentors used to inoculate sequentially at a 1-20% volume / volume ratio. The pre-fermentors are operated at 10-50°C and a pH of 4-6 until the nutrient source is completely depleted. Depending on the inoculum size and activity, this process takes 12-240 hours.

[0181] Finally, the main fermentation takes place for up to 400m 3The fermentation is carried out at a scale of 1000 kJ / s, under controlled conditions (10-50°C, pH 3-8, controlled dissolved oxygen, controlled aeration rate, and agitation rate) using the above medium. The discontinuous mode fermentation continues until the carbon source is exhausted, and depending on the inoculum size and activity, the process takes 12-240 hours. In one embodiment, the preferred pH of the fermenter related to the present invention is in the range of 3-8, more preferably 4-6, and even more preferably 4-5. In one embodiment, the preferred pH of the fermenter related to the present invention is preferably about 4, 4.2, 4.3, 4.5, 4.7, 4.8, or 5. Preferably, the pH is controlled by an acid / base titrant. In an alternative embodiment, the pH is maintained via a buffer solution, preferably selected from phosphate buffer or citrate buffer. Both the pH and the C / N ratio are important factors for obtaining the desired growth and, therefore, the properties listed in Table 3 (protein content, protein composition, fiber content, and therefore, the effect on EUC). pH can significantly affect protein composition and content, as observed in the literature (Calsamiglia S, Ferret A, Devant M. Effects of pH and pH fluctuations on microbial fermentation and nutrient flow from a dual-flow continuous culture system. J Dairy Sci. 2002 Mar;85(3):574-9. doi:10.3168 / jds.S0022-0302(02)74111-8. PMID:11949862.). The C / N ratio is determined by the carbon and nitrogen sources present in the medium, which varies in media A, B, and C. It is understood that this composition, related to the fermentation conditions of pH, leads to different osmotic stresses in the fungal strains, which in turn leads to different pools of total / free amino acids and, therefore, different protein content and composition, which results in different EUCs.The trends observed for protein and fiber content for the Pleurotus species exemplified in this invention show surprising trends that differ from those reported for the same and other Pleurotus species, both grown in the same medium and at pH values ​​in both publications that are not reported to reach yields of at most 30% (Reference 1: Food Chemistry 85 (2004) 101-105; Reference 2: Carbohydrate Polymers 87 (2012) 368-376, both using the medium detailed in Reference 1).

[0182] The defined medium preferably contains minerals, vitamins, and trace elements selected from the compounds listed above and is sterilized by either heat sterilization (at or above 121°C for at least 20 minutes) or microfiltration as known to those skilled in the art, before being added to the fermenter. At the end of the main fermentation period, the biomass is harvested by simply separating it from the cultivation supernatant using a liquid-solid separation method, such as centrifugation, filtration, or sieving.

[0183] In one embodiment for all media of the present invention, the separated biomass is washed with acidic water, preferably with an acidic water having a pH of 3 to 7, more preferably 3 to 6. It is understood that this washing step treats the biomass to substantially remove or inhibit the formation of any bitter compounds subsequently formed, thus affecting the EUC concentration. The acidity is preferably adjusted with citric acid, sulfuric acid, phosphoric acid, or hydrochloric acid.

[0184] It is understood that the biomass yield is calculated according to the ratio of the amount of biomass produced to the amount of carbon substrate consumed.

[0185] It is understood that according to the method of the invention for a medium corresponding to (and usable in the preparation of) component A (defined medium with one complex nitrogen source), the biomass yield is preferably at most 65%, preferably at most 60% (based on the conversion of the carbon sources available in the medium). In one embodiment, the biomass yield of this medium corresponding to component A is preferably at least 45%, more preferably at least 50%.

[0186] It is understood that according to the method of the invention for a medium corresponding to (and usable in the preparation of) component B (a synthetic medium containing at least one amino acid essential for mycelial growth), the biomass yield is preferably at most 55%, preferably at most 50% (based on the conversion of the carbon source available in the medium). In one embodiment, the biomass yield of this medium corresponding to component B is preferably at least 40%, more preferably at least 45%.

[0187] It is understood that the method of the present invention for the medium corresponding to (and usable in the preparation of) component C (a natural complex medium containing a complex carbon source from a sidestream extract) provides a biomass yield of up to 100%, preferably up to 99%, and more preferably up to 85-95%. It is understood that the biomass yield is calculated according to the ratio of the amount of biomass produced to the amount of substrate (e.g., C5 sugars) consumed.

[0188] The fungal strains may be grown on natural complex media, i.e., extracts prepared using side streams or optionally pre-processed side streams. It is particularly preferred that the side stream is brewer's grains. The carbon to nitrogen ratio (medium) in this complex medium ranges from 5 to 50, preferably from 10 to 40, and most preferably from 10 to 25. In one embodiment, the C:N ratio in this natural complex medium ranges from 2 to 18, preferably from 2 to 16, and more preferably from 2 to 14.

[0189] In one embodiment, the C:N ratio in the natural complex medium ranges from 2 to 18, preferably from 6 to 16, and more preferably from 6 to 14. In one embodiment, the C:N ratio in the natural complex medium ranges from 2 to 13, and most preferably from 4 to 8. In one embodiment, the C:N ratio of the natural complex medium is about 2, about 3, about 4, about 6.5, about 7, about 8, about 9, about 10, about 11, about 13, about 13.5, or about 14, or about 20, or about 22, or about 25, depending on the C:N ratio of the side stream extract used as the carbon source in the natural complex medium.

[0190] In one embodiment, the carbon-to-nitrogen ratio of the side stream extract used as the carbon source of the natural complex medium, i.e., the C:N ratio in the side stream extract, is preferably in the range of 1 to 60, more preferably 5 to 25, most likely 10 to 20, and even more preferably 10 to 18. This C:N ratio of the side stream extract will therefore affect the C:N ratio of the final natural complex medium. In a preferred embodiment, the carbon-to-nitrogen ratio of the side stream extract from brewer's grains used as the C5 sugar carbon source in the natural complex medium is preferably in the range of 1 to 50, more preferably 1 to 25, most preferably 5 to 25, and even more preferably 5 to 18.

[0191] The side stream may be pre-processed, for example, by using heat or enzyme treatment. It is understood that the pre-processing step is optional, and the side stream may also be used as is without any further pre-processing.

[0192] Preferably, the protein content of the extract is in the range of 1-200 g / L, 1-150 g / L, 1-100 g / L, 1-90 g / L, 1-80 g / L, 1-70 g / L, 1-60 g / L, 1-50 g / L, 1-40 g / L, 1-30 g / L, 1-25 g / L, 1-20 g / L, 1-15 g / L, 1-10 g / L, or 1-5 g / L. In a preferred embodiment, the protein content of at least one extract for a complex medium comprises 1-200 g / L of protein, preferably 1-100 g / L, more preferably 1 to about 50 g / L, most preferably about 5-45 g / L, and more preferably 7-30 g / L. Such a protein content is essential for obtaining a high EUC in the resulting biomass. In a further preferred embodiment, the protein content of the at least one extract for the complex medium comprises a protein content of 10 to 30 g / l, more preferably 10 to 25 g / l. Preferably, the protein content of the at least one extract for the complex medium comprises a protein content of about 10 g / l, more preferably about 15 g / l, even more preferably about 20 g / l.

[0193] According to the inventors, an increase in EUC is observed at high protein concentrations in the extracts used in fungal culture, ie, between 7 g / L and 30 g / L.

[0194] Preferably, the glutamic acid content of the side stream extract used in the culture medium is at least 400 mg / L, more preferably at least 500 mg / L. More preferably, the glutamic acid content of the side stream extract used in the culture medium is in the range of 400 to 4500 mg / L, more preferably 400 to 2250 mg / L, and even more preferably 500 to 2000 mg / L. Preferably, the glutamic acid content of the side stream extract is at least 150 mg / L, more preferably at least 200 mg / L. More preferably, the aspartic acid content of the side stream extract is in the range of 150 to 1500 mg / L, more preferably 150 to 750 mg / L, and even more preferably 220 to 650 mg / L.

[0195] The range of high glutamate concentrations achieved in the extract, which directly influences EUC, preferably depends on the residence time of the extraction in the extraction method used, as described below in connection with steam pretreatment or liquid extraction of lignocellulosic material, preferably liquid extraction with the addition of acid. In one embodiment, the residence time of the extraction is in the range of 1 to 25 minutes, preferably 5 to 15 minutes, to achieve a glutamic acid content in the extract surprisingly higher than the aspartic acid content in the extract obtainable by the extraction method under acidic conditions. Thus, in this embodiment, a brewer's grain C5 sugar extract can be obtained by extraction under acidic conditions for 1 to 25 minutes, preferably 5 to 15 minutes. In another embodiment, the residence time of the extraction is in the range of 1 to 25 minutes, preferably 1 to 20 minutes, to achieve a glutamic acid content in the extract surprisingly higher than the aspartic acid content in the extract obtainable by the extraction method under base. Thus, in this embodiment, a brewer's grain C5 sugar extract can be obtained by extraction under acidic conditions for 1 to 25 minutes, preferably 1 to 20 minutes. The observed trends suggest that a longer residence time may result in a higher glutamic acid content, which in turn results in a higher EUC. Preferably, acidic conditions herein refer to, for example, at least 0.1 wt% H2SO4, not more than 1.6 wt% H2SO4, not more than 1.0% wt% H2SO4, and preferably about 0.9 wt% H2SO4.

[0196] The fermentation medium can also be supplemented or processed to reach the desired C:N ratio (medium) through material crystallization or precipitation if the protein is not present in the extract, or through side stream combination.

[0197] Preferably, as understood herein, when referring to a numerical value, the term about refers to the value in question ±10% of the value in question, more preferably the term about refers to the value in question ±5% of the value in question, even more preferably the term about refers to the value in question ±1% of the value in question, and even more preferably the term about refers to the value in question.

[0198] Sidestreams as understood herein may also preferably include lignocellulosic materials, in particular lignocellulosic materials originating from industrial and / or agricultural sidestreams. Lignocellulosic materials are preferably defined herein as materials comprising dry plant matter. Preferably, the lignocellulosic materials comprise cellulose, hemicellulose, and lignin. Preferably, the at least one lignocellulosic material is at least one industrial and / or agricultural sidestream as defined herein. More preferably, the lignocellulosic materials are preferably solid or processed into a powder before use. As understood herein, lignocellulosic materials are preferably characterized by a particular color, density, and / or mesh size distribution. Examples of lignocellulosic materials are beer spent grains, brewer's grains, cereal bran, bagasse, cotton cake and pressed oil cake from sunflowers, hazelnuts, shells and husks from tree nuts, grass and leaf waste, wood chips, coffee grounds, coffee husks, coffee silverskin, by-products from the rapeseed and soybean industries such as soybean pulp ("okara"), banana leaves, banana peels, chicory root, cassava skin, citrus pulp, cocoa, cocoa bean shells, cocoa mucilage, cocoa pod husks, coconut fiber, coconut husks, coconut shells, coffee pulp, corn cobs, corn stover, cotton, cottonseed meal, cottonseed, almonds, the waste product is selected from the group consisting of peanut hulls, hop waste, pea by-products, peanut hulls, peanut meal, peanuts, potato rawhide, potato tubers, eucalyptus bark, lantana grass, switchgrass, rice bran, rice hulls, rice stalks, sugar beet meal, sugar beet pulp, sawdust, sugarcane wet bagasse, walnut husks, wheat bran, wheat distillers grains, wheat germ, wheat stalks, lupin seeds, chickpea bran, chickpea pod hulls, chickpea stalks, olive waste, grape marc, pear pulp, sorghum bran, sorghum germ, sorghum stalks, sorghum stalks, sunflower waste, and / or tea waste. Additionally, the skins or waste or fruit or pomace of the following side streams are also preferably included as lignocellulosic materials encompassed by the present invention: oats, pine wood, dates, apples, apricots, barley meal, broccoli, cabbage, carrots, turnips, eggplant, kiwi, melon, alfa alfa, pineapple, pomegranate, plum, watermelon, zucchini, asparagus, beetroot, cauliflower, garlic, onion, pumpkin, squash, and / or tomato. The side streams referred to herein may also be understood as side streams from the agri-food industry. Examples of non-lignocellulosic materials, such as protein materials, are preferably palm oil, sugarcane viscous, molasses, whey, whey permeate, wool, and silk.

[0199] It will be apparent to one skilled in the art that in addition to lignocellulosic and / or proteinaceous materials, all of the side streams listed herein contain sugars, minerals, and / or vitamins.

[0200] A particularly preferred side stream is brewer's grains.

[0201] The use of brewer's spent grains of different particle sizes will yield different results and will depend on the application. In certain embodiments, the mode particle size in the particle size distribution of the brewer's spent grains is in the range of 8-10 mm, preferably 6-10 mm, more preferably 4-6 mm, more preferably 2-4 mm, more preferably 1-2 mm, and more preferably 0.2-1 mm. In a preferred embodiment, the particle sizes present in the distribution are about 0.2 mm, 0.4 mm, 0.6 mm, 0.7 mm, 1 mm, 2 mm, 4 mm, 6 mm, and 10 mm. In another preferred embodiment, the brewer's grains suitable for use in the method of the present invention are characterized by a particle size distribution determined by using different sets of sieves, the particle size distribution comprising a maximum of 2-4 mm, which is up to 60% by weight, preferably up to 50% by weight, more preferably 30-50% by weight, most preferably 35-45% by weight, and most preferably at least 35% by weight, followed by a second maximum of 1-2 mm, which is preferably up to 35% by weight, more preferably 15-30% by weight, and most preferably 20-30% by weight, followed by a third maximum of 0.25-1 mm, which is preferably up to 20% by weight, more preferably 5-15% by weight, and most preferably 8-15% by weight. Sieving was carried out by air jet sieving according to DIN 10765 2016-07, after removing particles with a particle size greater than 4 mm using an automatic sieving tower, i.e., a vibration sieving method.

[0202] In an exemplary embodiment, the sieving residue after the vibratory sieving process is 80%, the fraction greater than 2.00 mm (2-4 mm) is 54.21 g / 100 g, the fraction greater than 1.00 mm is 33.58 g / 100 g, the fraction greater than 0.25 mm is 10 g / 100 g, and the sieving residue is 1.94 g / 100 g.

[0203] The preparation of the fungal component for fermentation may include a further preliminary step of pre-processing the side stream. The following steps are described herein using the example of brewer's spent grains. Preferably, the pre-processing of the brewer's spent grains includes modifying the grain size or other mechanical properties of the brewer's spent grains. For example, the pre-processing of the brewer's spent grains may include grinding the brewer's spent grains. This step is conventional and known to those skilled in the art. The brewer's spent grains may further be subjected to a preliminary treatment, for example, to increase its accessible surface area by, for example, breaking down particles by grinding, crushing, pulverizing, etc. Such steps, which are disclosed herein as optional steps, are known to those skilled in the art. In addition, as partially discussed in the literature (Ozturk et al., J. Inst. Brew. 108(1):23-27, 2002), the brewer's spent grains may be sieved through a set of sieves with openings of 850, 425, and 212 μm during grinding. Depending on the fraction, brewer's grain preparations can be considered to be coarse (425-850 μm), medium (212-425 μm), and fine (less than 212 μm), although brewer's grain preparations can also be ground or modified to have larger particle sizes, for example, larger particle sizes including up to 0.3-1 mm, preferably what is referred to by those skilled in the art as a coarse particle size, preferably up to 0.4-0.8 mm, depending on the application to be used.

[0204] Brewer's Spent Grain (BSG) or brewer's grains is preferably understood as a residue or by-product of the brewing industry. Preferably, brewer's grains is the material remaining after the grinding process and preferably has a dry matter content of 10% to 30%. However, the dry matter contents listed herein are not meant to be limiting, and as the skilled person will recognize, the dry matter content can be increased in preliminary processing, for example by pressing, drying, or other methods known to the skilled person. Furthermore, brewer's grains originating from other industries (e.g., brewer's grains that can be obtained as a by-product of the production of foodstuffs) can also be used within the scope of the present invention. The skilled person will be in a position to extend the method by including further side streams from the above list of lignocellulosic materials.

[0205] Preferably, the BSG contains 20% to 25% w / w cellulose (preferably 22%), 23 to 28% w / w hemicellulose (preferably 25.8%), and / or 20% to 30% w / w protein (preferably 25% w / w protein). These figures are understood to refer to the BSG content relative to its dry mass. BSG is characterized by a final moisture content of 50% to 75% w / w. Accordingly, this final moisture content can be achieved by dehydrating the BSG, for example, by pressing the material to a final moisture content of 50 to 75% w / w using methods known to those skilled in the art. However, this is not meant to be construed as limiting, and other means and methods for dehydrating the BSG may also be applied herein, as will be further apparent to those skilled in the art.

[0206] The extraction used for the present invention to produce the mycelium component is performed by extracting C5 sugars from the lignocellulosic material contained in the BSG by steam pretreatment, followed by a washing step with liquid water at a temperature below 50°C, and combining the extract so obtained with at least one non-carbohydrate nutrient for fungal cultivation.

[0207] Preferably, during the steam pretreatment step, the lignocellulosic material contained in the BSG is contacted with steam at a temperature of 130°C to 180°C, preferably at a temperature of 160°C to 180°C, more preferably at a temperature of 165°C to 175°C. Preferably, during the steam pretreatment step, the lignocellulosic material contained in the BSG is contacted with steam for a period of up to 30 minutes, preferably for a period of up to 15 minutes. Preferably, during the steam pretreatment step, the lignocellulosic material contained in the BSG is contacted with steam at a temperature of 165°C to 175°C and / or for a period of up to 15 minutes, preferably for a period of up to 12.5 minutes, more preferably for a period of up to 10 minutes, even more preferably for a period of up to 7.5 minutes, even more preferably for a period of up to 5 minutes, even more preferably for a period of up to 2.5 minutes, even more preferably for a period of up to 2 minutes, even more preferably for a period of up to 1 minute. More preferably, during the steam pretreatment step, the lignocellulosic material contained in the BSG is contacted with steam at a temperature of 165°C to 175°C for a time period of up to 15 minutes. More preferably, during the steam pretreatment step, the lignocellulosic material contained in the BSG is contacted with steam at a temperature of 165°C to 175°C for a time period of up to 7.5 minutes. More preferably, during the steam pretreatment step, the lignocellulosic material contained in the BSG is contacted with steam at a temperature of 165°C to 175°C for a time period of up to 2 minutes. Preferably, a temperature of 165°C to 175°C relates to a temperature of about 175°C, more preferably, a temperature of 165°C to 175°C relates to a temperature of 170°C. Preferably, the time period for steam pretreatment is at most 5 minutes, preferably at most about 4 minutes, more preferably at most about 3 minutes.

[0208] The water used for steam prehydrolysis may contain a dilute acid, for example, up to 1% w / w of the acid. Preparations of 0.2% w / w or 0.4% w / w H2SO4 are particularly suitable. In a particular embodiment, the concentration of the acid (preferably H2SO4) is 1.1-1.6% w / w. This is particularly applicable to embodiments in which the lignocellulosic material is brewer's spent grains. In one embodiment of the present invention, the water used for steam prehydrolysis may contain a dilute base, for example, up to 1% w / w of the base. 0.2% w / w NaOH is particularly suitable. Alternatively, the water used for steam prehydrolysis may be replaced with a phosphate buffer solution at pH 5.5.

[0209] The steam pretreatment is followed by a washing step with liquid water at a temperature below 50° C., preferably below 40° C., even more preferably below 30° C., even more preferably below 25° C. Preferably, the liquid water used in the washing step is at room temperature, i.e. at a temperature between 20° C. and 25° C., preferably at a temperature of about 22° C., more preferably at a temperature of 22° C.

[0210] Therefore, the aqueous extract from BSG used to grow the mycelium component may be further supplemented with a nitrogen source, a carbon source, trace elements, vitamins, and / or a protein composition. The nitrogen source as defined herein is preferably selected from ammonia, urea, yeast extract, malt extract, corn steep liquor, and peptone. More preferably, the nitrogen source is ammonia and / or urea. The carbon source is preferably selected from glucose, fructose, sucrose, lactose, maltose, xylose, galactose, dextrose, glycerol, and molasses, and more preferably, the carbon source is glucose. Preferably, no carbon source other than that derived from BSG is added to the medium of the present invention. Trace elements as defined herein may include, for example, iron(III) salts, copper(II) salts, zinc salts, manganese(II) salts, molybdenum salts, and / or cobalt(II) salts. Vitamins as defined herein preferably include vitamins beneficial for the growth of fungi on a medium obtainable according to the method of the present invention, such as folic acid, riboflavin, pantothenic acid or biotin. The protein composition may further be used to supplement the aqueous extract.

[0211] In another embodiment, the fungal component may be produced using a different method, in which C5 sugars are extracted from the lignocellulosic material contained in the BSG by a liquid extraction treatment with water at a temperature of 145°C to 155°C and / or for a time period of up to 70 minutes, preferably up to 50 minutes, preferably at a pressure of 30 to 50 bar, and the extract so obtained is combined with at least one non-carbohydrate nutrient for fungal cultivation as discussed above.

[0212] The extraction is carried out at a temperature of 140° C. to 180° C., preferably at a temperature of 145° C. to 175° C., more preferably at a temperature of 145° C. to 170° C., even more preferably at a temperature of 145° C. to 160° C., even more preferably at a temperature of 145° C. to 155° C., even more preferably at a temperature of about 150° C., and / or the extraction is carried out for a time period of up to 70 minutes, preferably up to 50 minutes. Furthermore, the extraction is preferably carried out at a pressure of 30 to 50 bar.

[0213] In one embodiment of the present invention, the water used for aqueous liquid extraction may contain a dilute acid, for example, up to 1% w / w of the acid. Preparations of 0.2% w / w or 0.4% w / w H2SO4 are particularly suitable. In a particular embodiment, the concentration of the acid (preferably H2SO4) is 1.1-1.6% w / w. This is particularly applicable to embodiments in which the lignocellulosic material is brewer's spent grains. In one embodiment of the present invention, the water used for aqueous liquid extraction may contain a dilute base, for example, up to 1% w / w of the base. 0.2% w / w NaOH is particularly suitable. Alternatively, the water used here may be replaced with a phosphate buffer solution at pH 5.5.

[0214] Preferably, the process of aqueous extraction of lignocellulosic material, preferably industrial and / or agricultural by-streams, according to the present invention is carried out with water at a pH of 2.0 to 12.0, preferably 3.0 to 10.0, more preferably 4.0 to 8.0, and even more preferably 5.0 to 8.0. The pH values ​​understood herein are measured at a pressure of 1.0 bar and a temperature of 25°C, even if the extraction itself, as disclosed herein, is carried out under different conditions. Preferably, the pH is adjusted before the water is placed in contact with at least one lignocellulosic material, preferably an industrial and / or agricultural by-stream. It is further understood herein that the addition of acids or bases to the water described herein to a final concentration of 1% w / w or less is preferably avoided.

[0215] As defined herein, C5 sugars preferably refer to fractions in which at least 80% w / w of the total sugar content is composed of pentoses (sugars containing polysaccharides composed of sugar subunits of 5 carbon atoms). It should be noted that C5-sugars, as defined herein as sugar-containing fractions, may contain other sugars, in particular C6 sugars (sugars with 6 carbon atoms, also called hexoses), as monomers and / or contained within polysaccharides and / or oligosaccharides. Thus, sugars other than pentoses may also be extracted.

[0216] Thus, as referred to herein, brewer's grain C5 sugar extract may refer to a composition in which at least 80% by weight of the total sugar content comprises pentoses, and more preferably, at least 80% by weight of the carbon-containing compounds (carbon source) comprise pentoses.

[0217] In a further embodiment, the present invention relates to the use of the fungal biomass of the present invention in the production of a fungal-based food product. Accordingly, the present invention also relates to a fungal-based food product obtainable as described herein. The fungal-based food product of the present invention can be prepared in any form known to those skilled in the art. For example, the fungal-based food product of the present invention can be in the form of balls (i.e., meatball substitutes), dumplings, vegetarian sausages, meat substitute steaks, meat substitute ground meat products, meat substitute products for preparing sandwiches, etc.

[0218] The food product according to the present invention may be, for example, a nutritional supplement. The nutritional supplement may be in the form of a liquid or a solid, for example, a pill, a lozenge, or a tablet. For example, the nutritional supplement of the present invention may be a protein supplement, a carbohydrate supplement, and / or a functional drink or beverage.

[0219] Food products as understood herein may be dairy products, such as cheese, yogurt, drinking yogurt, milk drinks, and ice cream. Food products as understood herein may also relate to different embodiments of seafood products, such as crab cakes, fish cakes, tuna, salmon, or shrimp, as well as various desserts, confectionery, or bakery items, flour, starch, bread, eggs, pasta, including chocolates, brownies, or cookies.

[0220] The food may be a textured food or a textured food. Thus, the food of the present invention comprises all amino acids required for the daily intake of humans that cannot be synthesized de novo. Furthermore, the textured food of the present invention is preferably heat-resistant, boil-resistant, and suitable for cooking. For example, the fungus-based food product of the present invention may be a meat substitute product, as described herein. It is noted that preferably, the meat substitute product is a textured food or a textured food. It is further noted that the structure of the textured food improves the acceptability of the textured food by consumers. It is further noted that the unique fibrous texture of the fungal biomass of the present invention may be beneficial for producing textured food products, or for producing textured food products without using traditional texturizing methods, such as extrusion.

[0221] The fungus-based food products of the present invention may be further processed and / or supplemented. For example, the fungus-based food products of the present invention may be further supplemented with water, salt, oil and / or spices according to protocols known to those skilled in the art. Further processing may also include heating and high-pressure treatment of the food (particularly useful for high-pressure pasteurization), brewing, boiling, baking, frying, fermenting, and / or drying. As known to those skilled in the art, preservatives may be added to extend the shelf life of the food products of the present invention.

[0222] Preferably, the food product of the present invention may be further supplemented with a compositional component, which is defined as a certain content of a substance / compound that can be described as a compositional component for forming a fungal product together with the fungal component of the present invention.

