Fungus ingredients and derivative products

Abstract

The present invention relates to a mycelium component derived from submerged fermentation of at least one fungal strain in three different media: a defined component medium producing component A, a synthetic medium producing component B, or a composite natural medium producing component C comprising a sidestream extract derived from an agricultural food sidestream. This unique mycelium composition is characterized by chemical, biological, physical, morphological, nutritional and organoleptic properties. The three different fibrous mycelia of the edible fungi obtained from at least one fungus strain are further used to produce characteristic food products, including meat analogs, fish analogs, dairy product analogs, beverages, or other food products. The three new mycelium components are useful in the manufacture of foods, foods, beverages, pharmaceuticals, cosmetics, nutraceuticals, biomaterials, and feed and industrial applications.

Description

Technical Field

[0001] The present invention relates to mycelial components derived from submerged fermentation of at least one fungal strain in three different culture media: a defined medium producing component A, a synthetic medium producing component B, or a complex natural medium comprising a lateral stream extract selected from an agricultural food lateral stream producing component C. This unique mycelial composition is characterized by chemical, biological, physical, morphological, nutritional and organoleptic characteristics. The three different fibrous mycelia of edible fungi obtained from at least one fungal strain are further used to produce characteristic foods, including meat analogs, fish analogs, dairy analogs, beverages or other foods. These three new mycelial components can be used to manufacture foods, foods, beverages, pharmaceuticals, cosmetics, nutritional products, biomaterials and feed and industrial applications. Background Art

[0002] Over the past 10 years, several food Scandinavians 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 identified as the main driver of an outbreak of Enteroaggregative Escherichia coli in budding bacteria that resulted in several deaths in Europe and related product recalls around the world. Later, in 2013, large amounts of horse meat were found in products advertised as beef in Europe, with potential health concerns associated with contamination with phenylbutazone, a common analgesic for horses. That same year, Muslim and Jewish groups were affected when pork was found in beef products. In 2017, eggs contaminated with Finopril, a common insecticide, were found in several European and Asian countries. In all of these cases, identifying the source of the contamination and the subsequent coordination of the removal of the spoiled food is hampered by the complexity of current global supply and distribution channels, thus leaving consumers at risk for a long time.

[0003] At a different level, the COVID-19 pandemic has also clearly highlighted that the global food system does not provide the required level of resilience with regard to food security. The first year of the pandemic led to an increase in global food prices of approximately 20%, and the World Food Program (WFP) estimates that the number of people suffering from acute food insecurity increased from 135 to 272 million per year (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 reviewed on October 17, 2022). Similarly, the food system is vulnerable to global disasters.

[0004] At the same time, one third of all food produced worldwide is either lost before it reaches the consumer or wasted afterwards. This represents approximately 1.3bn tonnes of food, much of which could potentially be recovered with optimal production and distribution logistics. Increased consumer awareness has led to the emergence of new purchasing trends, where local, natural, healthy and sustainable products are preferred over ultra-processed, unbalanced foods. Modern consumers expect to be able to track the origin of what they buy and understand the ingredients on the packaging, as well as the impact of the product on their health and our planet.

[0005] Environmental awareness has increased in recent years as the unsustainability and (in some cases) difficulties in industrial production methods and practices for meat and fish have been highlighted by several published studies cited in this article (https: / / doi.org / 10.3390 / foods9091227 and https: / / doi.org / 10.3390 / foods9091151). Although plant-based alternatives can significantly reduce CO2 emissions compared to conventional meat production, 2 Emissions and improve animal welfare, but they do not fully address the challenges of local production, as these products are mainly derived from three monocrop cultivations (soybeans, peas and rice) that grow only in a few countries and need to be exported worldwide. In addition, their cultivation requires large land areas that are unfortunately often obtained through logging, and their effective production still relies heavily on chemical agents, such as pesticides and fertilizers, which contribute to soil and water pollution and have a lasting impact on biodiversity. In addition, only concentrates and isolates from crops are used in the production of meat substitutes, so a large amount of waste is generated in this method. Finally, these plant proteins have a strong bitter taste and no inherent texture. Therefore, their application in food requires further processing steps and a large amount of ingredients. Therefore, plant-based substitutes cannot solve all consumer issues about food traceability and sustainability.

[0006] While plant-based meat alternatives are being developed, some traditional foods, such as mushrooms, are also receiving increasing attention 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 taste, and certain variations between species enable the production of products with taste characteristics close to meat or other savory foods without adding large amounts of ingredients. In addition, their fruiting bodies or fruit caps also have a meat-like texture and can be further improved with minimal processing for specific applications. Nutritionally, mushrooms contain up to 40% complete protein, but also contain prebiotic fiber that is often lacking in the Western diet. Mushrooms also contain large amounts of key minerals, such as iron, zinc, calcium, potassium or magnesium, as well as B vitamins; they can be considered one of the rare foods that provide a complete and balanced nutritional profile.

[0007] The cultivation of mushrooms is a long process, although it is highly environmentally friendly. In nature, mushrooms are equipped with a variety of unique enzymes that enable them to remove waste materials present in forest soils, such as fallen leaves or wood residues. They can degrade complex plant compounds, which are otherwise nutrients that are usually unavailable to other biological groups. This feature makes mushrooms an ideal candidate for waste from the agricultural food industry, which is usually highly unstable and is usually used as feed, burned or simply discarded, although they have residual nutrients (for example, straw or rice husks, etc.). To date, only about 144,000 fungal species have been described, and it is estimated that there are more than 10 million species on the earth, including a wide range of unknown edible mushroom species. Many of these undiscovered species may provide new ways of experiencing new experiences and opportunities for the renewal of waste materials. Despite their attractive attributes as food, mushrooms are relatively slow-growing, and the production cycle usually takes at least 6 weeks. In addition, traditional production methods are very basic and use difficult-to-scale technologies, such as growing on forest trees and / or in bags containing lignocellulose materials. In recent years, more modern techniques involving the use of culture chambers or hydroponics in which temperature and humidity are strictly controlled have enabled significant improvements in process standardization and yield, but they require large investments and do not fully address the scalability issue.

[0008] The use of fermentation to produce mushroom mycelium in this paper presents advantages in terms of sustainability, food safety, and traceability. Fermenters are sterile containers that operate under controlled conditions; therefore, the risk of spoilage is minimized. They can be scaled vertically, therefore allowing for a smaller factory footprint and the potential to produce food locally using by-products from farms or food processors, eliminating the need for long supply or distribution chains. Furthermore, producing mushroom mycelium in fermenters is also more sustainable than producing traditional plant-based alternatives. Water consumption, land area requirements, energy consumption, CO 2 Emissions are estimated to be lower than traditional meat alternatives derived from soy.

[0009] Therefore, in order to avoid the above problems and limitations, three new mushroom mycelium components or fungal biomass components have been developed in liquid culture medium by liquid or submerged fermentation, wherein the taste, composition (carbohydrate, fat, protein, nutrients, vitamins, fiber content, amino acids, etc.), texture and structure are controlled during the fermentation process according to the fermentation conditions used (e.g., medium used, species, process configuration and conditions), resulting in unique biological, physical and chemical properties for each mycelium component. In addition, when cooked, the biomass retains a typical umami flavor, which can also be controlled at the fermentation level, so that food products prepared with the mycelium meat or components developed and disclosed in the present invention, such as meat analogs and dairy analogs or other food products developed in the present invention, require minimal processing and a very short list of ingredients that can be easily communicated to the consumer. Summary of the invention

[0010] The present invention solves the above problems by introducing three new raw materials or ingredients into the production of meat, fish and dairy analogs or other foods, namely the mycelium of edible mushrooms. These three new ingredients can also be used to make food, food, beverages, medicines, cosmetics, nutritional products, biomaterials and feed and industrial applications. Mycelium has been widely used in meat substitute products due to its filamentous structure (GB2137226A) and as a fat-simulating substance in dairy drinks or yogurt (WO2002090527A1).

[0011] CN103184246A discloses a preparation method of thioneine, which utilizes the liquid culture medium of wild oyster mushroom, pleurotus eryngii or flower face oyster mushroom to produce thioneine with the low yield of 51mg / L, and the incubation time is 10 days.For example, CN110283856A discloses the method for producing thioneine by the fungal strain oyster mushroom of fermentation 3210, and the productive rate is 300mg / L, but the method grows 15 days on PD in mycelium, and then needs at least 25 days between the fermented glutinous rice days 10 days.It is observed in the patent literature that co-fermentation can produce thioneine (CN112195215 or CN114214387) of higher content more than one fungal strain.Finally, CN109939027A discloses a method for producing thioneine by fermenting Hericium erinaceus with glucose and peptone, and the productive rate is 331mg / L, but the production cost of substrate is high, and the process needs a long time (about 25 days).

[0012] CN212786880 reported that the fiber content of Pleurotus pulmonarius fruiting bodies was low, thus supporting the review published in 2021 (Fungal Biotec 1(2):65–87(2021)), which outlined that the fiber content of Pleurotus spp. fruiting bodies ranged from 2.97 wt% to a maximum of 31 wt%, specifically that Pleurotus pulmonarius fruiting bodies contained 4 wt% to 9 wt% fiber based on dry weight. CN105054261 also disclosed the following findings: When Pleurotus pulmonarius was mixed with other strains, crude fiber was degraded, specifically on oyster mushrooms (Pleurotus spp.), thereby reducing the fiber content by 2.3 wt% to 20.25 wt%. Another study in 2020 (Int J Med Mushrooms. 2020; 22 (7): 651-657. doi: 10.1615 / Int J Med Mushrooms. 2020035449) also revealed that the mycelium of oyster mushrooms (Pleurotus pulmonaria) contains 22% by weight of insoluble fiber. Document US2020 / 270559 discloses a specific method for producing an edible filamentous fungal biomaterial preparation and shows nutritional data from two Fusarium filamentous fungi with a total fiber content of up to 25% by weight and a fat content between 7% and 12% by weight.

[0013] On the other hand, the RNA level of currently available mycelial components is typically actively reduced by a process involving a final processing step, which includes 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% (based on dry matter), thereby reducing undesirable associated health risks and bitterness and meeting regulatory requirements, as disclosed in the methods developed in these publications US4041189, US4501765, WO201802579.

[0014] Jeng-Leun Mau (2015) summarized the equivalent umami concentration (EUC) values ​​of fruiting bodies and showed that it varied widely and ranged from a maximum value of 4465% (Volvariella volvaacea fl at the crown) to a minimum value of 0.12% (Auricularia polytricha). EUC values ​​were divided 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 (International Journal of Medicinal Mushroom, Vol. 7, pp. 119-125 (2005)). The EUC value of Pleurotus ostreatus is the highest (the second level is 511%), followed by the descending order of Pleurotus eryngii small fruiting bodies (97.9%), Pleurotus vesicularis (85.2%), and Annona (48.0%). Based on the form of fruiting bodies in culture bottles or plastic bags (logs), the EUC values ​​of Pleurotus eryngii are 68.7, 97.9, and 32.1% for large fruiting bodies and small fruiting bodies, respectively. In a separate study, Pleurotus pulmonary fruiting bodies were grown on three types of forestry waste (pine, poplar, and honeysuckle), showing that the EUC values ​​of Pleurotus pulmonary fruiting bodies were between 72.31% and 116.73% (Food Chemistry 397 (2022) 133714). It should be understood that EUC is preferably expressed in 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 based on patented inventions (eg US2693664) to enhance the taste of mycelium derived from submerged fermentation (J. Sci. Food Agric. 91958).

[0016] WO2022 / 107388 discloses a method for providing an umami-enhanced composition of Pyrola mushrooms by enzyme treatment, achieving a reported maximum equivalent umami concentration (EUC) value of 9.3 g / 100 g.

[0017] CN114027089 discloses a method for improving the flavor of edible mushrooms, wherein the equivalent umami concentration value of Pleurotus eryngii obtained by adding fermented broth of edible fungal roots increases EUC from 4.48 g / 100 g to 10.8 g MSG / 100 g.

[0018] CN109156702 A discloses a soaking method for Hericium erinaceus, wherein the original EUC is reduced from 1131 g MSG / 100 g to the following values ​​by various treatments, namely 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). Similar studies were reported in 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, resulting in different EUC concentrations of the fruiting bodies.

[0019] KR101535985 relates to a method comprising the steps of inducing a Maillard reaction between powder of one or more mushrooms selected from the group consisting of shiitake mushrooms, shiitake (Linguine), (Oyster) and Enoki mushrooms, with other flavoring ingredients to achieve an EUC value of 176 mg MSG / 100 g. Note that the measurements performed relate to the fruiting bodies of the fungi in question, not their mycelium.

[0020] Provided herein are three novel mycelial ingredients, A, B and C, which are chemically, biologically, physically, nutritionally and sensory unique (ie, 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 culture media: a defined medium producing component A, a synthetic medium producing component B, or a natural medium comprising a lateral stream extract selected from an agricultural food lateral stream producing component C.

[0022] In one embodiment of the present invention, mycelia are provided with different carbon to nitrogen ratios, similar chitin content, but with different sugar contents (sugars, polysaccharides, oligosaccharides).

[0023] In another embodiment of the present invention, mycelium is provided with low intrinsic RNA levels of at most 4 wt%, preferably at most 2 wt%, which avoids the need for additional treatments to actively reduce RNA levels.

[0024] In another specific embodiment of the present invention, the mycelium provided has an ergothioneine content of at most 800 mg / kg. Compared with the time reported in the prior art, the ergothioneine content can be achieved in a shorter time.

[0025] In another embodiment of the present invention, the provided mycelium has a specific / unique pore volume, pore size distribution and / or characteristic structure.

[0026] In another specific embodiment of the present invention, at least 20 wt. %, at least 30 wt. %, at least 40 wt. %, at least 50 wt. % or at least 60 wt. % in terms of its adjustable insoluble fiber content provides even higher health value for its high prebiotic insoluble fiber content which is essential for intestinal health.

[0027] In another specific embodiment of the present invention, with respect to its amino acid content, the amount of branched chain amino acids (BCAA) is at least about 19 wt%, at least about 20 wt% of the total amount of amino acids present (i.e., of the total protein).

[0028] In another embodiment of the present invention, the protein content thereof can be adjusted in the range of 10 wt % to 65 wt %.

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

[0030] In another specific embodiment, mycelium from at least one fungal strain is mixed with at least one protein-rich ingredient, at least one lipid-rich ingredient, and at least one constituent ingredient to produce a meat or dairy analog.

[0031] In another specific embodiment, the mycelium from at least one fungal strain has a very high equivalent umami concentration (EUC) up to at least 34%, more preferably at least 300%, even more preferably at least 2800%, which is about 24 to 40 times higher than mushroom fruiting bodies of the same species, and higher than the reported EUC values ​​of mycelium as discussed in the present invention.

[0032] The flavor can also be significantly improved, since the mycelium can bring a natural umami flavor typical of cheese without the use of additional flavoring agents, especially in the case where the mycelium is derived from a fruiting body-forming fungus, for example from the family Pleurotus, such as a fungus selected from the group consisting of Pleurotus pulmonatus, Pleurotus ostreatus, Pleurotus florida and Pleurotus rosa, and in particular from Pleurotus pulmonatus; or from a fungus selected from the group consisting of Morchella esculenta, Morchella angusticeps, Morchella deliciosa and Morchella rufibrunnea, preferably Morchella rufibrunnea. This is due to the presence of a large amount of glutamate in such species together with other amino acids (e.g. aspartic acid) and / or 5'-nucleotides, which largely play a role in transmitting the umami sensation.

[0033] In another specific 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 yet another specific embodiment, the present invention relates to a method 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 yet another specific embodiment, the present invention relates to a method for producing fungal biomass by submerged fermentation of at least one fungal strain, wherein more than one fungal strain is co-fermented.

[0036] In a particular embodiment, the present invention relates to fungal biomass produced according to the method for producing fungal biomass by submerged fermentation of at least one fungal strain of the present invention, wherein the fungal strain is selected from the family Pleurotus ostreatus, in particular wherein the fungal strain is Pleurotus pulmonaria, Pleurotus ostreatus, Pleurotus aurantii or Pleurotus roxburghii.

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

[0038] In an alternative embodiment, the edible fibrous mycelium is obtained from at least one fungal strain that preferably produces ergothioneine. The fungal strain is preferably selected from the group consisting of Basidiomycota, Ascomycota, Hymenochaetaceae, Agaricomycetes, Sordariomycetes, Tremellomycetes, wherein preferred species herein are from at least one fungal species selected from the group consisting of Cordyceps spp., Inonotus spp., Grifola spp., Pleurotus spp., Ganoderma spp., Lentinula spp., Treella spp., Trametes spp., Lepista spp., Trichooma spp., Aspergillus spp. and / or all fungal species.

[0039] The present invention also relates to methods of producing the above-mentioned edible non-animal dairy alternative products comprising the edible fibrous mycelial masses of the present invention. These methods are described in detail below.

[0040] The present invention also relates to the use of edible fibrous mycelium masses in the production of edible meat substitute products, wherein the edible meat substitute products are selected from the group consisting of products replacing meatballs, sausages, tartar, minced meat, meat pate, processed meat, Mett meat, goose liver, steak, beef jerky, hamburger patties, fish fillets, kernels, salami, whole slices, bacon, hot dogs, raw ham, dried meat and extruded products.

[0041] The present invention also relates to the use of edible fibrous mycelium masses for producing edible non-animal dairy alternative products, wherein the edible non-animal dairy alternative products are selected from products replacing milk, yogurt, fresh cheese, whey cheese, cream cheese, medium-hard cheese, hard cheese and soft cheese.

[0042] The present invention also relates to the use of edible fibrous mycelium for producing edible fish analogs or seafood products, such as crab cakes, kamaboko, tuna, salmon or shrimp.

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

[0044] In another embodiment, the present invention also relates to the use of the supernatant produced during the fermentation process in the development of a specific healthy beverage containing antioxidants and a specific aroma, taste and flavor produced during the fermentation process with edible mushrooms. For example, mushroom strains are known to produce pleasant volatiles and other compounds with apple or almond taste. They also produce compounds known to regulate blood sugar levels.

[0045] The invention also relates to the use of the supernatant to be further processed, for example for extracting specific components thereof, such as proteins, enzymes, polysaccharides, peptides, antioxidants etc. produced in particular by the microorganisms cultivated in the culture medium. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 Shown is the overlay of the pore size distribution curves (-dV / dlogD) of mycelial fractions A, B and C calculated from the normalized volume curves.

[0047] Figure 2 A thermogravimetric analysis curve of component A is shown, which shows the effect of temperature on sample weight. The weight is expressed as the percentage of the sample retained at a given temperature / time relative to the weight at the beginning of the experiment. The second y-axis on the graph represents data for the first derivative of the TGA curve. This is called a derivative thermogravimetric (DTG) curve and represents the rate of change of mass relative to temperature (e.g., % mass loss per degree Celsius).

[0048] Figure 3 A thermogravimetric analysis curve of component B is shown, which shows the effect of temperature on the weight of the sample. The weight is expressed as the percentage of the sample retained at a given temperature / time relative to the weight at the beginning of the experiment. The second y-axis on the graph represents data for the first derivative of the TGA curve. This is called a derivative thermogravimetric (DTG) curve and represents the rate of change of mass relative to temperature (e.g., % mass loss per degree Celsius).

[0049] Figure 4 A thermogravimetric analysis curve of component C is shown, which shows the effect of temperature on the weight of the sample. The weight is expressed as the percentage of the sample retained at a given temperature / time relative to the weight at the beginning of the experiment. The second y-axis on the graph represents data for the first derivative of the TGA curve. This is called a derivative thermogravimetric (DTG) curve and represents the rate of change of mass relative to temperature (e.g., % mass loss per degree Celsius).

[0050] Figure 5 Beef, chicken, real peas and soybeans are shown for comparative experiments. DETAILED DESCRIPTION

[0051] The present invention will be described in detail below. It should be understood that all disclosed features can be combined with each other, unless explicitly stated otherwise. In particular, the features disclosed in different embodiments can be combined with each other, unless explicitly stated otherwise.

[0052] The mycelial components A, B and C of the present invention have unique organoleptic and biological / physical / chemical properties.

[0053] In particular, the present invention provides an edible mycelial ingredient comprising undifferentiated mycelial biomass having an elemental composition of mycelial C:N ratio ranging from 2 to 12 (preferably 2 to 8, more preferably 2 to 6), and characterized by an EUC of at least 500 g MSG / 100 g. As understood herein, the undifferentiated mycelial biomass may be obtained, for example, in a submerged fermentation process.

[0054] This particularly preferred edible mycelium component of the present invention may also be referred to as component C.

[0055] Preferably, EUC is defined as:

[0056] EUC=∑aibi+1218(∑aibi)(∑ajbj),

[0057] where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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.

[0058] In an alternative embodiment, EUC is defined as:

[0059] EUC=∑aibi+1218(∑aibi)(∑ajbj),

[0060] where EUC of the sample is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP, 5'-XMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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, 0.61 for 5'-XMP, and 0.18 for 5'-AMP.

[0061] Thus, the present invention provides an edible mycelial ingredient characterized by a specific very high EUC value, in other words, the ingredient has a very strong umami taste profile. The ingredient of the present invention has an EUC of at least 500 g MSG / 100 g, preferably at least 1000 g MSG / 100 g, more preferably at least 1500 g MSG / 100 g, even more preferably at least 2000 g MSG / 100 g.

[0062] The composition of the present invention is also 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 does not contain 5'-IMP.

[0063] As understood herein, preferably, the organoleptic or morphological properties are selected from taste attributes, odor attributes, fragrance attributes, mouthfeel attributes, texture attributes, consistency, edibility and color. Although this list is interpreted as exemplary and preferred, it should not be interpreted as limiting. Those skilled in the art will be able to include additional one or more organoleptic or morphological properties by including additional one or more organoleptic properties or the method.