[0223] "Ingredients" are preferably understood herein as supplemented preservatives, antioxidants and acidity regulators, thickeners, stabilizers and emulsifiers, pH regulators and anti-caking agents, flavor enhancers, improvers, stabilizers, thickening agents, colors, glazing agents and sweeteners, additives, aromatic compounds, and / or nutrients.

[0224] Preferably, the preservatives include calcium carbonate, acetic acid, potassium acetate, sodium acetate, calcium acetate, lactic acid, sorbate, and malic acid.

[0225] Preferably, the antioxidants and acidity regulators include ascorbic acid, sodium ascorbate, calcium ascorbate, fatty acid esters of ascorbic acid, tocopherol-rich extracts, alpha tocopherol, gamma tocopherol, delta tocopherol, lecithin, sodium lactate, potassium lactate, calcium lactate, citric acid, sodium citrate, potassium citrate, calcium citrate, tartaric acid (L(+)), sodium tartrate, potassium tartrate, sodium potassium tartrate, sodium malate, potassium malate, calcium malate, calcium tartrate, and triammonium citrate.

[0226] Preferably, the thickening agents, stabilizers, and emulsifiers (or hydrocolloids) are selected from the group consisting of alginic acid, sodium alginate, potassium alginate, ammonium alginate, calcium alginate, agar, carrageenan, modified euchema seaweed, locust bean gum, guar gum, tragacanth, gum arabic (acacia gum), xanthan gum, tara gum, gellan gum, sorbitol, mannitol, glycerol, konjac, pectin, cellulose, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylmethylcellulose, sodium carboxy ... Includes cellulose, cellulose gum, enzymatically hydrolyzed carboxymethylcellulose, sodium-potassium and calcium salts of fatty acids, magnesium salts of fatty acids, mono- and diglycerides of fatty acids, acetate esters of mono- and diglycerides of fatty acids, lactate esters of mono- and diglycerides of fatty acids, citrate esters of mono- and diglycerides of fatty acids, tartrate esters of mono- and diglycerides of fatty acids, microcrystalline cellulose-cellulose gel, mono- and diacetyl tartaric acid esters of mono- and diglycerides of fatty acids, mixed acetate and tartrate esters of mono- and diglycerides of fatty acids, sorbitol and mannitol.

[0227] Preferably, the pH adjuster and anti-caking agent includes sodium carbonate, potassium carbonate, ammonium carbonate, magnesium carbonate, hydrochloric acid, potassium chloride, calcium chloride, magnesium chloride, sulfuric acid, sodium sulfate, potassium sulfate, calcium sulfate, sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonium hydroxide, magnesium hydroxide, calcium hydroxide, magnesium hydroxide, fatty acids, gluconic acid, glucono-delta-lactone, sodium gluconate, potassium gluconate, and calcium gluconate.

[0228] Preferably, the flavor enhancer includes glutamic acid, monosodium glutamate, monopotassium glutamate, calcium diglutamate, monoammonium glutamate, magnesium diglutamate, guanylic acid, disodium guanylate, dipotassium guanylate, calcium guanylate, inosinic acid, disodium inosinate, dipotassium inosinate, calcium inosinate, calcium 5'-ribonucleotides, disodium 5'-ribonucleotides, and glycine and its sodium salts.

[0229] Preferably, the improver includes L-cysteine.

[0230] Preferably, the stabilizers include invertase and polydextrose.

[0231] Preferably, the thickener includes psyllium husk, polydextrose, oxidized starch, phosphated starch, cross-linked starch, monoesterified cross-linked starch phosphate, acetylated cross-linked starch phosphate, and acetylated starch. Preferably, the thickener includes acetylated adipate cross-linked starch, hydroxypropyl starch, hydroxypropylated cross-linked starch phosphate, sodium starch octenyl succinate, a starch-based component, and acetylated oxidized starch. More preferably, the thickener includes psyllium husk and / or a starch-based component.

[0232] Preferably, the coloring agent includes riboflavin, chlorophyll and chlorophyllin, anthocyanin, betanin, lycopene, copper complexes of chlorophyll and chlorophyllin, terpene compounds such as carotene compounds and xanthophyll compounds, plain caramel, caustic sulfite caramel, ammonia caramel, sulfite ammonia caramel, vegetable carbon, calcium carbonate, iron oxide and iron hydroxide, curcumin, tartrazine, cellulose gel, cochineal, carminic acid, carmine, azorubine, carmoisine, lutein, cocoa powder (melanoidin), beet powder, tomato extract, duckweed powder, spirulina powder, paprika powder (capsanthin and / or capsorubin), turmeric powder, blueberry powder, strawberry powder, berry pigment powder, heme powder, lycopene powder, betanin powder, alfalfa powder, saffron powder, mint powder, and annatto extract.

[0233] Preferably, the glazing agents and sweeteners include isomalt, maltitol, acesulfame potassium, aspartame, cyclamate, saccharin, sucralose, alitame, steviol glycosides, neotame, lactitol, xylitol, and erythritol.

[0234] Preferably, the further additives defined to be understood as falling under the category of composition ingredients are selected from vitamin B12, vitamin B6, vitamin B2, vitamin B3 (also called niacin), riboflavin, thiamine, vitamin A, vitamin E, omega-3 fatty acids, vitamin D2, folic acid, iodized salts (NaCl, further comprising iodized salts in an amount of up to 5% w / w), enzymes (e.g. transglutaminase, amylase), minerals (e.g. salts containing calcium, iron and / or potassium, etc.), flavors or flavor components (salt, pepper, garlic, onion, mushroom fruiting body pieces, vegetable pieces, ginger, turmeric, curry, sugars (i.e. sucrose, glucose, mono- or disaccharides), oils, lemon juice, orange juice, herbs and spices, yeast flakes), texturizing vegetable proteins, and natural aromatic compounds. As defined herein, herbs and spices include natural aromatic compounds such as methyl acetate, linalool, limonene, vanillin, etc., or synthetic compounds such as aprifloren, cinnamyl propionate, cyclohexadecanolide, and ethyl levulinate. Such additional additives may improve optical visibility, flavor, nutritional value, and provide additional texture.

[0235] Preferably, the nutrients are selected from high protein ingredients (e.g., pea protein isolate, chickpea protein isolate, wheat gluten, egg white powder, and / or mung bean protein isolate), high carbohydrate / dietary fiber ingredients (e.g., cereal flours, cereal starches, legume starches, fruit fibers, polysaccharides, starch-based ingredients, psyllium husk, inulin, wheat starch, corn starch), high vitamin / mineral ingredients, and / or high lipid ingredients (e.g., all types of edible oils and butter). High fiber ingredients are preferably used to improve freeze-thaw stability and / or juiciness.

[0236] The list of compositional ingredients does not end here, and it should be noted that those skilled in the art will recognize that components or substances may fall into more than one category.

[0237] The sensory characteristics may also relate to odor attributes, preferably pungent, savory, floral, sour, ripe, musty, earthy, off-flavor; texture attributes; mouthfeel attributes related to juiciness and crunchy; taste attributes, preferably sweet, sour, salty, bitter, umami, metallic, astringent; and aroma attributes, preferably related to aroma complexity, aroma intensity, aroma roundness, and off-flavor.

[0238] Textural attributes are defined herein as the density, cutting strength, shear strength / force, hardness, springiness (i.e., elasticity), cohesiveness, gumminess, chewiness, adhesiveness, firmness, spreadability, stickiness, puncture force, water release, and water retention of the mycelial component or fungal-derived product, particularly the food product.

[0239] As referred to herein, cutting strength preferably describes the resistance of a food product to penetration by a cutting tool, preferably expressed in N.

[0240] As referred to herein, shear force preferably describes the ability of a biomass or product (especially a food product) to resist non-linear forces applied to the biomass or food product at different parts thereof, acting in different directions for the same weight. Preferably, the force is applied only from above. The applied force is collinear and is also known as compressive force. Preferably, it is expressed in N.

[0241] As referred to herein, hardness preferably refers to the force required to deform an article a given distance, preferably expressed in N.

[0242] As referred to herein, resilience (i.e., elasticity) preferably describes the resilience of a product, the more fractured the product is, the less resilient it is, i.e., the more it rebounds and returns to its structural integrity after being subjected to stress. Preferably, resilience is expressed as a percentage reflecting the degree of allowable deformation relative to the original size.

[0243] As referred to herein, product cohesion preferably describes the ability of a product to have structural integrity to withstand compressive or tensile stress. A product is cohesive when it sticks to itself under such stress. Preferably, cohesion is expressed as a percentage reflecting the degree of allowable deformation relative to the original size.

[0244] As referred to herein, viscosity preferably describes the energy required to disintegrate a food product into a swallowable state. Viscosity, expressed as N, is calculated as the product (i.e., multiplication) of hardness N and cohesiveness %.

[0245] As referred to herein, chewiness is preferably described as the energy required to chew a solid food. Chewiness, expressed in N, is the result of multiplying the viscosity N by the elasticity %.

[0246] As referred to herein, stickiness describes the level of stickiness of a food product. When a product is subjected to pressure deformation and the surface of the sample is sticky, a negative force is generated, which is calculated as the negative area under the curve. A large negative value is preferably interpreted as a sticky mouthfeel.

[0247] As referred to herein, firmness preferably describes the toughness of a sample, where the force (N) at the maximum penetration depth is interpreted as the sample firmness expressed in N. The area under the curve (Ns) represented the total amount of force required to carry out the shearing process. This is considered to be a good instrumental measurement of the spreadability of cream cheese and other spreadable products. A smaller value in this area indicates easier stretching. The force (N) of the maximum negative peak indicates the sample stickiness, expressed as a negative value in N. The lower the value, the stickier the sample.

[0248] As referred to herein, the penetration force from a penetration test preferably describes the energy required to penetrate a sample to a particular depth. Preferably, the penetration force is expressed as the area under the curve (Ns) of the positive area.

[0249] Water Holding Capacity (WHC) describes the ability of a material to hold water during processing, and Water Release (RW) describes the ability of a material to release water during processing.

[0250] "Edible" in the context of the present invention means that the edible product is safe for feed (fish, cattle), safe for use as pet food, and / or safe for human consumption. The food products of the present invention are products that are suitable as a substitute for dairy or meat products. The products may also be referred to as vegetarian products, or even vegan products. The suitability of the product for substitution is determined by the texture, mouthfeel, taste, nutritional content, water content, appearance, and other factors, which should be as similar as possible to the product being replaced.

[0251] In certain embodiments, the edible mycelium component has an elemental composition with a C to N ratio (mycelium) in the range of 1 to 50, preferably 1 to 20, preferably 1 to 19, more preferably 1 to 18, more preferably 1 to 17, more preferably 1 to 16, more preferably 1 to 15, more preferably 1 to 14, more preferably 1 to 13, more preferably 1 to 12, more preferably 1 to 11, more preferably 1 to 10, more preferably 1 to 9, more preferably 1 to 8, more preferably 1 to 7, more preferably 1 to 6, more preferably 1 to 5, more preferably 1 to 4, more preferably 1 to 3, more preferably 1 to 2.

[0252] In a preferred embodiment, the edible mycelium component A has an elemental composition with a C to N ratio (mycelium) in the range of 1-30, preferably 1-15, more preferably 5-10, most preferably 6-8.

[0253] In a preferred embodiment, the edible mycelium component B has an elemental composition with a C to N ratio (mycelium) in the range of 1-30, preferably 1-15, preferably 5-12, more preferably 8-12, most preferably 8-10.

[0254] In a preferred embodiment, the edible mycelium component C has an elemental composition with a C to N ratio (mycelium) in the range of 1-30, preferably 1-15, preferably 1-8, more preferably 2-6, most preferably 4-6.

[0255] In a further embodiment, the present invention relates to an edible fibrous mycelial mass as described herein. Preferably, the edible fibrous mycelial mass is derived from submerged fermentation.

[0256] In another embodiment, without any additional ribonucleic acid (RNA) reduction steps via additional heating or pH treatment in the component production process, the method of the present invention preferably results in component indigenous RNA levels of at most 4% by weight, preferably at most 2% by weight, on a dry basis for all disclosed mycelial components. This is applicable to each of Components A, B, and C.

[0257] In one embodiment, the RNA value for the edible mycelium component is 0.1-4% by weight, preferably 0.1-2% by weight, preferably 0.1-1.5% by weight, more preferably 0.1-1% by weight, more preferably 0.1-0.4% by weight. In a preferred embodiment, the indigenous RNA level for component A is 0.1-4% by weight, preferably 0.1-2% by weight, more preferably 0.4%-1.88% by weight, on a dry basis. In a preferred embodiment, the indigenous RNA level for component B is 0.1-4% by weight, preferably 0.1-2% by weight, more preferably 0.4%-1.7% by weight, on a dry basis. In a preferred embodiment, the indigenous RNA level for component C is 0.1-4% by weight, preferably 0.1-2% by weight, more preferably 0.5%-1.9% by weight, on a dry basis.

[0258] In another embodiment, the edible mushroom ingredient contains 1-35 wt% chitin by dry weight, preferably 1-20 wt%, more preferably 1-15 wt%, more preferably 1-14 wt%, more preferably 1-13 wt%, more preferably 1-12 wt%, more preferably 1-11 wt%, more preferably 1-10 wt%, more preferably 1-9 wt%, more preferably 1-8 wt%, more preferably 1-7 wt%, more preferably 1-6 wt%, more preferably 1-5 wt%, more preferably 1-4 wt%, more preferably 1-3 wt%, and more preferably 1-2 wt%. In a preferred embodiment, the edible mushroom ingredient contains 5-15 wt%, more preferably 6-12 wt%, and most preferably 6-9 wt% chitin by dry weight.

[0259] In certain embodiments, the mycelium component has an ergothioneine level in the range of 1 to 1000 mg / kg, preferably 1 to 900 mg / kg, more preferably 1 to 800 mg / kg, more preferably 1 to 700 mg / kg, more preferably 1 to 600 mg / kg, more preferably 1 to 500 mg / kg, more preferably 1 to 400 mg / kg, more preferably 1 to 300 mg / kg, more preferably 1 to 200 mg / kg, more preferably 1 to 100 mg / kg, more preferably 1 to 50 mg / kg, and more preferably 1 to 25 mg / kg. In a preferred embodiment, the mycelium component contains an ergothioneine level in the range of 25 to 1000 mg / kg, preferably 50 to 700 mg / kg, and more preferably 80 to 600 mg / kg. In a preferred embodiment, the mycelium component A contains an ergothioneine level in the range of 70 to 270 mg / kg, and this level is preferably achieved within 5 days. In a preferred embodiment, mycelium component B contains an ergothioneine value in the range of 100-350 mg / kg, which value is preferably achieved within 5 days. In a preferred embodiment, mycelium component C contains an ergothioneine value in the range of 300-800 mg / kg, which value is preferably achieved within 5 days, preferably an ergothioneine value in the range of 350-800 mg / kg, which value is preferably achieved within 5 days, and most preferably an ergothioneine value in the range of 400-800 mg / kg, which value is preferably achieved within 5 days. As understood herein, ergothioneine value preferably refers to the amount of ergothioneine expressed in mg per kg of mycelium component expressed on a dry mass basis.

[0260] As has been surprisingly found by the present inventors, when Pleurotus pulmonarius is used as the fungal strain in a fermentation system for 5 days, mycelium component A preferably has an ergothioneine content in the range of 70-100 mg / kg, component B preferably contains ergothioneine at a value in the range of 110-150 mg / kg, and component C exhibits the highest content of ergothioneine, with the content in the range of 380-455 mg / kg. The average value reached 433 mg / kg. This is a higher value of this functional substance compared to patents disclosed in the art that use Pleurotus pulmonarius to produce ergothioneine.

[0261] In a specific embodiment, the ergosterol content in the mycelium component is in the range of 0 to 50 mg / g, preferably 0 to 25 mg / g, more preferably 0 to 15 mg / g, more preferably 0 to 12.5 mg / g, more preferably 0 to 10 mg / g, more preferably 0 to 9 mg / g, more preferably 0 to 8 mg / g, more preferably 0 to 7 mg / g, more preferably 0 to 5 mg / g, more preferably 0 to 4 mg / g, more preferably 0 to 3 mg / g, more preferably 0 to 2 mg / g, and more preferably 0 to 1 mg / g. In a preferred embodiment, the ergosterol content in the mycelium component is in the range of 0 to 8 mg / g, more preferably 1 to 6 mg / g, and most preferably 2 to 6 mg / g. In an even more preferred embodiment, mycelium components A and B contain ergosterol in the range of 0 to 4 mg / g, more preferably 2 to 4 mg / g. In a further preferred embodiment, mycelium component C contains an ergosterol range of 0 to 8 mg / g, more preferably 2 to 7 mg / g, more preferably 4 to 7 mg / g, and most preferably 5 to 6 mg / g.

[0262] In certain embodiments, the flavor is measured relative to known-tasting 5'-nucleotides (5'-NMPs) in g / kg, including 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP), and 5'-adenosine monophosphate (AMP). In a preferred embodiment, IMP is absent and has a value of 0 g / kg. In a further embodiment, the umami flavor derived from 5'-nucleotides can be further enhanced by at least 30%, preferably at least, more preferably at least 50%, when treated with an enzyme, preferably 5'-adenylate deaminase, to convert AMP to IMP.

[0263] In one embodiment, the edible mushroom ingredient exhibits good richness of 5'NMP, which contains 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP), in the range of 0.1 to 40 g / kg, preferably 0.1 to 20 g / kg, preferably 0.1 to 15 g / kg, more preferably 0.1 to 12.5 g / kg, more preferably 0.1 to 10 g / kg, more preferably 0.1 to 8 g / kg, more preferably 0.1 to 6 g / kg, more preferably 0.1 to 5 g / kg, more preferably 0.1 to 4 g / kg, more preferably 0.1 to 3 g / kg, more preferably 0.1 to 2 g / kg.

[0264] In a preferred embodiment, this value of 5'NMP for component A ranges from 1 to 20 g / kg, more preferably from 2 to 6 g / kg, most preferably from 3 to 6 g / kg.

[0265] In a preferred embodiment, this value of 5'NMP for component B ranges from 0.1 to 15 g / kg, more preferably from 0.1 to 5 g / kg, and most preferably from 0.1 to 3 g / kg.

[0266] In a preferred embodiment, this value of 5'NMP for component C ranges from 0.1 to 40 g / kg, more preferably from 5 to 20 g / kg, most preferably from 8 to 20 g / kg.

[0267] In another preferred embodiment, this value of 5'NMP for mycelial component A is at most 40 g / kg, preferably at most 20 g / kg.

[0268] In another preferred embodiment, this value of 5'NMP for mycelial component B is at most 40 g / kg, preferably at most 20 g / kg.

[0269] In another preferred embodiment, this value of 5'NMP for mycelial component C is at most 40 g / kg, preferably at most 20 g / kg.

[0270] It is observed that mycelia grown on brewer's grain extract resulted in a 10-fold increase in the content of 5'NMP.

[0271] In another embodiment, mycelium component C exhibits a high concentration of uridine monophosphate (UMP), preferably in the range of 2.5 to 5 g / kg of UMP.

[0272] In certain embodiments, the mycelium components preferably contain all 20 essential amino acids, hi a preferred embodiment, the mycelium components include the amino acids alanine, asparagine, aspartate, arginine, tryptophan, glycine, glutamic acid, glutamine, methionine, phenylalanine, serine, valine, cystine, proline, leucine, tyrosine, threonine, isoleucine, histidine, and lysine.

[0273] In a preferred embodiment, the edible mycelium component contains at least about 10% by weight, preferably at least about 15% by weight, more preferably at least about 19% by weight, more preferably at least about 20% by weight, more preferably at least about 21% by weight, more preferably at least about 22% by weight, more preferably at least about 23% by weight of branched chain amino acids (BCAAs) of the total amount of amino acids present (i.e., of the total protein, since the amino acids present as referred to herein relate to free amino acids and the total content of amino acids contained within proteins and peptides).

[0274] In a preferred embodiment, the edible mycelium component contains at least about 10% by weight, preferably at least about 15% by weight, more preferably at least about 19% by weight, more preferably at least about 20% by weight, more preferably at least about 21% by weight, more preferably at least about 22% by weight, more preferably at least about 23% by weight of umami amino acids of the total amount of amino acids present (i.e., of the total protein, since the amino acids present as referred to herein relate to the total content of free amino acids and amino acids contained within proteins and peptides).

[0275] Essential amino acids are understood herein to be amino acids that cannot be synthesized efficiently enough by an organism to meet its needs and must be supplied from the diet.

[0276] In a preferred embodiment, the edible mycelium component contains at least about 30% by weight, preferably at least about 35% by weight, more preferably at least about 40% by weight of essential amino acids of the total amount of amino acids present (i.e., of the total protein, since the amino acids present referred to herein relate to the total content of free amino acids and amino acids contained within proteins and peptides).

[0277] In a preferred embodiment, the total amount of amino acids in component A corresponds to 150 to 300 mg / g, preferably 200 to 300 mg / g, more preferably 250 to 300 mg / g, and most preferably 250 to 280 mg / g.

[0278] In a preferred embodiment, the total amount of amino acids in component B grown in a minimal medium corresponds to 100 to 200 mg / g, preferably 120 to 180 mg / g, more preferably 150 to 180 mg / g.

[0279] In a preferred embodiment, the total amount of amino acids in component B grown on brewer's grain extract corresponds to 200 to 700 mg / g, preferably 250 to 550 mg / g, more preferably 300 to 500 mg / g, most preferably 350 to 450 mg / g.

[0280] In another preferred embodiment, in the edible mycelium component A, the amount of each BCAA and the amount of umami amino acid are each separately in the range of 40 to 100 mg / g, preferably 50 to 60 mg / g.

[0281] In another preferred embodiment, in the edible mycelium component B, the amount of each BCAA and the amount of umami amino acid are each separately in the range of 20 to 80 mg / g, preferably 30 to 40 mg / g.

[0282] In another preferred embodiment, in the edible mycelium component C, the amount of BCAA is in the range of 50 to 150 mg / g, preferably 65 to 75 mg / g, and the amount of umami amino acids is in the range of 50 to 150 mg / g, preferably 70 to 100 mg / g, most preferably 75 to 85 mg / g.

[0283] In certain embodiments, the EUC concentration of the mycelium component ranged from 1 to 25,000%. In preferred embodiments, the EUC concentration of the mycelium component ranged from 1 to 20,000%, preferably from 1 to 150,000%, more preferably from 1 to 13,000%, more preferably from 1 to 12,000%, more preferably from 1 to 10,000%, more preferably from 1 to 5,000%, more preferably from 1 to 2,500%, more preferably from 1 to 1,500%, more preferably from 1 to 1,000%, and more preferably from 1 to 600%.

[0284] In a preferred embodiment of mycelium component A, the EUC concentration ranges from 1 to 1000%, more preferably from 250 to 800%, and even more preferably from 250 to 500%. Preferably, the EUC concentration for mycelium component A is about 300%. In another preferred embodiment of mycelium component A, the EUC is 200 to 500 g MSG / 100 g.

[0285] Preferably, the mycelium component A is characterized by an insoluble fiber content of 25 to 45% w / w, more preferably 30 to 45% w / w, for example, 34% w / w, where % w / w refers to the insoluble fiber content on a dry mass basis.

[0286] In a preferred embodiment for mycelium component B, the EUC concentration is in the range of 1-600%, preferably 1-200%, more preferably 10-100%, and even more preferably 30-60%. Preferably, the EUC concentration for mycelium component B is about 34%. Alternatively, in a preferred embodiment for mycelium component B, the EUC is less than 200g MSG / 100g, more preferably less than 100g MSG / 100g. Alternatively, in one embodiment for mycelium component B, the EUC is preferably 30-200g MSG / 100g, and even more preferably 30-60g MSG / 100g.

[0287] Preferably, the mycelium component B is characterized by an insoluble fiber content of 30-60% w / w, preferably 40-60% w / w, for example 54% w / w, where % w / w refers to the insoluble fiber content on a dry mass basis.

[0288] In a preferred embodiment for mycelial component C, the EUC concentration is in the range of 1-10000%, more preferably 500-5000%, even more preferably 2000-4000%, and even more preferably 2500-3500%. Preferably, the EUC concentration for mycelial component C is about 2890%. In a preferred embodiment for mycelial component C, mycelial component C is characterized by an EUC concentration of at least 500%, more preferably at least 1000%, even more preferably 1500%, and even more preferably at least 2000%.

[0289] Preferably, the mycelium component C is characterized by an insoluble fiber content of at most 35% w / w, more preferably at most 30% w / w, e.g., 25% w / w, where % w / w refers herein to the insoluble fiber content by dry mass.

[0290] Note that the above EUC concentrations are achieved in 5 days.

[0291] The uronic acids analyzed in the mycelium component, particularly D-galacturonic acid and D-glucuronic acid, are highly valuable substances and can act as antioxidants and detoxifiers and inactivators of various substances in the human body. In certain embodiments, the content of both uronic acids in the mycelium component after acid hydrolysis is in the range of 0-50% by weight, preferably 0-25% by weight, more preferably 0-15% by weight, more preferably 0-10% by weight, more preferably 0-5% by weight, more preferably 0-2.5% by weight, and more preferably 0-2% by weight. In a preferred embodiment, the content of uronic acids in the mycelium component is in the range of 0.1-15% by weight, more preferably 0.1-5% by weight, and most preferably 1-2% by weight.

[0292] In certain embodiments, the total phenolic content of the mycelium component is in the range of 1-200 mg GAE / g, 1-150 mg GAE / g, 1-100 mg GAE / g, 1-50 mg GAE / g, 1-25 mg GAE / g, 1-10 mg GAE / g, or 1-5 mg GAE / g. In preferred embodiments, the total phenolic content of the mycelium component is in the range of 1-150 mg GAE / g, preferably 1-15 mg GAE / g, and more preferably 1-5 mg GAE / g.

[0293] In certain embodiments, the total flavonoid content of the mycelium component is in the range of 1-200 mg QE / g, 1-150 mg QE / g, 1-100 mg QE / g, 1-50 mg QE / g, 1-25 mg QE / g, 1-10 mg QE / g, or 1-5 mg QE / g. In preferred embodiments, the total flavonoid content of the mycelium component is in the range of 1-150 mg QE / g, preferably 1-15 mg QE / g, and more preferably 1-5 mg QE / g.

[0294] In a specific embodiment, mycelium component A has a total phenolic content in the range of 1-5 mg GAE / g, preferably about 3.2 mg GAE / g.

[0295] In a specific embodiment, mycelium component B has a total phenolic content in the range of 1-5 mg GAE / g, preferably about 2.7 mg GAE / g.

[0296] In a specific embodiment, mycelium component C has a total phenolic content in the range of 1-5 mg GAE / g, preferably about 4.5 mg GAE / g.

[0297] In a specific embodiment, mycelium component A has a total flavonoid content in the range of 1-5 mg QE / g, preferably about 1.7 mg QE / g.

[0298] In a specific embodiment, mycelium component B has a total flavonoid content in the range of 1-5 mg QE / g, preferably about 2.5 mg QE / g.

[0299] In a specific embodiment, mycelium component C has a total flavonoid content in the range of 1-5 mg QE / g, preferably about 3 mg QE / g.

[0300] It is clear that the extracts contained a lower total flavonoid content than the total phenolic content. The results suggest that compounds other than flavonoids are the major phenolic substances present in the tested strains, particularly for components A and B. In one embodiment, the flavonoid content of mycelium component A constitutes approximately 35% to 60% of the total phenolic content, preferably approximately 45% to 55%, and preferably approximately 55% of the total phenolic content. In one embodiment, the flavonoid content of mycelium component B constitutes approximately 70% to 95% of the total phenolic content, preferably approximately 80% to 95%, and preferably approximately 93% of the total phenolic content. In one embodiment, the flavonoid content of mycelium component C constitutes approximately 50% to 80% of the total phenolic content, preferably approximately 60% to 70%, and preferably approximately 65% ​​of the total phenolic content.

[0301] In one embodiment, the total polyphenols of the mycelium component ranges from 1 to 2000 mg / kg, 1 to 1000 mg / kg, 1 to 900 mg / kg, 1 to 800 mg / kg, 1 to 700 mg / kg, 1 to 600 mg / kg, 1 to 500 mg / kg, 1 to 400 mg / kg, 1 to 300 mg / kg, 1 to 200 mg / kg, 1 to 100 mg / kg, 1 to 50 mg / kg, or 1 to 25 mg / kg. In a preferred embodiment, the total polyphenols of the mycelium component ranges from 1 to 1500 mg / kg, preferably 25 to 1000 mg / kg, more preferably 50 to 900 mg / kg, and this value is preferably achieved within 5 days. This is applicable to Components A, B, and C.

[0302] In a specific embodiment, mycelium component A has a total polyphenol content in the range of 1 to 500 mg / kg, preferably 50 to 200 mg / kg, more preferably approximately 165 mg / kg. In an even more preferred embodiment, mycelium component A has a total polyphenol content in which approximately 35 to 45 wt% of the total polyphenols corresponds to catechin, approximately 35 to 45 wt% of the total polyphenols corresponds to protocatechuic acid, and the remaining approximately 1 to 10 wt% is each of quercetin, chlorogenic acid, and / or syringic acid.

[0303] In a specific embodiment, mycelium component B has a total polyphenol content in the range of 1 to 250 mg / kg, preferably 50 to 150 mg / kg, more preferably approximately 75 mg / kg. In an even more preferred embodiment, mycelium component B has a total polyphenol content in which approximately 25 to 35% by weight of the total polyphenols corresponds to catechin, approximately 35 to 45% by weight of the total polyphenols corresponds to protocatechuic acid, approximately 10 to 20% by weight of the total polyphenols corresponds to quercetin, and the remaining approximately 5 to 15% by weight of the total polyphenols corresponds to chlorogenic acid.

[0304] In a specific embodiment, mycelium component C has a total polyphenol content in the range of 1 to 1500 mg / kg, preferably 100 to 1000 mg / kg, more preferably 250 to 950 mg / kg, more preferably approximately 650 mg / kg. In a further preferred embodiment, mycelium component C has a total polyphenol content in which approximately 45 to 55% by weight of the total polyphenols corresponds to catechin, approximately 45 to 55% by weight of the total polyphenols corresponds to protocatechuic acid, and approximately 1 to 5% by weight of the total polyphenols corresponds to syringic acid.

[0305] Ethanol extracts of mycelial components were prepared using 96% ethanol in an accelerated solvent extractor, as described in [2020;8(7):803. https: / / doi.org / 10.3390 / pr8070803]. Antioxidant assays were performed to assess antioxidant activity, which was determined by the ability to scavenge stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals according to the scavenging activity protocol (A potential antioxidant resource: endophytic fungi from medicinal plants. Econ. Bot. 61, 14-30.). This is typically defined by the EC50 value, preferably defined as the concentration of antioxidant required for scavenging 50% of DPPH radicals within a specified time period.

[0306] In certain embodiments, the EC50 for the mycelial component ranges from 1 to 100 mg / ml, preferably from 1 to 50, more preferably from 1 to 15 mg / ml. In a preferred embodiment, the EC50 for mycelial component A ranges from 1 to 25 mg / ml, preferably about 10.5 mg / ml.

[0307] In a preferred embodiment, the EC50 for mycelial component B is in the range of 1 to 25 mg / ml, preferably about 13 mg / ml. In a preferred embodiment, the EC50 for mycelial component C is in the range of 1 to 25 mg / ml, preferably about 9 mg / ml.

[0308] In a preferred embodiment, mycelium component C has the highest DPPH radical scavenging activity, preferably with an EC50 of about 9 mg / ml, which correlates well with the phenolic content of component C, which has the highest phenolic content, and therefore phenolic content and antioxidant activity are well correlated.

[0309] In a preferred embodiment, mycelium component A has the second highest DPPH radical scavenging activity, preferably with an EC50 of about 10.5 mg / ml.

[0310] In a preferred embodiment, mycelium component B has second to third DPPH radical scavenging activity, preferably with an EC50 of about 13 mg / ml.

[0311] The following sugars were measured in the mycelial component after acid hydrolysis: glucan, xylan, arabinan, galactan, mannan, and rhamnan. In certain embodiments, the total lignocellulosic sugars detected from the hydrolysis of the mycelial component range from 1 to 100%, preferably 1 to 90%, more preferably 1 to 80%, more preferably 1 to 70%, more preferably 1 to 60%, more preferably 1 to 50%, more preferably 1 to 40%, more preferably 1 to 30%, more preferably 1 to 25%, more preferably 1 to 20%, more preferably 1 to 15%, more preferably 1 to 10%, and more preferably 1 to 5%. In a preferred embodiment, the total lignocellulosic sugars detected from the hydrolysis of mycelial component A range from 20 to 60%, more preferably 25 to 45%, with glucan being the most abundant sugar, constituting 80 to 90% of the total sugars measured (the glucan content is preferably 25 to 40% by weight in absolute terms).

[0312] In a further preferred embodiment, the total sugars detected from the hydrolysis of mycelial component B are in the range of 30-80%, more preferably 20-70%, and most preferably 40-60%, with the most abundant sugar being glucan, which constitutes 85-95% of the total sugars (the glucan content is preferably 40-50% by weight in absolute terms). In a further preferred embodiment, the total sugars detected from the hydrolysis of mycelial component C are in the range of 1-50%, more preferably 1-35%, and most preferably 1-30%, with the most abundant sugar being glucan, which constitutes 75-85% of the total sugars (the glucan content is preferably 10-25% by weight in absolute terms).

[0313] In another preferred embodiment, the most abundant sugar detected from the hydrolysis of mycelial components A, B, or C is glucan in the range of 70-95% of the total sugars.

[0314] As understood herein, the term "insoluble fiber" preferably refers to a portion of dietary fiber that does not dissolve in water. The insoluble fiber preferably comprises chitin and β-glucan, and is distinguishable from insoluble fiber of plant origin, which preferably comprises plant cellulose and / or hemicellulose but does not comprise chitin. Insoluble fiber is known to help the human body process waste better, improve intestinal health, and reduce the risk of colorectal conditions.

[0315] Preferably, the edible fibrous mycelium component has an insoluble fiber content of at least 20%, preferably at least 30%, preferably at least 40%, preferably at least 50%, preferably at least 60%. More preferably, the edible fibrous mycelium has an insoluble fiber content of 20% w / w to 60% w / w. Even more preferably, the edible fibrous mycelium has an insoluble fiber content of 40% w / w to 55% w / w.

[0316] In a preferred embodiment, the insoluble fiber content of the mycelium component A is in the range of 30 to 60% by weight, preferably 30 to 40% by weight.

[0317] In a preferred embodiment, the insoluble fiber content of mycelium component B is in the range of 40 to 60% by weight, preferably 40 to 50% by weight.

[0318] In a preferred embodiment, the insoluble fiber content of the mycelium component C is in the range of 10 to 40% by weight, preferably 20 to 30% by weight.

[0319] Preferably, it is contemplated that the mycelium used for dairy vs. meat vs. other products can be tailored to the specific requirements of the final product with respect to fiber content, protein content, nutrients, taste, etc.

[0320] In certain preferred embodiments, the insoluble fiber content of the edible fibrous mycelium used in the meat substitute product formulation is 20-60% by weight, preferably 40-50% by weight, and preferably about 45% by weight. In another preferred embodiment, the insoluble fiber used in the meat substitute is about 35% by weight. In another preferred embodiment, the insoluble fiber used in the meat substitute is about 25% by weight. In another preferred embodiment, the insoluble fiber used in the meat substitute is about 45% by weight. In one embodiment, the edible fibrous mycelium in the meat substitute has an insoluble fiber content of at least 20% by weight, preferably at least 30% by weight, preferably at least 45% by weight, and preferably at least 50% by weight.

[0321] In a further preferred embodiment, the insoluble fiber content of the edible fibrous mycelium used in dairy alternative products or other products (other than meat alternatives) is at least 20% by weight, more preferably at least 30% by weight, more preferably at least 40% by weight. More preferably, the edible fibrous mycelium has an insoluble fiber content of 40% to 60% by weight. Even more preferably, the edible fibrous mycelium has an insoluble fiber content of 40% to 55% by weight. Even more preferably, the edible fibrous mycelium has an insoluble fiber content of 40 to 50% by weight. Most preferably, the edible fibrous mycelium has an insoluble fiber content of about 45% by weight. Most preferably, the edible fibrous mycelium has an insoluble fiber content of at least 50% by weight. It should be understood that weights referred to herein are relative to the dry mass of the mycelium. In one embodiment, the edible fibrous mycelium has an insoluble fiber content of at least 40% by weight, preferably at least 50% by weight, more preferably at least 60% by weight.

[0322] The edible fibrous mycelium of the present invention preferably has a protein content of 10% to 65% by weight, most preferably 30% to 60% by weight, hi certain embodiments, the protein content is about at least 30%, at least 40%, or at least 60% by weight.

[0323] In a preferred embodiment, the protein content of the mycelium component A is in the range of 30 to 50% by weight, preferably 30 to 40% by weight.

[0324] In a preferred embodiment, the protein content of mycelium component B is in the range of 30 to 50% by weight, preferably 30 to 40% by weight.

[0325] In a preferred embodiment, the protein content of mycelium component C is in the range of 30 to 65% by weight, preferably 45 to 65% by weight.

[0326] The protein content of the mycelial mass can be adjusted by selecting fermentation conditions that control the protein content of the resulting mycelial mass. Such conditions are known to those skilled in the art and include adjusting the contents of the fermentation medium to control uptake and thus the composition of the resulting mycelial mass. For example, the ratio between the carbon source and the nitrogen source in the medium can be varied, such as providing nitrogen in various excesses. This affects the insoluble fiber content, and therefore, as shown in Table 3, higher protein means lower insoluble fiber content.

[0327] In one embodiment, the edible mycelium component has a carbohydrate content (non-fiber carbohydrates, i.e., carbohydrates that do not include fiber) of at most 5%, preferably at most 1%. In a preferred embodiment, the edible mycelium component has a carbohydrate content of at most 0.5%, most preferably less than 0.01%.

[0328] In one embodiment, the edible mycelium component has a beta-glucan content of at least 80% of the total glucans.

[0329] In one embodiment, the present invention relates to an edible mycelium component A, which has a total glucan content in the range of 20 to 35% by weight, preferably 25 to 35% by weight. In one embodiment, the present invention relates to an edible mycelium component B, which has a total glucan content in the range of 25 to 50% by weight, preferably 30 to 40% by weight. In one embodiment, the present invention relates to an edible mycelium component C, which has a total glucan content in the range of 10 to 35% by weight, preferably 10 to 20% by weight.

[0330] In one embodiment, the present invention relates to mycelial component A, B, or C, wherein at least 96% by weight of the polyunsaturated fatty acid content is linoleic acid (omega-6 fatty acid). In a particularly preferred embodiment, the present invention relates to mycelial component A, wherein at least 98% by weight of the polyunsaturated fatty acid content is linoleic acid (omega-6 fatty acid). In a particularly preferred embodiment, the present invention relates to mycelial component B, wherein at least 97% by weight of the polyunsaturated fatty acid content is linoleic acid (omega-6 fatty acid). In a particularly preferred embodiment, the present invention relates to mycelial component C, wherein at least 96% by weight of the polyunsaturated fatty acid content is linoleic acid (omega-6 fatty acid). In a further specific embodiment, the present invention relates to a mycelium component C having a concentrated omega-6 fatty acid (linoleic acid) content in the range of 1-9% by weight, more preferably 2-5% by weight, and most preferably about 3.5% by weight.

[0331] In one embodiment, mycelium components A, B, and C have a fat content of at most 15% by weight, preferably at most 10% by weight. In a specifically preferred embodiment, the present invention relates to mycelium component A, which has a fat content in the range of 0.5 to 5% by weight, preferably 0.1 to 3% by weight. In a specifically preferred embodiment, the present invention relates to mycelium component B, which has a fat content in the range of 0.5 to 5% by weight, preferably 0.1 to 3% by weight. In a specifically preferred embodiment, the present invention relates to mycelium component C, which has a fat content in the range of 1 to 10% by weight, preferably 3 to 10% by weight, more preferably 3 to 8% by weight. In a specifically preferred embodiment, the present invention relates to mycelium component C, which has a fat content of at most 8% by weight.

[0332] In one embodiment, the mycelium components are free of mycotoxins. Mycotoxins were analyzed via liquid chromatography-tandem mass spectrometry (SOP M 3650). Mycotoxins were not present above their detection limits in μg / kg. The mycotoxins are selected from aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, ochratoxin A, deoxynivalenol (DON), zearalenone, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, T-2 toxin, HT-2 toxin, 4,15-diacetoxyscirpenol, fusarenon-X, fumonisin B1, and fumonisin B2.

[0333] In another embodiment, polyaromatic hydrocarbons were analyzed using gas chromatography-mass spectrometry (SOP M 2920). Polyaromatic hydrocarbons were not detected above their detection limit in μg / kg. The polyaromatic hydrocarbons are selected from benzo(a)anthracene, benzo(c)fluorene, chrysene, cyclopenta(c,d)pyrene, 5-methylchrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(j)fluoranthene, benzo(a)pyrene, indeno(1,2,3-cd)pyrene, dibenzo(ah)anthracene, benzo(ghi)perylene, dibenzo(a,l)pyrene, dibenzo(a,e)pyrene, dibenzo(a,i)pyrene, and dibenzo(a,h)pyrene.

[0334] In certain embodiments, the calorific value of the mycelium component is in the range of 1 to 1000 Kcal / 100g, preferably 1 to 900 Kcal / 100g, more preferably 1 to 800 Kcal / 100g, more preferably 1 to 700 Kcal / 100g, more preferably 1 to 500 Kcal / 100g, more preferably 1 to 400 Kcal / 100g, more preferably 1 to 300 Kcal / 100g, more preferably 1 to 200 Kcal / 100g, more preferably 1 to 100 Kcal / 100g, more preferably 1 to 50 Kcal / 100g, more preferably 1 to 25 Kcal / 100g. In a preferred embodiment, the calorific value of the mycelium component is in the range of 1 to 800 Kcal / 100 g, preferably 200 to 800 / 500 Kcal / 100 g, more preferably 300 to 600 Kcal / 100 g, and most preferably about 300 to 500 Kcal / 100 g.

[0335] In a preferred embodiment, for pore diameters less than 1 mm, 55-65%, preferably about 58%, of the pore volume for edible mushroom component A corresponds to pore diameters of 1000-30 μm, with the modal pore diameter in the range of 85-185 μm. In a second range of 30-2 μm, corresponding to about 35-45%, preferably 42%, of the pore volume, the modal pore diameter peak is equal to 16 μm, which is also the modal pore diameter in the range of 1000-2 μm. The specific pore volume of component A is 9 cm3 / g, and the median pore diameter is 44.5 μm.

[0336] In a preferred embodiment, for pore diameters less than 1 mm, about 75-85%, preferably 81.5%, of the pore volume for edible mushroom component B corresponds to pore diameters of 1000-30 μm, with the modal pore diameter equal to 147 μm, which is also the modal pore diameter in the range of 1000-2 μm. In a second range of 30-2 μm, corresponding to about 15-25%, preferably 18.5%, of the pore volume, the modal pore diameter peak is equal to 15 μm. The specific pore volume of component B is 4.94 cm3 / g, and the median pore diameter is 143 μm.

[0337] In a preferred embodiment, for pore diameters less than 1 mm, 15% to 25%, preferably about 20%, of the pore volume for edible mushroom component C corresponds to pore diameters between 1000 and 20 μm, and 75% to 85%, preferably about 80%, of the pore volume corresponds to pore diameters between 20 and 2 μm, with the modal pore diameter peak being 5.5 μm. The specific pore volume for component B is 2.46 cm3 / g, and the median pore diameter is 7.1 μm.

[0338] Component C pore volume is much lower and the pores are smaller compared to A and B.

[0339] In a preferred embodiment, the BET surface areas of components A, B, and C were also measured using krypton, as krypton is a suitable adsorbate for measuring low surface areas. The surface areas of the components without further milling are preferably 0.79 m / g for A, 0.67 m / g for B, and 1.59 m / g for C.

[0340] In one embodiment, edible mycelium components A and B have a thermal stability of up to 210-220°C, preferably at most 220°C, under N2 atmosphere. This is shown in thermogravimetric analysis (TGA), determined at the point where all water content has been lost by mass. In one embodiment, edible mycelium component C has a thermal stability of up to 175-185°C, preferably at most 190°C, under N2 atmosphere. As shown in Figures 2, 3, and 4, it is observed that C has less thermal stability than A, and A has slightly less thermal stability than B, up to 1000°C.

[0341] The textural attributes of the mycelial components are analyzed herein in terms of shear strength, water retention, water release, and density.

[0342] The shear force is influenced by the morphology of the mycelial biomass and its ability to withstand the applied force. In certain embodiments, the shear force of the mycelial component is in the range of 1-200N, 1-150N, 1-100N, 1-90N, 1-80N, 1-70N, 1-60N, 1-50N, 1-40N, 1-30N, 1-25N, 1-20N, 1-15N, 1-10N, or 1-5N. In preferred embodiments, the shear force of the mycelial biomass is at least 10N, preferably at least 15N, preferably at least 25N, and preferably in the range of 25-150N, more preferably 30-120N, and most preferably 30-105N.

[0343] In certain embodiments, the mycelial biomass has a water holding capacity in the range of 1-100%, 1-90%, 1-80%, 1-70%, 1-60%, 1-50%, 1-40%, 1-30%, 1-25%, 1-20%, 1-15%, 1-10%, or 1-5%. In preferred embodiments, the water holding capacity is in the range of 20-90%, preferably 30-80%, and more preferably 40-70%.

[0344] In certain embodiments, the mycelial biomass has a water release in the range of 1-100%, 1-90%, 1-80%, 1-70%, 1-60%, 1-50%, 1-40%, 1-30%, 1-25%, 1-20%, 1-15%, 1-10%, or 1-5%. In another preferred embodiment, the water release of the mycelium is in the range of 25-70%, more preferably 30-55%.

[0345] In certain embodiments, the mycelial biomass is between 0.1 and 10 g / cm 3 , 0.1~9g / cm 3 , 0.1~8g / cm 3 , 0.1~7g / cm 3 , 0.1~6g / cm 3 , 0.1~5g / cm 3 , 0.1~4g / cm 3 , 0.1~3g / cm 3 , 0.1~2.5g / cm 3 , 0.1~2g / cm 3 , 0.1~1.5g / cm 3 , 0.1~1g / cm 3 , 0.1~0.8g / cm 3 , or 0.1 to 0.5 g / cm 3 In a preferred embodiment, the density of the mycelial biomass is in the range of 0.1 to 3, preferably 0.5 to 2.5, more preferably 0.5 to 1.5 g / cm. 3 Preferably, the density of the mycelial biomass is in the range of approximately 1 g / cm 3 is.

[0346] The textural attributes in fungal products derived from mycelium components are influenced by the compositional ingredients of the product, resulting in either soft or non-soft (i.e., hard) mycelium-based meat or milk substitutes.

[0347] In some embodiments, the meat substitute or meat-like food product is understood to have a similar consistency or similarity or taste to animal meat in all its forms (breasts, fillets, thighs, ribs, wings, chunks, steaks, etc.), preferably selected from beef, poultry, fish, chicken, duck, goose, turkey, beef, pheasant, lamb and mutton, white meat, pork, ham, veal, deer (or venison), seafood, shrimp, crab, salmon, cod, pangasius, sardines, mussels, and oysters.

[0348] In a preferred embodiment, the soft meat substitute is preferably understood as meatballs. In another preferred embodiment, the soft meat substitute is preferably meatballs, sausages, fish fingers, tartare, minced meat, meat spreads, processed meats, met meats, luncheon meats and foie gras.

[0349] In another preferred embodiment, non-tender meat substitutes are preferably understood as steaks, beef jerky, burger patties, tenderloins, nuggets, salami, whole cuts, bacon, hot dogs, prosciutto, dried meats and extruded products.

[0350] In another embodiment, the concepts of non-soft meat and soft meat may be interchangeable only if the ingredients used to produce the conventional non-soft meat result in a meat substitute that is softer in terms of consistency compared to the conventional definition.

[0351] In a preferred embodiment, the soft milk substitute is preferably understood as cream cheese. In another preferred embodiment, the soft milk substitute is preferably understood as cream cheese, cheese spread, processed cheese, whey cheese, pizza cheese, shredded mozzarella cheese, mozzarella cheese, soft cheese, semi-soft cheese, feta cheese, ricotta cheese, cottage cheese, Camembert cheese, Roquefort cheese, Gorgonzola cheese, Brie cheese, blue cheese, Bouchette cheese, goat cheese, quark, cream, coffee creamer, whipped cream, sour cream, milk chocolate spread, margarine, butter, dessert, custard. In another embodiment, the non-soft or hard milk substitute is preferably understood as hard cheese, semi-hard cheese, Cheddar cheese, Parmesan cheese, etc.

[0352] In certain embodiments, the cutting strength of the food product is in the range of 1 to 100 N, 1 to 90 N, 1 to 80 N, 1 to 70 N, 1 to 60 N, 1 to 50 N, 1 to 40 N, 1 to 30 N, 1 to 25 N, 1 to 20 N, 1 to 15 N, 1 to 10 N, or 1 to 5 N. In preferred embodiments, the cutting strength for soft meat substitutes is in the range of 1 to 25 N, most preferably 4 to 8 N. In preferred embodiments, the cutting strength for hard meat substitutes is in the range of 1 to 50 N, most preferably 10 to 30 N.

[0353] In certain embodiments, the hardness of the food product is in the range of 1 to 200 N, 1 to 150 N, 1 to 100 N, 1 to 90 N, 1 to 80 N, 1 to 70 N, 1 to 60 N, 1 to 50 N, 1 to 40 N, 1 to 30 N, 1 to 25 N, 1 to 20 N, 1 to 15 N, 1 to 10 N, or 1 to 5 N. In a preferred embodiment, the hardness for the soft meat substitute is in the range of 10 to 55 N, preferably 15 to 55 N, more preferably 20 to 50 N, and most preferably 20 to 45 N. In a preferred embodiment, the hardness for the hard meat substitute is in the range of 30 to 100 N, more preferably 50 to 100 N.

[0354] In certain embodiments, the springiness of the food product is in the range of 1-100%, 1-90%, 1-80%, 1-70%, 1-60%, 1-50%, 1-40%, 1-30%, 1-25%, 1-20%, 1-15%, 1-10%, or 1-5%. In a preferred embodiment, the springiness for the soft meat substitute is in the range of 35-85%, preferably 45-80%, more preferably 50-80%, and most preferably 55-75%. In a preferred embodiment, the springiness for the hard meat substitute is in the range of 20-70%, preferably 30-60%.

[0355] In certain embodiments, the cohesiveness of the food product is in the range of 1-100%, 1-90%, 1-80%, 1-70%, 1-60%, 1-50%, 1-40%, 1-30%, 1-25%, 1-20%, 1-15%, 1-10%, or 1-5%. In preferred embodiments, the cohesiveness for soft meat substitutes is in the range of 15-70%, preferably 20-60%, more preferably 25-50%, and most preferably 30-45%. In preferred embodiments, the cohesiveness for firm meat substitutes is in the range of 20-85%, preferably 30-40%.

[0356] In certain embodiments, the viscosity of the food product is in the range of 1 to 200N, 1 to 190N, 1 to 180N, 1 to 170N, 1 to 170N, 1 to 160N, 1 to 170N, 1 to 160N, 1 to 150N, 1 to 140N, 1 to 130N, 1 to 120N, 1 to 110N, 1 to 110N, 1 to 90N, 1 to 80N, 1 to 70N, 1 to 60N, 1 to 50N, 1 to 40N, 1 to 30N, 1 to 25N, 1 to 20N, 1 to 15N, 1 to 10N, or 1 to 5N. In a preferred embodiment, the viscosity for the soft meat substitute is in the range of 1 to 40N, preferably 3 to 33N, more preferably 5 to 25N, and most preferably 6 to 21N. In a preferred embodiment, the viscosity of the hard meat substitute is in the range of 6 to 85N, preferably 15 to 40N.