[0064] As preferably understood herein, the taste attributes and odor attributes can be determined by a tasting panel, which is composed of individuals who evaluate the taste and / or odor of the samples provided. Preferably, the tasting and olfactory panel is performed in parallel on several samples and includes certain reference samples for standardized evaluation. Therefore, in an exemplary manner of performing a tasting panel, each trained panelist is masked and receives samples continuously. They define the sensory attributes they identify in the sample, discuss these attributes together, and select common attributes that each panelist can associate with the same taste and fragrance of the sample and the reference sample to compare them. Then the second stage begins, and the panelists evaluate the meatballs according to the selected attributes and score each attribute between 0 and 5. This stage is repeated on different days to increase the statistical relevance of the data, and the average of the scores is calculated and plotted on a spider web graph.

[0065] Preferably, mouthfeel attributes and texture attributes are also determined by a specific panel.

[0066] The color referred to herein is preferably measured using an RGB system and a color analyzer at several locations on the sample, for example 20 different locations, and then the average of these measurements is used to compare the color of the meatballs. Preferably, a calibrated image capture device is used to determine the color.

[0067] 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. Although this list is interpreted as exemplary and preferred, it should not be interpreted as limiting. Those skilled in the art should be able to expand the method by including further one or more nutritional properties.

[0068] As understood herein, in the present invention, sugar content or carbohydrate content preferably refers to the %w / w or wt.% content of the biomass or product of the present invention, preferably expressed relative to the dry mass of the biomass or product. Information about sugar content can also include further details, such as the content of complex and simple carbohydrates, including the decomposition of pentoses or hexoses. As indicated herein, the content of different types of sugars / carbohydrates can also be expressed in %w / w or wt.% relative to the total sugar / carbohydrate content of the product or biomass.

[0069] As understood herein, the terms % w / w and wt. % are interchangeable unless otherwise stated.

[0070] 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 amino acid analysis methods, due to the hydrolysis conditions used in the method, it is impossible to distinguish between aspartic acid and asparagine and glutamic acid and glutamine. Therefore, the content of Asp / Asn and the content of Glu / Gln should be expressed as the total content of the two amino acids in each of these amino acid pairs.

[0071] As understood herein, umami amino acids are the aspartic acid (aspartate) and glutamic acid (glutamate) amino acids that contribute to flavor, while branched chain amino acids (BCAAs) are a group of three essential amino acids: leucine, isoleucine, and valine, which are responsible for muscle growth, athletic performance, weight loss, and fatigue reduction.

[0072] Umami or savory taste is also defined by 5-ribonucleotides or 5'NMPs, including the following 5'-nucleotides (5'NMPs): 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP), preferably expressed in g / kg mycelial component.

[0073] 5'-GMP is known to produce a meaty flavor and is a stronger flavorful seasoning than monosodium glutamate (MSG) (JHLitchfield, Morel mushroom mycelium as a food flavoring materialBiotechnology and Bioengineering, 9 (1967), pp.289-304). It has also been reported that a synergistic effect of umami 5'-nucleotides and umami amino acids may exist, thereby greatly increasing the umami taste 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) This synergistic effect of 5'-IMP or 5'-GMP on MSG was then obtained in a linear relationship, expressed as follows

[0074] EUC=A+1218(A)(N)

[0075] Wherein EUC (equivalent umami concentration) is the equivalent concentration of monosodium glutamate (MSG), which is a measure of the relative taste intensity of umami amino acids and 5'-nucleotides, because the intensity of umami amino acid taste and flavor nucleotides is shown to be proportional to the intensity of MSG relative to IMP or GMP, respectively. EUC is expressed in g MSG / 100g dry weight, and A and N are the concentrations of amino acids and nucleotides expressed in terms of the concentrations of MSG and GMP in the solution, respectively. Based on the concentration g / 100g used, 1218 is a positive constant when IMP is used. When GMP is used for standardization, the constant is 2800, and both will lead to the same results within an acceptable standard deviation (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 at an equivalent concentration of MSG.

[0076] In more detail, the calculation is further expressed as: EUC = ∑aibi + 1218 (∑aibi) (∑ajbj), where the EUC of the sample 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, 5'-AMP) (g / 100g), bi is the relative umami concentration (RUC) of each umami amino acid with MSG (Glu = 1 and Asp = 0.077), as reported in the literature; bj is the RUC of each umami 5'-nucleotide (5'-IMP = 1; 5'-GMP = 2.3; 5'-AMP = 0.18) and 1218 is the synergy constant based on the concentration used (g / 100g). EUC is a useful parameter for objectively benchmarking umami taste with a specific value.

[0077] 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 expressed as % w / w or wt.% of the dry mass of the biomass or product.

[0078] As understood herein, in the present invention, the content of metabolites refers to the amount of every kind of metabolites in biomass or product, expressed in mg / g biomass or product, respectively. For example, metabolite content can refer to any metabolite selected from those known to those skilled in the art. Usually, metabolites refer to the metabolites of fungal species as indicated herein. Exemplary metabolites include alcohols, amino acids, nucleotides, antioxidants, organic acids (e.g., acetic acid, lactic acid), polyols (e.g., glycerol) and vitamins. However, this enumeration is not meant to be interpreted as limiting.

[0079] As understood herein, in the present invention, mineral content refers to the content of any mineral considered essential for human nutrition, which can be expressed in mg / kg biomass or product for each mineral, referring to the dry mass of the biomass or the 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.

[0080] As understood herein, in the present invention, the vitamin content preferably refers to the content of a specific vitamin in μg / kg dry biomass or dry product. Vitamins are known to those skilled in the art and include vitamin A, vitamin B 12 , Vitamin B 1 , Vitamin B 3 , Vitamin B 6 , vitamin C, vitamin D, vitamin E, vitamin K and vitamin E, etc.

[0081] It should be understood that the final fungal-derived product based on the disclosed mycelial component can include the supernatant obtainable during the production process of the mycelial component of the present invention. Therefore, the final fungal-derived product based on the disclosed mycelial component can be the supernatant itself, or the biomass (i.e., the mycelial component) or a combination thereof, or any relevant extract from each individual product or a combination thereof. Therefore, it is obvious to those skilled in the art that the fungal component can be applied to at least one form of these products.

[0082] As understood herein, in the present invention, a fungal composition preferably relates to a species / strain composition of biomass or products derived therefrom (ie from the mycelial component disclosed in the present invention).

[0083] As understood herein, in the present invention, biomass dry weight is the weight of the biomass obtained after dehydration / water removal, preferably after washing away residual culture medium, preferably extrapolated to 0% w / w water content.

[0084] As understood herein, preferably whenever reference is made to any content or any ratio determined "on a dry weight basis" or "on a dry weight basis", this refers to the material after dehydration / water removal, preferably after washing away residual media, preferably extrapolated to 0% w / w water content.

[0085] As understood herein, in the present invention, the fiber content preferably refers to the % w / w content of dietary fiber in the dry biomass or dry product.

[0086] As understood herein, in the present invention, protein content preferably refers to the % w / w content of protein in dry biomass or dry product.Preferably, the measured protein content refers to the protein content determined by Kjeldahl method.

[0087] As understood herein, in the present invention, the fatty acid content preferably refers to the % w / w content of fatty acids in the dry biomass or dry product.

[0088] 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.

[0089] It is to be understood that the terms mycelial component or mushroom mycelial component or mycelial component or fungal component or mycelium / mycelial biomass are equivalent.

[0090] As understood herein, in the process of the present invention, product titer preferably refers to the concentration of the product obtained, preferably expressed in g / L. In the present context, the term product preferably refers to fungal biomass, fungal metabolites (i.e. compounds obtainable from the mycelium), colorants, nutraceuticals, active compounds, enzymes or cosmetics.

[0091] As understood herein, in the methods of the present invention, the culture or fermentation conditions include the data necessary for repeating the culture experiment, i.e., temperature, CO 2 Content / emissions / other emissions of production or volatile gases, agitation, humidity, pH, dissolved oxygen concentration, dissolved CO 2 concentrations, etc. This list is not meant to be limiting, as the culture conditions will be apparent to those skilled in the art.

[0092] As understood herein, in the method of the present invention, metabolic behavior preferably includes transcriptome information, metabolome information, proteome information, secretome information and / or fluxome information. It should be understood that the secretome preferably includes information about the structure and / or amount of compounds produced by fungal biomass and secreted outside the fungal cell, such as metabolites / proteins that can be secreted by the organism into the fermentation broth. As understood by those skilled in the art, it may include valuable compounds that can be used for products, such as vitamins, enzymes, pigments or mycelium-derived functional or active compounds. Therefore, information about the secretome can inform efforts to produce secretory compounds in the process involving fungal biomass.

[0093] Preferably, the molecular and metabolite information relates to metabolites present in or obtainable from a particular fungus. Therefore, a metabolite should preferably be understood as each node of the metabolic pathway map of the particular species under study. Information about the structure and content of a particular metabolite is preferably included herein.

[0094] As understood herein in the present invention, the fermentation medium preferably comprises at least one fungal strain, optionally other microorganisms to be co-cultivated with the fungal strain (preferably comprising algae, bacteria, plant cells, archaeal cells, animal cells, fat cells or a combination thereof), and / or a side stream from the agro-food industry.

[0095] As understood herein, chitin is a polysaccharide composed of linked N-acetylglucosamine subunits, with the chemical formula (C 8 H 13 O 5 N) n , where n is the number of subunits. It should be understood that n is a natural number. Chitin and its degradation products are sensed in the skin, lungs and digestive tract by the enzyme chitinase present in humans and other mammals, triggering an immune response with the potential to fight parasites. In addition, chitin is often used as a food additive to improve flavor and as an emulsifier. It also has anti-inflammatory properties, lowers cholesterol and is beneficial for weight loss and blood pressure. Chitin can also be used to produce biomaterials or biodegradable packaging materials, and can also be used in the food industry or other industries. It can also be a compound used as sausage casings, as chitin can be extracted from mushrooms or mycelium.

[0096] Preferably, the production of functional compounds includes information on compounds produced by mycelium. Preferably, the functional compound may also be referred to as an active compound, preferably any substance having a beneficial (proven or confirmed) effect on biological function, preferably an active compound derived from the mycelium selected from ergothioneine, lovastatin, ergosterol, resveratrol, glutathione, shiitake purine, shiitake polysaccharide and concanavalin A. However, this enumeration is not meant to be interpreted as being particularly limited, and other compounds produced in mycelium as 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).

[0097] Ergothioneine is a sulfur-based amino acid that is found primarily in mushrooms and red / black beans. Its antioxidant and anti-inflammatory properties are known to prevent chronic diseases of aging, such as heart or brain-related diseases, and to protect the body from cell and tissue damage. Due to its beneficial effects, it is sometimes considered a longevity vitamin. Preferably, ergothioneine is a compound of the formula:

[0098]

[0099] or a salt thereof.

[0100] Oyster mushrooms are a major source of nutraceuticals and are known to have a wide range of therapeutic properties, such as mediating anti-tumor, anti-angiogenic, immunomodulatory, antioxidant, and anti-diabetic effects. Such macromolecules are beta-glucans, alpha-glucans, ergosterol, linoleic acid, etc. It has also been shown that the intake of ergosterol can increase vitamin D concentrations in serum and liver.

[0101] Polyphenols are the most consumed antioxidants in the human diet. Total phenolic content (TPC) refers to phenolic compounds with redox properties, which are responsible for antioxidant activity and are known to have potential beneficial effects on human health. Similarly, phytonutrients such as flavonoids also have anti-inflammatory properties, and they can act as antioxidants and protect cells from oxidative damage that leads to diseases.

[0102] The total phenol content is measured by the Folin Ciocalteu assay and is preferably expressed here in mg GAE / g biomass, wherein GAE is gallic acid equivalent. The total flavonoid content (TFC) is measured by aluminum chloride colorimetric assay and is preferably expressed here in mg QE / g biomass, wherein QE is quercetin equivalent. The further distribution of polyphenols is measured by high performance liquid chromatography with diode array detection (HPLC-DAD) to check the biomass content in the presence of potential polyphenols such as catechins, 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.

[0103] In particular, catechins are natural antioxidants that help prevent cell damage and provide other benefits. Quercetin is a plant pigment (flavonoid) and can help reduce swelling, kill cancer cells and help prevent heart disease. Chlorogenic acid is mainly found in coffee and other foods and is widely studied in neurodegenerative diseases due to its anti-inflammatory activity. 3,4-dihydroxybenzoic acid (protocatechuic acid) is commonly found in green tea and is known to have mixed effects on normal cells and cancer cells. Protocatechuic acid is considered to be the active ingredient of some traditional Chinese herbal medicines, such as Chibotium barometz (L.) (Functional Foods in Health and Disease, Vol. 7, pp. 232-244, 2011). For example, Acai oil derived from Acaí palm fruit (Eutelrpe oleracea) is rich in protocatechuic acid (630 mg / kg) (Journal of Agricultural and Food Chemistry, vol. 56, no. 12, pp. 4631-4636, 2008).

[0104] As understood herein, the C:N ratio (mycelium) preferably relates to the ratio of the carbon to nitrogen content in the mycelium component, while the C:N ratio (medium) preferably relates to the ratio of the carbon to nitrogen content in the fermentation medium (i.e. in the fermentation broth). The ratio is preferably understood herein as a w / w ratio.

[0105] As understood herein, fermentation media can be divided into three categories according to their complexity: synthetic media, defined media, and complex media.

[0106] A synthetic medium is preferably a medium in which no complex compounds are present, ie the concentrations of all components are known. For example, a culture medium which does not contain a complex nitrogen source is a synthetic medium.

[0107] A defined medium is preferably a medium containing at most one coordinating compound (eg, yeast extract) in which the potential compound has an unknown concentration.

[0108] Complex media are preferably completely undefined compositions with more than one complex compound and unknown substances or natural extracts with unknown concentrations (e.g. compounds extracted from agricultural tributaries or waste streams from the food industry). Preferably, the complex medium is defined by its production process.

[0109] The Brunauer-Emmett Teller (BET) surface area is a method for the characterization of porous and finely divided solids by gas adsorption. In a gas adsorption process (according to DIN-ISO 9277 and DIN 66131, respectively), the specific surface area of ​​a solid substance is determined by default by nitrogen adsorption at 77.4 K using the BET method. The evaluation is carried out in the generally valid range of the BET method of p / p0=0.05-0.3, respectively, in the stated relative pressure range. For the determination of very small surfaces, krypton adsorption (at 77.4 K) is used. Since the expected surface areas are very low, krypton has been used, since krypton is a suitable adsorbate for measuring low surface areas. The specific surface area is expressed in m 2 / g indicates.

[0110] The median pore size means that 50% of the pore volume is from pores larger than the median pore size, and the other 50% of the pore volume is from pores smaller than the median pore size.

[0111] If the adsorbent is mesoporous or microporous, the specific pore volume can be determined from nitrogen adsorption measurements. For macroporous adsorbents with pore sizes greater than 1000 micrometers, 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 size and the pressure applied to a non-wetting liquid such as mercury.

[0112] The fermentation process (which may preferably be a biochemical process) is not meant to be particularly limited, and as will be appreciated by those skilled in the art, any process involving microorganisms may be included in the present invention. Particularly preferred are cultivation methods (e.g., production of biomass, in particular fungal biomass) and biological production methods (e.g., expression of enzymes in microbial cultures, biological production of ethanol, production of intracellular or extracellular compounds, such as colorants, flavoring compounds, antioxidants, enzymes, moisturizing compounds and the like).

[0113] Thus, the submerged fermentation in the method of the invention can be operated as a batch, fed-batch or continuous process. These three main fermentation processes are known to those skilled in the art and differ in the outflow and inflow of materials to / from the fermentation vessel.

[0114] Batch process is characterized in that no material flows into the fermentation vessel. In batch process, all nutrients are provided at the beginning of cultivation, and no nutrients are added in the subsequent bioprocess. During the whole bioprocess, except gas, acid and alkali, no additional nutrients are added. Then the bioprocess continues until nutrients are consumed. This strategy is suitable for rapid experiments, such as optimization of strain characterization or nutrient medium. The shortcoming of this convenient method is that biomass and product yields are limited. Because carbon source and / or oxygen transfer are usually limiting factors, microorganisms are not in exponential growth phase for a long time. After the bioprocess running in batch mode ends, only biomass or culture medium are harvested and appropriately treated to obtain desired products. From the perspective of bioreactor, the process is repeatedly interrupted by cleaning and sterilization steps, and biomass is only produced in stages.

[0115] In the fed-batch process, substrate, nutrients and other materials can be added to the fermentation vessel to extend possible culture time or increase productivity, etc. The advantage of feeding during the culture period is that it allows to achieve a higher overall product amount. Under specific growth conditions, microorganisms and / or cells constantly double and therefore follow an exponential growth curve. Therefore, in certain embodiments, the feed rate can also increase exponentially. Usually, for example, substrate is pumped to the culture vessel from a supply bottle by a silicone tube. The user can manually set the feeding (linear, exponential, pulse mode) at any time, or add nutrients when meeting specific conditions (such as when reaching a specific biomass concentration or when nutrients are exhausted). The fed-batch process provides a wide range of control strategies, and is also suitable for highly specialized applications. However, it can increase processing time and potentially lead to inhibition by the accumulation of toxic byproducts.

[0116] Preferably, in the method of the present invention, submerged fermentation is operated as a continuous process. After the batch growth phase, a balance (also referred to as steady state) about a specific component is established. Under these conditions, when fresh culture medium is removed, the same amount of fresh culture medium (chemostat) is added. These biological processes are called continuous culture, and are particularly suitable when excessive nutrients will cause inhibition due to, for example, acid or ethanol accumulation or overheating. After reaching steady state, it should be understood that the continuous mode is constantly operated during the exponential growth phase, wherein the cells are maintained at a constant concentration. The transient state is the state before the constant steady state condition of the continuous mode is reached, and is actually similar to the start of batch mode operation. Other advantages of the method include reduced product inhibition and improved space-time yield. When removing culture medium, harvest cells, which is the reason why the inflow and outflow rate must be less than the doubling time of microorganisms. Alternatively, cells can be retained in a variety of ways (for example, in a rotary filter), which is called perfusion. In a continuous process, compared with a fed-batch process, the space-time yield of a bioreactor can even be further improved. However, long culture periods also increase the risk of pollution and long-term changes in culture. The three most common types of continuous cultures are chemostat (the rate of addition of a single growth-limiting substrate controls cell proliferation), turbidostat (indirect measurement of cell number - turbidity or optical density, which requires additional sensors but is driven by real-time feedback, controlling the addition and removal of liquids), and perfusion (this type of continuous bioprocessing mode is based on retaining cells in the bioreactor or recycling cells back to the bioreactor; fresh medium is provided, and the cell-free supernatant is removed at the same rate).

[0117] Preferably, in the method of the present invention, the submerged fermentation is operated as a continuous process.

[0118] In a preferred embodiment, edible fiber mycelium mass is derived from deep fermentation. Deep fermentation allows mycelium to grow without the need for a substrate that structurally supports them as in solid-state fermentation. In addition, the growth rate is higher because nutrients can be transported to all points of the mycelium and it is easier to maintain aseptic conditions. Depending on the fermentation conditions, mycelium has the possibility of growing into small balls or threads in deep fermentation. This structure is maintained when harvesting, thereby giving the resulting product (such as cheese products during gentle processing) different texture characteristics.

[0119] Furthermore, submerged fermentation offers the additional advantage that a clean product is obtained which is not contaminated by residues of solid substrates which could bring about an unpleasant taste or subsequent allergies, in particular because washing of the mycelium grown in submerged fermentation can be easily achieved. Similarly, fermentation by-products, such as acids or alcohols, which could also alter the taste, can also be easily removed.

[0120] In submerged fermentation, it is also easier to better control fermentation conditions such as pH and oxygen content. Therefore, it is easier to achieve maximum growth of mycelium compared to substrate limitation in solid-state fermentation. Therefore, a more uniform product composition can be obtained, which also enables reduced batch-to-batch variation.

[0121] 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 a dairy alternative product. The mycelium obtained from solid-state fermentation is usually dry and therefore needs to be resuspended in water to obtain a milk-like substance.

[0122] In addition, in submerged fermentation, because the fermentation medium can be stirred, it is possible to easily and effectively prevent the spore formation or fruiting body formation that can cause toxin production. It is also apparent to those skilled in the art that if growth is carried out in submerged fermentation, it is preferred that no differentiation of mycelium to spores and / or fruiting bodies occurs. Therefore, it is apparent to those skilled in the art that the mycelium derived from submerged fermentation may also be referred to as non-differentiated mycelium biomass. Those skilled in the art will recognize other advantages of using submerged fermentation, such as better control of the texture and composition of biomass compared to solid-state fermentation.

[0123] In addition, submerged fermentation method is easier to scale up, because the second culture can be inoculated with the first culture, because both cultures are in liquid state. This is not easy to achieve in solid state fermentation, because the first culture can not flow. This shortcoming can be partially compensated by using a rotary drum reactor, wherein the reactor is rotated to mix solids and mycelium. However, known drum reactors have the huge problem of heat transfer causing different temperatures in the reactor during fermentation. There may be partial fermentation volume overheating at a certain point because heat cannot be properly removed, or the partial fermentation volume, particularly when fermentation begins, is not heated enough. In submerged fermentation, heat transfer will be more effective, because liquid is good in transferring heat, thereby causing more uniform heat distribution and therefore causing more controlled, more effective fermentation process.

[0124] Thus, preferably as encompassed by the present invention, the method is performed 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.

[0125] Preferably, in one embodiment, edible fungi currently discovered or to be discovered and co-cultivation or co-fermentation of such edible fungi with each other are used. In another embodiment, the aforementioned embodiment is further combined with the use of algae or bacteria or plants or archaea or animal cells / fat cells or a combination thereof.

[0126] Preferably, the edible fibrous mycelium mass is obtained from at least one fungal strain selected from the following: Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Pezizomycotina, Agaricaceae, Coleomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morelaceae, Truffleaceae, Pleurotaceae, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thin Poraceae, Polyporaceae, Strophariaceae, Pleurotus eryngii, Trichodermaceae, Omphalaceae, Cordyceps, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese.

[0127] In one embodiment, edible fungi currently discovered or to be discovered are used and co-cultivation or co-fermentation of such edible fungi with each other. In another embodiment, the foregoing embodiments are further combined with the use of algae or bacteria or plants or archaea or animal cells / fat cells or a combination thereof.

[0128] 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 the phylum Basidiomycota may be a fungal strain selected from the subphylum Agaricida. As defined herein, the fungal strain selected from the subphylum Agaricida may be a fungal strain selected from the class Agaricales. Preferably, the fungal strain selected from the class Agaricales may be a fungal strain selected from the orders Agaricales, Auriculariales, Boletales, Chanterelles, Polyporales and Russulares.

[0129] When the fungal strain is selected from the order Agaricales, the fungal strain is preferably selected from the family Agaricaceae, Fistulinaceae, Lyophyllaceae, Marasmiaceae, Omphalotaceae, Physalacriaceae, Pleurotaceae, Schizophyllaceae, Strophariaceae and Trichlomataceae.

[0130] The fungal strain selected from the family Agaricus may be Agaricus bisporus or Agaricus blazei, more preferably Agaricus bisporus.

[0131] The fungal strain selected from the family Bolognaceae is preferably Fistulina hepatica.

[0132] The fungus strain selected from the family Cynomorium is preferably Calocybe indica.

[0133] The fungus strain selected from the family Dermestidaceae is preferably Lentinula edodes.

[0134] The fungal strain selected from the family Omphalaceae is preferably Calvatia gigantea.

[0135] The fungal strain selected from the family of Cyperaceae is preferably Flammulina velutipes.

[0136] More preferably, at least one fungal strain selected from the order Agaricales may be a fungal strain selected from the family Pleurotus. Even more preferably, at least one fungal strain of the present invention is a fungal strain selected from the group consisting of Pleurotus pulmonatus, Pleurotus ostreatus, Pleurotus aurantii, Pleurotus florida and Pleurotus rosa, more preferably selected from the group consisting of Pleurotus pulmonatus or Pleurotus ostreatus, most preferably Pleurotus pulmonatus.

[0137] The fungal strain selected from the family Schizophyllum is preferably Schizophyllum commune.

[0138] The fungal strain selected from the family Strophariaceae is preferably a fungal strain selected from the group consisting of Agrocybe aegerita and Hypholoma capnoides.

[0139] The fungal strain selected from the family Tricholomataceae is preferably a fungal strain selected from the group consisting of Hypsizygus tesselatus and Clitocybe nuda.

[0140] Alternatively, the fungal strain selected from the class 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 family Auriculariaceae is Auricularia auricula-judae.

[0141] When the fungal strain is selected from the order Boletales, the fungal strain is preferably selected from the family Boletaceae and Sclerodermataceae. The fungal strain selected from the family Boletaceae is preferably Boletusedulis.

[0142] When the fungal strain is selected from the order Cantharellales, the fungal strain is preferably selected from the family Cantharellaceae and the family Hydnaceae. The fungal strain selected from the family Cantharellaceae may be Cantharelus cibarius. The fungal strain selected from the family Hydnaceae may be Hydnum repandum.

[0143] When the fungal strain is selected from the order Polyporaceales, the fungal strain is preferably selected from the order Ganodermataceae, Meripilaceae, Polyporaceae and Sparassidaceae.

[0144] The fungal strain selected from the family of Lactoporaceae is preferably Grifola frondosa. The fungal strain selected from the family of Polyporaceae may be from Polyporus umbellatus and Latiporus sulphureus. The fungal strain selected from the family of Hydrangeaceae may be Sparassis crispa. The fungal strain selected from the family of Meruliaceae is preferably selected from B. adusta and B. fumosa.

[0145] When the fungal strain is selected from the Russulares, the fungal strain may be selected from the Bondarzewiaceae and Hericiaceae. Preferably, the fungal strain selected from the Russulares is a fungal strain selected from the Hericiaceae, preferably selected from the Hericium erinaceus and Hericium coralloides. The fungal strain selected from the Bondarzewiaceae may be Bondarzewia berkeleyi.

[0146] According to the present invention, the at least one fungal strain may be selected from the subdivision Somnomidia. Preferably, the at least one fungal strain selected from the subdivision Somnomidia may be a fungal strain selected from the subdivision Pezizomycotina.

[0147] The fungal strain selected from the Pezizomycotina may be selected from the class Pezizomycotina. Preferably, the fungal strain selected from the class Pezizomycotina may be a fungal strain selected from the order Peziales. Preferably, the fungal strain selected from the order Peziales may be selected from the family Morchellaceae and the family Trepharaceae.

[0148] In a preferred embodiment, the mycelium is not derived from the family Sordaceae, in particular from Neurospora, such as from Neurospora crassa, or from Fusarium, such as from Fusarium venenatum.

[0149] Preferably, the fungal strain selected from the Morchellaceae family is Morchella septifolium, Morchella angustifolia, Morchella tenuifolia, Morchellasceptrifomtis, Morchella steppicola, Morchella puncripes, Morchella rubrum, Morchella importuna, Morchella Jaurentinaa or Morchella purpumscens, preferably Morchella septifolium, Morchella angustifolia or Morchella tenuifolia.

[0150] Preferably, the fungal strain selected from the family Tuberaceae is Tuber magnatum, T. estivum, T. uncinatum, T. indicum, T. rufum or T. melanosporum, more preferably T. melanosporum and T. melanosporum.

[0151] Alternatively, the at least one fungal strain selected from the subdivision Sorthomycetes may be a fungal strain selected from the class Sorthomycetes.

[0152] Preferably, the fungal strain selected from the class Sordaria may be a fungal strain selected from the order Hypocreales.

[0153] The fungal strain selected from the Hypocreales may be a fungal strain selected from the Cordycipitaceae. The fungal strain selected from the Cordycepsaceae may be a fungal strain selected from the Cordyceps militaris and Cordyceps sinensis.

[0154] Alternatively, the fungal strain selected from the Hypocreales may be a fungal strain selected from the family Hypocreaceae. The fungal strain selected from the family Hypocreaceae may be a Fusarium strain.

[0155] In another embodiment, the fungal strain selected from the class Sordariae may be a fungal strain selected from the family Sordaceae. The fungal strain selected from the family Sordaceae may be a strain of the genus Neurospora.

[0156] Preferably, the edible fibrous mycelium mass is obtained from at least one fungal strain selected from the following: Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Pezizomycotina, Agaricaceae, Coleomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morelaceae, Truffleaceae, Pleurotaceae, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thin Poraceae, Polyporaceae, Strophariaceae, Pleurotus eryngii, Trichodermaceae, Omphalaceae, Cordyceps, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese.

[0157] More preferably, the edible fibrous mycelium mass is obtained from at least one fungal strain selected from the group consisting of Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Agaricales, Agaricales, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morelaceae, Truffleaceae, Pleurotus, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus, Trichodermaceae, Omphalaceae, Pyriculariaceae, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese.

[0158] Even more preferably, the mycelial mass is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus floridus, Pleurotus aurantii, Pleurotus rosaceae, Morchella, Morchella black veined morel or Morchella edulis.

[0159] Even more preferably, the mycelial mass is obtained from Pleurotus pulmonarius, Pleurotus florida, Pleurotus aurantii, Pleurotus rosaceus, Morchella, Morchella black vein morel or Morchella edulis.

[0160] Most preferably, the mycelial mass is obtained from Pleurotus pulmonaria or Morchella rubrum.

[0161] In a preferred embodiment, the mycelial mass is obtained from Pleurotus pulmonatus. In another preferred embodiment, the mycelial mass is obtained from Morchella rubra. In a preferred embodiment, the mycelial biomass includes Pleurotus pulmonatus and Morchella rubra. In a preferred embodiment, the mycelial biomass includes Pleurotus pulmonatus and Morchella rubra.

[0162] In one embodiment, the mycelial biomass is obtained from Sophora solfataricus or Sophora nicotianae.

[0163] In a separate embodiment, the edible fibrous mycelium is obtained from at least one fungal strain selected from the group consisting of Basidiomycota, Ascomycota, Lepistaceae, Agaricomycetes, Polymycetes, Tremella, wherein preferred species herein are from at least one fungal species selected from the group consisting of Cordyceps spp., Inonotus spp., Grifola spp., Pleurotus spp., Ganoderma spp., Lentinula spp., Treella spp., Trametes spp., Lepista spp., Trichomonas spp., Aspergillus spp. and / or Panus spp. Therefore and preferably, the at least one fungal strain is a strain producing ergothioneine.

[0164] Typically, a constant temperature is maintained throughout the process, which temperature may be selected for optimal growth of a particular fungal strain, as known to those skilled in the art. For example, in the case of Pleurotus ostreatus, step (b) is preferably carried out at a temperature between 25°C and 30°C. Further preferably, the growth is carried out at a pH between 3.0 and 8.5. As will be appreciated by those skilled in the art, the choice of pH may depend on the fungal strain to be cultivated, or on potential contaminating strains to be excluded from the growth. Further preferably, step (b) is carried out for a time between 12 hours and 240 hours.

[0165] Therefore, the present invention also relates to a method for producing mycelial components by submerged fermentation, wherein the fermentation medium comprises 5-60 g / L carbon source, 0.1-60 g / L nitrogen source, 0.01-15 g / L minerals and 0.01-50 mg / L vitamins. Those skilled in the art will be able to scale up the method by adjusting the carbon source (i.e. total added sugar) and / or other parameters to meet the requirements of the reactor configuration used (batch mode vs. fed-batch mode vs. continuous mode).

[0166] In the method for producing mycelial components by deep fermentation, the fermentation medium preferably comprises 5-60g / L carbon source, 0.1-60g / L nitrogen source, 0.5-15g / L minerals and 0.1-10mg / L vitamins. Preferably, the carbon source is selected from potato dextrose agar, starch, cellulose, malt extract, beet sugar, corn sugar, 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, 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, cupric chloride, cupric sulfate, diammonium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate hydrate and / or sodium chloride. Preferably, the vitamin is selected from biotin, choline chloride, folic acid, inositol, nicotinamide, pantothenic acid, pyridoxal, riboflavin, thiamine, cobalamin and / or ascorbic acid. pH control is preferably controlled by a substance selected from sodium chloride, sodium hydroxide, potassium hydroxide, sulfuric acid, phosphoric acid, hydrochloric acid, citric acid, acetic acid, hypochlorous acid. It should be noted that other components may be used, such as agar or food grade defoamers, such as edible oils or others. Those skilled in the art will be able to expand the method by including other substances, metabolites and other types of carbon, nitrogen, minerals and vitamin sources.

[0167] In one embodiment, for nutritional supplementation of mycelial biomass, the culture medium is enriched with vitamin B12 (cobalamin) by adding B12 in pure chemical form. In another embodiment, the culture medium is enriched with vitamin B12 in the range of 1 μg / L-200 μg / L, preferably in the range of 10 μg / L-100 μg / L. In certain embodiments, the vitamin B12 accumulated in the biomass is at least 10% of the initial addition, preferably at least 5%, preferably at least 2.5%, preferably at least 1%, preferably at least 0.5%. 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%.

[0168] In one embodiment, the fungal strain can be grown on a defined medium. As defined herein, the mycelial fraction obtainable when grown on a defined medium may also be referred to as mycelial fraction A.

[0169] Thus, the present invention relates to an edible mycelial component (also referred to as mycelial component A) comprising a (preferably non-differentiated) mycelial biomass having an elemental composition of the mycelium with a C:N ratio of between 6 and 8 and characterized by an EUC of between 200 and 500 g MSG / 100 g. The mycelium is preferably Pleurotus pulmonaria mycelium. EUC is defined herein.

[0170] In another embodiment, the fungal strain may be grown on a synthetic medium. As defined herein, the mycelial fraction obtainable when grown on a synthetic medium may also be referred to as mycelial fraction B.

[0171] Thus, the present invention relates to an edible mycelial fraction (also referred to as mycelial fraction B) comprising non-differentiated mycelial biomass having an elemental composition of mycelial C:N ratio between 8 and 12, characterized in that the EUC is less than 200 g MSG / 100 g. EUC is defined herein.

[0172] In a preferred embodiment, the present invention provides an edible mycelium component comprising Pleurotus pulmonatus mycelial biomass (preferably non-differentiated biomass), having an elemental composition of the mycelium C:N between 2 and 12, and characterized by an equivalent umami concentration (EUC) of at least 30 monosodium glutamate g MSG / 100g. EUC is as defined herein. The edible mycelium component described herein can be obtained in a method for producing the edible mycelium component described herein by submerged fermentation, the method comprising the step of culturing Pleurotus pulmonatus in a fermentation medium, wherein the fermentation medium provided at the beginning of the fermentation is characterized by a C:N ratio of 1-50, preferably 5-50, wherein the fermentation medium provided at the beginning of the fermentation comprises 5-60 g / L of a carbon source, 0.1-60 g / L of a nitrogen source, 0.01-15 g / L of a mineral and 0.01-50 mg / L of a vitamin, characterized in that the medium comprises arginine and glutamic acid as the only amino acids. In one embodiment, the w / w ratio of arginine to glutamic acid 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 the specific mycelial component or food comprising the mycelial component. In a preferred embodiment, the w / w ratio of total arginine to glutamic acid is preferably 20:80, more preferably 50:50, even more preferably 40:60, most preferably 30:70. In a preferred embodiment, the culture medium comprises glutamic acid as the only amino acid. It should be understood that the present invention also provides an edible product, preferably a meat substitute product or a milk substitute product, comprising 1 to 99% by weight of the edible mycelial component described herein.

[0173] Preferably, the defined medium comprises more than one amino acid and more than one vitamin, preferably each with different concentrations. In this case, the biomass can have the possibility of growing on different sources. Preferably, amino acids and vitamins are derived from at least one complex nitrogen source, preferably selected from yeast extract, CSL and peptone, more preferably yeast extract. The carbon source is preferably derived from dextrose, glucose, maltose, galactose or malt extract or cellulose. In the medium determined, the carbon-nitrogen ratio (medium), i.e., the C:N ratio is 5-50, preferably 10-25, preferably 14-19. In one embodiment, the carbon-nitrogen ratio (medium) in the defined medium, i.e., the C:N ratio is between 10 and 25, preferably between 16 and 18, more preferably between 16.5 and 17.5.

[0174] Preferably, the synthetic medium comprises at least one amino acid and at least one type of vitamin, wherein (1) the amount of the at least one vitamin used is characterized in that the ratio of the at least one vitamin used to the total amount of all vitamins present in the defined medium (taking the preferred nitrogen complex as reference and expressing the ratio as w / w) is in the range of 0.01 to 10, preferably 0.1 to 10, more preferably 0.5 to 10, and (2) the amount of the at least one amino acid used is characterized in that the ratio of the at least one amino acid used to the total amount of all amino acids present in the defined medium is in the range of 0.01 to 10, preferably 0.1 to 10, more preferably 0.5 to 10 (wherein the ratio is expressed as a w / w ratio).

[0175] Preferably, the nitrogen source concentration ranges from 0.1-60 g / l, more preferably 1-40 g / l, even more preferably 1-30 g / l, most preferably 1-20 g / l. In another preferred embodiment, the nitrogen source concentration is 0.1-10 g / l, preferably about 6 g / l.

[0176] The carbon-nitrogen ratio (medium), i.e., the C:N ratio, in the synthetic medium is 1-50, preferably 5-50, more preferably 10-25, even more preferably 16-23. In one embodiment, the C:N ratio in the synthetic medium is 2-22, preferably 15-22, more preferably 20-25, even more preferably 20-22. In an alternative embodiment, the C:N ratio in the synthetic medium is 2-22, preferably 2-16, more preferably 2-14. In one embodiment, the C:N ratio in the synthetic medium is 1-5.

[0177] The at least one amino acid is preferably selected from alanine, asparagine, aspartic acid, arginine, tryptophan, glycine, glutamic acid, glutamine, methionine, phenylalanine, serine, valine, cystine, proline, leucine, tyrosine, threonine, isoleucine, histidine, lysine and selenocysteine ​​or a combination thereof.

[0178] 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 a combination thereof.

[0179] Alternatively, the at least one amino acid is preferably selected from 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 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.

[0180] 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. The list excludes umami amino acids. Preferably, no more than 5, preferably no more than 3, even more preferably no more than 2 types of amino acids are added to the culture medium. Most preferably, the at least one amino acid (preferably no more than 5 amino acid types, more preferably no more than 3 amino acid types, even more preferably no more than 2 amino acid types) is selected from alanine, arginine, cysteine, glycine, proline, serine, tyrosine and selenocysteine.

[0181] Therefore, the defined medium in the present invention can also be defined as a medium characterized by a C:N ratio of 1 to 50, and comprises at least one amino acid selected from alanine, arginine, cysteine, glycine, proline, serine, tyrosine and selenocysteine. Preferably, at least one amino acid as defined herein includes arginine. It is obvious to those skilled in the art that some amino acids are considered to be essential amino acids for mycelial growth, so the growth of mycelial may be impossible without adding them to the medium. The inventors were surprised to find that it is possible to grow mycelium in the presence of only one non-essential amino acid arginine, which allows obtaining mycelium with low umami content (as defined below and represented by EUC parameters). The minimum amount of the amino acid (preferably arginine) added is determined by the C:N ratio of the desired medium. In other words, the minimum amount of the amino acid added is preferably associated with the minimum amount of the carbon source required for growth, wherein the carbon source required for growth is at least 5g / L, more preferably at least 10g / L.

[0182] Preferably, the at least one non-essential amino acid added is no more than 5 (preferably no more than 4, even more preferably no more than 3, still more preferably no more than 2) amino acids, including arginine and (i) non-essential amino acids selected from alanine, cysteine, glycine, proline, serine, tyrosine and selenocysteine, and / or (ii) 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.

[0183] Furthermore, in one embodiment, the culture medium may optionally further comprise at least one umami amino acid, as defined below.

[0184] In one embodiment the culture medium comprises only one amino acid - arginine.It will be appreciated that the mycelial fraction obtainable using this culture medium is characterised by an EUC between 30 and 100 g MSG / 100 g, more preferably between 30 and 60 g MSG / 100 g.

[0185] Preferably, the C:N ratio is between 10 and 25, more preferably between 16 and 23. In one embodiment, the C:N ratio in the synthetic medium is 2-22, preferably 15-22, more preferably 20-25, even more preferably 20-22. In one embodiment, the C:N ratio ranges from 5 to 50. In one embodiment, the C:N ratio ranges from 1 to 5.

[0186] The inventors surprisingly found that by culturing mycelium using a culture medium disclosed herein, wherein the culture medium does not contain any umami amino acids, as described herein, an edible mycelium component can be obtained characterized by an EUC of less than 200 g MSG / 100 g, preferably characterized by an EUC of less than 100 g MSG / 100 g, even more preferably characterized by an EUC of 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, even more preferably at most 0.1 g / L, to obtain the desired mycelium EUC of less than 200 g MSG / 100 g, preferably 30 to 200 g MSG / 100 g, required for a specific food application.

[0187] The present inventors have surprisingly found that by culturing mycelium under the fermentation conditions of the present invention using a defined medium, an edible mycelial component can be obtained, which is preferably characterized by an EUC range between 200 and 500 g MSG / 100 g, preferably characterized by an EUC range between 250 and 450 g MSG / 100 g.

[0188] After one of the nutrients becomes limiting (e.g., carbon source, nitrogen source, oxygen) and is completely exhausted, the flask is used to inoculate another flask or a similarly sized pre-fermenter with a 10 to 100 times higher volume. Subsequently, the fermentation seed train consists of several pre-fermenters that are inoculated one after another at a volume / volume ratio of 1-20%. The pre-fermenter is operated at 10 to 50°C with a pH of 4 to 6 until the nutrient source is completely exhausted. Depending on the inoculum size and activity, the process will take 12 to 240 hours.

[0189] The main fermentation is finally carried out on a scale of up to 400m3 using the above culture medium under controlled conditions (10 to 50°C, pH 3 to 8, controlled dissolved oxygen, controlled aeration rate and stirring speed). Fermentation is continued in a discontinuous mode until the carbon source is exhausted, and the process will take 12 to 240 hours depending on the size and activity of the inoculum. In one embodiment of Ine, the preferred pH range of the fermenter to which the present invention relates is 3-8, more preferably 4-6, and even more preferably 4-5. In one embodiment of Ine, the preferred pH of the fermenter to which the present invention relates 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 buffer solution is maintained, preferably selected from a phosphate buffer or a citrate buffer. pH and C / N ratio are key factors in obtaining the desired growth and the properties (protein content, protein composition, fiber content and therefore the effect on EUC) listed in Table 3. As in the literature (Calsamiglia S, Mustela 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 March; 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 culture medium, which vary in medium A, B and C. This combination of fermentation conditions in combination with pH is understood to result in different osmotic pressures in the fungal strains, resulting in different total / free amino acid pools, and therefore in different protein contents and compositions, resulting in different EUCs. The observed trends in protein and fiber content observed for the P. ostreatus species exemplified in the present invention show surprisingly different trends from those reported for the same species and another P. ostreatus species, both grown on the same culture medium, where yields up to 30% were not reported in either publication (Ref. 1: Food Chemistry 85 (2004) 101-105; Ref. 2: Carbohydrate Polymers 87 (2012) 368-376, both using the culture medium detailed in Ref. 1).