[0357] In certain embodiments, the chewiness of the food product is in the range of 1 to 200N, 1 to 190N, 1 to 180N, 1 to 170N, 1 to 170N, 1 to 160N, 1 to 170N, 1 to 160N, 1 to 150N, 1 to 140N, 1 to 130N, 1 to 120N, 1 to 110N, 1 to 110N, 1 to 90N, 1 to 80N, 1 to 70N, 1 to 60N, 1 to 50N, 1 to 40N, 1 to 30N, 1 to 25N, 1 to 20N, 1 to 15N, 1 to 10N, or 1 to 5N. In a preferred embodiment, the chewiness for the soft meat substitute is in the range of 0.3 to 35N, preferably 1 to 27N, more preferably 2.5 to 20N, and most preferably 3 to 16N. In a preferred embodiment, the chewiness of the hard meat substitute is in the range of 1 to 60N, preferably 4 to 25N.

[0358] In certain embodiments, the stickiness of the developed meat substitutes is in the range of 0 to -100 N.s, 0 to -90 N.s, 0 to -80 N.s, 0 to -70 N.s, 0 to -60 N.s, 0 to -50 N.s, 0 to -40 N.s, 0 to -30 N.s, 0 to -20 N.s, 0 to -10 N.s, 0 to -5 N.s, 0 to -1 N.s, 0 to -0.01 N.s, or 0 to -0.001 N.s. In certain embodiments, the stickiness for mycelium-based meat substitutes is in the range of 0 to -0.3 N.s, more preferably -0.01 to -0.1 N.s, and most preferably -0.02 to -0.05 N.s, which is a low stickiness texture.

[0359] In certain embodiments, the firmness of the milk replacer, such as cream cheese or any mycelium-based spread, is in the range of 1 to 100N, 1 to 90N, 1 to 80N, 1 to 70N, 1 to 60N, 1 to 50N, 1 to 40N, 1 to 30N, 1 to 25N, 1 to 20N, 1 to 15N, 1 to 10N, or 1 to 5N. In another preferred embodiment, the firmness for soft milk replacer is in the range of 1 to 20N, more preferably 5 to 15N, and most preferably 5 to 10N. In a preferred embodiment, the firmness for hard milk replacer is in the range of 20 to 100N, preferably 20 to 50N, more preferably 20 to 40N, and most preferably 25 to 35N.

[0360] In certain embodiments, the spreadability of the milk replacer, such as cream cheese or any mycelium-based spread, is in the range of 1 to 100 N.s, 1 to 90 N.s, 1 to 80 N.s, 1 to 70 N.s, 1 to 60 N.s, 1 to 50 N.s, 1 to 40 N.s, 1 to 30 N.s, 1 to 25 N.s, 1 to 20 N.s, 1 to 15 N.s, 1 to 10 N.s, or 1 to 5 N.s. In another preferred embodiment, the spreadability for soft milk replacers is in the range of 1 to 20 N.s, more preferably 10 to 20 N.s. In a preferred embodiment, the spreadability for hard milk replacers is in the range of 30 to 100 N.s, preferably 40 to 80 N.s, more preferably 40 to 70 N.s, and most preferably 45 to 70 N.s.

[0361] In certain embodiments, the viscosity of the developed milk replacer, such as cream cheese or any mycelium-based spread, is in the range of -1 to -100N, -1 to -90N, -1 to -80N, -1 to -70N, -1 to -60N, -1 to -50N, -1 to -40N, -1 to -30N, -1 to -25N, -1 to -20N, -1 to -15N, -1 to -10N, or -1 to -5N. In another preferred embodiment, the viscosity for soft milk replacer is in the range of -1 to -14N, more preferably -3 to -10N, which is a low viscosity. In a preferred embodiment, the viscosity for hard milk replacer is in the range of -15 to -100N, preferably -15 to -50N, more preferably -15 to -30N, and most preferably -15 to -25N.

[0362] In certain embodiments, the puncture force of the milk replacer, such as cream cheese or any mycelium-based spread, results in an area under the curve in the range of 1 to 100 N.s, 1 to 90 N.s, 1 to 80 N.s, 1 to 70 N.s, 1 to 60 N.s, 1 to 50 N.s, 1 to 40 N.s, 1 to 30 N.s, 1 to 25 N.s, 1 to 20 N.s, 1 to 15 N.s, 1 to 10 N.s, or 1 to 5 N.s. In a preferred embodiment, this area for soft milk replacers is in the range of 1 to 30 N.s, preferably 5 to 25 N.s, more preferably 5 to 20 N.s, and most preferably 8 to 18 N.s. In another preferred embodiment, the area for the firm milk replacer is in the range of 40 to 100 N.s, more preferably 40 to 80 N.s, and most preferably 50 to 60 N.s, which indicates that a higher time and higher force is required to puncture the firm milk replacer.

[0363] In a specific exemplary embodiment illustrating the above embodiment, mycelium of Pleurotus Pulmonarius is grown at ambient temperature in a medium selected from three different media: a defined medium providing component A, a synthetically defined medium providing component B, or a natural medium based on an extract from brewer's grains, the brewer's grains being characterized by a particle size distribution determined by using different sets of sieves (DIN 10765 modified), which included a maximum distribution of 2-4 mm at about 40% by weight, followed by a second maximum distribution of 1-2 mm at about 26% by weight.

[0364] The mycelial components of the present invention were then analyzed for their ash content and their elemental composition (Carbon (C), Hydrogen (H), Nitrogen (N), Sulfur (S), Oxygen (O)), and the oxygen content was determined by difference. The ash contents of components A, B, and C are 9.39%, 9.73%, and 8.40%. Elemental analysis of the C, H, N, O, and S analyses of these components yields the following: For A: 44.45% (carbon C), 5.95% (H), 6.25% (N), 0.32% (S), 33.53% (O); Carbon to nitrogen ratio (mycelium): 7.11. For B: 43.92% (carbon C), 5.83% (H), 4.62% (N), 0.26% (S), 35.64% (O); Carbon to nitrogen ratio (mycelium): 9.5. For C: 46.90% (carbon C), 6.26% (H), 9.57% (N), 0.42% (S), 28.45% (O); Carbon to nitrogen ratio (mycelium): 4.9.

[0365] The lower nitrogen content in sample B indicates that the sample is richer in fiber content but has a lower protein content, while the brewer's grain-based sample C has the highest nitrogen content, indicating a high protein content and a lower fiber content. Sample A falls between the two. This indicates that the protein and fiber content can be tailored by fine-tuning the composition of the medium either synthetically or by using a nutrient side stream with an analyzed elemental composition, allowing for the production of tailored food products rich in either fiber or protein.

[0366] The higher heating value (HHV, often referred to as gross heating value) was determined directly using an oxygen bomb calorimeter as outlined in EN 14918:2009. The lower heating value (LHV, often referred to as net heating value) was calculated based on the HHV and elemental composition of the sample.

[0367] For A, the HHV and LHV are 18.4 MJ / kg (440 Kcal / 100 g) and 17.11 MJ / kg (410 Kcal / 100 g), respectively. For B, the HHV and LHV are 18.88 MJ / kg (450 Kcal / 100 g) and 17.61 MJ / kg (420 Kcal / 100 g), respectively. For C, the HHV and LHV are 19.73 MJ / kg (470 Kcal / 100 g) and 18.37 MJ / kg (440 Kcal / 100 g), respectively.

[0368] The most abundant sugars as a percentage of the total sugar content in the mycelial components after hydrolysis for A are 88% glucan, for B 92.5% glucan, and for C 78.6% glucan (glucan here is based on all glucose, including glucose derived from other polysaccharides, e.g., heteroglycans and / or exopolysaccharides).

[0369] The sugars in the aqueous extract or extract comprise sugars extracted from fresh mycelial components via water. Water extraction was performed in an 11 ml stainless steel extraction cell in an ASE 200 (Accelerated Solvent Extractor) using deionized water at 100°C and 1500 psi (heat time: 5 min, rest time: 7 min, flush volume: 150%, purge time: 180 s, rest cycles: 3). The total sugars in the aqueous extracts of components A, B, and C were 5.65%, 7%, and 3.26%, respectively. From these, the sugars in the aqueous extract of A, expressed as a % of total extract sugars, included disaccharides (20.1% trehalose), trehalose sugars (22% glucose, 0.5% fructose, 0.1% mannose, 1.4% galactose), pentose sugars (0.1% arabinose), and sugar alcohols (55% mannitol, 0.7% sorbitol). For B, the sugars in the aqueous extract, expressed as a % of total extract sugars, include disaccharides (19.2% trehalose), trehalose sugars (43.3% glucose, 0.4% fructose, 0.1% galactose), pentose sugars (0.1% arabinose), and sugar alcohols (35.7% mannitol, 1% sorbitol). For C, the sugars in the aqueous extract, expressed as a % of total extract sugars, include disaccharides (46% trehalose), trehalose sugars (16% glucose, 0.1% galactose), pentose sugars (0.03% arabinose and 0.15% xylose), and sugar alcohols (22.7% mannitol, 13.5% arabinitol, 1% xylitol, 0.64% sorbitol).

[0370] In one embodiment, the water extract of mycelium component A contains at least 50% by weight of mannitol of the extracted sugars (i.e., relative to the total sugar content), which is the highest amount of extracted sugar compared to the others.

[0371] In one embodiment, the water extract of mycelium component B contains at least 40% by weight of glucose of the extracted sugars (i.e., relative to the total sugar content), which is the highest amount of extracted sugars compared to the others.

[0372] In one embodiment, the water extract of mycelium component C contains at least 40% by weight of trehalose of the extracted sugars (i.e., relative to the total sugar content), which is the highest amount of extracted sugars by weight.

[0373] Flavors were measured for known flavor 5'-nucleotides (5'-NMPs) in g / kg, including 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP), and 5'-adenosine monophosphate (AMP). Component A exhibited a better enrichment of 5'-NMP at 3.56 g / kg compared to B, which exhibited 1.78 g / kg. However, the use of brewer's grain extract resulted in a 10-fold increase in component C compared to C, resulting in 10.43 g / kg of 5'-NMP. Additionally, component C exhibited the highest concentration of uridine monophosphate (UMP) at 3.56 g / kg compared to 1.26 g / kg and 0.71 g / kg for A and B, respectively.

[0374] In one embodied example, without any additional ribonucleic acid (RNA) reduction steps via additional heating or pH treatment in the component production process, the method of the present invention results in component specific RNA levels of 2% by weight or less for all disclosed mycelial components, specifically 1.88% by weight for A, 1.65% by weight for B, and 2% by weight for C.

[0375] Free bases include cytosine, uracil, guanine, hypoxanthine, and adenine. Total free bases measured for mycelium component A was 0.62 g / kg, compared to 0.48 g / kg for component B and 1.3 g / kg for component C. For component A, adenine and uracil were observed to be the highest, at 0.28 g / kg and 9.18 g / kg, respectively. For component B, the same observations were made as for component A, but adenine and uracil were present at lower concentrations, at 0.17 g / kg and 0.13 g / kg, respectively. For component C, higher amounts of adenine, uracil, and cytosine were observed, at 0.54 g / kg, 0.22 g / kg, and 0.49 g / kg, respectively.

[0376] Free purine nucleosides, including guanosine, inosine, and adenosine, were determined to be highest for component A (2.77 g / kg), followed by component C (2.24 g / kg), and then component B (1.11 g / kg).

[0377] The chitin content for the components was approximately 7.6 wt% for A, 6.9 wt% for B, and 7.5 wt% for C. The ergothioneine content was approximately 91 mg / kg for A, 127 mg / kg for B, and 433 mg / kg for C. An ergosterol content of 2.35 mg / g dry weight was determined for sample A, 2.24 mg / g dry weight for sample B, and 5.56 mg / g dry weight for sample C. (Oyster mushrooms have been reported to have an ergosterol content of 4.4 mg / g dry weight, and P. pulmonarius fruiting bodies grown on three forestry wastes (pine, poplar, and honeysuckle) in particular are known to have an ergosterol content of 2.9 to 3.3 mg / g, which is less than the content of ergosterol found in mycelial component C, indicating a higher concentration compared to the mushroom fruiting body itself.)

[0378] All components A, B, and C exhibit at least about 20% by weight of umami amino acids, more specifically, 21.48% by weight of umami amino acids for A, 19.5% by weight of umami amino acids for B, and 22.64% by weight of umami amino acids for C.

[0379] All components A, B, and C have a BCAA content of at least about 20% by weight, more specifically, A has a BCAA content of 21.73% by weight, B has a BCAA content of 23.1% by weight, and C has a BCAA content of 19.6% by weight.

[0380] All components A, B, and C have an essential amino acid content of at least about 40% by weight, more specifically, A has an essential amino acid content of 40.81% by weight, B has an essential amino acid content of 40.52% by weight, and C has an essential amino acid content of 39.61% by weight.

[0381] The total amino acid content in mg / g was richest in C (361.67 mg / g), followed by A (268.85 mg / g), and then B (163.43 mg / g).

[0382] Mycelial component A is grown in a medium containing a carbon-to-nitrogen ratio (medium) of 16-18, preferably about 17. Mycelial component B is grown in a medium containing a carbon-to-nitrogen ratio (medium) of 19-21, preferably about 20. Mycelial component B is grown in a medium containing a carbon-to-nitrogen ratio (medium) of 12-14, preferably about 13.

[0383] The insoluble fiber content increased from approximately 23% by weight to approximately 34% by weight to approximately 54% by weight from C to A to B, while the protein content decreased from approximately 60% by weight to approximately 39% by weight to approximately 32% by weight from C to A to B. This demonstrates the relationship between protein and fiber discussed above, in which protein and fiber are inversely proportional and can be controlled by varying the fermentation medium composition. However, this trend is not always present, as reported in the following study on submerged fermentation of Pleurotus tuber-regium, where a lower C / N ratio in the medium resulted in higher total dietary fiber in the mycelial cell walls (Food Chemistry 85 (2004) 101-105). However, the observed trend in the present invention indicates that fiber content increases with increasing C / N ratio. This indicates that such a trend may be highly dependent on medium composition, e.g., carbon source or nitrogen source, fermentation process conditions, e.g., pH and agitation rate, inoculum, and / or other factors.

[0384] The fat content of component A is about 3% by weight. The fat content of component B is about 2% by weight. The fat content of component C is approximately 7% by weight. As shown in the table below, this fat includes saturated fatty acids, monosaturated fatty acids, polyunsaturated fatty acids, and trans fatty acids. Note that the omega-6 fatty acid (linoleic acid) content is 0.77% by weight for component A, 0.46% by weight for component B, and 3.5% by weight for component C. All components exhibit a calculated carbohydrate content of less than 0.1% by weight. Component B is the richest in total glucan, with a total glucan value of 31% by weight. Mycelium component A has a total glucan value of approximately 26% by weight. Mycelium component C has a total glucan value of approximately 17% by weight.

[0385] The EUC for the mycelial components was calculated to be 302% for A, 34% for B, and 2892% for C. In a separate study, P. pulmonarius fruiting bodies were grown on three forestry wastes (pine, poplar, and honeysuckle), which showed EUC values ​​for P. pulmonarius fruiting bodies ranging from 72.31% to 116.73% (Food Chemistry 397(2022)133714). The disclosed EUC values ​​for mycelial components A, B, and C of the present invention are higher than the reported values ​​for fruiting bodies of the same fungal strain; for example, component C, the difference is 25-40 times higher for component C compared to the reported range for P. pulmonarius fruiting bodies. It is also known that the highest value of EUC in mushroom fruiting bodies was reported to be 4465% for (Volvariella volvacea), which is 2.35 times lower compared to component C.

[0386] It is noted that the reported mycelium (not fruiting bodies) had lower EUC values ​​than fruiting bodies for Pleurotus eryngii (30.9% EUC mycelium vs. 116% fruiting bodies), Agrocybe aegerita (19.2% EUC mycelium vs. 322% EUC fruiting bodies), and Lentinus edodes (16.7% EUC mycelium vs. 99.75% EUC fruiting bodies) (https: / / doi.org / 10.1080 / 10942912.2015.1089891), which highlights the importance and unique taste that the mycelium component of the present invention has through submerged fermentation. Upon further review of the literature, the EUC for mycelia was found to be for the following species: Termitomyces albuminosus, 460%, Wen (2003); Grifola frondosa, 375%, Wen (2003); Morchella esculenta, 363%, Wen (2003); Hypsizygus marmoreus, 128%, Lee (2003); Cordyceps militaris, 124%, Chang et al. (2001); Pleurotus citrinopileatus, 37.1%, Huang (2003); Antrodia camphorata, 21.2%, Chang et al. (2001); Ganoderma tsugae, 19.4%, Tseng et al. (2004); Agaricus brasiliensis, 1.92%, Chang et al. (2001); pleurotus eryngii, 9 to 144% by submerged fermentation (all of these documents are appropriately disclosed in Mau et al., International Journal of Medicinal Mushrooms, Vol. 7, pp. 119-125 (2005)). All of these can be considered as evidence that the mycelial component of the present invention exhibits an unprecedented high profile umami flavor for use in the food industry and food products, such as milk substitutes, meat substitutes, or other food products.

[0387] Additionally, by comparing mycelial components A and C with an unwashed mycoprotein product from Quorn, according to the data disclosed in WO2021234349 (Tables 1 and 2), the maximum EUC can be estimated to be at most 148% (it is known that in this calculation total amino acids were used instead of free amino acids due to lack of data, so the true value of EUC should be less than 148%).

[0388] Interestingly, the samples grown in the sidestream had the richest values ​​in terms of total amino acid content, umami flavor, ergothionene content, ergosterol content, and other properties, which indicates the great advantage of using natural sidestreams or waste streams to produce richer food and at the same time upcycle waste streams from other value chains, thereby contributing to sustainability. However, these ingredients have different properties that make them suitable for different applications. One example is the use of ingredient B for sports supplements, because ingredient B is low in fat, high in fiber content and BCAA amino acids, and has a sufficient protein content.

[0389] The present invention also relates to methods of producing edible milk substitute, meat substitute, and fish / seafood substitute products using the fungal components of the present invention.

[0390] In certain embodiments, the mycelium component may be used in the form of wet biomass (as is), washed biomass, dried biomass, ground into a powder with a specific particle size distribution, either fine or coarse or medium particle size.

[0391] A method for producing a soft or hard meat substitute composition comprising at least one mycelium component from at least one fungal strain, the method comprising a process for producing each mycelium component, and further comprising a step of preparing such meat substitute composition by mixing at least one mycelium component from the at least one fungal strain with a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and optionally at least one composition component.

[0392] In one embodiment, 50-95% by weight of the mycelium component (edible fibrous mycelium) is blended, then added to at most 1-40% by weight of each of the following ingredients: canola oil, salt, egg white, and wheat gluten, and then optionally added to at most 1-40% by weight of each of the following ingredients: methylcellulose, hydrocolloid, texturizing vegetable protein, starch-based ingredient, flavor component. The resulting dough is then shaped through a former and / or extruder, or a combination thereof, to obtain the final product as meatballs or sausages or extruded products.

[0393] A method for producing a milk substitute composition comprising at least one mycelium component from at least one fungal strain, the method comprising producing each mycelium component, and further comprising the steps of preparing such milk substitute composition by (1) forming a slurry comprising at least one mycelium component together with at least one protein-rich component, at least one plant-based lipid-rich component, and a composition comprising at least one composition component, and (2) mixing the slurry with at least one composition component, specifically a texturizing or thickening agent, or a carbohydrate-rich component.

[0394] In one embodiment, the method of producing an edible dairy substitute does not include an additional acidification step, but rather involves acidification via the formulation thereof below. (1) homogenizing 10 to 50% by weight of edible fibrous mycelium obtained by submerged fermentation from at least one fungal strain with about 55% by weight of drinking water, about 10 to 45% by weight of a plant-derived fat component, at most 2% by weight of a sodium chloride source, at most 2% by weight of yeast flakes, at most 5% by weight of a plant-derived monosaccharide or disaccharide source, and 5% by weight of a natural plant-derived acidity source; (2) Heating the homogenized slurry to a temperature of 75-95°C for a maximum of 60 seconds under constant blending. To coagulate the homogenized slurry, a pre-prepared coagulant solution is added during this preceding limited heating step as soon as the temperature reaches 25-40°C. This coagulant solution contains at most 15% by weight of each of the following: water, wheat starch, and hydrolyzed corn starch. (3) The resulting slurry is removed from the heat source, and then the mixture is allowed to stand at room temperature (21°C). Upon reaching room temperature, the solidified slurry is stored in a dark, cool environment, preferably at 4°C to 7°C.

[0395] In a further embodiment, the method for producing an edible non-animal dairy substitute comprises the following additional acidification step: (1) Homogenizing 10 to 50% by weight of edible fibrous mycelium obtained by submerged fermentation from at least one fungal strain with about 55% by weight of drinking water, about 10 to 45% by weight of a plant-derived fat component, at most 2% by weight of a sodium chloride source, at most 2% by weight of yeast flakes, and at most 5% by weight of a plant-derived monosaccharide or disaccharide source. (2) Heating the homogenized slurry to a temperature of 75-95°C for a maximum of 60 seconds under constant blending. To coagulate the homogenized slurry, a pre-prepared coagulant solution is added during this preceding limited heating step as soon as the temperature reaches 25-40°C. This coagulant solution contains at most 15% by weight of each of the following: water, wheat starch, and hydrolyzed corn starch. (3) The resulting slurry is removed from the heat source, and the mixture is then allowed to stand at room temperature (21°C). When the resulting mixture reaches a temperature of 40°C or less, 0.1 g to 0.25 g of acid-forming bacteria, particularly lactic acid bacteria, is added to induce microbial acidification. To provide a hospital environment for the microorganisms, the sample is placed in a controlled temperature environment (e.g., a water bath or incubator) at 28°C for 150 minutes. Depending on the intensity of the fermentation, the sample may be placed at a temperature of 20 to 45°C for 1 to 6 hours. After microbial fermentation is complete, the solidified slurry is stored in a dark, cool environment, preferably at 4 to 7°C.

[0396] In a third embodiment, mycelium from one fungal strain is mixed with mycelium from another fungal strain, or with algae, bacteria, plant cells, archaeal cells, fat cells, or combinations thereof. In a preferred embodiment, mycelium from one fungal strain is partially replaced by mycelium from another fungal strain in a ratio of total mycelium content ranging from 1:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, or 100:1 to produce a milk or meat substitute. In a preferred embodiment, when a second mycelium component is present, the ratio of the two mycelium components is 20:80, preferably 50:50, more preferably 40:60, and most preferably 30:70. In one embodiment, Pleurotus Pulmonarius edible filamentous mycelium is mixed with Morchella rufobrunnea edible filamentous mycelium, and 60-90% by weight of the total mycelium is from Pleurotus Pulmonarius mycelium and 10-40% by weight of the total mycelial components are from Morchella rufobrunnea mycelium (edible filamentous mycelium). In another embodiment, Pleurotus Pulmonarius edible filamentous mycelium is mixed with Morchella rufobrunnea edible filamentous mycelium, and 60-90% by weight of the total mycelium is from Morchella rufobrunnea mycelium and 10-40% by weight of the total mycelial components are from Pleurotus Pulmonarius mycelium (edible filamentous mycelium).

[0397] In another embodiment, the same method is extended to at least three fungal strains.

[0398] This results in an edible substitute cheese product selected from the group consisting of edible substitute products of whey cheese, cream cheese, medium hard cheese, hard cheese, and soft mold cheese, thereby resulting in an edible substitute cheese product that is cream cheese.

[0399] The addition of a texturizing agent is an optional step depending on the water content of the homogenized mycelial mass obtained in step a): if the water content is similar to that of the substitute product to be produced, the addition of a texturizing agent is not necessary to obtain the desired texture.

[0400] In a preferred embodiment, the product obtained by this method is an edible substitute fresh cheese product or cream cheese.The product can also be an edible substitute cheese product selected from the group including whey cheese, cream cheese, medium-hard cheese, hard cheese, and soft mold cheese edible substitute products, and the edible substitute cheese product can be obtained without a conventional coagulation step if the water content of the homogenized mycelium mass is suitably adjusted to be sufficiently low.The coagulation step of conventional cheese making is carried out to remove excess water provided by the starting material, which is milk.By using a starting material, i.e., a homogenized mycelium mass, that has a significantly lower water content than milk, these coagulation steps of conventional cheese making are not necessary.

[0401] In the process for producing edible dairy substitute products for fresh cheese, whey cheese, cream cheese, medium hard cheese, hard cheese, and soft mold cheese, the edible plant-based fat component is preferably selected from coconut oil, sunflower oil, rapeseed oil, palm oil, cottonseed oil, olive oil, canola oil, algae oil, or oleaginous yeast-derived oil.

[0402] In a further preferred embodiment, particularly when the edible dairy substitute is a yogurt or cheese substitute, the edible dairy substitute further comprises an edible plant or algae-based fat component, a fat component derived from fungi or yeast, preferably up to 60% by weight, more preferably up to 25% by weight, most preferably in the range of 1% to 5% by weight. The amount of fat component can be adjusted depending on the product being substituted.

[0403] The edible dairy substitute product of the present invention preferably comprises an edible fibrous mycelial mass content of 1% to 99% by weight, preferably 10% to 90% by weight, preferably 1% to 65% by weight, most preferably 10 to 50% by weight. When discussing the content of edible fibrous mycelium in the product of the present invention, preferably reference is made to the weight % in the product containing water (i.e., no normalization to dry mass content).

[0404] Preferably, the edible fibrous mycelium in the alternative dairy product has an insoluble fiber content of at least 20% by weight, more preferably at least 30% by weight, more preferably at least 40% by weight. Therefore, the insoluble fiber content is preferably 20% to 60% by weight. More preferably, the edible fibrous mycelium has an insoluble fiber content of 40% to 60% by weight. Even more preferably, the edible fibrous mycelium has an insoluble fiber content of 40% to 55% by weight. Even more preferably, the edible fibrous mycelium has an insoluble fiber content of 40 to 50% by weight. Most preferably, the edible fibrous mycelium has an insoluble fiber content of about 45% by weight. Most preferably, the edible fibrous mycelium has an insoluble fiber content of at least 50% by weight. It should be understood that weights referred to herein are relative to the dry mass of the mycelium. In one embodiment, the edible fibrous mycelium has an insoluble fiber content of at least 40% by weight, preferably at least 50% by weight, more preferably at least 60% by weight.