[0190] The defined medium contains minerals, vitamins and trace elements, preferably selected from the compounds listed above, and is sterilized by heat sterilization (≥121° C. for at least 20 minutes) or microfiltration known to those skilled in the art before being added to the fermentor. At the end of the main fermentation period, the biomass is harvested by simply separating the biomass from the culture supernatant using a liquid-solid separation method such as centrifugation, filtration or sieving.

[0191] In one embodiment of all media of the present invention, the separated biomass is washed with acidic water, preferably at a pH of 3-7, more preferably 3-6. It should be understood that this washing step treats the biomass and substantially removes any bitter compounds formed or inhibits their formation, thus affecting EUC concentration. Preferably, the acidity is adjusted with citric acid, sulfuric acid, phosphoric acid or hydrochloric acid.

[0192] It will be appreciated that biomass yield is calculated based on the ratio of the amount of biomass produced to the amount of substrate (eg, C5 sugars) consumed.

[0193] It should be understood that according to the method of the present invention, for the culture medium corresponding to (and useful for preparing) component A (defined culture medium with a source of nitrogen complex), the biomass yield is preferably up to 65%, preferably up to 60% (based on the conversion rate of the carbon source available in the culture medium). In one embodiment, the biomass yield of the culture medium corresponding to component A is preferably at least 45%, more preferably at least 50%.

[0194] It should be understood that according to the method of the present invention, for a culture medium corresponding to (which can be used to prepare) component B (a synthetic culture medium comprising at least one amino acid necessary for mycelial growth), the biomass yield is preferably up to 55%, preferably up to 50% (based on the conversion of the carbon source available in the culture medium). In one embodiment, the biomass yield of this culture medium corresponding to component B is preferably at least 40%, more preferably at least 45%.

[0195] It will be appreciated that the method according to the invention produces a biomass yield of up to 100%, preferably up to 99%, more preferably up to 85 to 95%, for a culture medium corresponding to (that can be used to prepare) component C (a natural complex culture medium comprising a complex carbon source from a lateral flow extract). It will be appreciated that the biomass yield is calculated based on the ratio of the amount of biomass produced to the amount of substrate (e.g., C5 sugars) consumed.

[0196] The fungal strains can be grown on a natural composite medium, i.e. an extract prepared using a sidestream or an optionally pretreated sidestream. It is particularly preferred that the sidestream is brewer's grains. The carbon to nitrogen ratio (medium) in the composite medium is 5-50, preferably 10-40, most preferably 10-25. In one embodiment, the C:N ratio in the natural composite medium ranges from 2-18, preferably 2-16, more preferably 2-14.

[0197] In one embodiment, the C:N ratio in the natural complex medium is in the range of 2-18, preferably 6-16, more preferably 6-14. In one embodiment, the C:N ratio in the natural complex medium is in the range of 2-13, most preferably 4-8. 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.

[0198] In one embodiment, the carbon to nitrogen ratio of the side stream extract used as a carbon source for the natural composite medium, i.e., the C:N ratio in the side stream extract is preferably between 1 and 60, more preferably between 5 and 25, most likely between 10 and 20, even more preferably between 10 and 18. Thus, the C:N ratio of the side stream extract affects the C:N ratio of the final natural composite medium. In a preferred embodiment, the carbon to nitrogen ratio of the side stream extract from spent grains used as a C5-sugar carbon source for the natural composite medium is preferably 1-50, more preferably 1-25, most preferably 5-25, even more preferably 5-18.

[0199] For example, the side stream may be pretreated by using a heat treatment or an enzyme treatment. As the pretreatment step is optional, it will be appreciated that the side stream may also be used as is, without any further pretreatment.

[0200] Preferably, the protein content of the extract ranges from 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 of the composite culture medium comprises 1 to 200 g / l protein, preferably 1 to 100 g / l, more preferably 1 to about 50 g / l, most preferably about 5 to 45 g / l, more preferably 7 to 30 g / l. Such a protein content is necessary for obtaining high EUC in the resulting biomass. In a further preferred embodiment, the protein content of at least one extract of the composite culture medium comprises 10 to 30 g / l, more preferably 10 to 25 g / l of protein content. In a further preferred embodiment, the protein content of the at least one extract of the complex culture medium comprises a protein content of about 10 g / l, more preferably about 15 g / l, even more preferably about 20 g / l.

[0201] According to the present inventors, an increase in EUC was observed at high protein concentrations (ie, 7 g / L to 30 g / L) in the extract used for fungal cultivation.

[0202] Preferably, the glutamic acid content of the lateral 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 lateral stream extract used in the culture medium is preferably in the range of 400 to 4500 mg / L, more preferably 400 to 2250 mg / L, even more preferably 500 to 2000 mg / L. Preferably, the glutamic acid content of the side stream extract is above 150 mg / L, more preferably above 200 mg / L. More preferably, the aspartic acid content of the lateral stream extract is preferably in the range of 150 to 1500 mg / L, more preferably 150 to 750 mg / L, even more preferably 220 to 650 mg / L.

[0203] The range of high glutamate concentrations obtained in the extract, which directly affects EUC, depends on the residence time of the extraction in the extraction method used below, which involves steam pretreatment or liquid extraction of lignocellulosic material, preferably with the addition of acid. In one embodiment, the residence time of the extraction ranges from 1 to 25 minutes, preferably 5 to 15 minutes, to achieve a glutamate content in the extract obtained in the extraction method under acidic conditions that is surprisingly higher than the aspartic acid content. Thus, in this embodiment, the vinasse C5-sugar extract can be obtained by extracting under acidic conditions for 1 to 25 minutes, preferably 5 to 15 minutes. In another embodiment, the residence time of the extraction is between 1 to 25 minutes, preferably between 1 to 20 minutes, to achieve a glutamate content in the extract obtained in the extraction method using alkali that is surprisingly higher than the aspartic acid content. Thus, in this embodiment, the vinasse C5-sugar extract can be obtained by extracting under acidic conditions for 1 to 25 minutes, preferably 1 to 20 minutes. The observed trend teaches that longer residence times can lead to higher glutamate contents and thus to higher EUC. Preferably, acidic conditions here refer to, for example, at least 0.1% by weight of H 2 SO 4 , not more than 1.6 wt% H 2 SO 4 , not more than 1.0%wt.% H 2 SO 4 , preferably about 0.9 wt % H 2 SO 4 .

[0204] The fermentation medium may also be supplemented or treated by crystallization or precipitation of material or by a combination of side streams to achieve the desired C:N ratio (medium) in the absence of protein in the extract.

[0205] As preferably understood herein, when referring to a numerical value, the term about refers to the stated value ±10% of the stated value, more preferably it refers to the stated value ±5% of the stated value, even more preferably it refers to the stated value ±1% of the stated value, even more preferably it refers to the stated value.

[0206] As understood herein, the side stream may also preferably comprise lignocellulosic material, in particular lignocellulosic material derived from an industrial and / or agricultural side stream. Lignocellulosic material is preferably defined herein as a material comprising dry plant matter. Preferably, the lignocellulosic material comprises cellulose, hemicellulose and lignin. Preferably, the at least one lignocellulosic material is at least one industrial and / or agricultural side stream as defined herein. Further preferably, the lignocellulosic material is preferably solid or is processed into a powder before use. As understood herein, the lignocellulosic material is preferably characterized by a specific color, density and / or mesh size distribution.

[0207] Thus, it will be appreciated by those skilled in the art that in the process of the present invention, brewer's spent grains may be replaced by a side stream selected from the group consisting of spent spent grains, spent cereals (from industries other than breweries), cereal bran, bagasse, cotton and oil cake from sunflower, hazelnuts, shells and husks from nuts, grass and leaf waste, sawdust, coffee grounds, coffee husks, coffee silver skins, rapeseed and by-products from the soy industry such as okara ("residue"), banana leaves, banana peels, chicory roots, cassava peels, citrus pulp, cocoa, cocoa bean shells, cocoa mucilage, cocoa pod shells, coconut fiber, coconut shells, coconut shells , coffee pulp, corn cobs, corn stover, cotton, cottonseed meal, cottonseed, hemp, spent hops, pea by-products, peanut shells, peanut meal, peanuts, raw potato peels, potato tubers, eucalyptus bark, lantana grass, switchgrass, rice bran, rice husks, rice straw, waste sugar beets, sugar beet pulp, sawdust, wet sugarcane bagasse, walnut shells, wheat bran, wheat distiller's grains, malt extract, wheat straw, lupin seeds, chickpea bran, chickpea pod husks, chickpea straw, olive waste, wine pomace, pear pulp, sorghum bran, sorghum germ, sorghum stalks, sorghum stover, sunflower waste, and tea waste. In addition, as lignocellulosic materials included in the present invention, it is also preferred to include peels or waste or pulp or residues of the following side streams: oats, pine, dates, apples, apricots, barley, broccoli, cabbage, carrots, kelp, eggplant, kiwi, melon, alfaalfa, pineapple, pomegranate, plum, watermelon, zucchini, asparagus, red turnip, cauliflower, garlic, onions, pumpkin, squash and / or tomatoes.

[0208] Side streams referred to herein may also be understood as side streams from the agri-food industry.Examples of non-lignocellulosic materials, such as proteinaceous materials, are preferably palm oil, sugar cane bagasse, molasses, whey, whey permeate, wool and silk.

[0209] It will be apparent to one skilled in the art that all side streams described herein comprise sugars, minerals and / or vitamins in addition to lignocellulosic material and / or proteinaceous material.

[0210] A particularly preferred side stream is spent brewery grains.

[0211] The use of spent particles of different particle sizes leads to different results and is determined by the application. In certain embodiments, the most abundant particle size in the particle size distribution of spent grains is between 8-10 mm, preferably between 6-10 mm, more preferably between 4-6 mm, more preferably between 2-4 mm, more preferably between 1-2 mm, and more preferably between 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 spent grains suitable for the process of the present invention are characterized by a particle size distribution determined by using different sets of sieves, comprising a maximum of 2-4 mm up to 60 wt.-%, preferably up to 50 wt.-%, more preferably 30-50 wt.-%, most preferably 35-45 wt.-%, most preferably at least 35 wt.-%, followed by a second maximum of 1-2 mm preferably up to 35 wt.-%, more preferably 15-30 wt.-%, most preferably 20-30 wt.-%, followed by a third particle size distribution of 0.25-1 mm, preferably up to 20 wt.-%, more preferably 5-15 wt.-%, most preferably 8-15 wt.-%. The sieving is performed by air jet sieving following DIN 10765 2016-07, after using an automatic sieving tower, i.e. a vibrating sieving method.

[0212] In an exemplary embodiment, the screening residue from the following vibration screening method is 80%, the portion larger than 2.00 mm (between 2-4 mm) is 54.21 g / 100 g, the portion larger than 1.00 mm is 33.58 g / 100 g, the portion larger than 0.25 mm is 10 g / 100 g, and the screening residue is 1.94 g / 100 g.

[0213] Preparation for the fermentation of the fungal component may include a pretreatment step for further pretreatment of the side stream. Herein, the following steps are explained using the example of brewer's grains. Preferably, the preprocessing of the brewer's grains includes changing the particle size or other mechanical properties of the spent grains. For example, the preprocessing of the spent grains may include grinding of the spent grains. This step is conventional and is known to those skilled in the art. It may be further subjected to pretreatment, such as by, for example, comminuting the particles (e.g., by grinding, crushing, pulverizing, etc.) to increase its accessible surface area. Such steps disclosed herein as optional steps are known to those skilled in the art. Thus, as discussed in the literature (Ozturk et al., Journal of the Brewing Society (J. Inst. Brew. 108(1):23–27, 2002), the spent grains can be sieved after grinding through a series of sieves with openings of 850 μm, 425 μm and 212 μm. Depending on the fraction, the spent grain preparation can be considered coarse (425-850 μm), medium (212-425 μm) and fine (<212 μm), however, depending on the application used, it can also be ground or modified to have a larger particle size, for example including a maximum value between 0.3 and 1 mm, preferably a particle size referred to by those skilled in the art as a coarse particle size, preferably having a maximum value between 0.4 and 0.8 mm.

[0214] Spent grains are preferably understood as residues or by-products of the brewing industry. Preferably, spent grains are the material remaining after the mashing step and have a dry matter content preferably between 10% and 30%. However, the dry matter content as described herein is not meant to be limiting, as it is clear to those skilled in the art that the dry matter content can be increased in pre-processing, for example by pressing, by drying or by other methods known to those skilled in the art. In addition, spent grains from other industries (for example spent grains obtainable as by-products of food production) can also be used within the scope of the present invention. Those skilled in the art will be able to expand the method by including additional side streams from the above list of lignocellulosic materials.

[0215] Preferably, the BSG comprises between 20% w / w and 25% w / w cellulose (preferably 22%), between 23% w / w and 28% w / w hemicellulose (preferably 25.8%), and / or between 20% w / w and 30% w / w protein (preferably 25% w / w protein). These figures are understood to refer to the content of BSG relative to its dry mass. BSG is characterized by a final moisture content of between 50% and 75% by weight. Therefore, the final moisture content can be achieved by dehydrating the BSG using methods known to those skilled in the art by pressing the material to a final moisture content of between 50% and 75% by weight. However, this is not meant to be understood as limiting, as it will be more obvious to those skilled in the art that other means and methods for dehydrating the BSG may also be applied hereto.

[0216] In one embodiment, a method for producing a fungal fermentation medium from brewer's spent grains (BSG) comprises the following steps: (a1) extracting C5 sugars from lignocellulosic material contained in BSG via steam pretreatment, followed by a washing step with liquid water at a temperature not exceeding 50°C, and (b) combining the extract thus obtained with at least one non-carbohydrate nutrient for fungal cultivation.

[0217] 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 in step (a1), the lignocellulosic material contained in the BSG is contacted with steam for a time of at most 30 minutes, preferably for a time of at most 15 minutes. Preferably, during the steam pretreatment in step (a1), the lignocellulosic material contained in the BSG is contacted with steam at a temperature between 165°C and 175°C, and / or for a time of at most 15 minutes, preferably for a time of at most 12.5 minutes, more preferably for a time of at most 10 minutes, even more preferably for a time of at most 7.5 minutes, even more preferably for a time of at most 5 minutes, even more preferably for a time of at most 2.5 minutes, even more preferably for a time of at most 1 minute. More preferably, during the steam pretreatment in step (a1), the lignocellulosic material contained in the BSG is contacted with steam at a temperature between 165°C and 175°C for a time of up to 15 minutes. More preferably, during the steam pretreatment in step (a1), the lignocellulosic material contained in the BSG is contacted with steam at a temperature between 165°C and 175°C for a time of up to 15 minutes. More preferably, during the steam pretreatment in step (a1), the lignocellulosic material contained in the BSG is contacted with steam at a temperature between 165°C and 175°C for a time of up to 15 minutes. Preferably, the temperature between 165°C and 175°C relates to a temperature of about 170°C, more preferably, the temperature between 165°C and 175°C relates to a temperature of 170°C. Preferably, the time of steam pretreatment is at most 5 minutes, preferably at most about 4 minutes, more preferably at most about 3 minutes.

[0218] In one embodiment of the invention, the water used for prehydrolysis with steam may contain a dilute acid, for example not more than 1% w / w of the acid. Particularly suitable are 0.2% w / w or 0.4% w / w H 2 SO 4 In one embodiment, the acid (preferably H 2 SO 4 ) is between 1.1% w / w and 1.6% w / w. This is particularly applicable to embodiments in which the lignocellulosic material is brewer's grains. In one embodiment of the invention, the water used for prehydrolysis with steam may contain a dilute alkali, for example not more than 1% w / w of said alkali. Alternatively, 0.2% w / w NaOH is particularly suitable. Alternatively, the water used for prehydrolysis with steam may be replaced by a phosphate buffer at pH 5.5.

[0219] The steam pretreatment is followed by a washing step with liquid water at a temperature not exceeding 50° C., preferably not exceeding 40° C., even more preferably not exceeding 30° C., even more preferably not exceeding 25° C. Preferably, the liquid water used for the washing step is at room temperature, i.e. between 20° C. and 25° C., preferably at a temperature of about 22° C., more preferably at a temperature of 22° C.

[0220] Therefore, the aqueous extract of step (a1) obtained according to the present invention can be further supplemented with nitrogen source, carbon source, trace elements, vitamins and / or protein composition. 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. 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, in addition to the carbon source derived from BSG, no carbon source is added to the culture 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 that are beneficial to fungi growing on culture medium obtainable according to the method of the present invention, such as folic acid, riboflavin, pantothenic acid or biotin. Protein composition can be further used to supplement the aqueous extract of the present invention (a1).

[0221] In a second embodiment, the present invention relates to a method for producing a fungal fermentation medium from brewer's spent grains (BSG), the method comprising: (a2) extracting C5 sugars from the lignocellulosic material contained in BSG via a liquid extraction treatment with water at a temperature between 145°C and 155°C and / or for a period of up to 70 minutes, preferably for a period of up to 50 minutes, preferably at a pressure of 30 bar to 50 bar, and (b) combining the extract thus obtained with at least one non-carbohydrate nutrient for fungal cultivation.

[0222] 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 of up to 70 minutes, preferably for a time of up to 50 minutes. Furthermore, the extraction can be carried out at a pressure of 30 bar to 50 bar.

[0223] In one embodiment of the invention, the water used for the liquid extraction process may contain a dilute alkali, for example not more than 1% w / w of said alkali. Particularly suitable is 0.2% w / w or 0.4% w / w H 2 SO4 In one embodiment, the acid (preferably H 2 SO 4 ) is between 1.1% w / w and 1.6% w / w. This is particularly applicable to embodiments in which the lignocellulosic material is brewer's grains. In one embodiment of the invention, the water used for the liquid extraction process with water may contain a dilute alkali, for example not more than 1% w / w of said alkali. Alternatively, 0.2% w / w NaOH is particularly suitable. Alternatively, the water used here may be replaced by a phosphate buffer at pH 5.5.

[0224] Preferably, step (a2) of aqueous extraction of lignocellulosic material, preferably industrial and / or agricultural sidestream, according to the present invention is carried out with water at a pH between 2.0 and 12.0, preferably between 3.0 and 10.0, more preferably between 4.0 and 8.0, even more preferably between 5.0 and 8.0. As understood herein, the pH value is measured at a pressure of 1.0 bar and a temperature of 25°C, even if the extraction itself is carried out under different conditions as disclosed herein. Preferably, the pH is adjusted before the water is contacted with at least one lignocellulosic material, preferably an industrial and / or agricultural sidestream. It is also understood herein that the addition of acid or base to the water to a final concentration of more than 1% w / w as described herein is preferably avoided.

[0225] C5 sugars as defined herein preferably refer to fractions in which at least 80% w / w of the total sugar content constitutes pentoses (sugars including polysaccharides consisting of sugar subunits of five carbon atoms). Note that the C5 sugars defined herein as a fraction comprising sugars may contain other sugars, in particular C6 sugars (sugars having 6 carbon atoms, also called hexoses) as monomers and / or contained in polysaccharides and / or oligosaccharides. Thus, other sugars than pentoses may also be extracted.

[0226] Thus, as referred to herein, a distillers grains C5-sugar extract may refer to a composition wherein at least 80 wt% of the total sugar content constitutes pentoses, more preferably at least 80 wt% of the carbon-containing compounds (carbon sources) constitute pentoses.

[0227] In another embodiment, the present invention relates to the use of the fungal biomass of the present invention in the production of fungi-based foods. Therefore, the present invention also relates to fungi-based foods that can be obtained as described herein. The fungi-based foods of the present invention can be prepared in any form known to those skilled in the art. For example, the fungi-based foods of the present invention can take the form of balls (i.e., meatball substitutes), dumplings, vegetarian sausages, meat substitute steaks, meat substitute minced meat products, meat substitute products for preparing sandwiches, etc.

[0228] The food according to the present invention can be, for example, a nutritional supplement. The nutritional supplement can be in the form of a liquid or solid, such as a pill, lozenge or tablet. For example, the nutritional supplement of the present invention can be a protein supplement and / or a carbohydrate supplement.

[0229] Food as understood herein may be dairy products, such as yogurt, milk drinks and ice cream. Food as understood herein may also relate to different embodiments of seafood products, such as crab cakes, fish cakes, tuna, salmon or shrimp, and to different desserts, confectionery or baked goods, including chocolate, brownies or cookies, flour, starch, bread, eggs, pasta.

[0230] The food can be an organized food or a textured food. Therefore, the food of the present invention contains all amino acids that are necessary for daily intake by people and cannot be resynthesized. In addition, the textured food product of the present invention is preferably heat-resistant, boiling-resistant and suitable for cooking. For example, as described herein, the fungus-based food of the present invention can be a meat substitute product. It should be noted that, preferably, the meat substitute product is an organized food or a textured food. It is also noted that the structure of the textured food improves the acceptability of the textured food by consumers. It is further noted that the inherent fiber structure of the fungal biomass of the present invention may be beneficial for producing textured foods or organized foods without using conventional texturing methods (such as extrusion).

[0231] The fungus-based foods of the present invention may be further processed and / or supplemented. For example, according to protocols known to those skilled in the art, the fungus-based foods of the present invention may be further supplemented with water, salt, oil and / or spices. Further processing may also include heat and high pressure treatment of the food (particularly for high pressure pasteurization), brewing, boiling, baking, frying, fermentation and / or drying. As known to those skilled in the art, preservatives may be added to extend the shelf life of the food of the present invention.

[0232] Preferably, the food product of the present invention may be further supplemented with a constituent component defined as the content of substances / compounds that can be described as constituent components of the fungal ingredient of the present invention to form a fungal product.

[0233] "Constituents" are preferably understood in this context as supplementary preservatives, antioxidants and acidity regulators, thickeners, stabilizers and emulsifiers, pH regulators and anti-caking agents, flavor enhancers, improvers, stabilizers, thickeners, colorants, glazing agents and sweeteners, additives, aroma compounds and / or nutrients.

[0234] Preferably, the preservatives include calcium carbonate, acetic acid, potassium acetate, sodium acetate, calcium acetate, lactic acid, sorbate and malic acid.

[0235] Preferably, the antioxidant and acidity regulator include ascorbic acid, sodium ascorbate, calcium ascorbate, fatty acid esters of ascorbic acid, tocopherol-rich extracts, α-tocopherol, γ-tocopherol, δ-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.

[0236] Preferably, the thickener, stabilizer and emulsifier (or hydrocolloid) include alginic acid, sodium alginate, potassium alginate, ammonium alginate, calcium alginate, agar, carrageenan, processed Eucheuma seaweed, locust bean gum, guar gum, tragacanth gum, gum arabic (acacia gum), xanthan gum, tara gum, gellan gum, sorbitol, mannitol, glycerin, konjac, pectin, cellulose, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, ethyl methyl cellulose, carboxymethyl cellulose sodium cellulose, cellulose gum, enzymatically hydrolyzed carboxymethyl cellulose, sodium-potassium and calcium salts of fatty acids, magnesium salts of fatty acids, mono- and diglycerides of fatty acids, acetic esters of mono- and diglycerides of fatty acids, citrates of mono- and diglycerides of fatty acids, tartaric esters of mono- and diglycerides of fatty acids, microcrystalline cellulose-cellulose gel, mono- and diacetyl tartaric esters of mono- and diglycerides of fatty acids, mixed acetic and tartaric esters of mono- and diglycerides of fatty acids, sorbitol and mannitol.

[0237] Preferably, the pH adjuster and anti-caking agent include 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 oxide, magnesium oxide, fatty acids, gluconic acid, glucono delta-lactone, sodium gluconate, potassium gluconate and calcium gluconate.

[0238] 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'-ribonucleotide, disodium 5'-ribonucleotide and glycine and its sodium salt.

[0239] Preferably, the improving agent includes L-cysteine.

[0240] Preferably, the stabilizer comprises invertase and polydextrose.

[0241] Preferably, the thickener comprises husk, polydextrose, oxidized starch, monostarch phosphate, distarch phosphate, distarch phosphate, acetylated distarch phosphate, acetylated starch. Preferably, the thickener comprises acetylated distarch adipate, hydroxypropyl starch, hydroxypropyl distarch phosphate, sodium starch octenyl succinate, starch-based ingredients, and acetylated oxidized starch. More preferably, the thickener comprises flaxseed hulls and / or starch-based ingredients.

[0242] Preferably, the pigment comprises riboflavin, chlorophyll and chlorophyllin, anthocyanin, betanin, lycopene, copper complexes of chlorophyll and chlorophyllin, terpene compounds such as carotene compounds and lutein compounds, ordinary caramel, caustic sulfite caramel, ammonia caramel, ammonia sulfite caramel, vegetable carbon, calcium carbonate, iron oxide and iron hydroxide, curcumin, tartrazine, cellulose gel, carmine, carminic acid, carmine, azo red, carmonidine, lutein, cocoa powder (melanin), beet powder, tomato extract, duckweed powder, spirulina powder, dream detective powder (capsicum red and / or capsicum red), turmeric powder, blueberry powder, strawberry powder, berry pigment powder, heme powder, lycopene powder, betanin powder, alfalfa powder, saffron powder, mint powder and annatto extract.

[0243] Preferably, the glazing agent and the sweetener include isomalt, maltitol, acesulfame potassium, aspartame, sodium cyclamate, saccharin, sucralose, alitame, steviol glycosides, neotame, lactitol, xylitol and erythritol.

[0244] Preferably, the other additives defined as understood under the composition ingredients are selected from vitamin B12, vitamin B6, vitamin B2, vitamin B3 (also known as niacin), riboflavin, thiamine, vitamin A, vitamin E, omega-3 fatty acids, vitamin D2, folic acid, iodized salt (NaCl, also including iodized salt 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.), flavoring agents or flavoring components (salt, pepper, garlic, onion, mushroom fruiting body pieces, vegetable pieces, ginger, turmeric, curry, sugar (i.e. sucrose, glucose, monosaccharides or disaccharides), oils, lemon juice, orange juice, herbs and spices, yeast pieces), texturized vegetable proteins and natural flavor compounds. As defined herein, herbs and spices include natural flavor compounds such as methyl acetate, linalool, limonene, vanillin, etc., or synthetic compounds such as aprifloren, cinnamyl propionate, 16-hexadecanoic acid and ethyl levulinate. Such additional additives may improve optical visibility, flavor, nutrition, and provide additional texture.

[0245] Preferably, the nutrients are selected from ingredients rich in protein (e.g., pea protein isolate, chickpea protein isolate, gluten, egg white powder and / or mung bean protein isolate), ingredients rich in carbohydrates / dietary fiber (e.g., cereal-based flour, cereal-based starch, legume-based starch, fruit-based fiber, polysaccharides, starch-based ingredients, pea hulls, inulin, wheat starch, corn starch), ingredients rich in vitamins / minerals and / or ingredients rich in lipids (e.g., all types of edible oils and butters). Fiber-rich ingredients are preferably used to improve freeze-thaw stability and / or juiciness.

[0246] Note that the list of constituents ends here and it is known to those skilled in the art that a component or substance may be classified into more than one category.

[0247] The organoleptic properties may preferably also relate to odor attributes of pungency, savory, floral, sour, aged, musty, earthy, etc.; - texture attributes; - mouthfeel attributes related to juiciness and crispness; - taste attributes of sweetness, sourness, saltiness, bitterness, umami, metallic, astringency; the aroma attributes preferably relate to aroma complexity, aroma intensity, aroma roundness and off-flavors.

[0248] Texture attributes are defined herein as density, cutting strength, shear strength / force, hardness, elasticity (i.e., springiness), cohesiveness, stickiness, chewiness, adhesion, firmness, spreadability, stickiness, piercing force, water release, water retention capacity of a mycelial component or fungus-derived product (particularly food).

[0249] The cutting strength referred to herein preferably describes the resistance of a food to the intrusion of a cutting tool. Preferably, it is expressed in N.

[0250] The shear force as mentioned herein preferably describes the ability of a biomass or a product, in particular a food, to resist non-aligned forces applied to the biomass or the food at different parts thereof and acting in different directions for the same weight. Preferably, the force is applied only from the top. The applied force is collinear, also referred to as a compressive force. Preferably, it is expressed in N.

[0251] The hardness referred to herein preferably describes the force required to deform a product to a given distance. Preferably, it is expressed in N.

[0252] As referred to herein, elasticity (i.e., resilience) preferably describes the elasticity of a product, wherein the more the product is damaged, the less elastic it will exhibit, i.e., springing back its structural integrity after being subjected to stress. Preferably, it is expressed in %, which reflects the acceptable degree of deformation relative to the original size.

[0253] As mentioned herein, the cohesion of a product preferably describes the ability of a product whose structural integrity withstands compressive or tensile stress. A product is cohesive when it adheres to itself under such stress. Preferably, it is expressed in %, which reflects the acceptable degree of deformation relative to the original dimensions.

[0254] As referred to herein, tackiness preferably describes the energy required to disintegrate a food product into an easily swallowable state. Tackiness expressed in N is calculated as the product of hardness N and % cohesion (ie, the product).

[0255] As referred to herein, chewiness is preferably described as the energy required to chew solid food. The chewiness expressed in N is the product of the multiplication of the stickiness N and the elasticity %.

[0256] Adhesion as referred to herein describes the level of stickiness of a food. If the product is subjected to pressure deformation and if the surface of the sample is sticky, a negative force will be generated, which is calculated as the negative area under the curve. Large negative values ​​are preferably interpreted as a sticky mouthfeel.

[0257] The hardness referred to herein preferably describes the toughness of the sample, and the force (N) at the maximum penetration depth is used as the sample hardness, expressed in N. The area under the positive curve (Ns) represents the total amount of force required for the shearing process, and it is considered to be a good instrumental measurement of the spreadability in cream cheese and other spreadable products. The smaller the value of this area, the easier it is to spread. The force (N) of the maximum negative peak indicates the sample viscosity expressed as a negative value of N. The lower the value, the stickier the sample.

[0258] As referred to herein, the puncture force from a penetration test preferably describes the energy required to penetrate a sample at a certain depth. Preferably, it is expressed as the area under the curve of the positive area (Ns).

[0259] The water holding capacity (WHC) describes the ability of a material to retain water during processing, whereas the released water (RW) describes the ability of a material to release water during processing.

[0260] In the context of the present invention, "edible" 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 of the present invention is a product suitable for replacing dairy or meat products. It can also be called a vegetarian product, or even a vegan product. The suitability of a dairy substitute is determined by texture, mouthfeel, taste, nutritional content, water content, appearance and other factors, which should be as similar as possible to the product to be replaced.

[0261] In certain embodiments, the elemental composition of the edible mycelium ingredient has a C to N ratio (mycelium) ranging from 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 13, more preferably 1 to 12, more preferably 1 to 11, 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.

[0262] In a preferred embodiment, the edible mycelium component A has an elemental composition with a C to N ratio (mycelium) of 1-30, preferably 1-15, more preferably 5-10, most preferably 6-8.

[0263] In a preferred embodiment, the edible mycelium component B has an elemental composition with a C to N ratio (mycelium) of 1-30, preferably 1-15, preferably 5-12, more preferably 8-12, most preferably 8-10.

[0264] In a preferred embodiment, the edible mycelium component C has an elemental composition with a C to N ratio (mycelium) of 1-30, preferably 1-15, preferably 1-8, more preferably 2-6, most preferably 4-6.

[0265] In another embodiment, the invention relates to edible fibrous mycelial mass, as described herein. Preferably, the edible fibrous mycelial mass is derived from submerged fermentation.

[0266] In another embodiment, without any additional ribonucleic acid (RNA) reduction step during the production of the ingredient by additional heating or pH treatment, the method of the present invention preferably results in an intrinsic RNA level of the ingredient of at most 4 wt%, preferably at most 2 wt%, based on a dry basis of all disclosed mycelial ingredients. This applies to each of ingredients A, B and C.

[0267] In one embodiment, the RNA value of 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, based on dry weight. In a preferred embodiment, the intrinsic RNA level of component A is between 0.1 and 4% by weight, preferably between 0.1% by weight and 2% by weight, more preferably between 0.4% by weight and 1.88% by weight. In a preferred embodiment, the intrinsic RNA level of component B is between 0.1 and 4% by weight, preferably between 0.1% by weight and 2% by weight, more preferably between 0.4% by weight and 1.7% by weight. In a preferred embodiment, the intrinsic RNA level of component C is between 0.1 and 4% by weight, preferably between 0.1% by weight and 2% by weight, more preferably between 0.5% by weight and 1.9% by weight.

[0268] In another embodiment, the edible mushroom component contains 1-35 wt%, 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 1-10 wt%, more preferably 1-6 wt%, more preferably 1-5 wt%, more preferably 1-4 wt%, more preferably 1-3 wt%, more preferably 1-2 wt% chitin on a dry weight basis. In a preferred embodiment, the edible mushroom component contains 5-15 wt%, more preferably 6-12 wt%, most preferably 6-9 wt% chitin on a dry weight basis.

[0269] In certain embodiments, the mycelium component has an ergothioneine value of 1-1000mg / kg, preferably 1-900mg / kg, more preferably 1-800mg / kg, more preferably 1-700mg / kg, more preferably 1-600mg / kg, more preferably 1-500mg / kg, more preferably 1-400mg / kg, more preferably 1-300mg / kg, more preferably 1-100mg / kg, more preferably 1-50mg / kg, more preferably 1-25mg / kg. 200 In a preferred embodiment, the mycelium component contains an ergothioneine value of 25-1000mg / kg, preferably 50-700mg / kg, more preferably 80-600mg / kg. In a preferred embodiment, mycelium component A contains an ergothioneine value of 70-270mg / kg, which is preferably reached within 5 days. In a preferred embodiment, mycelium component B contains an ergothioneine value of 100-350mg / kg, which is preferably reached within 5 days. In a preferred embodiment, mycelial component C contains an ergothioneine value of 300 to 800 mg / kg, preferably within 5 days, preferably within the range of 350 to 800 mg / kg, preferably within 5 days, most preferably within the range of 400 to 800 mg / kg, preferably within 5 days. As understood herein, the ergothioneine value preferably refers to the amount of ergothioneine expressed in mg / kg mycelial component, expressed on a dry mass basis.

[0270] As the inventors surprisingly found, when Pleurotus pulmonatus is used as a fungal strain in a fermentation system for 5 days, mycelium component A preferably has an ergothioneine content of 70 to 100 mg / kg, component B preferably contains an ergothioneine content of 110 to 150 mg / kg, and component C shows a maximum ergothioneine content of 380 to 455 mg / kg, with an average value of 433 mg / kg, which is a higher value than the patent disclosed in the art for producing ergothioneine using Pleurotus pulmonatus.

[0271] In certain embodiments, the content of ergosterol in the mycelial component ranges from 0-50 mg / g, preferably 0-25 mg / g, more preferably 0-15 mg / g, more preferably 0-12.5 mg / g, more preferably 0-10 mg / g, more preferably 0-8 mg / g, more preferably 0-7 mg / g, more preferably 0-5 mg / g, more preferably 0-3 mg / g, more preferably 0-2 mg / g, more preferably 0-1 mg / g. 0 0 In a preferred embodiment, the content of ergosterol in the mycelial component is 0-8 mg / g, more preferably 1-6 mg / g, and most preferably 2-6 mg / g. In a further preferred embodiment, mycelial components A and B contain 0-4 mg / g, more preferably 2-4 mg / g of ergosterol. In a further preferred embodiment, mycelial component C contains 0-8 mg / g, more preferably 2-7 mg / g, more preferably 4-7 mg / g, and most preferably 5-6 mg / g of ergosterol.

[0272] In certain embodiments, the flavoring agent in the form of 5' nucleotides (5'NMP) of known flavoring agents, including 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP) is measured in g / kg. In a preferred embodiment, there is no 0 g / kg of IMP. In another embodiment, after enzymatic treatment (preferably with 5'-adenylate deaminase) to convert AMP to IMP, the umami taste from the 5'-nucleotide can be further enhanced by at least 30%, preferably at least more preferably at least 50%.

[0273] In one embodiment, the edible mushroom ingredients show a good abundance of 5'NMP containing 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP), which ranges from 0.1-40 g / kg, preferably 0.1-20 g / kg, preferably 0.1-15 g / kg, more preferably 0.1-12.5 g / kg, more preferably 0.1-10 g / kg, more preferably 0.1-8 g / kg, more preferably 0.1-6 g / kg, more preferably 0.1-5 g / kg, more preferably 0.1-4 g / kg, more preferably 0.1-3 g / kg, more preferably 0.1-2 g / kg.

[0274] In a preferred embodiment, the value of 5'NMP of component A is 1-20 g / kg, more preferably 2-6 g / kg, most preferably 3-6 g / kg.

[0275] In a preferred embodiment, the value of 5'NMP of component B is 0.1-15 g / kg, more preferably 0.1-5 g / kg, most preferably 0.1-3 g / kg.

[0276] In a preferred embodiment, the value of 5'NMP of component C is 0.1-40 g / kg, more preferably 5-20 g / kg, most preferably 8-20 g / kg.

[0277] In another preferred embodiment, for the mycelial component A, the value of 5'NMP is at most 40 g / kg, preferably at most 20 g / kg.

[0278] In another preferred embodiment, the 5'NMP value of the mycelium component B is at most 40 g / kg, preferably at most 20 g / kg.

[0279] In another preferred embodiment, for the mycelium component C, the value of 5'NMP is at most 40 g / kg, preferably at most 20 g / kg.

[0280] It was observed that mycelia grown on vinasse extract resulted in a 10-fold increase in the 5'NMP component.

[0281] In another embodiment, the mycelial fraction C exhibits a high concentration of uridine monophosphate (UMP), preferably in the range of 2.5 to 5 g / kg.

[0282] In certain embodiments, the mycelial component preferably contains all 20 essential amino acids. In a preferred embodiment, the mycelial component comprises the amino acids alanine, asparagine, aspartic acid, arginine, tryptophan, glycine, glutamic acid, glutamine, methionine, phenylalanine, serine, valine, cystine, proline, leucine, tyrosine, threonine, isoleucine, histidine and lysine.

[0283] 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, and more preferably at least about 23% by weight of branched chain amino acids (BCAAs), based on the total amount of amino acids present (i.e., the total amount of total protein, as referred to herein, amino acids present relate to the total content of free amino acids as well as amino acids contained in proteins and peptides).

[0284] 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, and more preferably at least about 23% by weight of umami amino acids based on the total amount of amino acids present (i.e., total protein, as referred to herein, the amino acids present relate to the total content of free amino acids and amino acids contained in proteins and peptides).

[0285] It is understood herein that essential amino acids are amino acids that cannot be synthesized efficiently enough by an organism to supply its requirements and must be supplied by the diet.

[0286] In a preferred embodiment, the edible mycelium component contains at least about 30% by weight, preferably at least about 35% by weight, and more preferably at least about 40% by weight of essential amino acids based on the total amount of amino acids present (i.e., total protein, as referred to herein, amino acids present refers to the total content of free amino acids as well as amino acids contained in proteins and peptides).

[0287] 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, most preferably 250 to 280 mg / g.

[0288] In a preferred embodiment, the total amount of amino acids in component B grown on minimal medium corresponds to 100-200 mg / g, preferably 120-180 mg / g, more preferably 150-180 mg / g.

[0289] In a preferred embodiment, the total amount of amino acids in component B grown on spent grains 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.

[0290] In another preferred embodiment, the edible mycelium component A, wherein the amount of BCAA and the amount of umami amino acids are 40-100 mg / g, respectively, preferably 50-60 mg / g, respectively.

[0291] In another preferred embodiment, the edible mycelium component B, wherein the amount of BCAA and the amount of umami amino acids are 20-80 mg / g, respectively, preferably 30-40 mg / g, respectively.

[0292] In another preferred embodiment, the edible mycelium component C, wherein the amount of BCAA is 50-150 mg / g, preferably 65-75 mg / g, and the amount of umami amino acids is 50-150 mg / g, preferably 70-100 mg / g, and most preferably 75-85 mg / g.

[0293] In certain embodiments, the EUC concentration of the mycelial component ranges from 1 to 25,000%. In a preferred embodiment, the EUC concentration of the mycelial component is 1 to 20,000%, preferably 1 to 150,000%, more preferably 1 to 13,000%, more preferably 1 to 12,000%, more preferably 1 to 10,000%, more preferably 1 to 5,000%, more preferably 1 to 2,500%, more preferably 1 to 1,500%, more preferably 1 to 1,000%, more preferably 1 to 600%.

[0294] In a preferred embodiment of mycelium component A, the EUC concentration ranges from 1-1000%, more preferably 250-800%, even more preferably 250-500%. Preferably, the EUC concentration of mycelium component A is about 300%. In another preferred embodiment of mycelium component A, the EUC is 200-500 g MSG / 100 g.

[0295] Preferably, the mycelial fraction A is characterized by an insoluble fiber content of 25-45% w / w, more preferably 30-45% w / w, such as 34% w / w. In this context, % w / w refers to the insoluble fiber content in dry matter.

[0296] In a preferred embodiment of mycelial component B, the EUC concentration ranges from 1-600%, preferably 1-200%, more preferably 10-100%, and even more preferably 30-60%. Preferably, the EUC concentration of mycelial component B is about 34%. Alternatively, in a preferred embodiment of mycelial component B, the EUC is less than 200 g MSG / 100 g, more preferably the EUC is less than 100 g MSG / 100 g. Alternatively, in an embodiment of mycelial component B, the EUC is preferably between 30 and 200 g MSG / 100 g, even more preferably between 30 and 60 g MSG / 100 g.

[0297] Preferably, the mycelial fraction B is characterized by an insoluble fiber content of 30-60% w / w, preferably 40-60% w / w, such as 54% w / w. In this context, % w / w refers to the content of insoluble fiber in dry matter.

[0298] In a preferred embodiment of the mycelium component C, the EUC concentration ranges from 1-10000%, more preferably 500-5000%, even more preferably 2000-4000%, even more preferably 2500-3500%. Preferably, the EUC concentration of the mycelium component C is about 2890%. In a preferred embodiment of the mycelium component C, it is characterized in that the EUC concentration is at least 500%, more preferably at least 1000%, even more preferably 1500%, even more preferably at least 2000%.

[0299] Preferably, the mycelial fraction C is characterized by an insoluble fiber content of at most 35% w / w, more preferably at most 30% w / w, such as 25% w / w. In this context, % w / w refers to the insoluble fiber content in dry matter.

[0300] Note that the above EUC concentrations were achieved within 5 days.

[0301] The uronic acids analyzed in the mycelial component, especially D-galacturonic acid and D-glucuronic acid, are very valuable substances that can be used as antioxidants and detoxifying and inactivating agents for various substances in the human body. In certain embodiments, the content of the two uronic acids in the mycelial component after acid hydrolysis ranges from 0 to 50% by weight, preferably 0 to 25% by weight, more preferably 0 to 15% by weight, more preferably 0 to 10% by weight, more preferably 0 to 5% by weight, more preferably 0 to 2.5% by weight, and more preferably 0 to 2% by weight. In a preferred embodiment, the content of uronic acid in the mycelial component is 0.1 to 15% by weight, more preferably 0.1 to 5% by weight, and most preferably 1 to 2% by weight.

[0302] In certain embodiments, the total phenolic content of the mycelial component ranges from 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, 1-5 mg GAE / g. In a preferred embodiment, the total phenolic content of the mycelial component is 1-150 mg GAE / g, preferably 1-15 mg GAE / g, and more preferably 1-5 mg GAE / g.

[0303] In certain embodiments, the total flavonoid content of the mycelial component ranges from 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, 1-5 mg QE / g. In a preferred embodiment, the total flavonoid content of the mycelial component is 1-150 mg QE / g, preferably 1-15 mg QE / g, more preferably 1-5 mg QE / g.