[0405] The protein or fiber content of the mycelial mass can be adjusted by selecting fermentation conditions that control the protein or fiber content of the resulting mycelial mass. For example, the ratio between the carbon and nitrogen sources in the medium can be varied, for example, by providing nitrogen in various degrees of excess, as contemplated in the present invention.

[0406] In some embodiments, particularly when the edible dairy substitute is a yogurt or cheese substitute, the edible dairy substitute can further comprise a texturing agent, such as agar, food starch, guar gum, locust bean gum, wheat gluten, cellulose, or a derivative thereof. The texturing agent provides structure to the product, making it suitable as a yogurt or cheese substitute. However, the texturing agent is an optional component for the edible dairy substitute of the present invention. If the moisture content of the water-homogenized mycelium is kept low by using pressed or partially dried mycelium with the desired moisture content, the resulting mass will have the desired structure similar to that of yogurt or cheese without the need to add an external texturing agent. Those skilled in the art will understand how to appropriately adjust the moisture content of the mycelial mass to suit the substitute product to be produced.

[0407] In one embodiment, the ingredient-mediated acidification is achieved by homogenizing 10-50% by weight of Pleurotus Pulmonarius mycelium (edible fibrous mycelium) with approximately 55% by weight of potable water, approximately 0-7% by weight of cashew nuts (a plant-derived fat component), 10-40% by weight of Cocos nucifera oil (a plant-derived fat component), at most 2% by weight of table salt (a sodium chloride source), at most 2% by weight of yeast flakes, at most 5% by weight of sucrose (a plant-derived sugar source), at most 5% by weight of lemon juice (a natural plant-derived acidity source), and at most 5% by weight of citric acid. While blending continuously, the homogenized slurry is heated to a temperature of 75-95°C for up to 60 seconds. A pre-prepared coagulant solution is added during this prior limited heating step, as soon as the temperature reaches 25-40°C. The coagulant solution contains at most 15% by weight each of water, wheat starch, and hydrolyzed corn starch, and the resulting slurry is removed from the heat source, after which the mixture is allowed to stand at room temperature (21° C.). Upon reaching room temperature, the coagulated slurry is stored in a dark, cool environment, preferably at 4° C. to 7° C., to obtain a mycelium-based cream cheese product.

[0408] In one embodiment, acidification via microbial fermentation is achieved by homogenizing 10-50% by weight of Pleurotus Pulmonarius mycelium (edible fibrous mycelium) with approximately 55% by weight of potable water, approximately 0-7% by weight of cashew nuts (a plant-derived fat component), 10-40% by weight of Cocos nucifera oil (a plant-derived fat component), at most 2% by weight of table salt (a sodium chloride source), at most 2% by weight of yeast flakes, and at most 5% by weight of sucrose (a plant-derived sugar source). The homogenized slurry is heated to a temperature of 75-95°C for up to 60 seconds while blending continuously. A pre-prepared coagulant solution is added during this preceding limited heating step, as soon as the temperature reaches 25-40°C. This coagulant solution contains at most 15% by weight of each of water, wheat starch, and hydrolyzed corn starch. The resulting slurry is removed from the heat source and then allowed to stand at room temperature (21°C). When the resulting mixture reaches a temperature of 40°C or less, 0.1 g to 0.25 g of acid-forming bacteria, particularly lactic acid bacteria, is added to induce microbial acidification. The sample is placed in a controlled temperature environment (e.g., a water bath or incubator) at 28°C for 150 minutes. Depending on the intensity of fermentation, the sample may be placed at a temperature of 20 to 45°C for 1 to 6 hours. After microbial fermentation is complete, the coagulated slurry is stored in a dark, cool environment, preferably at 4 to 7°C, to obtain a mycelium-based cream cheese product.

[0409] The adjustable composition of the mycelial mass of the present invention provides the advantage of versatility, allowing the composition of the mycelial mass to be easily adjusted to suit the requirements of the resulting substitute product. For example, if the protein content of the mycelial mass is increased, the amount of mycelial mass in the substitute product can be reduced without changing the protein content of the substitute product. If the substitute product is a specific type of cheese characterized by a specific amount of calcium, phosphorus, and / or zinc, these nutrients can be provided by the mycelial mass itself, eliminating the need to supplement the substitute product with additional nutrients from outside. This reduces the cost of the production method and the resulting product for the purpose of producing edible dairy substitute products with clean labels due to easier handling, fewer production steps, and reduced raw material costs. Most importantly, the nutrients from the mycelium are bioavailable, making metabolic digestion predictable and easy, especially compared to products containing nutrients supplemented from outside.

[0410] In one embodiment, this mixture of at least two mycelial components is used to produce a milk substitute. In another embodiment, this mixture of at least two mycelial components is used to produce a meat substitute. In a preferred embodiment, this mixture of at least two mycelial components is used to produce a milk and a meat substitute. In a preferred embodiment, this mixture of at least two mycelial components is used to produce a fish substitute. In one embodiment, this mixture of mycelial components is used to produce other food products as defined above.

[0411] In a further embodiment, a method of producing an edible vegetarian meat substitute product comprises: (1) Blending 1 to 99% by weight, preferably 50 to 95% by weight, of an edible mycelium component and adding it to a mixture of at least one protein-rich component and at least one plant-based lipid-rich component. (2) Optionally, adding at least one compositional ingredient, such ingredients including methylcellulose, hydrocolloid, texturizing vegetable protein, starch-based ingredient, fiber-rich ingredient, and flavor component for seasoning. (3) The obtained dough is shaped through an extruder and / or a molding machine to obtain a final product as a meatball or sausage or extruded product, wherein step 3 includes: (i) cold extruding the obtained dough into a rope having a thickness of 1.5 to 5 cm, preferably 1.5 to 3.5 cm, by a cold extruder or a vacuum filling machine filling molding machine; (ii) Forming the product into a desired shape (iii) As an alternative to step (ii), in the case of sausage products, stuffing the casing. (iv) boiling the product in water at a temperature ranging from 60 to 100°C for a duration of 1 to 30 minutes to increase the mycelium product texture due to protein denaturation and increase the product shelf life stability. (v) Alternatively, step (iv) may be replaced by deep frying the product followed by steam or hot air cooking at a temperature in the range of 50 to 150°C. (vi) Storing the finished product in a freezer

[0412] The edible meat substitute product of the present invention preferably comprises an edible fibrous mycelium mass content of 1% to 99% by weight, preferably 5% to 99% by weight, preferably 10% to 95% by weight, preferably 20% to 95% by weight, more preferably 40% to 95% by weight, and most preferably 60% to 95% by weight. Preferably, the edible fibrous mycelium in the meat substitute product has an insoluble fiber content of 20% to 60% by weight, preferably 40% to 50% by weight, and preferably about 45% by weight. In another preferred embodiment, the insoluble fiber used in the meat substitute is about 35% by weight. In another preferred embodiment, the insoluble fiber used in the meat substitute is about 25% by weight. In another preferred embodiment, the insoluble fiber used in the meat substitute is about 45% by weight. In one embodiment, the edible fibrous mycelium in the meat substitute has an insoluble fiber content of at least 20% by weight, preferably at least 30% by weight, preferably at least 45% by weight, and preferably at least 50% by weight.

[0413] The protein or fiber content of the mycelial mass can be adjusted by selecting fermentation conditions that control the protein or fiber content of the resulting mycelial mass. For example, the ratio between the carbon and nitrogen sources in the medium can be varied, for example, by providing nitrogen in various degrees of excess, as contemplated in the present invention.

[0414] In one embodiment, the at least one plant-based lipid-rich ingredient comprises at most 40%, at most 20%, at most 15%, at most 10%, at most 9%, at most 8%, at most 7%, at most 6%, at most 5%, at most 4%, at most 3%, at most 2%, or at most 1% by weight of the total recipe of the alternative meat composition. In a preferred embodiment, the at least one plant-based lipid-rich ingredient of the alternative meat composition comprises 1-40%, preferably 1-20%, more preferably 1-10% by weight.

[0415] In one embodiment, the at least one protein-rich ingredient of the alternative meat composition each constitutes at most 95%, at most 80%, at most 60%, at most 40%, at most 20%, at most 15%, at most 10%, at most 9%, at most 8%, at most 7%, at most 6%, at most 5%, at most 4%, at most 3%, at most 2%, or at most 1% by weight of the total recipe of the alternative meat composition. In a preferred embodiment, the at least one protein-rich ingredient of the alternative meat composition each constitutes 0.1 to 50% by weight, preferably 0.1 to 30% by weight, more preferably 0.1 to 20% by weight.

[0416] In one embodiment, at least one component of the alternative meat composition each comprises at most 80%, at most 60%, at most 40%, at most 20%, at most 15%, at most 10%, at most 9%, at most 8%, at most 7%, at most 6%, at most 5%, at most 4%, at most 3%, at most 2%, or at most 1% by weight of the total recipe of the alternative meat composition. In a preferred embodiment, at least one component of the alternative meat composition each comprises 0.1 to 50%, preferably 0.1 to 30%, more preferably 0.1 to 20% by weight.

[0417] In one embodiment, the protein-rich component comprises wheat gluten. In one embodiment, the protein-rich component comprises wheat gluten and egg white. In one embodiment, the composition component comprises a flavor component. In one embodiment, the composition component comprises a flavor component and a starch-based component. In one embodiment, the composition component comprises a flavor component, a starch-based component, and methylcellulose. In one embodiment, the composition component comprises a flavor component and methylcellulose. In one embodiment, the composition component comprises a flavor component, a starch-based component, and a hydrocolloid. In one embodiment, the composition component comprises a flavor component and a hydrocolloid. In one embodiment, the composition component comprises a flavor component, a starch-based component, and a texturized vegetable protein. In one embodiment, the composition component comprises a flavor component and a texturized vegetable protein. In one embodiment, the composition component comprises a flavor component and a fiber-rich component. In one embodiment, the composition component comprises a flavor component, a fiber-rich component, and a starch-based component. In one embodiment, the composition component comprises a flavor component, a starch-based component, a fiber-rich component, and methylcellulose. In one embodiment, the composition ingredients include a flavor component, methylcellulose, and a fiber-rich component. In one embodiment, the composition ingredients include a flavor component, a starch-based component, a fiber-rich component, and a hydrocolloid. In one embodiment, the composition ingredients include a flavor component, a fiber-rich component, and a hydrocolloid. In one embodiment, the composition ingredients include a component, a starch-based component, a fiber-rich component, and a texturizing vegetable protein. In one embodiment, the composition ingredients include a flavor component, a fiber-rich component, and a texturizing vegetable protein. In one embodiment, the composition ingredients include methylcellulose, a hydrocolloid, a texturizing vegetable protein, a starch-based component, a fiber-rich component, and a flavor component for seasoning. In one embodiment, the flavor component, which is also understood to be under the category of composition ingredient, includes at least one of the following ingredients: salt, pepper, garlic, onion, mushroom fruiting body pieces, ginger, turmeric, curry, sugar (i.e., sucrose, glucose, monosaccharides or disaccharides), oil, lemon juice, orange juice, herbs and spices, yeast flakes.In one embodiment, the fiber-rich ingredient comprises at least one fiber-rich ingredient selected from the following ingredients: grain-based flour, grain-based starch, legume-based starch, fruit-based fiber, polysaccharides, starch-based ingredients, psyllium husk, inulin, wheat starch, and corn starch. Note that the fiber-rich ingredient can also be a starch or carbohydrate-rich ingredient, since a starch-based ingredient or a carbohydrate-rich ingredient can also be a fiber-rich ingredient.

[0418] In one embodiment, a method for producing the same alternative meat product by making it vegan comprises replacing egg white with more of the mycelium component of the present invention, with the dry mycelium component being increased by 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% by weight. In a preferred embodiment, egg white can be replaced by at most 15-20% by weight, preferably at most 7-10% by weight, of the dry mycelium component. This is primarily determined by the quality and type of biomass, preferably to replace any milk-derived or vegetarian product, making such a product vegan.

[0419] In a further embodiment, in the method for producing a vegan meat substitute, 1 wt.% of egg white is preferably replaced with about 20 wt.%, 19 wt.%, 18 wt.%, 17 wt.%, 16 wt.%, 15 wt.%, 14 wt.%, 13 wt.%, 12 wt.%, 11 wt.%, 10 wt.%, 9 wt.%, 8 wt.%, 7.5 wt.%, 7 wt.%, 6 wt.%, 5.5 wt.%, 5 wt.%, 4 wt.%, 3.5 wt.%, 3 wt.%, 2.5 wt.%, 2 wt.%, 1.5 wt.%, or 1 wt.% of dry mycelial biomass, or an equivalent amount of dry mycelial biomass. In a preferred embodiment, in the method for producing a vegan meat substitute, 1 wt. % of egg white is preferably replaced with at most 10 wt. %, preferably at most 5% or an equivalent amount of dried mycelium component, more preferably an equivalent amount of 2.5 wt. % to 4 wt. %, most preferably an equivalent amount of 1.5 to 2 wt. % of dried mycelium component.

[0420] In a preferred embodiment, in the method for producing a vegan soft meat substitute, 1 wt. % of egg white is replaced with preferably at most 10 wt. %, preferably at most 5% or an equivalent amount of dried mycelium component, more preferably an equivalent amount of 2.5-4 wt. %, most preferably an equivalent amount of 1.5-2 wt. %, and the cutting strength of the original vegetarian product and that of the vegan product are in the same range of 1-25N, most preferably 4-8N.

[0421] As an example, for a mycelium-based meatball composition, the same cutting strength of 4.8 N is achieved when each 1% by weight of egg white is replaced with about 1-2% by weight, preferably about 1.5% by weight, of dry biomass. Varying such proportions results in different textures, i.e., different cutting strengths.

[0422] In a preferred embodiment, in the method for producing a vegan hard meat substitute, 1 wt. % of egg white is replaced with preferably at most 10 wt. %, preferably at most 5% or an equivalent amount of dried mycelium component, more preferably an equivalent amount of 2.5-4 wt. %, most preferably an equivalent amount of 1.5-2 wt. %, and the cutting strength of the original vegetarian product and that of the vegan product are in the same range of 1-50 N, most preferably 10-30 N.

[0423] In another embodiment, 50-95% by weight of Pleurotus pulmonarius mycelia or any edible fibrous mycelia are blended and then added to at most 1-40% by weight of each of the following ingredients: canola oil, salt, and wheat gluten. The missing egg white is accounted for by adding additional dry mycelia at an equivalent ratio of 1-1.4, respectively. Optionally, at most 1-40% by weight of each of the following ingredients is added: methylcellulose, hydrocolloid, texturizing vegetable protein, starch-based ingredient, and flavor component for seasoning. The resulting dough is then shaped through a former and / or extruder, or a combination thereof, to obtain the final product as meatballs or sausages or extruded products.

[0424] In a further embodiment, the resulting dough or formulation, if shaping is required, may be subjected to an extrusion process, either before or after shaping, to form a meat product or other extrudable food product, which extrusion may be either cold extrusion, low temperature extrusion, or high temperature extrusion.

[0425] In one embodiment, cold extrusion is used, and the extrusion temperature is in the range of preferably -20 to -10°C, preferably -10 to 0°C, preferably 1 to 20°C, preferably 4 to 15°C, preferably 10 to 15°C, preferably 15 to 25°C, preferably 25 to 40°C, preferably 40 to 100°C, preferably 40 to 80°C, preferably 50 to 75°C, at a pressure in the range of 1 to 100 bar, more preferably 1 to 70 bar. In a preferred embodiment, the extrusion is carried out at freezing or cold or room temperature, preferably at a temperature of at most -20°C, more preferably 1 to 20°C, most preferably 4 to 15°C, and at a pressure in the range of 1 to 100 bar, more preferably 1 to 70 bar.

[0426] In one embodiment, molding is used and the molding temperature is in the range of preferably 1 to 20° C., preferably 4 to 15° C., preferably 10 to 15° C., preferably 15 to 25° C., preferably 25 to 40° C., preferably 40 to 100° C., preferably 40 to 80° C., preferably 50 to 75° C., at a pressure in the range of 1 to 100 bar, more preferably 1 to 70 bar. In a preferred embodiment, molding is carried out at cold or room temperature, preferably at a temperature of 1 to 25° C., most preferably 4 to 15° C., at a pressure in the range of 1 to 100 bar, more preferably 1 to 70 bar.

[0427] In one embodiment, low temperature extrusion is used, with extrusion temperatures preferably ranging from 30 to 100°C, more preferably from 40 to 80°C, more preferably from 50 to 75°C, at pressures ranging from 5 to 25 bar.

[0428] In a second embodiment, high temperature extrusion is used, the extrusion temperature is preferably in the range of 100-190°C, more preferably 120-180°C, more preferably 130-180°C, at a pressure in the range of 100-300 bar, and the moisture content of the extrudate is preferably less than 45% by weight, preferably less than 40% by weight, preferably less than 35% by weight.

[0429] In a third embodiment, high temperature extrusion is used, the extrusion temperature is preferably in the range of 100-190°C, more preferably 120-180°C, more preferably 130-180°C, at a pressure in the range of 100-300 bar, and the moisture content of the extrudate is preferably greater than 40% by weight, preferably greater than 45% by weight, preferably greater than 50% by weight.

[0430] In one embodiment, the extruded dough rope has a thickness in the range of 1 to 8 cm, preferably 1.5 to 5 cm, more preferably 1.5 to 3.5 cm.

[0431] In further embodiments, the product is frozen and stored at temperatures of -40°C, -30°C, -20°C, -15°C, or -5°C, 0°C, 4°C, 5°C. In a preferred embodiment, the product is frozen at a temperature of -40 to 5°C, more preferably -20 to 5°C, most preferably -5°C to 5°C. In another preferred embodiment, the product is stored at approximately -20°C. In another preferred embodiment, the product is stored at approximately -4°C. In another embodiment, supercooling of the food product is applied and the product is stored at a temperature of -0.5 to -5°C.

[0432] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of 30-200g MSG / 100g.

[0433] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of less than 200g MSG / 100g.

[0434] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of 200-500g MSG / 100g.

[0435] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and at least 30g MSG / 100g EUC.

[0436] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 500g MSG / 100g.

[0437] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 1000g MSG / 100g.

[0438] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 1500g MSG / 100g.

[0439] In one embodiment, the edible meat substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 2000g MSG / 100g.

[0440] In one embodiment, the edible dairy substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of 30-200g MSG / 100g.

[0441] In one embodiment, the edible dairy substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of less than 200g MSG / 100g.

[0442] In one embodiment, the edible dairy substitute comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and 200-500g MSG / 100g EUC.

[0443] In one embodiment, the edible dairy substitute comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and at least 30g MSG / 100g EUC.

[0444] In one embodiment, the edible dairy substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and at least 500g MSG / 100g EUC.

[0445] In one embodiment, the edible dairy substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 1000g MSG / 100g.

[0446] In one embodiment, the edible dairy substitute comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 1500g MSG / 100g.

[0447] In one embodiment, the edible dairy substitute product comprises a mycelium component in the range of 1-99% by weight, the mycelium component characterized by an insoluble fiber content of 20-60% by weight and an EUC of at least 2000g MSG / 100g.

[0448] In a preferred embodiment, the edible product preferably comprises a mycelium component obtained from a fungus selected from Pleurotus species (Pleurotaceae or Pleurotus fungi), such as Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus citrinopileatus, Pleurotus florida, and Pleurotus salmoneostramineus, preferably Pleurotus pulmonarius; or a mycelium component obtained from a fungus selected from Morchella esculenta, Morchella angusticeps, Morchella deliciosa, and Morchella rufobrunnea, and Morchella rufobrunnea.

[0449] In one preferred embodiment, the edible product comprises a mycelium component obtained from Pleurotus pulmonarius. In another preferred embodiment, the edible product comprises a mycelium component obtained from Morchella rufobrunnea. In a further embodiment, the edible product comprises a mycelium component obtained from a combination of Pleurotus pulmonarius and Morchella rufobrunnea. In another embodiment, the edible product comprises a mycelium component obtained from L. sulphureus. In another embodiment, the edible product comprises a mycelium component obtained from B. adusta.

[0450] The meat and milk substitutes may be vegetarian or vegan. The above embodiments also apply to products that are not meat or milk substitutes, which may include, but are not limited to, fish substitutes, confectioneries, baked goods, flours (including bread or pasta or noodles), sweets and desserts, snacks, grain products, alcoholic and non-alcoholic beverages, spice blends, ready-to-eat meals, frozen meals, colloidal foods, protein supplements, and extruded / puffed products.

[0451] Various modifications and variations of this invention will be apparent to those skilled in the art without departing from the scope of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the relevant fields are intended to be covered by the present invention.

[0452] The following examples are merely illustrative of the present invention and should not be construed in any way as limiting the scope of the invention as defined by the appended claims. [Example]

[0453] For all constituents determined via wet chemical analysis, each sample was analyzed at least in duplicate, and percentages are expressed on a dry basis. The following examples were performed on fungal strains derived from Pleurotus pulmonarius. It is understood that Pleurotus species, and in particular Pleurotus pulmonarius, are preferred fungal strains in the methods and products of the present invention. However, the present invention is not limited to this strain and may be practiced with other fungal strains.

[0454] Ingredient production example As an example, Medium A (defined medium) contains the following composition for use in producing Component A: 0.45% by weight C, 0.026% by weight N, 1.24% by weight O, and 0.326% by weight P, with the nitrogen source being CSL (10 g / L) and the carbon source being dextrose.

[0455] Medium B (synthetic medium) for use in producing component B contains the following composition: 0.41% by weight C, 0.02% by weight N, 1.2% by weight O, and 0.326% by weight P; the amino acid is selected from arginine (0.62 g / L); the vitamins are 1 mg / L D-biotin, 1 mg / L folic acid, 1 mg / L niacinamide, 1 mg / L D-pantothenic acid (hemicalcium), 1 mg / L pyridoxal HCl, 0.1 mg / L riboflavin, and 1 mg / L thiamine HCl. The carbon source is dextrose.

[0456] Medium C (natural complex medium) contained a brewer's grain extract extracted using a 3-minute steam pretreatment at 170°C for less than 5 minutes, followed by nutrient recovery via washing in water; the final protein content of the extract used in fermentation was approximately 10 g / L, the total glutamate concentration was 566 mg / L, and the total aspartate concentration was 216 mg / L; Medium C had the following composition by weight: 0.772% C, 0.059% N, 1.6% O, and 0.326% P. The brewer's grains had a particle size distribution in which 2-4 mm constituted the highest range of the distribution, accounting for approximately 35% by weight of the total particle size distribution. The carbon to nitrogen ratio of the extract is approximately 11 (extract composition based on CHNO analysis: 1 wt% C, 0.093 wt% N, 0.81 wt% O, and 0.12 wt% H).

[0457] All media contained the following salts based on potassium (0.1 g / l), sodium (2 g / l), iron (0.001 g / l), copper (0.01 g / l), magnesium (0.09 g / l), calcium (0.009 g / l), manganese (0.09 g / l), and zinc (0.05 g / l).

[0458] Mycelium production is divided into two distinct lines: a seed line and specific main fermentations in the respective media. The seed line is divided into three distinct steps. First, the mycelium is grown on Petri dishes on PDA medium. The mycelium is then cultured at 28°C for 7 days in one of media A, B, or C. The entire culture is then cultured at ambient temperature. For each respective main fermentation, one culture from the seed line is used and grown aerobically in a fermentation system cultivated in a fermenter for approximately 5-6 days until complete carbon source consumption. In all three cases (A, B, and C), fermentation is carried out aerobically at ambient temperature (25°C) and a pH in the range of 4-5. In these examples, the pH of the fermentation medium was approximately 5. It is also common to control dissolved oxygen in fermentation. The ingredients are then harvested and washed with acidic water at pH 3.5 before being prepared for further processing. Unless expressly indicated to the contrary, it is understood that the medium compositions used herein (e.g., the specific compositions of media A, B, and C) correspond to the medium composition at the start of the fermentation. As indicated above, it will be apparent to those skilled in the art that the composition of the medium may change over time, e.g., certain components, e.g., the carbon source, may be at least partially consumed.

[0459] Method for freeze-drying components A, B, and C Freeze-drying was carried out in a "Christ Alpha 1-2" freeze-dryer connected to an external vacuum pump, and each sample (A, B, or C) was freeze-dried at -50°C with a direct application of 6 mbar vacuum. All samples were pre-frozen at -80°C. Samples were freeze-dried for 72 hours.

[0460] BET surface area by gas adsorption (according to DIN-ISO 9277, respectively DIN 66131) During gas adsorption (according to DIN-ISO 9277, respectively DIN 66131), the specific surface area of ​​a solid material is determined using the BET method, by default with nitrogen adsorption at 77.4 K. Sample preparation is performed at each specific temperature. The sample surface must not change during this time. In the statistical-volume method, a specific amount of the measurement gas is introduced onto the sample placed in a vacuum. The determination of the adsorbed gas amount is based on the gas equation and pressure measurements in a volumetric calibration system. In the dynamic method, the determination of the adsorbed gas amount is carried out using a thermal conductivity sensor that detects changes in the gas composition in an N2 / He mixture. From the measurement readings (adsorbed volume Va vs. relative pressure p / p0), the molecular weight in the monolayer on the surface of the solid material is calculated. Evaluations are carried out in the general range of validity of the BET method, p / p0 = 0.05 to 0.3, respectively, in the relative pressure ranges indicated. For the determination of very small surfaces, krypton adsorption (at 77.4 K) is used. Because the expected surface areas are very low, the sample cell is completely filled with sample (approximately 0.5-0.6 g per sample - see attached data sheet), and krypton is used because it is a suitable adsorbate for measuring low surface areas. Prior to analysis, samples were degassed under vacuum at room temperature for approximately 65 hours. Analysis is performed on a Quantachrome Quadrasorb.