[0304] In a specific embodiment, the mycelial fraction A has a total phenolic content of 1-5 mg GAE / g, preferably about 3.2 mg GAE / g.

[0305] In a specific embodiment, the mycelial fraction B has a total phenolic content of 1-5 mg GAE / g, preferably about 2.7 mg GAE / g.

[0306] In a specific embodiment, the mycelial fraction C has a total phenolic content of 1-5 mg GAE / g, preferably about 4.5 mg GAE / g.

[0307] In a specific embodiment, the mycelial fraction A has a total flavonoid content of 1-5 mg QE / g, preferably about 1.7 mg QE / g.

[0308] In a specific embodiment, the mycelial fraction B has a total flavonoid content of 1-5 mg QE / g, preferably about 2.5 mg QE / g.

[0309] In a specific embodiment, the mycelial fraction C has a total flavonoid content of 1-5 mg QE / g, preferably about 3 mg QE / g.

[0310] Apparently, the extract contains less total flavonoids than total phenolics. The results indicate that compounds other than flavonoids are the major phenolics present in the tested strains, particularly for fractions A and B. In one embodiment, the mycelial fraction A contains about 35% to 60% flavonoids, preferably about 45 to 55% of the total phenolics, preferably about 55% of the total phenolics. In one embodiment, the mycelial fraction B contains about 70-95% flavonoids, preferably about 80-95% of the total phenolics, preferably about 93% of the total phenolics. In one embodiment, the mycelial fraction C contains about 50-80% flavonoids, preferably 60-70% of the total phenolics, preferably about 65%.

[0311] In one embodiment, the total polyphenols of the mycelial component ranges from 1-2000 mg / kg, 1-1000 mg / kg, 1-900 mg / kg, 1-800 mg / kg, 1-700 mg / kg, 1-600 mg / kg, 1-500 mg / kg, 1-400 mg / kg, 1-300 mg / kg, 1-200 mg / kg, 1-100 mg / kg, 1-50 mg / kg or 1-25 mg / kg. In a preferred embodiment, the total polyphenols of the mycelial component ranges from 1-1500 mg / kg, preferably 25-1000 mg / kg, more preferably 50-900 mg / kg, said values ​​preferably being reached within 5 days. This applies to components A, B and C.

[0312] In a specific embodiment, mycelial fraction A has 1-500 mg / kg, preferably 50-200 mg / kg, more preferably about 165 mg / kg of total polyphenols. In a further preferred embodiment, mycelial fraction A has total polyphenols, wherein about 35-45% by weight of the total polyphenols correspond to catechins, about 35-45% by weight of the total polyphenols correspond to protocatechuic acid, and the remainder is about 1-10% by weight of each of quercetin, chlorogenic acid and / or syringic acid.

[0313] In a specific embodiment, mycelial fraction B has 1-250 mg / kg, preferably 50-150 mg / kg, more preferably about 75 mg / kg of total polyphenols. In a further preferred embodiment, mycelial fraction B has total polyphenols, wherein about 25-35% by weight of the total polyphenols correspond to catechins, about 35-45% by weight of the total polyphenols correspond to protocatechuic acid, about 10-20% by weight of the total polyphenols correspond to quercetin, and the remaining about 5-15% by weight correspond to chlorogenic acid.

[0314] In a specific embodiment, the mycelial fraction C has 1-1500 mg / kg, preferably 100-1000 mg / kg, more preferably 250-950 mg / kg, more preferably about 650 mg / kg of total polyphenols. In a further preferred embodiment, the mycelial fraction C has total polyphenols, wherein about 45 to 55% by weight of the total polyphenols correspond to catechins, about 45 to 55% by weight of the total polyphenols correspond to protocatechuic acid, and about 1 to 5% by weight of the total polyphenols correspond to syringic acid.

[0315] Ethanol extracts of mycelial fractions were prepared with an accelerated solvent extractor using 96% ethanol as performed in Methods. 2020;8(7):803. https: / / doi.org / 10.3390 / pr8070803Antioxidant assays were performed and antioxidant activity was assessed by scavenging stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals according to a scavenging activity protocol (Potential antioxidant sources: Endophytic fungi from medicinal plants. Econ. Bot. 61·14-30 This is usually defined by the EC50 value, preferably defined as the concentration of antioxidant required to scavenges 50% of DPPH radicals within a specific time period.

[0316] In certain embodiments, the EC50 of the mycelial component is 1-100 mg / ml, preferably 1-50 mg / ml, more preferably 1-15 mg / ml. In a preferred embodiment, the EC50 range of mycelial component A is 1-25 mg / ml, preferably about 10.5 mg / ml.

[0317] In a preferred embodiment, the EC50 of mycelial fraction B is 1-25 mg / ml, preferably about 13 mg / ml. In a preferred embodiment, the EC50 of mycelial fraction C is 1-25 mg / ml, preferably about 9 mg / ml.

[0318] In a preferred embodiment, mycelial fraction C has the highest DPPH radical scavenging activity, preferably with an EC50 of about 9 mg / ml. This is well correlated with the phenol content of fraction C having the highest phenol content, and thus phenol content and antioxidant activity are well correlated.

[0319] In a preferred embodiment, mycelial component A has the second highest DPPH radical scavenging activity, preferably having an EC50 of about 10.5 mg / ml.

[0320] In a preferred embodiment, mycelial component B has a second or third DPPH radical scavenging activity, preferably with an EC50 of about 13 mg / ml.

[0321] The following sugars are measured in the mycelial component after acid hydrolysis: glucan, xylan, arabinan, galactan, mannan, rhamnan. In certain embodiments, the total lignocellulose 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%, more preferably 1 to 5%. In a preferred embodiment, the total lignocellulose sugars detected from the hydrolysis of mycelial component A are 20-60%, more preferably 25-45%, wherein the most abundant sugar is glucan accounting for 80-90% of the total measured sugars (the glucan content is preferably 25-40% by weight in absolute terms).

[0322] In another preferred embodiment, the total sugars detected from the hydrolysis of mycelial component B are 30-80%, more preferably 20-70%, and most preferably 40-60%, wherein the most abundant sugar is glucan accounting for 85-95% of the total sugars (the absolute value of the glucan content is preferably 40-50% by weight). In a further preferred embodiment, the total sugars detected from the hydrolysis of mycelial component C are 1-50%, more preferably 1-35%, and most preferably 1-30%, wherein the most abundant sugar is glucan accounting for 75-85% of the total sugars (the glucan content is preferably 10-25% by weight in absolute value).

[0323] In another preferred embodiment, the most abundant sugar detected from the hydrolysis of mycelial fraction A, B or C is glucan which accounts for 70-95% of the total sugars.

[0324] As understood herein, the term "insoluble fiber" preferably refers to a portion of dietary fiber, which is insoluble in water. The insoluble fiber preferably comprises chitin and beta-glucan and is preferably distinguished from insoluble fibers of plant origin which comprise plant cellulose and / or hemicellulose but do not comprise chitin. Insoluble fiber is known to help the human body process waste better, improve intestinal health and reduce the risk of colorectal disorders.

[0325] 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 between 39% and 60% w / w. Even more preferably, the edible fibrous mycelium has an insoluble fiber content of between 40% and 55% w / w.

[0326] In a preferred embodiment, the insoluble fiber content of the mycelium component A is 30-60% by weight, preferably 30-40% by weight.

[0327] In a preferred embodiment, the insoluble fiber content of the mycelium component B is 40-60% by weight, preferably 40-50% by weight.

[0328] In a preferred embodiment, the insoluble fiber content of the mycelium component C is 10-40% by weight, preferably 20-30% by weight.

[0329] It is preferably envisioned that the mycelium used in dairy and meat products and other products can be tailored to the specific requirements of the final product in terms of fiber content, protein content, nutrition, taste, etc.

[0330] In certain preferred embodiments, the edible fibrous mycelium used in the meat analog product formulation has an insoluble fiber content of 20-60% by weight, preferably 40-50% by weight, preferably about 45% by weight. In another preferred embodiment, the insoluble fiber used in the meat analog is about 35% by weight. In another preferred embodiment, the insoluble fiber used in the meat analog is about 25% by weight. In another preferred embodiment, the insoluble fiber used in the meat analog is about 45% by weight. In one embodiment, the edible fibrous mycelium in the meat analog has an insoluble fiber content of at least 20% by weight, preferably at least 30% by weight, preferably at least 45% by weight, preferably at least 50% by weight.

[0331] In further preferred embodiments, the insoluble fiber content of the edible fibrous mycelium for use in dairy analog products or other products (other than meat analogs) is at least 20 wt%, more preferably at least 30 wt%, more preferably at least 40 wt%. More preferably, the edible fibrous mycelium has an insoluble fiber content between 39 wt / wt% and 60 wt / wt%. Even more preferably, the edible fibrous mycelium has an insoluble fiber content between 40 wt / wt% and 55 wt / wt%. Even more preferably, the edible fibrous mycelium has an insoluble fiber content between 40 wt / wt% and 55 wt / wt%. Most preferably, the edible fibrous mycelium has an insoluble fiber content of about 45 wt / wt%. Most preferably, the edible fibrous mycelium has an insoluble fiber content of about 45 wt / wt%. It should be understood that the weight referred to herein relates to the dry mass of the mycelium. In one embodiment, the edible fibrous mycelium has an insoluble fiber content of at least 40 wt / wt%, preferably at least 50 wt / wt%, more preferably at least 60 wt / wt%.

[0332] 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. In certain embodiments, the protein content is about at least 30%, at least 40%, at least 60% by weight.

[0333] In a preferred embodiment, the protein content of the mycelium component A is 30-50% by weight, preferably 30-40% by weight.

[0334] In a preferred embodiment, the protein content of the mycelium component B is 30-50% by weight, preferably 30-40% by weight.

[0335] In a preferred embodiment, the protein content of the mycelium component C is 30-65% by weight, preferably 45-65% by weight.

[0336] 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 involve adjusting the content of the fermentation medium to control the uptake and thus the composition of the mycelial mass to be obtained. For example, the ratio between the carbon source and the nitrogen source in the medium can be changed, such as providing different excess levels of nitrogen. This will affect the insoluble fiber content accordingly, i.e., higher protein means lower insoluble fiber content, as shown in Table 3.

[0337] In one embodiment, the edible mycelium component has a carbohydrate content of at most 5% (non-fiber carbohydrates, i.e., carbohydrates that do not contain fiber), 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%.

[0338] In one embodiment, the edible mycelium component has a beta-glucan content of at least 80% of the total glucans.

[0339] In one embodiment, the present invention relates to edible mycelium component A, wherein the total glucan content is 20-35% by weight, preferably 25-35% by weight. In one embodiment, the present invention relates to edible mycelium component B, wherein the total glucan content is 25-50% by weight, preferably 30-40% by weight. In one embodiment, the present invention relates to edible mycelium component C, wherein the total glucan content is 10-35% by weight, preferably 10-20% by weight.

[0340] In one embodiment, the present invention relates to mycelial component AB or C, wherein at least 96% by weight of its polyunsaturated fatty acid content is linoleic acid (ω-6 fatty acid). In a specific preferred embodiment, the present invention relates to mycelial component A, wherein at least 98% by weight of its polyunsaturated fatty acid content is linoleic acid (ω-6 fatty acid). In a specific preferred embodiment, the present invention relates to mycelial component B, wherein at least 97% by weight of its polyunsaturated fatty acid content is linoleic acid (ω-6 fatty acid). In a specific preferred embodiment, the present invention relates to mycelial component C, wherein at least 96% by weight of the polyunsaturated fatty acids comprise linoleic acid (ω-6 fatty acid). In another specific embodiment, the present invention relates to mycelial component C, which has an enriched ω-6 fatty acid (linoleic acid) of 1-9% by weight, more preferably 2-5% by weight, and most preferably about 3.5% by weight.

[0341] In one embodiment, the mycelial components A, B and C have a fat content of up to 15% by weight, preferably up to 10% by weight. In a particularly preferred embodiment, the present invention relates to mycelial component A, wherein the fat content is 0.5-5% by weight, preferably 0.1-3% by weight. In a particularly preferred embodiment, the present invention relates to mycelial component B, wherein the fat content is 0.5-5% by weight, preferably 0.1-3% by weight. In a particularly preferred embodiment, the present invention relates to mycelial component C, wherein the fat content is 1-10% by weight, preferably 3-10% by weight, more preferably 3-8% by weight. In a particularly preferred embodiment, the present invention relates to mycelial component C, wherein the fat content is up to 8% by weight.

[0342] In one embodiment, the mycelium component does not contain mycotoxins. Mycotoxins are analyzed by liquid chromatography with tandem mass spectrometry (SOP M 3650). In μg / kg, no mycotoxins are present above their detection limit. Mycotoxins are selected from aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, ochratoxin A, deoxy-annular alcohol (DON), zearalenone, 3-acetyl-deoxy-annular alcohol, 15-acetyl-deoxy-annular alcohol, annular alcohol, T-2 toxin, HT-2 toxin, 4,15-diacetyl-fusarone, Fusarone-X, fumonisin B1 and fumonisin B2.

[0343] In another embodiment, polyaromatic hydrocarbons are analyzed using gas chromatography-mass spectrometry (SOP M 2920). Polyaromatic hydrocarbons are not detected above their detection limit (μg / kg). Polyaromatic hydrocarbons are selected from benzo(a)anthracene, benzo(c)fluorene, cyclopentadienyl(c,d)pyrene, 5-methyl, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(j)fluoranthene, benzo(a)pyrene, indeno(123-cd)pyrene, dibenzo(ah)anthracene, benzo(ghi)perylene, dibenzo(a,l)pyrene, dibenzo(a,e)pyrene, dibenzo(a,i)pyrene, dibenzo(a,h)pyrene.

[0344] In certain embodiments, the calorific value of the mycelium component ranges from 1 to 1000 Kcal / 100 g, preferably 1 to 900 Kcal / 100 g, more preferably 1 to 800 Kcal / 100 g, more preferably 1 to 700 Kcal / 100 g, more preferably 1 to 500 Kcal / 100 g, more preferably 1 to 400 Kcal / 100 g, more preferably 1 to 300 Kcal / 100 g, more preferably 1 to 100 Kcal / 100 g, more preferably 1 to 50 Kcal / 100 g, more preferably 1 to 25 Kcal / 100 g. In a preferred embodiment, the calorific value of the mycelium component is 1-800 Kcal / 100 g, preferably 200-800 500 Kcal / 100 g, more preferably 300-600 Kcal / 100 g, and most preferably about 300-500 Kcal / 100 g.

[0345] In a preferred embodiment, for pore sizes less than 1 mm, for the edible mushroom component A, 55-65%, preferably about 58%, of the pore volume corresponds to pore sizes of 1000-30 μm, with the most common pore size being in the range of 85-185 μm. In the second range of 30 to 2 μm, corresponding to about 35 to 45%, preferably 42%, of the pore volume, the most common pore size peak is equal to 16 μm, which is also the most common pore size in the range of 1000-2 μm. The specific pore volume of component A is 9 cm3 / g and the median pore size is 44.5 μm.

[0346] In a preferred embodiment, for pore sizes less than 1 mm, for the edible mushroom component B, about 75-85%, preferably 81.5%, of the pore volume corresponds to pore sizes between 1000-30 μm, with the most common pore size equal to 147 μm, which is also the most common pore size in the range of 1000-2 μm. In the second range of 30 to 2 μm, corresponding to about 15 to 25%, preferably 18.5%, of the pore volume, the most common pore size peak is equal to 15 μm. The specific pore volume of component B is 4.94 cm3 / g, and the median pore size is 143 μm.

[0347] In a preferred embodiment, for pore sizes less than 1 mm, for the edible mushroom component C, 15% to 25%, preferably about 20%, of the pore volume corresponds to pore sizes between 1000 and 20 μm, and 75% to 85%, preferably about 80%, of the pore volume corresponds to pore sizes between 20 and 2 μm, with the most common pore size peak being 5.5 μm. The specific pore volume of component B is 2.46 cm3 / g and the median pore size is 7.1 μm.

[0348] Compared to A and B, component C has a much lower pore volume and smaller pores.

[0349] In a preferred embodiment, the BET surfaces of ingredients A, B and C are also measured using krypton, as it is a suitable adsorbate for measuring low surface areas. The surface areas of the ingredients without further grinding are preferably: A is 0.79 m2 / g, B is 0.67 m2 / g, and C is 1.59 m2 / g.

[0350] In one embodiment, the edible mycelium components A and B are in N 2 The edible mycelium component C has a thermal stability of up to 210-220°C, preferably up to 220°C, in a N atmosphere. This is visible in thermogravimetric analysis (TGA), measured at the point where all moisture content has been lost in a mass manner. 2 It has a thermal stability of up to 175-185°C, preferably up to 190°C, under an atmosphere of Figure 2 , 3 As shown in Figures 4 and 5, it is observed that up to 1000°C, the thermal stability of C is lower than that of A, and the thermal stability of A is slightly lower than that of B.

[0351] In this paper, the textural properties of mycelial components were analyzed with respect to shear force, water holding capacity, water release, and density.

[0352] The shear force is affected 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 ranges from 1-200 N, 1-150 N, 1-100 N, 1-90 N, 1-80 N, 1-70 N, 1-60 N, 1-50 N, 1-40 N, 1-30 N, 1-25 N, 1-20 N, 1-15 N, 1-10 N or 1-5 N. In a preferred embodiment, the shear force of the mycelial biomass is at least 10 N, preferably at least 15 N, preferably at least 25 N, preferably 25-150 N, more preferably 30-120 N, and most preferably 30-105 N.

[0353] In certain embodiments, the mycelial biomass has a water holding capacity 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 water holding capacity ranges from 20-90%, preferably 30-80%, more preferably 40-70%.

[0354] In certain embodiments, the mycelial biomass has a water release 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 25-70%, more preferably 30-55%.

[0355] In certain embodiments, the mycelial biomass has a density of 0.1-10 g / cm. 3 0.1 and 9 g / cm 3 0.1 and 8 g / cm 3 0.1 and 7 g / cm 3 0.1 and 6 g / cm 3 0.1 and 5 g / cm 3 0.1 and 4 g / cm 3 0.1 and 3 g / cm 3 0.1 and 2.5 g / cm 3 0.1 and 2 g / cm 3 0.1 and 1.5 g / cm 3 0.1 and 1 g / cm 3 0.1 and 0.8 g / cm 3 or 0.1 and 0.5 g / cm 3 In a preferred embodiment, the density of the mycelial biomass is in the range of 0.1-3, preferably 0.5-2.5, more preferably 0.5-1.5 g / cm 3The preferred density of mycelial biomass is about 1 g / cm 3 .

[0356] The textural attributes of fungal products derived from mycelial components are influenced by the composition of the product, resulting in either soft or non-soft (ie, hard) mycelial-based meat analogs or dairy analogs.

[0357] In some embodiments, the meat analog or meat-like food is understood to preferably have a consistency or similarity or taste similar to the following animal meat in all forms (breast, fillet, thigh, rib, wing, chunk, steak, etc.), selected from: beef, poultry, fish, chicken, duck, goose, turkey, beef, pheasant, lamb and mutton, chicken breast, pork, ham, veal, deer or venison, seafood, Japanese prawns, crab, salmon, cod, pangasius, sardines, mussels and oysters.

[0358] In a preferred embodiment, the soft meat analog is preferably understood as meatballs. In another preferred embodiment, the soft meat analog is preferably meatballs, sausages, fish fingers, tartar, minced meat, meat pate, processed meat, Mett meat, luncheon meat, goose liver.

[0359] In another preferred embodiment, non-soft meat analogs are preferably understood to be steaks, beef jerky, hamburger patties, slices, kernels, salami, whole cuts, bacon, hot dogs, raw ham, dried meat and extruded products.

[0360] In another embodiment, the concepts of non-tender meat and tender meat may be interchangeable only when the ingredients used to produce traditional non-tender meat result in a meat analog that is softer in consistency than the traditional definition.

[0361] In a preferred embodiment, soft dairy analogs are preferably understood as cream cheese. In another preferred embodiment, soft dairy analogs are preferably understood as cream cheese, cheese spreads, processed cheese, ricotta 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, Buchette cheese, goat cheese, quark, cream, coffee creamer, whipped cream, sour cream, tapioca spread, margarine, butter, dessert, custard. Cream chocolate In another embodiment, non-soft or hard dairy analogs are preferably understood as hard cheese, semi-hard cheese, cheddar cheese, Parmesan cheese, etc.

[0362] In certain embodiments, the cutting strength of the food product ranges from 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 cutting strength of the soft meat analog is 1-25 N, most preferably 4-8 N. In a preferred embodiment, the cutting strength of the hard meat analog is 1-50 N, most preferably 10-30 N.

[0363] In certain embodiments, the hardness of the food product ranges from 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 of the soft meat analog ranges from 10-55 N, preferably 15-55 N, more preferably 20-50 N, and most preferably 20-45 N. In a preferred embodiment, the hardness of the hard meat analog ranges from 30-100 N, more preferably 50-100 N.

[0364] In certain embodiments, the food product has an elasticity ranging from 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 soft meat analog has an elasticity ranging from 35-85%, preferably 45-80%, more preferably 50-80%, and most preferably 55-75%. In a preferred embodiment, the hard meat analog has an elasticity ranging from 20-70%, preferably 30-60%.

[0365] In certain embodiments, the cohesiveness of the food product ranges from 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 cohesiveness of the soft meat analog ranges from 15-70%, preferably 20-60%, more preferably 25-50%, and most preferably 30-45%. In a preferred embodiment, the cohesiveness of the hard meat analog ranges from 20-85%, preferably 30-40%.