[0461] Mercury porosimetry (DIN-ISO 15901-1, respectively, according to DIN 66133). Pore ​​analysis by mercury porosimetry is performed using a Quantachrome Poremaster-60GT. The method is based on the Washburn equation, which describes the relationship between pore diameter and the pressure applied to a non-wetting liquid, such as mercury. Using the Poremaster-60GT, the penetrometer is filled in a horizontal position before analysis. This also avoids hydrostatic mercury pressure on the sample as undetected pore filling. The resulting intrusion curve is plotted as pressure versus pore diameter. Because the measurement starts at low pressure, larger pores fill first; therefore, typically, pore size decreases from left to right on the chart. The upper method limit is a pore diameter of approximately 1 mm, meaning that detection of larger pores was not the goal of this experiment.

[0462] Free bases, nucleosides, and 5' NMP nucleotides The lyophilized components were dissolved in water at 25 mg / mL, heated at 100° C. for 10 min, and homogenized. For protein precipitation, TCA was added to a final concentration of 5% and incubated at −20° C. for 16 h. Five μL of the deproteinized extract was then injected and analyzed by ion-pairing HPLC.

[0463] RNA quantification method To quantify RNA, nuclease was added to the solubilized sample (25 mg / mL) and incubated for 16 hours at 60° C. to convert the RNA to 5′NMP, and for protein precipitation, TCA was added to a final concentration of 5% and incubated for 16 hours at −20° C. Five μL of the deproteinized extract was then injected and analyzed by ion-pairing HPLC.

[0464] Flavor was measured for known flavor 5'-nucleotides (5'-NMPs) in g / kg, including 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP), and 5'-adenosine monophosphate (AMP). Component A exhibited a better enrichment of 5'-NMP at 3.56 g / kg compared to B, which exhibited 1.78 g / kg. However, the use of brewer's grain extract resulted in a 10-fold increase in component C compared to C, resulting in 10.43 g / kg of 5'-NMP. Additionally, component C exhibited the highest concentration of uridine monophosphate (UMP) at 3.56 g / kg compared to 1.26 g / kg and 0.71 g / kg for A and B, respectively.

[0465] In one embodied example, without any additional ribonucleic acid (RNA) reduction steps via additional heating or pH treatment in the component production process, the method of the present invention results in component specific RNA levels of less than 2% by weight for all disclosed mycelial components, specifically 1.88% by weight for A, 1.65% by weight for B, and 2% by weight for C.

[0466] [Table 1]

[0467] Amino acid quantification methods and data Amino acid analysis is by acid hydrolysis of the sample followed by HPAEC-IPAD. Tryptophan is not detected by this method because it is subject to degradation during acid hydrolysis of the sample. All components A, B, and C exhibit at least about 20% by weight of umami amino acids, more specifically, 21.48% by weight of umami amino acids for A, 19.5% by weight of umami amino acids for B, and 22.64% by weight of umami amino acids for C.

[0468] All components A, B, and C have a BCAA content of at least about 20% by weight, more specifically, A has a BCAA content of 21.73% by weight, B has a BCAA content of 23.1% by weight, and C has a BCAA content of 19.6% by weight.

[0469] All components A, B, and C have an essential amino acid content of at least about 40% by weight, more specifically, A has an essential amino acid content of 40.81% by weight, B has an essential amino acid content of 40.52% by weight, and C has an essential amino acid content of 39.61% by weight.

[0470] The total amino acid content in mg / g was the richest in C (361.67 mg / g), followed by A (268.85 mg / g), and then C (163.43 mg / g).

[0471] [Table 2]

[0472] Biomass characterization and EUC concentration Mycelial component A is grown in a medium containing a carbon to nitrogen ratio (medium) of about 17. Mycelial component B is grown in a medium containing a carbon to nitrogen ratio (medium) of about 20. Mycelial component B is grown in a medium containing a carbon to nitrogen ratio (medium) of about 13.

[0473] The insoluble fiber content increases from approximately 23% to approximately 34% to approximately 54% by weight from C to A to B, and the protein content decreases from approximately 60% to approximately 39% to approximately 32% by weight from C to A to B. This illustrates the relationship discussed above between protein and fiber, which are inversely proportional and can be controlled by varying the fermentation medium composition.

[0474] The fat content of component A is about 3% by weight. The fat content of component A is about 2% by weight. The fat content of component C is approximately 7% by weight. As shown in the table below, this fat includes saturated, monosaturated, polyunsaturated, and trans fatty acids. Mushroom mycelium is highest in saturated fatty acids (maximum 1.5g / 100g for components A and B, maximum 2g / 100g for component C), while component C is rich in polyunsaturated fatty acids (3.63g / 100g). The components contain minimal amounts of trans fatty acids (maximum 0.02-0.03g / 100g). Note that the omega-6 fatty acid (linoleic acid) content is 0.77% by weight for component A, 0.46% by weight for component B, and 3.5% by weight for component C. All components exhibit a calculated carbohydrate content of less than 0.1% by weight. Component B is the richest in total glucan, with a total glucan value of 31% by weight. Mycelium component A has a total glucan value of approximately 26% by weight. Mycelial component C has a total glucan value of about 17% by weight.

[0475] [Table 3]

[0476] The EUC for the mycelial components was calculated to be 302% for A, 34% for B, and 2892% for C.

[0477] In a separate study, P. pulmonarius fruiting bodies were grown on three forestry wastes (pine, poplar, and honeysuckle), which showed that the EUC values ​​of P. pulmonarius fruiting bodies were 72.31% to 116.73% (Food Chemistry 397(2022)133714). The disclosed EUC values ​​of mycelial components A, B, and C of the present invention are higher than the reported values ​​for fruiting bodies of the same fungal strain; for example, component C, the difference is 25-40 times higher for component C compared to the reported range for P. pulmonarius fruiting bodies. It is also known that the highest value of EUC in mushroom fruiting bodies was reported to be 4465% for (Volvariella volvacea), which is 1.54 times lower compared to component C.

[0478] Additionally, the EUC concentrations of beef, chicken, green peas, and soybeans (see Figure 5 for the exact products used in the comparison) were measured and compared to mycelial components A, B, and C, and the EUC found was found to be 78.77%, 134.7%, 65%, and 1%, respectively.

[0479] Total sugar determination method Approximately 300 mg of ingredient is added to a pressure tube. 3 mL of 72% H2SO4 is added using an autotitrator, and the weight of the acid added is recorded. The sample is thoroughly mixed with the acid using a glass rod. The tube is transferred to a water bath maintained at 30°C. These steps are repeated for duplicate samples. Every 10 minutes, the glass rod for each pressure tube is agitated, allowing the acid to reach all parts of the sample and resulting in complete hydrolysis. Exactly one hour after placing the tube in the water bath, the pressure tube is removed, placed on a balance, and 84 mL of water is added (the weight of the water added is recorded). At this point, the water is used to remove the acid / sample from the rod. The lid is screwed onto the tube, and the tube is inverted several times to ensure complete mixing of the acid. Two Sugar Recovery Solution (SRS) pressure tubes are prepared to monitor sugar loss associated with the second stage of hydrolysis. This involves the following steps: (a) 348 microliters of 72% H2SO4 is added to a test tube containing a solution containing a known weight (approximately 10 g) of sugar standard. This standard should be of a sugar composition similar to that expected for the sample being analyzed. The acid and sugar solution are mixed thoroughly. (b) The sugar-acid mixture is transferred to a pressure tube, which is then sealed. All SRS and sample pressure tubes are placed in an autoclave operated at 121 °C for 60 minutes. After the temperature inside the autoclave drops below 80 °C, the tubes are removed and left (closed) in the laboratory until room temperature is reached. The hydrolysate is then filtered (using vacuum suction) through a filtration crucible of known weight, and the resulting filtrate is stored. Any residual solids are washed away from the tube using deionized water until all acid-insoluble residue is present on the filtration crucible. Following the acid hydrolysis step, the hydrolysate (filtrate from vacuum filtration) is diluted 5-fold using a fucose solution in water. Fucose is the internal standard used in the chromatographic analysis of the hydrolysate. Following this dilution, the sample is immediately subjected to the chromatographic system. The instruments used are a NIR spectrophotometer (FOSS XDS NIR) and an ion chromatograph (ICS-3000).

[0480] Sugars in water extract data The ingredients were hydrolyzed to their sugar (glucan, xylan, arabinan, galactan, mannan, rhamnan) content using acid. However, this hydrolysis was performed on the original samples from which extractives (e.g., ethanol or water-soluble components) had not been removed. All data are expressed as a percentage of the total dry mass.

[0481] [Table 4]

[0482] The most abundant sugar as a percentage of total sugar content in A is 88% glucan, in B 92.5% glucan, and in C 78.6% glucan (glucan here is based on all glucose, including glucose derived from other polysaccharides, e.g., heteroglycans and / or exopolysaccharides).

[0483] The sugars in the water extract or extract shown in the table above, including sugars extracted from fresh mycelium components via water, were extracted in an 11 ml stainless steel extraction cell in an ASE 200 (Accelerated Solvent Extractor) using deionized water at 100°C and 1500 psi (heat time: 5 min, rest time: 7 min, flush volume: 150%, purge time: 180 s, rest cycle: 3). The total sugars in the water extracts of components A, B, and C were 6.25%, 7%, and 3.26%, respectively. From these, the sugars in the water extract of A, expressed as a % of the total extract sugars, included disaccharides (20.1% trehalose), trehalose sugars (22% glucose, 0.5% fructose, 0.1% mannose, 1.4% galactose), pentose sugars (0.1% arabinose), and sugar alcohols (55% mannitol, 0.7% sorbitol). For B, the sugars in the aqueous extract, expressed as a % of total extract sugars, include disaccharides (19.2% trehalose), trehalose sugars (43.3% glucose, 0.4% fructose, 0.1% galactose), pentose sugars (0.1% arabinose), and sugar alcohols (35.7% mannitol, 1% sorbitol). For C, the sugars in the aqueous extract, expressed as a % of total extract sugars, include disaccharides (46% trehalose), trehalose sugars (16% glucose, 0.1% galactose), pentose sugars (0.03% arabinose and 0.15% xylose), and sugar alcohols (22.7% mannitol, 13.5% arabinitol, 1% xylitol, 0.64% sorbitol).

[0484] Elemental analysis, ash content, and gross calorific value determination methods Final (elemental) analysis of biomass provides the mass concentrations of the major elements (carbon, oxygen, hydrogen, nitrogen, and sulfur) in the sample. The carbon, hydrogen, nitrogen, and sulfur content of the sample was measured according to the procedures outlined in European Standard EN 15104:2011 ("Solid biofuels - Determination of total content of carbon, hydrogen, and nitrogen - Instrumental methods"). An Elementar Vario MACRO Cube elemental analyzer was used. The differential oxygen content follows the formula: oxygen (%) = 100 - carbon (% dry basis) - hydrogen (% dry basis) - nitrogen (% dry basis) - sulfur (% dry basis) - ash (% dry basis). To perform this calculation, the ash content of the sample was also measured in a Nabertherm L-240H1SN muffle furnace heated to a maximum of 575°C. The sample was then weighed and the ash content calculated. The higher heating value (HHV, often referred to as gross heating value) was determined directly using an oxygen bomb calorimeter as outlined in EN 14918:2009. The lower heating value (LHV, often referred to as net heating value) was calculated based on the HHV and elemental composition of the sample.

[0485] Elemental analysis, ash content, and gross calorific value data The samples were also analyzed for their ash content and their elemental composition (carbon C, hydrogen H, nitrogen N, sulfur S, oxygen O), and the oxygen content was determined by difference. The ash contents of components A, B, and C are 9.39%, 9.73%, and 8.40%.

[0486] Elemental analysis of these components for C, H, N, O, S analysis gives the following: For A: 44.45% (carbon C), 5.95% (H), 6.25% (N), 0.32% (S), 33.53% (O), 0.0815% (chlorine); Carbon to nitrogen ratio: 7.11. For B: 43.92% (carbon C), 5.83% (H), 4.62% (N), 0.26% (S), 35.64% (O), 0.0788% (chlorine); Carbon to nitrogen ratio: 9.5. For C: 46.90% (carbon C), 6.26% (H), 9.57% (N), 0.42% (S), 28.45% (O), 0.0703% (chlorine); Carbon to nitrogen ratio: 4.9.

[0487] The lower nitrogen content in Sample B indicates that the sample is richer in fiber content but has a lower protein content, while the brewer's grain-based Sample C has the highest nitrogen content, indicating a high protein content and a lower fiber content. Sample A falls between the two. This indicates that the protein and fiber content can be fine-tuned by adjusting the composition of the medium either synthetically or by using a nutrient side stream with a known elemental composition, allowing for the production of tailored food products rich in either fiber or protein.

[0488] The higher heating values ​​(HHV, often referred to as gross heating values) were determined directly using an oxygen bomb calorimeter as outlined in EN 14918:2009. The lower heating values ​​(LHV, often referred to as net heating values) were calculated based on the HHV and elemental composition of the samples: for A, 18.4 MJ / kg and 17.11 MJ / kg; for B, 18.88 MJ / kg and 17.61 MJ / kg; for C, 19.73 MJ / kg and 18.37 MJ / kg.

[0489] Other methods of determination Chitin Analysis The chitin content was determined using the method described in the following paper (https: / / doi.org / 10.1155 / 2020 / 5084036). The maximum recovery of glucosamine was ensured. The glucosamine content was estimated by ion chromatography. Chitosan was used as a control.

[0490] Ergothioneine analysis For quantification of ergothioneine, approximately 1.25 g of sample was weighed into a 25 mL volumetric flask. The volumetric flask was filled to the mark with 70% cold ethanol, to which 10 mM dithiothreitol was added, and sonicated for 15 minutes. The sample was then centrifuged at 4000 rpm for 20 minutes to separate insoluble material. 2 mL of the resulting supernatant was evaporated to dryness. The residue was resuspended in 1 mL of water and centrifuged at 10,000 rpm for 1 minute. Finally, the supernatant was filtered through a 0.45 μm filter and further diluted with water prior to HPLC injection.

[0491] HPLC analysis of ergothioneine was performed on a Prominence system (Shimadzu, Duisburg, Germany) equipped with an LC-20AD high-performance liquid chromatography (HPLC) pump, a SIL-20AC HT autosampler, an SPD-M20A diode array detector (DAD), a CBM-20A communication bus module, and a LabSolutions Multi LC Data System Manager. An EC 250 / 4 Nucleosil 100-5 C18 column (Macherey-Nagel, Duren, Germany) was used with a matching precolumn. The injection volume was 10 μL. The mobile phase used was 3% acetonitrile and 0.1% acetic acid in water. The separation was performed under isocratic elution at a flow rate of 0.7 mL / min and ambient temperature for 15 min, and detection was performed at 254 nm. External calibration with ergothioneine dissolved in water (5 μg mL−1–100 μg mL−1) was used for quantification.

[0492] Ergosterol analysis Ergosterol was quantified by internal calibration using 7-dehydrocholesterol as an internal standard. For ergosterol analysis, 150–200 mg of each sample was weighed into a derivatization tube and mixed with 50 mg of sodium ascorbate and 250 μL of 7-dehydrocholesterol stock solution (5 mg mL in 2-butanone). After adding 5 mL of sodium hydroxide solution in methanol and homogenizing by vortexing, the tube was incubated in a water bath at 80 °C for a total of 1 h. Shaking was performed every 20 min. After cooling the samples to room temperature in the dark, each sample was membrane filtered and transferred to a new derivatization tube. Subsequently, three extractions with hexane were performed. The hexane phases were combined in a 15 mL volumetric flask, and the flask was filled to the mark with hexane. After drying with sodium sulfate, 6 mL was transferred to a new derivatization tube. The solvent was then removed under a stream of nitrogen. The residue was taken up in 0.5 mL of tetrahydrofuran (THF) and 0.5 mL of N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) and dissolved in an ultrasonic bath. The solution was then heated in a water bath at 70 °C for 2 min. After each 1 min, homogenization was performed by vortexing. Silylation was carried out overnight. After transfer to vials, the samples were analyzed by gas chromatography (GC) and flame ionization detection (FID). A calibration series containing 1 mL of each 7-dehydrocholesterol stock and 0.25–4.5 mL of ergosterol stock (2 mg mL in 2-butanone) was prepared in a 10 mL volumetric flask. 2-butanone was used to fill to the mark, and 1 mL was transferred to a derivatization tube and dried under a stream of nitrogen. A calibration series was then recorded and silylated in THF and MSTFA similar to the samples. An ergosterol content of 2.35 g / g dry weight was determined for sample A, 2.24 g / g dry weight for sample B, and 5.56 g / g dry weight for sample C.

[0493] Further analysis Protein content is determined according to DIN EN ISO 16634-1, 2009-07, (N * 6.25) DUMAS. Fat content was determined according to Weibull-Stoldt. Fiber content was determined according to enzymatic gravimetric method ASU L 00.00-18. Carbohydrate content was determined via HPLC according to SOP M 2569. Vitamin content was determined by a Dionex ICS-3000 ion chromatography system equipped with electrochemical, conductivity, and UV-visible detectors. Megazyme's beta-glucan assay kit for yeast and mushrooms was used for alpha / beta-glucan determination. Thermogravimetric analysis (TGA) was performed on a TA Instruments Q500 TGA unit, which allows monitoring of sample weight loss under a nitrogen atmosphere up to 1000°C (ramp used: equilibration at 35°C, 2-minute isothermal, equilibration at 105°C, 5-minute isothermal, 20°C / min ramp to 900°C).

[0494] Trade names of the products purchased for EUC calculation: beef and chicken (METZGERFRISCH, Rinder-SupppenFleisch and Hanschen-Innenfilets, respectively), green peas (Sunat), and soybeans (Rapunzel).

[0495] Density measurement The density of mycelium component samples was measured according to Archimedes' principle using an analytical density balance, KERN EMB-V (KERN & SOHN GmbH, Balingen, Germany), and a YDB-01 set (KERN & SOHN GmbH, Balingen, Germany). Tests were performed at room temperature (approximately 21 °C) and 50-60% relative humidity and were run in triplicate. The kit provided a weighing plate, platform, beaker, and immersion basket for lowering and suspending solid materials. Samples were weighed in air and distilled water (or other suitable liquids for densities less than 1 g / cm3), and the density was calculated directly in g / cm3. In one example, the component density was found to be 1.012 g / cm3. The density of the mycelium component ranges from 0.1 to 2.5 g / cm3, most preferably between 0.7 and 1.2 g / cm3.

[0496] [Table 5]

[0497] shear force methodology Shear force was measured using a TA.XTplus100 texture analyzer (Stable Micro Systems, Surrey, United Kingdom). 50 kg and 100 kg load cells were used for testing. Samples were analyzed at ambient temperature (approximately 21 °C). Due to the heterogeneity of the biomass, 5 g samples were sheared, compressed, and bulk extruded using a Miniature Kramer Shear / Ottowa Cell (HDP / MKS5). Results were obtained as the average force (N) required to break the sample. In addition to force, the area under the curve (i.e., shear effort) was also calculated (N·s).

[0498] [Table 6]

[0499] Water retention (WHC) and water release (RW) Water-holding capacity (WHC) describes the ability of a material to retain water during processing. Among the many procedures for measuring WHC, in this study, WHC was determined using a method modified from van der Sman et al. and Liu et al. (Van der Sman, R.G.M.; Paudel, E.; Voda, A.; Khalloufi, S. Hydration properties of vegetable foods explained by Flory-Rehner theory. Food Research International 2013, 54, 804-811; and Liu, C.; Li, W.; Lin, B.; Yi, S.; Ye, B.; Mi, H.; Li, J.; Wang, J.; Li, X. Comprehensive analysis of ozone water rinsing on the water-holding capacity of grass carp surimi gel. Food Science and Technology 2021, 150, 111919). After separation of the biomass and supernatant, 3 g of wet biomass was placed in a 50 ml falcon tube consisting of a cotton ball and a filter paper layer at the bottom. The sample was centrifuged at 4500 rpm for 10 minutes at 23°C. During centrifugation, water was removed from the mycelium particles through the filter and collected at the bottom. After centrifugation, the biomass was carefully removed and weighed. The falcon tube with the cotton and filter paper was also weighed for comparison of the results. WHC (%) was calculated according to the following formula:

[0500]

number

[0501] Released water was determined in triplicate according to the work of Seon-Tea Joo (Joo, S. Determination of water-holding capacity of porcine musculature based on released water method using optimal load. Korean Journal for Food Science of Animal Resources 2018, 38(4), 823.). Approximately 1.0 g of released biomass was weighed and carefully placed on two pre-dried and weighed thin plastic films (PP5) and filter paper (MN 615, 12.5 cm diameter). A 100 g weight was applied for 5 minutes using a metal circular plate as a weight. After accurately removing the compressed biomass sample, the wet filter paper and two plastic films were weighed. RW (%) was defined according to the following equation:

[0502]

number

[0503] [Table 7]

[0504] Puncture test and spreadability of milk substitute (cream cheese) Puncture test Measurements were performed on a TA.XTplus100 texture analyzer (Stable Micro Systems, Surrey, United Kingdom) equipped with a 5 kg load cell. Samples were prepared in polypropylene plastic boxes (inner diameter = 7.8 cm, height = 4.4 cm) and subjected to a perforation test using a cylindrical Perspex probe (P / 20P) with a diameter of 20 mm. The probe perforated the cheese samples to a depth of 20 mm at a test speed of 1 mm s-1, a pre-test speed of 1 mm s-1, and a post-test speed of 10 mm s-1. Samples were analyzed at 16 °C to minimize differences in solid fat content. Three measurements were performed for each of three replicates. The deformation level was appropriately selected to ensure complete penetration of the probe into the coextrudate. Force (N) versus time (s) data was recorded and several mechanical parameters were determined using Exponent software (Stable Micro Systems, Surrey, UK). The parameters maximum positive force (N), maximum negative force (N), positive area (N·s), and negative area (N·s) were determined. Maximum force is highly correlated with the firmness of the cheese samples (Journal of Texture Studies, 32:41-55).

[0505] Cream cheese spreadability The spreadability test was performed using a TA.XTplus100 texture analyzer (Stable Micro Systems, Surrey, United Kingdom). A 5 kg load cell was used for the test. Samples were analyzed at 16°C to minimize differences in solid fat content. The TTC spreadability fixture (HDP / SR) included a male 90° cone probe and five precisely fitted female Perspex cone-shaped product holders. Cream cheese samples were filled with a spatula and then flattened to a flat surface. The sample holders were stored at the temperatures listed above before testing the samples. The degree of spreadability was calculated when the product was forced to flow outward between the male and female cone surfaces at a 45° angle during the test. A force-time curve was recorded (see figure below), and the force (N) at the maximum penetration depth was taken as the sample firmness. The area under the curve (N·s) represented the total amount of force required to carry out the shearing process. This is considered a good instrumental measure of spreadability for cream cheese and other spreadable products. Smaller values ​​in this area indicate easier spreadability. The maximum negative peak force (N) indicates sample stickiness, and the maximum negative area (N·s) represents adhesive behavior (Bayarri, S.; Carbonell, I.; Costell, E. Viscoelasticity and texture of spreadable cheeses with different fat contents at refrigeration and room temperatures. Journal of Dairy Science 2012, 95(12), pp. 6926-6936.).

[0506] Comparison between two mycelium-based cream cheeses Sample 1 is an example of a milk substitute containing mycelium component A, 8.0% pea starch, and 2.5% corn starch. Sample 2 is an example of a milk substitute containing mycelium component A, approximately 3.0% wheat starch, and 2.5% corn starch. Spreadability was measured and a puncture test was performed. It was observed that the lower the starch content, the softer the texture and therefore the lower the maximum force required to spread the cream cheese milk substitute, i.e., the lowest force (N) required to spread, i.e., the lowest surface area (Ns) and the lowest puncture force (N) required, and these values ​​increase with increasing starch content. However, this usually depends on the starch type and starch combination used.

[0507] [Table 8]

[0508] [Table 9]

[0509] Cutting strength methodology The cutting strength was determined using a TA.XTplus100 texture analyzer (Stable Micro System, Surrey, United Kingdom). A 50 kg load cell was used for the test. The samples were analyzed at a cooled temperature (approximately 7°C). The meatballs were cut by a blade at 80% of their original height. The results were obtained as the average of the maximum force (N) required to cut the sample. In addition to the force, the area under the curve (N·s) was also calculated. The force-time curve was recorded, and the maximum force was taken as the maximum cutting strength. The positive area under the curve (Ns) represented the total amount of force.

[0510] [Table 10]

[0511] Texture Profile Analysis (TPA) method TPA was performed using a TA.XTplus100 texture analyzer (Stable Micro System, Surrey, United Kingdom). A 50 kg load cell was used for the test. Samples were analyzed at cooled temperature (approximately 7°C). Unfried meatballs cooked at approximately 85°C for 5-10 minutes were compressed at 60% of their original height twice consecutively at a test speed of 1 mm / s. Therefore, a compression plate (SMS P / 75) was used. Results were obtained as the average of the maximum force (N) required to break the sample. The force-time curve was recorded, and the maximum force (N) at the first peak was taken as the sample hardness. The area under the first negative peak (Ns) represented the sample's adhesiveness. The height recovery represented the sample's elasticity (%), and the second area divided by the first area (%) demonstrated the sample's cohesiveness. The value of viscosity (N) was obtained by multiplying the viscosity (N) by the cohesiveness (%), and the chewiness (N) was the result of multiplying the viscosity (N) by the elasticity (%).

[0512] [Table 11]

[0513] Recipe preparation for mycelium-based cream cheese via three different methods: Acidification by ingredients 10 to 50% by weight of Pleurotus Pulmonarius mycelium (edible fibrous mycelium) is homogenized with approximately 55% by weight of potable water, approximately 0 to 7% by weight of cashew nuts (a plant-derived fat component), 10 to 40% by weight of Cocos nucifera oil (a plant-derived fat component), at most 2% by weight of table salt (a sodium chloride source), at most 2% by weight of yeast flakes, at most 5% by weight of sucrose (a plant-derived sugar source), at most 5% by weight of lemon juice (a natural plant-derived acidity source), and at most 5% by weight of citric acid.

[0514] Under constant blending, the homogenized slurry is heated to a temperature of 75-95° C. for a maximum of 60 seconds. To coagulate the homogenized slurry, a pre-prepared coagulant solution is added during this preceding limited heating step as soon as the temperature reaches 25-40° C. The coagulant solution contains at most 15% by weight of each of the following: water, wheat starch, and hydrolyzed corn starch.

[0515] The resulting slurry is quickly removed from the heat source, and the mixture is then allowed to settle to room temperature (21° C.) Upon reaching room temperature, the solidified slurry is stored in a dark, cool environment, preferably at 4° C. to 7° C.

[0516] Acidification by microbial fermentation 10 to 50% by weight of Pleurotus Pulmonarius mycelium (edible fibrous mycelium) is homogenized with approximately 55% by weight of drinking water, approximately 0 to 7% by weight of cashew nuts (a plant-derived fat component), 10 to 40% by weight of Cocos nucifera oil (a plant-derived fat component), at most 2% by weight of table salt (a sodium chloride source), at most 2% by weight of yeast flakes, and at most 5% by weight of sucrose (a plant-derived sugar source).

[0517] Under constant blending, the homogenized slurry is heated to a temperature of 75-95° C. for a maximum of 60 seconds. To coagulate the homogenized slurry, a pre-prepared coagulant solution is added during this preceding limited heating step as soon as the temperature reaches 25-40° C. The coagulant solution contains at most 15% by weight of each of the following: water, wheat starch, and hydrolyzed corn starch.

[0518] The resulting slurry is quickly removed from the heat source, and the mixture is then allowed to settle to room temperature (21°C). When the resulting mixture reaches a temperature of 40°C or less, 0.1 g to 0.25 g of acid-forming bacteria, particularly lactic acid bacteria, is added to induce microbial acidification. To provide a hospital environment for the microorganisms, the sample is placed in a controlled temperature environment (e.g., a water bath or incubator) at 28°C for 150 minutes. Depending on the intensity of the fermentation, the sample may be placed at a temperature of 20 to 45°C for 1 to 6 hours. After microbial fermentation is complete, the solidified slurry is stored in a dark, cool environment, preferably at 4 to 7°C.

[0519] Cream cheese based on two fungal strains Following the same instructions as shown in Examples 1 and 2, only replacing a fraction of the Pleurotus Pulmonarius edible filamentous mycelium with Morchella rufobrunnea edible filamentous mycelium. Assuming the amounts listed above, 60-90% by weight of the total mycelium is from Pleurotus Pulmonarius mycelium and 10-40% by weight of the total mycelium components is from Morchella rufobrunnea mycelium (edible filamentous mycelium).

[0520] Recipe Preparation for Vegetarian Mycelium-Based Meat Substitute Composition: 50-95% by weight of Pleurotus pulmonarius mycelia (edible fibrous mycelia) are blended and added to at most 1-40% by weight of each of the following ingredients: canola oil, salt, egg white, and wheat gluten. Optionally, at most 1-40% by weight of each of the following ingredients: methylcellulose, hydrocolloid, texturizing vegetable protein, starch-based ingredient, and flavor component. The resulting dough is then shaped through a former and / or extruder, or a combination thereof, to obtain a final product as meatballs or sausages or an extruded product.

[0521] Recipe Preparation for Vegan Mycelium-Based Meat Substitute Composition: 50-95% by weight of Pleurotus pulmonarius mycelia (edible fibrous mycelia) are blended and added to at most 1-40% by weight of each of the following ingredients: canola oil, salt, and wheat gluten. Missing egg white is accounted for by adding additional dry mycelia at an equivalent ratio of 1-1.4, respectively. Optionally, at most 1-40% by weight of each of the following ingredients: methylcellulose, hydrocolloid, texturizing vegetable protein, starch-based ingredient, and flavor component is added, and the resulting dough is then shaped through a former and / or extruder, or a combination thereof, to obtain a final product as meatballs or sausages or an extruded product.

[0522] Further examples and embodiments of the present invention are disclosed in the following numbered items: 1. An edible mycelium component A obtained by submerged fermentation, wherein the mycelium has an elemental composition with a C:N ratio in the range of 6 to 8. 2. An edible mycelium component B obtained by submerged fermentation, the edible mycelium component B having an elemental composition of the mycelium with a C:N ratio in the range of 8 to 12. 3. An edible mycelium component C obtained by submerged fermentation, the mycelium having an elemental composition with a C:N ratio in the range of 2 to 6. 4. The edible mycelium ingredient according to item 1, wherein the indigenous RNA level is less than 1.88% by weight on a dry basis without further process steps to reduce the RNA. 5. The edible mycelium ingredient according to item 2, wherein the indigenous RNA level is less than 1.65% by weight on a dry basis without further process steps to reduce the RNA. 6. An edible mycelium ingredient according to items 1 to 3, wherein the intrinsic RNA level is at most 2% by weight on a dry basis without further process steps to reduce the RNA. 7. The edible mycelium ingredient according to item 1, wherein the mycelium has an ergothioneine content in the range of 70 to 100 mg / kg. 8. The edible mycelium ingredient according to item 2, wherein the mycelium has an ergothioneine content in the range of 110 to 150 mg / kg. 9. The edible mycelium ingredient according to item 3, wherein the mycelium has an ergothioneine content in the range of 380 to 455 mg / kg. 10. The edible mycelium ingredient according to items 1 to 3, wherein the content of 5'-inosine monophosphate (IMP) is 0 g / kg. 11. The edible mycelium ingredient according to item 1, wherein the content of umami 5' nucleotides (5' NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) is up to 6 g / kg. 12. The edible mycelium ingredient according to item 2, wherein the content of umami 5' nucleotides (5' NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) is up to 3 g / kg. 13. The edible mycelium ingredient according to item 3, wherein the content of umami 5'-nucleotides (5'-NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) is up to 20 g / kg. 14. The edible mycelium ingredient according to any one of items 1 to 3, wherein the umami taste can be further enhanced by at least 60% upon enzymatic treatment of AMP to IMP. 15. The edible mycelium ingredient according to item 3, wherein the concentration of uridine monophosphate (UMP) is 3.56 g / kg. 16. An edible mycelium ingredient according to any one of items 1 to 3, wherein the amount of branched chain amino acids (BCAAs) is at least about 19% by weight of the total amount of amino acids present. 17. An edible mycelium ingredient according to any one of items 1 to 3, wherein the amount of umami amino acids is at least about 19% by weight of the total amount of amino acids present. 18. An edible mycelium ingredient according to items 1 to 3, wherein the amount of essential amino acids is at least about 40% by weight of the total amount of amino acids present. 19. An edible mycelium ingredient according to item 1, wherein the amount of BCAA and the amount of umami amino acids are each in the range of 40 to 100 mg / g. 20. The edible mycelium ingredient according to item 2, wherein the amount of BCAA and the amount of umami amino acids are each in the range of 20 to 80 mg / g. 21. An edible mycelium ingredient according to item 3, wherein the amount of BCAA is in the range of 50 to 100 mg / g and the amount of umami amino acids is in the range of 70 to 100 mg / g. 22. An edible mycelium component according to item 1, having an EUC concentration of at least 500%. 23. An edible mycelium component according to item 2, having an EUC concentration of at least 400%, preferably about 460%. 24. An edible mycelium ingredient according to item 3, having an EUC concentration of at least 500%, more preferably at least 5000%, and most preferably at least 10000%. 25. An edible mycelium component according to item 1 or 2, having a thermal stability of at most 220°C under an N2 atmosphere. 26. An edible mycelium component according to item 3, having a thermal stability of at most 190°C under an N2 atmosphere. 27. The edible mycelium ingredient according to any one of items 1 to 3, having a calorific value in the range of 300 to 600 Kcal / 100 g. 28. An edible mycelium component according to item 1, having an insoluble fiber content of 30 to 60% by weight, preferably 30 to 40% by weight. 29. An edible mycelium ingredient according to item 2, having an insoluble fiber content of 40 to 60% by weight, preferably 40 to 50% by weight. 30. An edible mycelium ingredient according to item 3, having an insoluble fiber content of 10 to 40% by weight, preferably 20 to 30% by weight. 31. An edible mycelium ingredient according to item 1, having a protein content of 30 to 50% by weight, preferably 30 to 40% by weight. 32. An edible mycelium ingredient according to item 2, having a protein content of 30 to 50% by weight, preferably 30 to 40% by weight. 33. An edible mycelium ingredient according to item 3, having a protein content of 30 to 65% by weight, preferably 45 to 65% by weight. 34. An edible mycelium ingredient according to item 1 or 2, having an ergosterol content of 2 to 4 mg / g. 35. The edible mycelium ingredient according to item 3, having an ergosterol content of 4 to 7 mg / g. 36. An edible mycelium component according to any one of items 1 to 3, having a carbohydrate content of at most 1%. 37. An edible mycelium component according to any one of items 1 to 3, having a uronic acid content of 0.1 to 5% by weight. 38. An edible mycelium ingredient according to any one of items 1 to 3, wherein the chitin content is in the range of 6 to 11% by weight. 39. An edible mycelium ingredient according to any one of items 1 to 3, wherein the beta-glucan content is at least 80% of the total glucan. 40. The edible mycelium ingredient according to item 1, wherein the total glucan content is in the range of 20 to 35% by weight. 41. An edible mycelium ingredient according to item 2, wherein the total glucan content is in the range of 25 to 50% by weight. 42. The edible mycelium ingredient according to item 3, wherein the total glucan content is in the range of 10 to 20% by weight. 43. An edible mycelium component according to any one of items 1 to 3, wherein at least 96% by weight of the polyunsaturated fatty acids of the edible mycelium component are linoleic acid. 44. An edible mycelium ingredient according to item 3, having a concentrated omega-6 fatty acid (linoleic acid) in the range of 2 to 5% by weight. 45. An edible mycelium component according to item 1 or 2, having a fat content of at most 3% by weight. 46. ​​The edible mycelium component according to item 3, wherein the edible mycelium component has a concentrated fat content, the fat content being at most 8% by weight. 47. An edible mycelium component according to any one of items 1 to 3, wherein the edible mycelium component has a total phenolic content (TPC) in the range of 1 to 15 GAE / g and a total flavonoid content (TFC) in the range of 1 to 15 mg QE / g, the flavonoid content of mycelium component A comprising approximately 35% to 60% of the total phenolic content, the flavonoid content of mycelium component B comprising approximately 70 to 95% of the total phenolic content, and the flavonoid content of mycelium component C comprising approximately 50 to 80% of the total phenolic content. 48. The edible mycelium component described in item 1, when freeze-dried, has a specific pore volume of 9 cm3 / g, a median pore diameter of 44.5 μm, and a BET surface area of ​​0.79 m2 / g. 49. The edible mycelium component described in item 2, when freeze-dried, has a specific pore volume of 4.94 cm3 / g, a median pore diameter of 143 μm, and a BET surface area of ​​0.67 m2 / g. 50. The edible mycelium component described in item 2, when freeze-dried, has a specific pore volume of 2.46 cm3 / g, a median pore diameter of 7.1 μm, and a BET surface area of ​​1.59 m2 / g. 51. The edible mycelium component according to item 1, wherein, when freeze-dried, for pore diameters of less than 1 mm, 58% of the pore volume of the edible mycelium component corresponds to pore diameters of 1000 to 30 μm, the modal pore diameter is in the range of 85 to 185 μm, 42% of the pore volume corresponds to pore diameters of 30 to 2 μm, the modal pore diameter peak is equal to 16 μm, and 16 μm is the modal pore diameter in the range of 1000 to 2 μm. 52. An edible mycelium component according to item 2, wherein, when freeze-dried, for pore diameters of less than 1 mm, 81.5% of the pore volume of the edible mycelium component corresponds to pore diameters of 1000 to 30 μm, the modal pore diameter being in the range of 147 μm, 147 μm also being the modal pore diameter in the range of 1000 to 2 μm, and 18.5% of the pore volume corresponds to pore diameters of 30 to 2 μm, the modal pore diameter peak being equal to 15 μm. 53. An edible mycelium component according to item 3, wherein, when freeze-dried, for pore diameters of less than 1 mm, 20% of the pore volume of the edible mycelium component corresponds to pore diameters of 1000 to 20 μm, 80% of the pore volume corresponds to pore diameters of 20 to 2 μm, and the most frequent pore diameter peak is equal to 5.5 μm. 54. An edible mycelium ingredient according to any one of items 1 to 3, wherein the mycelium has a shear strength of at least 15 N, a water retention capacity in the range of 20 to 90%, and a water release capacity of 25 to 70%. 55. A method for producing the mycelium component according to any one of items 1 to 3 by submerged fermentation, wherein the fermentation medium contains 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins. 56. The method according to Item 55, wherein the production of component A according to Item 1 comprises culturing at least one fungal species in a defined medium, the defined medium comprising a complex nitrogen source and at least one carbon source, a plurality of amino acids, a vitamin source, and minerals, and the carbon-to-nitrogen ratio in the medium is in the range of 14 to 19. 57. The method according to Item 55, wherein the production of component B according to Item 2 comprises culturing at least one fungal species in a minimal synthetic medium, the minimal synthetic medium containing one amino acid and one vitamin, and the carbon-to-nitrogen ratio in the medium is in the range of 16 to 23. 58. The method according to Item 55, wherein the production of component C according to Item 3 comprises culturing at least one fungal species in a natural synthetic medium, wherein the natural synthetic medium comprises a brewer's grain C5 sugar extract, the carbon-to-nitrogen ratio in the medium is in the range of 10 to 25, and at least 35% by weight of the extracted brewer's grains is in the range of 2 to 4 mm. 59. At least one fungal species is Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Pezizomycetes, Agaricomycetes, Sordariomycetes, Pezizales, Boletales, Cantharelal es, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hy 59. The method according to any one of items 55 to 58, wherein the family is selected from the group consisting of: Dnaceae, Boletaceae, Meripillaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinacea. 60. The method of any one of items 55 to 58, wherein at least one fungal species is combined with another edible fungus, algae, bacteria, plant cells, archaeal cells, animal cells, fat cells, or a combination thereof. 61. The method of any one of items 55 to 58, further comprising the step of recovering the supernatant from the culture medium. 62. The method according to item 61, wherein the recovery comprises a step of crystallizing or precipitating the obtained supernatant. 63. A method for producing a soft or hard meat substitute composition comprising at least one of the mycelium components described in any one of items 1 to 3, comprising a method for producing each of the mycelium components described in any one of items 55 to 58, and further comprising the step of preparing such meat substitute composition by mixing at least one of the mycelium components described in any one of items 1 to 3 from at least one fungal strain with a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and optionally at least one composition component. 64. A method for producing a milk replacer composition comprising at least one of the mycelium components described in any one of items 1 to 3 from at least one fungal strain, including a method for producing each of the mycelium components described in any one of items 55 to 58, the method further comprising the steps of preparing such a milk replacer composition by: (1) forming a slurry comprising at least one of the mycelium components described in any one of items 1 to 3 together with a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and at least one composition component; and (2) mixing the slurry with at least one composition component, specifically a texturizing or thickening agent, or a carbohydrate-rich component. 65. The edible product according to item 63 or 64, further comprising an edible plant- or algae-based fat component, a fat component derived from a fungus or yeast, in a range of up to 25% by weight, preferably in a range of 1 to 5% by weight. 66. The edible product according to item 64 or 64, wherein the edible plant-based fat-rich ingredient component is selected from tree nuts, coconut oil, sunflower oil, rapeseed oil, palm oil, cottonseed oil, olive oil, canola oil, algae oil, and / or oleaginous yeast-derived oil. 67. The edible product according to item 64, wherein the at least one protein-rich ingredient or at least one composition ingredient is selected from agar, egg white, yeast flakes, food starch, guar gum, locust bean gum, wheat gluten, a sugar source, cellulose or a derivative thereof, lemon juice, a colorant, and / or a flavoring agent, such as salt, extracts, and / or spices. 68. The obtained meat substitute according to item 63, comprising mycelial mass in the range of 1% to 99% by weight and water in the range of up to 99% by weight, wherein the edible fibrous mycelium has an insoluble fiber content of 40 to 50% by weight, preferably about 45% w / w. 69. The resulting milk replacer according to item 64, comprising mycelial mass in the range of 1% to 99% by weight and water in the range of up to 99% by weight, wherein the edible fibrous mycelium has an insoluble fiber content of at least about 40% by weight. 70. The edible meat substitute according to Item 64, comprising 50% to 95% by weight of fibrous mycelium mass. 71. The edible milk replacer according to Item 63, comprising 1% to 50% by weight of fibrous mycelium mass. 72. Product according to item 63 or 64, characterized in that it can have a soft or non-soft / harder texture. 73. The food product according to item 63, characterized by a soft texture, preferably having a hardness of 10-55 N, a springiness of 35-85%, a cohesiveness of 15-70%, a viscosity of 1-40 N, a chewability of 0.3-35 N, a cutting strength of 1-25 N, and a stickiness of 0 N.s to -0.3 N.s. 74. The food product according to item 63, characterized by a non-soft or harder texture, preferably having a hardness of 30 to 100 N, a springiness of 20 to 70%, a cohesiveness of 20 to 85%, a viscosity of 6 to 85 N, a chewability of 1 to 60 N, and a cutting strength of 1 to 50 N. 75. The food product according to item 64, characterized by a soft texture, preferably having a hardness of 1 to 20 N, a spreadability of 30 to 100 N.s, a stickiness of -15 to -100 N, and a puncture force of 1 to 30 N.s. 76. The food product according to item 64, characterized by a non-soft or harder texture, preferably having a firmness of 20 to 100 N, a spreadability of 1 to 20 N.s, a stickiness of -1 to -14 N, and a puncture force of 40 to 100 N.s.

[0523] Further embodiments of the present invention are disclosed in the following numbered clauses: 1. An edible mycelium component C obtained by submerged fermentation, the mycelium having an elemental composition with a C:N ratio in the range of 2 to 6. 2. an intrinsic RNA level of at most 4% by weight, preferably at most 2% by weight, on a dry basis, without further process steps to reduce the RNA, and / or a content of umami 5' nucleotides (5' NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) of at most 40 g / kg, preferably at most 20 g / kg; and / or 5'-inosine monophosphate (IMP) is 0 g / kg, and / or the umami taste derived from 5'-nucleotides can be further enhanced by at least 40% when treated with an enzyme, preferably 5'-adenylate deaminase, to convert AMP to IMP; and / or the amount of branched chain amino acids (BCAAs) is at least about 19% by weight of the total amount of amino acids present; and / or the amount of essential amino acids is at least about 40% by weight of the total amount of amino acids present; and / or 2. The edible mycelium ingredient according to clause 1, wherein the amount of BCAA is in the range of 50 to 150 mg / g and the amount of umami amino acids is in the range of 70 to 100 mg / g. 3. 3. The edible mycelium ingredient according to clause 1 or 2, wherein the mycelium has an ergothioneine content in the range of 350 to 800 mg / kg. 4. The edible mycelium ingredient according to clause 1 or 3, wherein the amount of umami amino acids is at least about 19% by weight of the total amount of amino acids present. 5. An edible mycelium ingredient according to any one of clauses 1 to 4, having an EUC concentration of at least 500%, more preferably at least 5000%, most preferably at least 10000%. 6. At least 96% by weight of the polyunsaturated fatty acids of the edible mycelium component constitute linoleic acid, preferably having a concentrated omega-6 fatty acid (linoleic acid) content in the range of 2-5% by weight, and / or a concentrated fat content of at most 8% by weight; and / or 6. The edible mycelium ingredient according to any one of clauses 1 to 5, having a DPPH radical scavenging activity in the range of 1 to 15 mg / ml relative to TPC, and / or having TPC, TFC and polyphenol content in the ranges of 1 to 15 mg GAE / g, 1 to 15 mg QE / g and 100 to 1000 mg / kg, respectively, wherein the flavonoid content constitutes approximately 70 to 95% of the total phenolic content, and catechin and protocatechuic acid each constitute approximately 45 to 55% of the total polyphenols. 7. The calorific value is in the range of 300-600 Kcal / 100 g, and the ingredients have a thermal stability of at most 190°C under N2 atmosphere, and / or an insoluble fiber content of 10 to 40% by weight, preferably 20 to 30% by weight, and / or a protein content of 30 to 65% by weight, preferably 45 to 65% by weight, and / or an ergosterol content of 4 to 7 mg / g and / or a carbohydrate content of at most 5% by weight, and / or The content of uronic acid is 0.1 to 5% by weight, and / or the chitin content is in the range of 6-11% by weight, and / or the beta-glucan content is at least 80% of the total glucan, preferably the total glucan content is in the range of 10-20% by weight. 8. when freeze-dried, for pore diameters of less than 1 mm, about 15-25%, preferably about 20%, of the pore volume of the edible mycelium component corresponds to pore diameters of 1000-20 μm, and about 75-85%, preferably about 80% of the pore volume corresponds to pore diameters of 20-2 μm, with the most frequent pore diameter peak being equal to 5.5 μm; and / or 8. The edible mycelium ingredient of any one of clauses 1 to 7, wherein the mycelium has a shear strength of at least 15 N and a water retention capacity in the range of 20-90% and a water release capacity of 25-70%. 9. A method for producing a mycelial component according to any one of clauses 1 to 8 by submerged fermentation, wherein the fermentation medium comprises 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins, and the production of component C comprises cultivating at least one fungal species in a natural synthetic medium, wherein the natural synthetic medium comprises a brewer's grain C5 sugar extract, the carbon to nitrogen ratio in the medium being in the range of 10 to 25, and at least 35% by weight of the extracted brewer's grains being in the range of 2 to 4 mm, and preferably the particle size distribution is determined by sieving, which is carried out in an automatic sieving tower, i.e., by air jet sieving according to DIN 10765 2016-07, after removing particles with a particle size greater than 4 mm using a vibration sieving method. 10. At least one fungal species is selected from Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Pezizomycetes, Agaricomycetes, Sordariomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, Boletaceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinacea, and preferably, at least one fungal strain is from Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Agaricomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, Boletaceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae,the mycelial mass is selected from Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae and Fistulinacea, preferably the mycelial mass is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus florida, Pleurotus citrinopileatus, Pleurotus salmoneostramineus, Morchella esculenta, Morchella angusticeps, Morchella deliciosa and / or Morchella rufobrunnea, preferably the mycelial mass is obtained from Pleurotus pulmonarius and / or Morchella rufobrunnea; 10. The method of clause 9, wherein optionally, the at least one fungal species is combined with another edible fungus, algae, bacteria, plant cells, archaeal cells, animal cells, fat cells, or combinations thereof. 11. The method of clause 9 or 10, wherein the method further comprises recovering a supernatant from the culture medium or portion thereof, and optionally, the recovering comprises crystallizing or precipitating the obtained supernatant. 12. An edible mycelium component obtainable by the method according to any one of clauses 9 to 11. 13. A method for producing a soft or hard meat substitute composition comprising the mycelium component of any one of clauses 1-8 or 12, the method comprising the step of preparing the meat substitute composition by adding the mycelium component to a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and optionally at least one composition component. 14. A method for producing a milk replacer composition comprising the mycelium component of any one of clauses 1-8 or 12, the method comprising the steps of preparing the milk replacer composition by: (1) forming a slurry comprising the mycelium component together with a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and at least one composition component; and (2) mixing the slurry with at least one composition component, specifically a texturizing or thickening agent, or a carbohydrate-rich component. 15. An edible product obtainable by the method described in clause 13 or 14, Preferably, the food product according to clause 13 characterized by a soft texture preferably has a hardness of 10-55 N, a springiness of 35-85%, a cohesiveness of 15-70%, a viscosity of 1-40 N, a chewability of 0.3-35 N, a breaking strength of 1-25 N and a stickiness of 0 N.s to -0.3 N.s, and / or Preferably, the food product according to clause 13, characterized by a non-soft or harder texture, preferably has a hardness of 30-100 N, a springiness of 20-70%, a cohesiveness of 20-85%, a viscosity of 6-85 N, a chewiness of 1-60 N, a cutting strength of 1-50 N, and / or Preferably, the food product according to clause 14 characterized by a soft texture preferably has a hardness of 1 to 20 N, a spreadability of 30 to 100 N.s, a stickiness of -15 to -100 N and a puncture force of 1 to 30 N.s, and / or 15. Preferably, the food product according to clause 14 characterized by a non-soft or harder texture, preferably having a hardness of 20-100 N, a spreadability of 1-20 N.s, a stickiness of -1 to -14 N and a puncture force of 40-100 N.s.

[0524] Further embodiments of the present invention are disclosed in the following numbered paragraphs. 1. An edible mycelium component obtained from submerged fermentation, the mycelium having an elemental composition with a C:N ratio in the range of 6-12. 2. The edible mycelium component according to Item 1, wherein the edible mycelium component is edible mycelium component A obtained by submerged fermentation in a defined medium, and the defined medium has an elemental composition with a C:N ratio in the range of 6 to 8 of the mycelium. 3. The edible mycelium component according to Item 1, wherein the edible mycelium component is an edible mycelium component B obtained by submerged fermentation in a synthetic medium, and the synthetic medium has an elemental composition with a C:N ratio in the range of 8 to 12 of the mycelium. 4. the level of indigenous RNA is at most 2% by weight on a dry basis without further process steps to reduce the RNA; and / or the mycelium has an ergothioneine content in the range of 70-270 mg / kg, achieved within 5 days; and / or a content of umami 5' nucleotides (5' NMP) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) of at most 20 g / kg, and / or The amount of BCAA and the amount of umami amino acids are each in the range of 40 to 100 mg / g, and / or Calorific value is in the range of 300-600 Kcal / 100g, and / or the edible mycelium component has an insoluble fiber content of 30-60% by weight, preferably 30-40% by weight, and / or the edible mycelium component has a protein content of 30-50% by weight, preferably 30-40% by weight, and / or a total glucan content in the range of 20-35% by weight, and / or when freeze-dried, for pore diameters of less than 1 mm, 55-65%, preferably about 58%, of the pore volume of the edible mycelium component corresponds to a pore diameter of 1000-30 μm, the modal pore diameter being in the range of 85-185 μm, 35-45%, preferably about 42% of the pore volume corresponds to a pore diameter of 30-2 μm, the modal pore diameter peak being equal to 16 μm, 16 μm being the modal pore diameter in the range of 1000-2 μm, and / or the flavonoid content constitutes approximately 35-60% of the total phenolic content, and / or Item 3. An edible mycelium ingredient according to Item 2, wherein catechin and protocatechuic acid each account for approximately 35 to 45% of total polyphenols. 5. The edible mycelium ingredient according to item 2 or 4, having an EUC concentration of at least 500%. 6. the level of indigenous RNA is at most 2% by weight on a dry basis without further process steps to reduce the RNA; and / or the mycelium has an ergothioneine content in the range of 100-350 mg / kg, and / or a content of umami 5' nucleotides (5' NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) of at most 15 g / kg; and / or The amount of BCAA and the amount of umami amino acids are each in the range of 20 to 80 mg / g, and / or the edible mycelium component has an insoluble fiber content of 40-60% by weight, preferably 40-50% by weight, and / or the edible mycelium component has a protein content of 30-50% by weight, preferably 30-40% by weight, and / or a total glucan content in the range of 25-50% by weight, and / or when freeze-dried, for pore diameters of less than 1 mm, 75-85% of the pore volume of the edible mycelium component corresponds to pore diameters of 1000-30 μm, preferably about 81.5%, the modal pore diameter being in the range of 147 μm, which 147 μm is also the modal pore diameter in the range of 1000-2 μm, 15-25% of the pore volume, preferably about 18.5%, corresponds to pore diameters of 30-2 μm, the modal pore diameter peak being equal to 15 μm, and / or the flavonoid content constitutes approximately 70-95% of the total phenolic content, and / or Item 4. An edible mycelium ingredient according to Item 3, wherein catechin and protocatechuic acid account for approximately 25 to 35% and approximately 35 to 45% of total polyphenols, respectively. 7. The edible mycelium ingredient according to any one of items 1 to 6, wherein the DPPH radical scavenging activity, total phenol content (TPC), total flavonoid conte...