[0366] In certain embodiments, the food has a viscosity ranging from 1 to 200 N, 1 to 190 N, 1 to 180 N, 1 to 170 N, 1 to 170 N, 1 to 160 N, 1 to 170 N, 1 to 160 N, 1 to 140 N, 1 to 130 N, 1 to 120 N, 1 to 110 N, 1 to 110 N, 1 to 90 N, 1 to 80 N, 1 to 60 N, 1 to 50 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 viscosity of the soft meat analog ranges from 1-40 N, preferably 3-33 N, more preferably 5-25 N, and most preferably 6-21 N. In a preferred embodiment, the viscosity of the hard meat analog ranges from 6-85 N, preferably 15-40 N.

[0367] In certain embodiments, the chewiness of the food product ranges from 1 to 200 N, 1 to 190 N, 1 to 180 N, 1 to 170 N, 1 to 170 N, 1 to 160 N, 1 to 170 N, 1 to 160 N, 1 to 150 N, 1 to 140 N, 1 to 130 N, 1 to 120 N, 1 to 110 N, 1 to 110 N, 1 to 90 N, 1 to 80 N, 1 to 70 N, 1 to 60 N, 1 to 50 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 chewiness of the soft meat analog is 0.3-35 N, preferably 1-27 N, more preferably 2.5-20 N, and most preferably 3-16 N. In a preferred embodiment, the chewiness of the hard meat analog is 1-60 N, preferably 4-25 N.

[0368] In certain embodiments, the viscosity of the developed meat analogs 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 viscosity of the mycelium-based meat analogs is between 0 and -0.3 Ns, more preferably between -0.01 and -0.1 Ns, and most preferably between -0.02 and -0.05, which is a low sticky mouthfeel.

[0369] In certain embodiments, the hardness of the dairy analog (e.g., cream cheese or any mycelium-based spread) ranges from 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 another preferred embodiment, the hardness of the soft dairy analog ranges from 1-20 N, more preferably 5-15 N, and most preferably 5-10 N. In another preferred embodiment, the hardness of the hard dairy analog ranges from 20-100 N, more preferably 20-50 N, and most preferably 25-35 N.

[0370] In certain embodiments, the spreadability of dairy analogs such as cream cheese or any mycelium-based spreads is 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 of soft dairy analogs is 1-20 N.s, more preferably 10-20 N.s. In a preferred embodiment, the spreadability of hard dairy analogs is 30-100 N.s, preferably 40-80 N.s, more preferably 40-70 N.s, and most preferably 45-70 N.s.

[0371] In certain embodiments, the viscosity of the developed dairy analogs (e.g., cream cheese or any mycelium-based spread) ranges between -1 and -100N, -1 and -90N, -1 and -80N, -1 and -70N, -1 and -60N, -1 and -50N, -1 and -40N, -1 and -30N, -1 and -25N, -1 and -20N, -1 and -15N, -1 and -10N or -1 and -5N. In another preferred embodiment, the viscosity of the soft dairy analogs ranges from -1 to -14N, more preferably -3 to -10N, which is low viscosity. In a preferred embodiment, the viscosity of the hard dairy analogs ranges from -15 to -100N, preferably -15 to -50N, more preferably -15 to -30N, and most preferably -15 to -25N.

[0372] In certain embodiments, the piercing force of the dairy analog (e.g., cream cheese or any mycelium-based spread) results in an area under the curve ranging from 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, the area of ​​the soft dairy analog ranges from 1-30 N.s, preferably 5-25 N.s, more preferably 5-20 N.s, and most preferably 8-18 N.s. In another preferred embodiment, the area of ​​the hard dairy analog ranges between 40 and 100 N.s, more preferably between 40 and 80 N.s, most preferably between 50 and 60 N.s, which indicates that higher time and higher force are required to pierce the hard dairy analog.

[0373] In a specific exemplary embodiment illustrating the above embodiment, mycelium of Pleurotus pulmonis is cultivated at ambient temperature in a culture medium selected from three different culture media, namely in a defined culture medium for the production of component A, in a completely synthetic culture medium for the production of component B, or in a natural culture medium based on an extract from wine lees, characterized in that the particle size distribution determined by using different sets of sieves (DIN 10765 mod.) includes a maximum distribution of about 40% by weight between 2-4 mm, followed by a second maximum distribution of about 26% by weight between 1-2 mm.

[0374] The mycelial components of the 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 content of components A, B and C was 9.39%, 9.73% and 8.40%. The elemental analysis C, H, N, O, S analysis of these components resulted in:

[0375] For A: 44.45% (carbon C), 5.95% (H), 6.25% (N), 0.32% (S), 33.53% (O);

[0376] Carbon to nitrogen ratio (mycelium): 7.11.

[0377] For B: 43.92% (carbon C), 5.83% (H), 4.62% (N), 0.26% (S), 35.64% (O);

[0378] Carbon to nitrogen ratio (mycelium): 9.5.

[0379] For C: 46.90% (carbon C), 6.26% (H), 9.57% (N), 0.42% (S), 28.45% (O);

[0380] Carbon to nitrogen ratio (mycelium): 4.9.

[0381] The lower nitrogen content in sample B indicates a sample rich in fiber content but with lower protein content, while sample C (based on spent grains) has the highest nitrogen content, indicating a high protein content with lower fiber content. Sample A is in between. This suggests that protein and fiber content can be tailored by adjusting the composition of the synthetic culture medium or by using a nutrient side stream with an analyzed elemental composition, allowing the production of customized foods rich in fiber or protein.

[0382] The higher heating value (HHV, often referred to as gross calorific value) is determined directly using an oxygen bomb calorimeter as outlined in EN 14918:2009. The lower heating value (LHV, often referred to as net calorific value) is calculated based on the HHV and the elemental composition of the sample.

[0383] For A, the HHV and LHV were 18.4 MJ / kg (440 Kcal / 100 g) and 17.11 MJ / kg (410 Kcal / 100 g), respectively.

[0384] For B, HHV and LHV are 18.88 MJ / kg (450 Kcal / 100 g) and 17.61 MJ / kg (420 Kcal / 100 g), respectively.

[0385] 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.

[0386] As a percentage of the total sugar content in the mycelial fraction after hydrolysis, the most abundant sugars were A 88% glucan, B 92.5% glucan and C 78.6% glucan (here glucan is based on all glucose including glucose derived from other polysaccharides such as heteropolysaccharides and / or exopolysaccharides).

[0387] Sugars in one or more water extracts include sugars extracted from fresh mycelial components by water. Water extraction was performed using ASE 200 (accelerated solvent extractor) in an 11 ml stainless steel extraction cell, using deionized water at 100°C and 1500 psi (heating time 5 minutes, static time: 7 minutes, flushing volume: 150%, purge time: 180 seconds, static cycle: 3). The total sugars in the water extracts of components A, B and C were 5.65%, 7% and 3.26%, respectively, wherein for A, the sugars in the water extract expressed as % of the total extracted sugars included disaccharides (trehalose 20.1%), hexoses (glucose 22%, fructose 0.5%, mannose 0.1%, galactose 1.4%), pentoses (arabinose 0.1%) and sugar alcohols (mannitol 55%, sorbitol 0.7%). For B, the sugars in the water extract expressed by the % of sugars in the water extract include disaccharides (trehalose 19.2%), hexoses (glucose 43.3%, fructose 0.4%, galactose 0.1%), pentoses (arabinose 0.1%), and sugar alcohols (mannitol 35.7%, sorbitol 1%). For C, the sugars in the water extract expressed by the % of sugars in the water extract include disaccharides (trehalose 46%), hexoses (glucose 16%, galactose 0.1%), pentoses (arabinose 0.03% and xylose 0.15%), and sugar alcohols (mannitol 22.7%, arabitol 13.5%, xylitol 1%, sorbitol 0.64%).

[0388] In one embodiment, the aqueous extract of mycelial component A contains at least 50% by weight of mannitol in the extracted sugars (ie, relative to the total sugar content), which is the maximum amount of sugar extracted compared to the others.

[0389] In one embodiment, the aqueous extract of mycelial component B contains at least 40% by weight of glucose in the extracted sugars (ie, relative to the total sugar content), which is the maximum amount of sugar extracted compared to the others.

[0390] In one embodiment, the aqueous extract of mycelial fraction C contains at least 40 wt% trehalose in the extracted sugars (ie, relative to the total sugar content), which is the highest amount of sugar extracted compared to the others.

[0391] The known flavoring agents in the form of 5' nucleotides (5'NMP) were measured in g / kg, including 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP). Ingredient A showed a good abundance of 3.56 g / kg of 5'NMP, while B showed 1.78 g / kg. However, the use of spent grain extract resulted in a 10-fold increase in ingredient C compared to C, resulting in 10.43 g / kg of 5'NMP. In addition, ingredient C showed the highest concentration of 3.56 g / kg of uridine monophosphate (UMP), compared to 1.26 g / kg and 0.71 g / kg for A and B, respectively.

[0392] In the absence of any additional ribonucleic acid (RNA) reduction steps during the production of the ingredients by additional heating or pH treatment, the method of the present invention results in an inherent RNA level of the ingredients equal to or below 2 wt%, specifically 1.88 wt% for A, 1.65 wt% for B and 2 wt% for C in one specific embodiment.

[0393] Free bases include cytosine, uracil, guanine, hypoxanthine and adenine. The total free base of mycelium component A measured was 0.62 g / kg, while component B was 0.48 g / kg and component C was 1.3 g / kg, wherein for component A, adenine and uracil were observed to be the highest, 0.28 g / kg and 9.18 g / kg, respectively, and for component B, component A was observed to be the same but with lower concentrations, 0.17 g / kg and 0.13 g / kg, wherein for component C, higher amounts of adenine, uracil and cytosine were observed, 0.54 g / kg, 0.22 g / kg and 0.49 g / kg, respectively.

[0394] Component A (2.77 g / kg), component C (2.24 g / kg), followed by component C (g / kg), and then component B (1.11 g / kg) were the highest.

[0395] The chitin content of the components is about 7.6% by weight for A, about 6.9% by weight for B, and about 7.5% by weight for C. The ergothioneine content is about 91 mg / kg (for A), 127 mg / kg (for B), and 433 mg / kg. The ergosterol content of sample A was determined to be 2.35 mg / g dry weight, the ergosterol content of sample B was 2.24 mg / g dry weight, and the ergosterol content of sample C was 5.56 mg / g dry weight (it is known that oyster mushrooms have been reported to have 4.4 mg / g dry weight, especially for P. The fruiting bodies of Pleurotus pulmonarius grown on three types of forestry waste (pine, poplar, and honeysuckle) have an ergosterol content of 2.9-3.3 mg / g, which is lower than the ergosterol content found in mycelial component C, showing a higher enrichment than the mushroom fruiting bodies themselves.

[0396] All ingredients A, B and C exhibit at least about 20 wt% umami amino acids, more specifically A at 21.48 wt%, B at 19.5 wt%, and C at 22.64 wt%.

[0397] All ingredients A, B and C show a BCAA content of at least about 20 wt. %, more specifically 21.73 wt. % for A, 23.1 wt. % for B and 19.6 wt. % for C.

[0398] All ingredients A, B and C exhibit an essential amino acid content of at least about 40 wt %, more specifically 40.81 wt % for A, 40.52 wt % for B and 39.61 wt % for C.

[0399] 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).

[0400] Mycelial component A is grown on a medium containing a carbon to nitrogen ratio (medium) of 16-18, preferably about 17. Mycelial component B is grown on a medium containing a carbon to nitrogen ratio (medium) of 19-21, preferably about 20. Mycelial component B is grown on a medium containing a carbon to nitrogen ratio (medium) of 12-14, preferably about 13.

[0401] Insoluble fiber content increases from about 23 % by weight to about 34 % by weight to about 54 % by weight from C to A to B, while protein content decreases from about 60 % by weight to about 39 % by weight to about 32 % by weight from C to A to B. This shows the relationship between protein and fiber discussed above, which are inversely proportional and can be controlled by changing the fermentation medium composition. However, this trend may not always exist, as reported in the following research on the submerged fermentation of Pleurotus sclerotiorum, wherein the lower C / N ratio in the culture medium causes a higher total dietary fiber (Food Chemistry 85 (2004) 101-105) in the mycelial cell wall, and the trend observed by the present invention shows that fiber content increases with the increase of the C / N ratio. This shows that this trend may be highly dependent on culture medium composition (for example carbon source or nitrogen source), fermentation process conditions (for example pH and stirring speed), inoculum and / or other factors.

[0402] The fat content of component A is about 3% by weight. The fat content of component B is about 2% by weight.

[0403] The fat content of component C is about 7% by weight. The fat contains saturated fatty acids, monosaturated fatty acids, polyunsaturated fatty acids and trans fatty acids, as shown in the table below. It is worth noting that the content of ω-6 fatty acids (linoleic acid) is 0.77% by weight for A, 0.46% by weight for B and 3.5% by weight for C. All components show a calculated carbohydrate content of less than 0.1% by weight. B is the most abundant in all glucans, with a value of 31% by weight. Mycelial component A has a total glucan value of about 26% by weight. Mycelial component C has a total glucan value of about 17% by weight.

[0404] The EUC of the mycelial components was calculated to be 302% for A, 34% for B, and 2892% for C. In a separate study, Pleurotus pulmonarius fruiting bodies were grown on three types of forestry waste (pine, poplar, and honeysuckle), showing that the EUC values ​​of Pleurotus pulmonarius fruiting bodies were between 72.31% and 116.73% (Food Chemistry 397 (2022) 133714). The disclosed EUC values ​​of the mycelial components A, B, and C of the present invention are higher than the reported values ​​of the fruiting bodies of the same fungal strains. Taking component C as an example, the difference of component C is 25-40 times higher than the reported range of Pleurotus pulmonarius fruiting bodies. It is also known that the highest EUC value of mushroom fruiting bodies is reported to be 4465% (Strawberry), which is 2.35 times lower than component C.

[0405] It was noted that the mycelium (non-fruiting body) was reported to have a lower EUC value than Pleurotus eryngii fruiting bodies (30.9% EUC mycelium vs. 116% fruiting bodies), Agrocybe edulis fruiting bodies (19.2% EUC mycelium vs. 322% EUC fruiting bodies), and Lentinula edodes fruiting bodies (16.7% EUC mycelium vs. 99.75% EUC fruiting bodies) (https: / / doi.org / 10.1080 / 10942912.2015.1089891), which highlights the significant and unique flavor of the mycelium component of the present invention through submerged fermentation. When further researching the literature, EUCs of mycelia of the following species were reported: Termitomyces albuminosus 460% Wen (2003), Grifola frondosa 375% Wen (2003), Grifola frondosa 363% Wen (2003), Diplosporus maguei 128% Lee (2003), Sphaerotheca pombe 124% Chang et al. (2001), Pleurotus ostreatus 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 submerged fermented 9 to 144% (all these references are correspondingly disclosed in Mau et al., International Journal of Medicinal Mushroom, Vol. 7, pp. 119-125 (2005)), which can all be used as a basis to show that the mycelium component of the present invention shows an unprecedented high-spectrum umami taste for use in the food industry and foods (such as dairy analogs, meat analogs or other foods).

[0406] In addition, according to the data disclosed in WO2021234349 (Tables 1 and 2), comparing mycelial components A and C with the unwashed fungal protein product from Quorn, the maximum EUC can be approximated to be at most 148% (it is known that the actual value of EUC should be lower than 148% because in this calculation, total amino acids are used instead of free amino acids due to lack of data).

[0407] It is interesting to note that the samples grown on the side stream have the most abundant values ​​in terms of total amino acid content, umami, ergothioneine content, ergosterol content and other properties, which shows a great advantage in using natural side streams or waste streams to produce more abundant food, while recycling waste streams from other value chains, which helps sustainability. However, these ingredients are different in their properties, which makes them suitable for different applications. For example, ingredient B is used for sports supplements because it has a low fat content and a high fiber content, and the BCAA amino acids have sufficient protein content.

[0408] The present invention also relates to methods of producing edible dairy substitutes, meat substitutes and fish / seafood substitutes using the fungal ingredients of the present invention.

[0409] In certain embodiments, the mycelial component can be used in the form of wet biomass (as is), washed biomass, dried biomass, ground into a powder having a specific particle size distribution, whether having a fine particle size or a coarse particle size or a medium particle size.

[0410] A method for producing a soft or hard meat analog composition comprising at least one mycelial component from at least one fungal strain, the method comprising a method for producing the corresponding mycelial component, further comprising the step of preparing such a meat analog composition by mixing at least one of the at least one mycelial component from at least one fungal strain with a composition comprising at least one protein-rich ingredient, at least one vegetable-based lipid-rich ingredient and optionally at least one constituent ingredient.

[0411] In one embodiment, 50 to 95% by weight of mycelium component (edible fibrous mycelium) is mixed, then up to 1 to 40% by weight of each of the following ingredients are added: rapeseed oil, salt, egg white and wheat gluten, then optionally up to 1 to 40% by weight of each of the following ingredients are added: methylcellulose, hydrocolloid, texturized vegetable protein, starch-based ingredients, flavoring components. The obtained dough is then shaped by a molding machine and / or an extruder or a combination thereof to have a final product as a meat ball or sausage or an extruded product.

[0412] A method for producing a dairy analog composition comprising at least one mycelial component from at least one fungal strain, the method comprising a method for producing the corresponding mycelial component, and further comprising the step of preparing such dairy analog composition by the steps of: (1) forming a slurry comprising the at least one mycelial component with a composition comprising at least one protein-rich ingredient, at least one vegetable-based lipid-rich ingredient and at least one constituent ingredient, and (2) mixing the slurry with at least one constituent ingredient, in particular a texturizing agent or thickening agent or a carbohydrate-rich ingredient.

[0413] In one embodiment, the method of producing an edible milk substitute product does not comprise an additional acidification step, but rather is acidified by formulating:

[0414] (1) about 55 wt% of drinking water, about 10-45 wt% of a fat component of plant origin, up to 2 wt% of a sodium chloride source, up to 2 wt% of yeast flakes, up to 5 wt% of a monosaccharide or disaccharide source of plant origin, 5 wt% of an acidic source of natural plant origin,

[0415] 10-50% by weight of edible fibrous mycelium derived from submerged fermentation of at least one fungal strain is homogenized.

[0416] (2) Heat the homogenized slurry at a temperature of 75 to 95°C with constant mixing for up to 60 seconds.

[0417] In order to coagulate the homogenized slurry, during the heating step defined previously, once

[0418] Having reached a temperature of 25° C. to 40° C., a previously prepared coagulant solution is applied. The coagulant solution contains up to 15% by weight of each of: - water, wheat starch and hydrolyzed corn starch.

[0419] (3) Remove the obtained slurry from the heat source and then allow the mixture to settle to room temperature (21°C). Once it reaches room temperature, store the coagulated slurry in a dark and cooled environment, preferably at 4°C.

[0420] to 7°C.

[0421] In another embodiment, a method of producing an edible non-animal dairy alternative product comprises:

[0422] (1) using about 55% by weight of drinking water, about 10-45% by weight of a vegetable-derived fat component,

[0423] Up to 2% by weight of a sodium chloride source, up to 2% by weight of yeast flakes, up to 5% by weight of a mono- or disaccharide source of plant origin, and 10-50% by weight of a source of at least one fungus.

[0424] Submerged fermentation of edible fibrous mycelium homogenized from the strain.

[0425] (2) Heat the homogenized slurry at a temperature of 75 to 95°C with constant mixing for up to 60 seconds.

[0426] In order to coagulate the homogenized slurry, during the heating step defined previously, once

[0427] Having reached a temperature of 25° C. to 40° C., a previously prepared coagulant solution is applied. The coagulant solution contains up to 15% by weight of each of: - water, wheat starch and hydrolyzed corn starch.

[0428] (3) The obtained slurry is removed from the heat source, and the mixture is subsequently allowed to settle to room temperature (21°C). Once the obtained mixture reaches a temperature of 40°C or lower, 0.1 g to 0.25 g of acid-producing bacteria, particularly lactic acid bacteria, is added to induce acidification of the microorganisms. In order to provide a hospital environment for microorganisms, the sample is placed in a controlled temperature environment (e.g., a water bath or an incubator) at 28°C.

[0429] 150 minutes. Depending on the intensity of the fermentation, the sample can be placed at a temperature of 20 to 45°C for 1 to 6 hours. After the microbial fermentation is complete, the coagulated slurry is stored in a dark and cooled environment, preferably between 4°C and 7°C.

[0430] 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 a combination thereof. In a preferred embodiment, mycelium from one fungal strain is partially replaced with mycelium from another fungal strain to produce a dairy or meat analog, and the total mycelium content is in a ratio of 1:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 or 100:1. In a preferred embodiment, if 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, the edible fibrous mycelium of Pleurotus pulmonarius is mixed with the edible fibrous mycelium of Morchella edulis, wherein 60-90% by weight of the total mycelium is derived from the mycelium of Pleurotus pulmonarius, and 10-40% by weight of the total mycelium component is derived from the mycelium of Morchella edulis (edible fibrous mycelium). In another embodiment, the edible fibrous mycelium of Pleurotus pulmonarius is mixed with the edible fibrous mycelium of Morchella edulis, wherein 60-90% by weight of the total mycelium is derived from the mycelium of Morchella edulis, and 10-40% by weight of the total mycelium component is derived from the mycelium of Morchella edulis (edible fibrous mycelium).

[0431] In another embodiment, the same method is extended to at least three fungal strains.

[0432] Thereby an edible non-animal cheese substitute product is obtained, the edible non-animal cheese substitute product being selected from the group comprising: edible non-animal substitute products for whey cheese, cream cheese, medium-hard cheese, hard cheese and soft cheese. Thereby an edible cheese substitute product being cream cheese is obtained.

[0433] Adding 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 alternative product to be produced, it is not necessary to add a texturizing agent to obtain the texture.