Claims

1. 1. An edible mycelial ingredient comprising undifferentiated mycelial biomass, said undifferentiated mycelial biomass having an elemental composition of mycelium with a C:N ratio in the range of 2-12, said edible mycelial ingredient being characterized by an equivalent umami concentration (EUC) of at least 500g monosodium glutamate MSG / 100g, wherein the EUC is: [Equation 1] is defined as wherein the EUC is expressed in g MSG / 100g, ai is the concentration of each umami amino acid, Asp or Glu (g / 100g), aj is the concentration of each umami 5'-nucleotide, 5'-IMP, 5'-GMP, or 5'-AMP (g / 100g), bi is the relative umami concentration (RUC) of each umami amino acid relative to MSG, defined as 1 for Glu and as 0.077 for Asp, and bj is the RUC of each umami 5'-nucleotide, defined as 1 for 5'-IMP, as 2.3 for 5'-GMP, and as 0.18 for 5'-AMP.

2. 2. The edible mycelium ingredient of claim 1, wherein said biomass has an elemental composition of mycelium with a C:N ratio ranging from 2 to 8.

3. 3. An edible mycelium ingredient according to claim 1 or 2, wherein the biomass has an elemental composition of the mycelium with a C:N ratio in the range of 2-6.

4. An edible mycelium component according to any one of claims 1 to 3 characterized by at least 1000g MSG / 100g EUC.

5. An edible mycelium ingredient according to any one of claims 1 to 4, characterized by at least 1500g MSG / 100g EUC.

6. An edible mycelium component according to any one of claims 1 to 5, characterized by at least 2000g MSG / 100g EUC.

7. 7. An edible mycelium ingredient according to any one of claims 1 to 6, wherein the indigenous RNA level is at most 4% by weight, preferably at most 2% by weight, on a dry basis, without further process steps to reduce the RNA.

8. 8. The edible mycelium ingredient according to any one of claims 1 to 7, wherein the content of umami 5' nucleotides (5' NMPs) containing 5'-guanosine monophosphate (5'-GMP) and 5'-adenosine monophosphate (5'-AMP) is at most 40 g / kg, preferably at most 20 g / kg.

9. An edible mycelium ingredient according to any one of claims 1 to 8, characterized by a 5'-AMP content of 3.5 to 10 g / kg and / or a 5'-GMP content of 3.5 to 10 g / kg.

10. An edible mycelium ingredient according to any one of claims 1 to 9, characterized by a 5'-AMP content of 4.5 to 7 g / kg and / or a 5'-GMP content of 4.5 to 7 g / kg.

11. The edible mycelium ingredient according to any one of claims 1 to 10, which is substantially free of 5'-IMP.

12. 12. An edible mycelium ingredient according to any one of claims 1 to 11, wherein the umami taste derived from 5'-nucleotides can be further enhanced by at least 40% when treated with an enzyme, preferably 5'-adenylate deaminase, to convert 5'-AMP to 5'-IMP.

13. 13. An edible mycelium ingredient according to any one of claims 1 to 12, wherein the amount of branched chain amino acids (BCAAs) is at least about 19% by weight of the total amount of amino acids present.

14. An edible mycelium ingredient according to any one of claims 1 to 13, wherein the amount of essential amino acids is at least about 40% by weight of the total amount of amino acids present.

15. 15. The edible mycelium ingredient according to any one of claims 1 to 14, wherein the amount of BCAAs is in the range of 50 to 150 mg / g and the amount of umami amino acids is in the range of 70 to 100 mg / g.

16. 16. An edible mycelium ingredient according to any one of claims 1 to 15, wherein the mycelium has an ergothioneine content in the range of 350 to 800 mg / kg.

17. 17. An edible mycelium ingredient according to any one of claims 1 to 16, wherein the amount of umami amino acids is at least about 19% by weight of the total amount of amino acids present.

18. 18. An edible mycelium ingredient according to any one of claims 1 to 17, wherein at least 96% by weight of the polyunsaturated fatty acids of said edible mycelium ingredient constitute linoleic acid, preferably having a concentrated omega-6 fatty acid (linoleic acid) in the range of 2-5% by weight, and / or having a concentrated fat content, the fat content being at most 8% by weight.

19. 19. An edible mycelium ingredient according to any one of claims 1 to 18, having a DPPH radical scavenging activity in the range of 1 to 15 mg / ml relative to TPC.

20. 20. The edible mycelium ingredient according to any one of claims 1 to 19, wherein the TPC, TFC and polyphenol content are in the ranges of 1-15 mg GAE / g, 1-15 mg QE / g and 100-1000 mg / kg, respectively, the flavonoid content constitutes approximately 70-95% of the total phenolic content, and catechin and protocatechuic acid each constitute approximately 45-55% of the total polyphenols.

21. The calorific value is in the range of 300 to 600 Kcal / 100 g, and the component is N ２ 21. An edible mycelium component according to any one of claims 1 to 20, which has a thermal stability of at most 190°C under atmospheric conditions.

22. An edible mycelium ingredient according to any one of claims 1 to 21, wherein the insoluble fibre content is 10 to 40% by weight, preferably 20 to 30% by weight.

23. An edible mycelium component according to any one of claims 1 to 22, wherein said mycelium component is characterized by an insoluble fibre content of up to 35% by weight.

24. An edible mycelium ingredient according to any one of claims 1 to 23, having a protein content of 30 to 65% by weight, preferably 45 to 65% by weight.

25. The edible mycelium ingredient according to any one of claims 1 to 24, wherein the ergosterol content is 4 to 7 mg / g.

26. An edible mycelium ingredient according to any one of claims 1 to 25, wherein the carbohydrate content is at most 5% by weight.

27. 27. The edible mycelium ingredient according to any one of claims 1 to 26, wherein the content of uronic acid is 0.1 to 5% by weight.

28. 28. An edible mycelium ingredient according to any one of claims 1 to 27, wherein the chitin content is in the range of 6 to 11% by weight.

29. 29. An edible mycelium ingredient according to any one of claims 1 to 28, wherein the beta-glucan content is at least 80% of the total glucan, preferably the total glucan content is in the range of 10-35% by weight, preferably 10-20% by weight.

30. 30. An edible mycelium component according to any one of claims 1 to 29, wherein when freeze-dried, for pore diameters of less than 1 mm, about 15-25%, preferably about 20%, of the pore volume of said edible mycelium component corresponds to pore diameters of 1000-20 μm, and about 75-85%, preferably about 80% of the pore volume corresponds to pore diameters of 20-2 μm, with the most frequent pore diameter peak being equal to 5.5 μm.

31. 31. An edible mycelium ingredient according to any one of claims 1 to 30, wherein the mycelium has a shear strength of at least 10 N and a water retention in the range of 20-90% and a water release of 25-70%.

32. comprising at least one fungal strain selected from Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Pezizomycetes, Agaricomycetes, Sordariomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, Boletaceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinaceae, Preferably, the at least one fungal strain is selected from the group consisting of Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Agarcomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polypolares, Russulares, Auriculares, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, Boleta ceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinacea.

33. 33. The edible mycelium ingredient of claim 32, wherein the mycelium is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus florida, Pleurotus citrinopileatus, Pleurotus salmoneostramineus, Morchella esculenta, Morchella angusticeps, Morchella deliciosa, and / or Morchella rufobrunnea.

34. 34. The edible mycelium ingredient of claim 33, wherein the mycelium is obtained from Pleurotus pulmonarius and / or Morchella rufobrunnea.

35. 32. The edible mycelium ingredient according to any one of claims 1 to 31, wherein the mycelium is obtained from L. sulphureus or B. adusta.

36. 36. A method for producing an edible mycelial ingredient according to any one of claims 1 to 35 by submerged fermentation, comprising culturing at least one fungal species in a fermentation medium, wherein the fermentation medium provided at the start of fermentation comprises 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins, the medium comprising a brewer's spent grains C5 sugar extract, the carbon to nitrogen ratio in the medium being in the range of 10 to 25, and at least 35% by weight of the extracted brewer's spent grains being characterized by a particle size of 2 to 4 mm.

37. 37. The method of claim 36, wherein the brewer's grain C5 sugar extract is the sole carbon source.

38. 38. The method of claim 36 or 37, wherein the carbon to nitrogen ratio in the fermentation medium ranges from 2 to 18.

39. 39. The method according to any one of claims 36 to 38, wherein the particle size distribution is determined by sieving, said sieving being carried out in an automatic sieving tower, i.e. by air jet sieving according to DIN 10765 2016-07, after removing particles of more than 4 mm using a vibration sieving method.

40. 40. The method of any one of claims 36 to 39, wherein the carbon to nitrogen ratio of the brewer's spent grain C5 sugar extract is in the range of 5 to 25, preferably 5 to 18.

41. 41. The method according to any one of claims 36 to 40, wherein the protein content of the brewer's grain C5 sugar extract is in the range of 7 to 30 g / l protein.

42. the at least one fungal species is selected from Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Pezizomycetes, Agaricomycetes, Sordariomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polyporales, Russulales, Auriculariales, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, Boletaceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinaceae, Preferably, the at least one fungal strain is selected from the group consisting of Basidiomycota, Ascomycota, Pezizomycotina, Agaromycotina, Agarcomycetes, Pezizales, Boletales, Cantharellales, Agaricales, Polypolares, Russulares, Auriculares, Hypocreales, Morchellaceae, Tuberaceae, Pleurotaceae, Agaricaceae, Marasmiaceae, Cantharellaceae, Hydnaceae, Bole 42. The method of any one of claims 36 to 41, wherein the phylum is selected from the group consisting of: Taceae, Meripilaceae, Polyporaceae, Strophariaceae, Lyophyllaceae, Tricholomataceae, Omphalotaceae, Physalacriaceae, Schizophyllaceae, Sclerodermataceae, Ganodermataceae, Sparassidaceae, Hericiaceae, Bondarzewiaceae, Cordycipitaceae, Auriculariaceae, and Fistulinacea.

43. 43. The method of claim 42, wherein the mycelium is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus florida, Pleurotus citrinopileatus, Pleurotus salmoneostramineus, Morchella esculenta, Morchella angusticeps, Morchella deliciosa, and / or Morchella rufobrunnea.

44. 44. The method of claim 43, wherein the mycelial mass is obtained from Pleurotus pulmonarius and / or Morchella rufobrunnea.

45. 42. The method of any one of claims 36 to 41, wherein the mycelium is obtained from L. sulphureus or B. adusta.

46. 46. ​​The method of any one of claims 42 to 45, wherein the at least one fungal species is combined with another edible fungus, algae, bacteria, plant cells, archaeal cells, animal cells, fat cells, or combinations thereof.

47. 47. The method of any one of claims 36 to 46, further comprising the step of recovering a supernatant from the culture medium or a portion thereof.

48. 48. The method of claim 47, wherein said recovering comprises crystallizing or precipitating the obtained supernatant.

49. 49. The method of any one of claims 36 to 48, wherein the pH inside the fermenter is maintained at 4.0 to 5.

0.

50. 50. The method according to any one of claims 36 to 49, wherein upon separation of the biomass, the separated biomass is washed with water having a pH of 3 to 7, preferably 3 to 6.

51. An edible mycelium ingredient obtainable by the method according to any one of claims 36 to 50.

52. 52. A method for producing a soft or hard meat substitute composition comprising the mycelium component of any one of claims 1 to 35 or 51, the method comprising preparing the meat analogue composition by adding the mycelium component to a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and optionally at least one compositional component.

53. 53. An edible product obtainable by the method of claim 52.

54. 54. An edible product according to claim 53, wherein the product is characterized by a soft texture, preferably having a hardness of 10-55N, a springiness of 35-85%, a cohesiveness of 15-70%, a viscosity of 1-40N, a chewiness of 0.3-35N, a cutting strength of 1-25N and a stickiness of 0 N.s to -0.3 N.s, and / or wherein the product is characterized by a non-soft or harder texture, preferably having a hardness of 30-100N, a springiness of 20-70%, a cohesiveness of 20-85%, a viscosity of 6-85N, a chewiness of 1-60N and a cutting strength of 1-50N.

55. 52. A method for producing a milk substitute composition comprising a mycelium component according to any one of claims 1 to 35 or 51, the method comprising the steps of preparing the milk analogue composition by: (1) forming a slurry comprising the mycelium component together with a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component, and at least one composition component; and (2) mixing the slurry with at least one composition component, specifically a texturizing or thickening agent, or a carbohydrate-rich component.

56. 56. An edible product obtainable by the method of claim 55.

57. 57. The edible product of claim 56, wherein the food product is characterized by a soft texture and / or has a hardness of 10-55 N, a springiness of 35-85%, a cohesiveness of 15-70%, a viscosity of 1-40 N, a chewiness of 0.3-35 N, a cutting strength of 1-25 N, and a stickiness of 0 N.s to -0.3 N.s, and / or wherein the food product is characterized by a non-soft or harder texture and / or has a hardness of 30-100 N, a springiness of 20-70%, a cohesiveness of 20-85%, a viscosity of 6-85 N, a chewiness of 1-60 N, a cutting strength of 1-50 N.

58. 58. An edible product according to claim 56 or 57, wherein the product is characterised by a soft texture and / or has a firmness of 1 to 20 N, a spreadability of 30 to 100 N.s, a stickiness of -1 to -414 N and a puncture force of 1 to 30 N.s, and / or wherein the product is characterised by a non-soft or harder texture, preferably having a firmness of 20 to 100 N, a spreadability of 1 to 20 N.s, a stickiness of -15 to -100 N and a puncture force of 40 to 100 N.s.

59. 1. An edible mycelial ingredient comprising undifferentiated mycelial biomass, said undifferentiated mycelial biomass having an elemental composition of mycelium with a C:N ratio in the range of 8-12, said edible mycelial ingredient being characterized by an EUC of less than 200g MSG / 100g, wherein the EUC is: [Equation 2] is defined as wherein the EUC is expressed in g MSG / 100g, ai is the concentration of each umami amino acid, Asp or Glu (g / 100g), aj is the concentration of each umami 5'-nucleotide, 5'-IMP, 5'-GMP, or 5'-AMP (g / 100g), bi is the relative umami concentration (RUC) of each umami amino acid relative to MSG, defined as 1 for Glu and as 0.077 for Asp, and bj is the RUC of each umami 5'-nucleotide, defined as 1 for 5'-IMP, as 2.3 for 5'-GMP, and as 0.18 for 5'-AMP.

60. 60. The edible ingredient of claim 59, wherein the ingredient is characterized by less than 100g MSG / 100g EUC.

61. 61. An edible ingredient according to claim 59 or 60, wherein the ingredient comprises a Pleurotus fungus, preferably Pleurotus pulmonarius, or the ingredient comprises Morchella rufobrunnea, or the ingredient comprises L. sulphureus, or the ingredient comprises B. adusta.

62. 62. The edible ingredient of any one of claims 59 to 61, wherein the mycelium has an ergothioneine content in the range of 110 to 150 mg / kg.

63. 63. The edible ingredient of any one of claims 59 to 62, having a content of umami 5'-nucleotides (5'NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) of up to 3 g / kg.

64. 64. The edible ingredient of any one of claims 59 to 63, wherein the amount of BCAAs and the amount of umami amino acids are each in the range of 20 to 80 mg / g, respectively.

65. 65. An edible ingredient according to any one of claims 59 to 64 having an insoluble fibre content of 40 to 60% by weight, preferably 40 to 50% by weight.

66. 66. An edible ingredient according to any one of claims 59 to 65 having a protein content of 30 to 50% by weight, preferably 30 to 40% by weight.

67. 67. The edible ingredient of any one of claims 59 to 66, wherein the total glucan content is in the range of 25 to 50% by weight.

68. 1. A method for producing an edible mycelial ingredient characterized by less than 200 g MSG / 100 g EUC by submerged fermentation, comprising culturing at least one fungal species in a fermentation medium, wherein the fermentation medium provided at the start of the fermentation is characterized by a C:N ratio ranging from 1 to 50, preferably from 5 to 50, and wherein the fermentation medium provided at the start of the fermentation contains 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins, and contains up to five amino acids including arginine, and at most four further amino acids selected from alanine, cysteine, glycine, proline, serine, tyrosine, and selenocysteine, or selected from phenylalanine, valine, tryptophan, threonine, isoleucine, methionine, histidine, leucine, and lysine.

69. 69. The method of claim 68, wherein the medium comprises arginine as the only amino acid.

70. 70. The method of claim 68 or 69, wherein the C:N ratio is from 10 to 25, more preferably from 16 to 23.

71. 70. The method of claim 68 or 69, wherein the C:N ratio is from 2 to 22, preferably from 15 to 22, more preferably from 20 to 25, even more preferably from 20 to 22.

72. 72. The method of any one of claims 68 to 71, wherein the EUC is less than 100g MSG / 100g, preferably less than 50g MSG / 100g.

73. 1. An edible mycelial ingredient comprising Pleurotus pulmonarius mycelial biomass, wherein the Pleurotus pulmonarius mycelial biomass has an elemental composition of mycelium with a C:N ratio in the range of 6-8, the edible mycelial ingredient being characterized by 200-500g MSG / 100g EUC, wherein the EUC is: [Equation 3] is defined as wherein the EUC is expressed in g MSG / 100g, ai is the concentration of each umami amino acid, Asp or Glu (g / 100g), aj is the concentration of each umami 5'-nucleotide, 5'-IMP, 5'-GMP, or 5'-AMP (g / 100g), bi is the relative umami concentration (RUC) of each umami amino acid relative to MSG, defined as 1 for Glu and as 0.077 for Asp, and bj is the RUC of each umami 5'-nucleotide, defined as 1 for 5'-IMP, as 2.3 for 5'-GMP, and as 0.18 for 5'-AMP.

74. 74. The edible mycelium ingredient of claim 73, wherein the indigenous RNA level is less than 1.88% by weight on a dry basis without further process steps to reduce the RNA.

75. 75. An edible mycelium ingredient according to claim 73 or 74, wherein the mycelium has an ergothioneine content in the range of 70 to 100 mg / kg.

76. 76. The edible mycelium ingredient according to any one of claims 73 to 75, wherein the content of umami 5' nucleotides (5' NMPs) containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) is up to 6g / kg.

77. 77. The edible mycelium ingredient according to any one of claims 73 to 76, wherein the amount of BCAA and the amount of umami amino acids are each in the range of 40 to 100 mg / g, respectively.

78. 78. An edible mycelium ingredient according to any one of claims 73 to 77, having an insoluble fibre content of 25 to 45% by weight, or an insoluble fibre content of 30 to 60% by weight.

79. An edible mycelium ingredient according to any one of claims 73 to 78, having a protein content of 30 to 50% by weight, preferably 30 to 40% by weight.

80. 80. An edible mycelium ingredient according to any one of claims 73 to 79, wherein the total glucan content is in the range of 20 to 35% by weight.

81. 80. A method for producing an edible mycelial component according to any one of claims 73 to 79 by submerged fermentation, comprising culturing at least one fungal species in a fermentation medium, wherein the fermentation medium provided at the start of fermentation comprises 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins, the carbon to nitrogen ratio in the medium being in the range of 10 to 25, the pH inside the fermenter being maintained at 4.0 to 5.0, and upon separation of the biomass, the separated biomass is washed with water having a pH of 3 to 7, preferably 3 to 6.

82. 82. The method of claim 81, wherein the carbon to nitrogen ratio in the medium is in the range of 16 to 18, preferably 16.5 to 17.

5.

83. 1. An edible product, preferably a meat substitute product or a dairy substitute product, comprising a mycelium component in the range of 1-99% by weight, said mycelium component being characterized by an insoluble fiber content of 20-60% by weight and an EUC of 200-500g MSG / 100g, wherein the EUC is: [Equation 4] is defined as wherein the EUC is expressed in g MSG / 100g, ai is the concentration (g / 100g) of each umami amino acid, Asp or Glu, aj is the concentration (g / 100g) of each umami 5'-nucleotide, 5'-IMP, 5'-GMP, or 5'-AMP, bi is the relative umami concentration (RUC) of each umami amino acid relative to MSG, defined as 1 for Glu and 0.077 for Asp, and bj is the RUC of each umami 5'-nucleotide, defined as 1 for 5'-IMP, 2.3 for 5'-GMP, and 0.18 for 5'-AMP.

84. 84. The edible product of claim 83, wherein the mycelium component is characterized by about 300g MSG / 100g EUC.

85. 85. An edible product according to claim 83 or 84, wherein the mycelium component is characterized by an insoluble fibre content of 30-40% by weight.

86. 1. An edible product, preferably a meat substitute product or a dairy substitute product, comprising a mycelium component in the range of 1-99% by weight, said mycelium component being characterized by an insoluble fiber content of 20-60% by weight and an EUC of less than 200g MSG / 100g, wherein the EUC is: [Equation 5] is defined as wherein the EUC is expressed in g MSG / 100g, ai is the concentration (g / 100g) of each umami amino acid, Asp or Glu, aj is the concentration (g / 100g) of each umami 5'-nucleotide, 5'-IMP, 5'-GMP, or 5'-AMP, bi is the relative umami concentration (RUC) of each umami amino acid relative to MSG, defined as 1 for Glu and 0.077 for Asp, and bj is the RUC of each umami 5'-nucleotide, defined as 1 for 5'-IMP, 2.3 for 5'-GMP, and 0.18 for 5'-AMP.

87. 87. The edible product of claim 86, wherein the mycelium component is characterized by an EUC of less than 100g MSG / 100g, preferably less than 50g MSG / 100g.

88. 88. An edible product according to claim 86 or 87, wherein the mycelium component is characterized by an insoluble fibre content of 30-60% by weight, preferably 40-60% by weight.

89. 89. The edible product of any one of claims 83 to 88, wherein the ingredient comprises a Pleurotus fungus, preferably Pleurotus pulmonarius.

90. 89. The edible product of any one of claims 83 to 88, wherein the ingredients include Morchella rufobrunnea.

91. 89. The edible product of any one of claims 83 to 88, wherein the ingredient comprises L. sulphureus.

92. 89. The edible product of any one of claims 83 to 88, wherein the ingredient comprises B. adusta.

93. 1. An edible mycelial ingredient comprising Pleurotus pulmonarius mycelial biomass, wherein the Pleurotus pulmonarius mycelial biomass has an elemental composition of mycelium with a C:N ratio ranging from 2 to 12, and wherein the edible mycelial ingredient is characterized by an equivalent umami concentration (EUC) of at least 30 g monosodium glutamate MSG / 100 g, wherein the EUC is: [Equation 6] is defined as wherein the EUC is expressed in g MSG / 100g, ai is the concentration of each umami amino acid, Asp or Glu (g / 100g), aj is the concentration of each umami 5'-nucleotide, 5'-IMP, 5'-GMP, or 5'-AMP (g / 100g), bi is the relative umami concentration (RUC) of each umami amino acid relative to MSG, defined as 1 for Glu and as 0.077 for Asp, and bj is the RUC of each umami 5'-nucleotide, defined as 1 for 5'-IMP, as 2.3 for 5'-GMP, and as 0.18 for 5'-AMP.

94. 94. The edible mycelium ingredient of claim 93, wherein the insoluble fiber content is 20 to 60% by weight.

95. 95. An edible mycelium ingredient according to claim 93 or 94, having a protein content of 10 to 65% by weight.

96. 96. An edible mycelium ingredient according to any one of claims 93 to 95, wherein the indigenous RNA level is at most 2% by weight on a dry basis without further process steps to reduce the RNA.

97. 97. A method for producing an edible mycelial component according to any one of claims 93 to 96 by submerged fermentation, comprising the step of culturing at least one fungal strain in a fermentation medium, wherein said fermentation medium provided at the start of fermentation is characterized by a C:N ratio ranging from 1 to 50, preferably from 5 to 50, and wherein said fermentation medium provided at the start of fermentation comprises 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01 to 15 g / L of minerals, and 0.01 to 50 mg / L of vitamins, and wherein said medium comprises arginine and glutamate as the only amino acids.

98. 98. The method of claim 97, wherein the medium comprises glutamate as the only amino acid.

99. 99. An edible product, preferably a meat substitute or dairy substitute product, comprising an edible mycelium component according to claims 93-98 in the range of 1-99% by weight.

100. 100. The edible product of any one of claims 93 to 99, wherein the at least one fungal strain comprises a Pleurotus fungus, preferably Pleurotus pulmonarius, or the at least one fungal strain comprises Morchella rufobrunnea, or the at least one fungal strain comprises L. sulphureus, or the at least one fungal strain comprises B. adusta.

101. 36. An edible product, preferably a meat or dairy substitute product, comprising an edible mycelium component according to any one of claims 1 to 35 in the range of 1 to 99% by weight, said mycelium component being characterized by an insoluble fibre content of 20 to 60% by weight.

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