[0434] In a preferred embodiment, the product obtained with this method is an edible fresh cheese substitute or cream cheese. It can also be an edible non-animal cheese substitute product selected from the group comprising whey cheese, cream cheese, medium-hard cheese, hard cheese and soft cheese, which can be obtained without the traditional coagulation step if the water content of the homogenized mycelial mass is appropriately adjusted to be low enough. The coagulation step of traditional cheese making is carried out to remove the excess water provided by the raw material (which is milk). These coagulation steps of traditional cheese making are unnecessary due to the raw material, i.e. the homogenized mycelial mass, having a significantly lower water content than milk.

[0435] In the method for producing an edible non-animal dairy alternative product for fresh cheese, whey cheese, cream cheese, medium-hard cheese, hard cheese and soft 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, algal oil or oil derived from oleaginous yeast.

[0436] In another preferred embodiment, in particular when the edible non-animal dairy alternative product is a yogurt or cheese alternative product, the edible non-animal dairy alternative product further comprises an edible plant- or algae-based fat component, a fat component derived from fungi or yeast. The fat component is preferably in the range of up to 60 wt%, preferably up to 25 wt%, and most preferably 1 wt% to 5 wt%. The amount of the fat component can be adjusted depending on the product to be replaced.

[0437] The edible non-animal dairy alternative product of the present invention preferably comprises edible fibrous mycelial mass having a protein content of between 1 wt.% and 99 wt.%, preferably between 10 wt.% and 90 wt.%, and most preferably between 1 wt.% and 65 wt.% of the dry mass of the fibrous mycelial mass. When discussing the content of edible fibrous mycelial mass in the product of the present invention, reference is preferably made to the wt.% in the aqueous product (i.e. not standardized to dry matter content).

[0438] Preferably, the edible fiber mycelium in the milk analog product has an insoluble fiber content of at least 20 wt%, more preferably at least 30 wt%, more preferably at least 40 wt%. Therefore, preferably the insoluble fiber content is 20 wt% to 60 wt%. More preferably, the edible fiber mycelium has an insoluble fiber content between 39 wt / wt% and 60 wt / wt%. Even more preferably, the edible fiber mycelium has an insoluble fiber content between 40 wt / wt% and 55 wt / wt%. Even more preferably, the edible fiber mycelium has an insoluble fiber content between 40 wt / wt% and 55 wt / wt%. Most preferably, the edible fiber mycelium has an insoluble fiber content of about 45 wt / wt%. Most preferably, the edible fiber mycelium has an insoluble fiber content of about 45 wt / wt%. It should be understood that the weight referred to herein relates to the dry mass of the mycelium. In one embodiment, the edible fibrous mycelium has an insoluble fiber content of at least 40 wt / wt%, preferably at least 50 wt / wt%, more preferably at least 60 wt / wt%.

[0439] The protein content of the mycelial mass can be adjusted by selecting fermentation conditions that control the protein content of the resulting mycelial mass. For example, the ratio between the carbon source and the nitrogen source in the culture medium can be varied, such as providing different excess levels of nitrogen.

[0440] In some embodiments, particularly when the edible non-animal dairy substitute product is a substitute for yogurt or cheese, the edible non-animal dairy substitute product may also include a texturizing agent, such as agar, edible starch, guar gum, locust bean gum, wheat gluten, cellulose or a derivative thereof. The texturizing agent imparts structure to the product, making it suitable as a substitute for yogurt or cheese. However, the texturizing agent is an optional component for the edible non-animal dairy substitute product of the present invention. If the water content of the mycelium homogenized with water is kept low by using pressed or partially dried mycelium with the desired water content, the resulting material will have a desired structure similar to yogurt or cheese without the need to add an external texturizing agent. Those skilled in the art will know how to appropriately adjust the water content of the mycelium mass depending on the substitute product to be produced.

[0441] In one embodiment, the acidification by ingredients is accomplished by homogenizing 10-50 wt% of Pleurotus pulmonarius mycelia (edible fibrous mycelia) with about 55 wt% of drinking water, about 0 to 7 wt% of cashew nuts (vegetable fat component), 10 to 40 wt% of coconut oil (vegetable fat component), up to 2 wt% of common salt (sodium chloride source), up to 2 wt% of yeast flakes, up to 5 wt% of sucrose (vegetable sugar source), up to 5 wt% of lemon juice (natural vegetable acid source), up to 5 wt% of citric acid. The homogenized slurry is heated at a temperature of 75-95° C. for a maximum of 60 seconds under constant mixing. Once the temperature has reached 25° C. to 40° C., a previously prepared coagulant is applied during this previously defined heating step. The coagulant solution contains up to 15 wt% of each of: - water, wheat starch and hydrolyzed corn starch, the slurry obtained is removed from the heat source and the mixture is subsequently allowed to settle to room temperature (21° C.). Once it reaches room temperature, the coagulated slurry is stored in a dark and cooled environment, preferably between 4°C and 7°C, to obtain a mycelium based creme cheese product.

[0442] In one embodiment, the acidification by microbial fermentation is carried out by homogenizing 10-50% by weight of Pleurotus pulmonarius mycelia (edible fibrous mycelia) with about 55% by weight of drinking water, about 0-7% by weight of cashew nuts (vegetable fat component), 10-40% by weight of coconut oil (vegetable fat component), up to 2% by weight of common salt (sodium chloride source), up to 2% by weight of yeast flakes, up to 5% by weight of sucrose (vegetable sugar source). The homogenized slurry is heated at a temperature of 75-95° C. for up to 60 seconds under constant mixing. Once the temperature has reached 25° C. to 40° C., a previously prepared coagulant is applied during this previously defined heating step. The coagulant solution contains up to 15% by weight of each of: - water, wheat starch and hydrolyzed corn starch, the obtained slurry is removed from the heat source and the mixture is subsequently allowed to settle to room temperature (21° C.). Once the obtained mixture reaches a temperature of 40°C or less, 0.1 g to 0.25 g of acid-producing 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 the fermentation, the sample may be placed at a temperature of 20 to 45°C for 1 to 6 hours. After the microbial fermentation is complete, the coagulated slurry is stored in a dark and cooled environment, preferably between 4°C and 7°C, to obtain a mycelium-based cream cheese product.

[0443] The above-mentioned adjustability of the composition of the mycelial mass of the present invention provides the advantage of versatility, wherein the composition of the mycelial mass can be easily adjusted to the requirements of the alternative product to be obtained. If the protein content of the mycelial mass is increased, for example, the amount of mycelial mass in the alternative product can be reduced without changing the protein content of the alternative product. If the alternative 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, thereby eliminating the need to supplement the alternative product with additional nutrients from the outside. This reduces the cost of the production method and the resulting product because it is easier to handle, has fewer production steps, and reduces the cost of raw materials for the production of edible non-animal dairy alternative products with a clean label. Most importantly, the nutrients from the mycelium are bioavailable, making metabolic digestion predictable and easy, especially compared to products containing externally supplemented nutrients.

[0444] In one embodiment, the mixture of at least two mycelial components is used to produce dairy analogs. In another embodiment, the mixture of at least two mycelial components is used to produce meat analogs. In a preferred embodiment, the mixture of at least two mycelial components is used to produce dairy and meat analogs. In a preferred embodiment, the mixture of at least two mycelial components is used to produce fish analogs. In one embodiment, the mixture of mycelial components is used to produce other food products as defined above.

[0445] In another embodiment, a method of producing an edible non-animal dairy alternative product comprises:

[0446] (1) 1-99 wt %, preferably 50-95 wt % of an edible mycelium component is mixed and added to a mixture of at least one protein-rich component and at least one vegetable lipid-rich component.

[0447] (2) Optionally adding at least one ingredient, such ingredients include methylcellulose, hydrocolloids, textured vegetable protein, starch-based ingredients, fiber-rich ingredients, and flavor components for seasoning.

[0448] (3) shaping the obtained dough by an extruder and / or a shaping machine so that the final product is a meat ball or a sausage or an extruded product, wherein step 3 comprises:

[0449] (i) The obtained dough is cold-extruded 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 of a filling forming machine.

[0450] (ii) Forming the product into the desired shape

[0451] (iii) as an alternative to step (ii), in the case of sausage products, filling the casing

[0452] (iv) Boiling the product in water at a temperature of 60-100°C for 1-30 minutes to increase the texture of the mycelial product due to protein denaturation and shelf life stability of the product.

[0453] (v) Alternatively, step (iv) may be replaced by frying and then steaming or hot air cooking the product at a temperature of 50-150°C.

[0454] (vi) Store the final product in a refrigerator

[0455] The edible meat substitute products of the present invention preferably contain an edible fibrous mycelial material 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, wherein the edible fibrous mycelium in the meat analog product preferably has an insoluble fiber content of 20% to 60% by weight, preferably 40% to 50% by weight, preferably about 45% by weight. In another preferred embodiment, the insoluble fiber used in the meat analog is about 25% by weight. In another preferred embodiment, the insoluble fiber used in the meat analog is about 45% by weight. In one embodiment, the edible fibrous mycelium in the meat analog 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.

[0456] The protein content of the mycelial mass can be adjusted by selecting fermentation conditions that control the protein content of the resulting mycelial mass. For example, the ratio between the carbon source and the nitrogen source in the culture medium can be varied, such as providing different excess levels of nitrogen.

[0457] In one embodiment, the at least one vegetable lipid-rich ingredient comprises at most 40 wt%, at most 20 wt%, at most 15 wt%, at most 10 wt%, at most 9 wt%, at most 8 wt%, at most 7 wt%, at most 6 wt%, at most 5 wt%, at most 4 wt%, at most 3 wt%, at most 2 wt% or at most 1 wt% of the total formulation of the meat analog composition. In a preferred embodiment, the at least one vegetable lipid-rich ingredient of the meat analog composition is 1-40 wt%, preferably 1-20 wt%, more preferably 1-10 wt%.

[0458] In one embodiment, the at least one protein-rich ingredient of the meat analog composition each accounts for up to 95% by weight, up to 80% by weight, up to 60% by weight, up to 40% by weight, up to 20% by weight, up to 15% by weight, up to 10% by weight, up to 9% by weight, up to 8% by weight, up to 7% by weight, up to 6% by weight, up to 5% by weight, up to 4% by weight, up to 3% by weight, up to 2% by weight, or up to 1% by weight of the total formula of the meat analog composition. In a preferred embodiment, the at least one protein-rich ingredient of the meat analog composition is each 0.1-50% by weight, preferably 0.1-30% by weight, and more preferably 0.1-20% by weight.

[0459] In one embodiment, at least the constituent components of the meat analog composition each account for up to 80 wt%, up to 60 wt%, up to 40 wt%, up to 20 wt%, up to 15 wt%, up to 10 wt%, up to 9 wt%, up to 8 wt%, up to 7 wt%, up to 6 wt%, up to 5 wt%, up to 4 wt%, up to 3 wt%, up to 2 wt% or up to 1 wt% of the total formulation of the meat analog composition. In a preferred embodiment, at least one constituent component of the meat analog composition is each 0.1-50 wt%, preferably 0.1-30 wt%, more preferably 0.1-20 wt%.

[0460] In one embodiment, the protein-rich ingredients include wheat gluten. In one embodiment, the protein-rich ingredients include wheat gluten and egg white. In one embodiment, the composition ingredients include flavoring components. In one embodiment, the composition ingredients include flavor components and starch-based ingredients. In one embodiment, the composition ingredients include flavor components, starch-based ingredients and methylcellulose. In one embodiment, the composition ingredients include flavoring components and methylcellulose. In one embodiment, the composition ingredients include flavor components, starch-based ingredients and hydrocolloids. In one embodiment, the composition ingredients include flavor components and hydrocolloids. In one embodiment, the composition ingredients include flavor components, starch-based ingredients and organized vegetable proteins. In one embodiment, the composition ingredients include flavor components and organized vegetable proteins. In one embodiment, the composition ingredients include flavor components and fiber-rich ingredients. In one embodiment, the composition ingredients include flavor components, fiber-rich ingredients and starch-based ingredients. In one embodiment, the composition ingredients include flavor components, starch-based ingredients, fiber-rich ingredients and methylcellulose. In one embodiment, the composition ingredients include flavor components and fiber-rich methylcellulose ingredients. In one embodiment, the composition ingredients include flavor components, starch-based ingredients, fiber-rich ingredients and hydrocolloids. In one embodiment, the composition comprises flavor components, fiber-rich ingredients and hydrocolloids. In one embodiment, the composition comprises components, starch-based ingredients, fiber-rich ingredients and textured plant proteins. In one embodiment, the composition comprises flavor components, fiber-rich ingredients and textured plant proteins. In one embodiment, the composition comprises methylcellulose, hydrocolloids, textured plant proteins, starch-based ingredients, fiber-rich ingredients and flavor components for seasoning. In one embodiment, the spice component (also understood as under the composition) comprises at least one of the following ingredients: salt, pepper, garlic, onion, mushroom fruiting body slices, 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: cereal-based flour, cereal-based starch, bean-based starch, fruit-based fiber, polysaccharides, starch-based ingredients, oil husks, inulin, wheat starch and corn starch. It should be noted that a fiber-rich ingredient may also be a starch or carbohydrate-rich ingredient, as a starch-based ingredient or a carbohydrate-rich ingredient may also be a fiber-rich ingredient.

[0461] In one embodiment, the method of producing the same meat analog product by expanding it comprises replacing egg white with more mycelium component of the present invention, wherein the dry mycelium component will increase by 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 wt%, 6 wt%, 5 wt%, 4 wt%, 3 wt%, 2 wt% or 1 wt%. In a preferred embodiment, egg white is replaced by up to 15 to 20 wt%, preferably up to 7 to 10 wt% of dry mycelium component. This is mainly determined by the quality and type of biomass to preferably replace any dairy derived product or vegetarian product to make such a product palatable.

[0462] In a further embodiment, in the method of producing a veggie meat analog, 1 wt% of egg white is preferably replaced or equivalent to 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% dry mycelial biomass. In a preferred embodiment, in the method of producing a veggie meat analog, 1 wt% of egg white is preferably replaced or equivalent to up to 10 wt%, preferably up to 5 wt%, more preferably 2.5 wt% to 4 wt% equivalent, most preferably 1.5 to 2 wt% equivalent dry mycelial component.

[0463] In a preferred embodiment, in the method for producing a soft meat analog, 1 wt% of egg white is preferably replaced or equivalent by up to 10 wt%, preferably up to 5 wt%, more preferably 2.5 to 4 wt% equivalent, most preferably 1.5 to 2 wt% equivalent of dry mycelium component, wherein the cutting strength of the original vegetarian product and the cutting strength of the rough product are in the same range of 1 to 25N, most preferably 4 to 8N.

[0464] For example, for a mycelium-based meatball composition, when about 1 to 2 wt%, preferably about 1.5 wt% of dry biomass is used to replace every 1 wt% of egg white, the same cutting strength of 4.8 N is achieved. Changing this ratio will result in a different texture, i.e. a different cutting strength.

[0465] In a preferred embodiment, in the method for producing a tough meat analog, 1 wt% of egg white is preferably replaced by or equivalent to up to 10 wt%, preferably up to 5 wt%, more preferably 2.5-4 wt%, most preferably 1.5-2 wt% of dry mycelium component, wherein the cutting strength of the original vegetarian product and the tough meat product is in the same range of 1-50N, most preferably 10-30N.

[0466] In another embodiment, 50-95% by weight of Pleurotus pulmonarius mycelium or any edible fibrous mycelium is mixed and then up to 1-40% by weight of each of the following ingredients is added: rapeseed oil, salt and gluten. The missing egg white is calculated by adding additional dry mycelium, and the equivalent ratio is 1 to 1.4, respectively. Optionally, up to 1 to 40% by weight of each of the following ingredients is added: methylcellulose, hydrocolloid, texturized vegetable protein, starch-based ingredients and flavoring components for seasoning. The obtained dough is then shaped by a molding machine and / or an extruder or a combination thereof to have a final product as a meat ball or sausage or an extruded product.

[0467] In another embodiment, if shaping is required, the obtained dough or formulation can be subjected to an extrusion process before or after shaping to form a meat product or other extrudable food product, wherein the extrusion is a cold extruder, low temperature extrusion or high temperature extrusion.

[0468] In one embodiment, cold extrusion is employed, wherein the extrusion temperature is 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, and the pressure range is 1 to 100 bar, more preferably 1 to 70 bar. In a preferred embodiment, the extrusion is carried out at a temperature of freezing conditions or at low temperatures or at room temperature, preferably at a temperature of up to -20°C, more preferably 1-20°C, most preferably 4-15°C, at a pressure of 1-100 bar, more preferably 1-70 bar.

[0469] In one embodiment, the shaping is carried out, wherein the shaping temperature is preferably 1-20° C., preferably 4-15° C., preferably 10-15° C., preferably 15-25° C., preferably 25-40° C., preferably 40-100° C., preferably 40-80° C., preferably 50-75° C., at a pressure of 1-100 bar, more preferably 1-70 bar. In a preferred embodiment, the shaping is carried out at low temperature or room temperature, preferably at a temperature of 1-25° C., most preferably 4-15° C., at a pressure of 1-100 bar, more preferably 1-70 bar.

[0470] In one embodiment, low temperature extrusion is employed, wherein the extrusion temperature is preferably 30-100°C, more preferably 40-80°C, more preferably 50-75°C at a pressure of 5-25 bar.

[0471] In a second embodiment, high temperature extrusion is used, wherein the extrusion temperature is preferably between 100 and 190°C, more preferably between 120 and 180°C, more preferably between 130 and 180°C, at a pressure of 100 to 300 bar, wherein the extruded moisture is preferably less than 45% by weight, preferably less than 40% by weight, preferably less than 35% by weight.

[0472] In a third embodiment, high temperature extrusion is used, wherein the extrusion temperature is preferably between 100 and 190°C, more preferably between 120 and 180°C, more preferably between 130 and 180°C, at a pressure of 100 to 300 bar, wherein the extruded moisture is preferably higher than 40% by weight, preferably higher than 45% by weight, preferably higher than 50% by weight.

[0473] In one embodiment, the rope of extruded dough has a thickness of 1-8 cm, preferably 1.5-5 cm, more preferably 1.5-3.5 cm.

[0474] In another embodiment, the product is frozen and stored at a temperature 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 between -40°C and 5°C, more preferably between -20°C and 5°C, most preferably between -5°C and 5°C. In another preferred embodiment, the product is stored at about -20°C. In another preferred embodiment, the product is stored at about -4°C. In another embodiment, supercooling of the food is applied, wherein the product is stored at a temperature between -0.5 and -5°C.

[0475] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of 30-200 g MSG / 100 g.

[0476] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of less than 200 g MSG / 100 g.

[0477] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of 200-500 g MSG / 100 g.

[0478] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of at least 30 g MSG / 100 g.

[0479] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of at least 500 g MSG / 100 g.

[0480] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of at least 1000 g MSG / 100 g.

[0481] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of at least 1500 g MSG / 100 g.

[0482] In one embodiment, the edible meat substitute product comprises 1-99 wt% mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20-60 wt% and an EUC of at least 2000 g MSG / 100 g.

[0483] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of 30 to 200 g MSG / 100 g.

[0484] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of less than 200 g MSG / 100 g.

[0485] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of 200 to 500 g MSG / 100 g.

[0486] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of at least 30 g MSG / 100 g.

[0487] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of at least 500 g MSG / 100 g.

[0488] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of at least 1000 g MSG / 100 g.

[0489] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of at least 1500 g MSG / 100 g.

[0490] In one embodiment, the edible dairy alternative product comprises 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of at least 2000 g MSG / 100 g.

[0491] In a preferred embodiment, the edible product preferably comprises a mycelial component obtained from the genus Pleurotus (Pleurotus ostreatus family or Pleurotus genus), for example a fungus selected from the group consisting of Pleurotus pulmonatus, Pleurotus ostreatus, Pleurotus ostreatus, Pleurotus ostreatus and Pleurotus rosaceus, preferably Pleurotus pulmonatus; or derived from a fungus selected from the group consisting of Morchella esculenta, Morchella angusticeps, Morchella deliciosa and Morchella chellarufobrunnea, preferably Morchella esculenta.

[0492] In a preferred embodiment, the edible product comprises mycelial components obtained from Pleurotus pulmonarius. In another preferred embodiment, the edible product comprises mycelial components obtained from Morchella rubra. In a further embodiment, the edible product comprises mycelial components obtained from a combination of Pleurotus pulmonarius and Morchella rubra. In another embodiment, the edible product comprises mycelial components obtained from L. sulphur. In another embodiment, the edible product comprises mycelial components obtained from Morchella fusca.

[0493] The meat and dairy analogs may be vegetarian or vegan. The above embodiments are also applicable to non-meat or dairy analog products, which may include, but are not limited to, fish substitutes, confectionery, bakery products, flour (including bread or pasta or noodles), sweets and desserts, snacks, cereal products, alcoholic and non-alcoholic beverages, spice mixes, ready meals, frozen meals, gelled foods, protein supplements, and extruded / expanded products.

[0494] Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope of the present invention. Although the present invention has been described in conjunction with specific preferred embodiments, it should be understood that the claimed invention should not be unduly limited to these specific embodiments. In fact, the present invention is intended to encompass various modifications of the described modes for implementing the present invention that are apparent to those skilled in the relevant art.

[0495] The following examples are merely illustrative of the present invention and should not be construed as limiting in any way the scope of the present invention as defined by the appended claims.

[0496] Example

[0497] For each component determined by wet chemical analysis, each sample was analyzed at least in duplicate, with percentages expressed on a dry basis. The following specific examples were performed on a fungal strain from Pleurotus pulmonatus. It should be understood that in the methods and products of the present invention, Pleurotus ostreatus, in particular Pleurotus pulmonatus, is a preferred fungal strain. However, the present invention is not limited to this strain and may be implemented with other fungal strains.

[0498] Example of ingredient production

[0499] For example, medium A (defined medium) comprises the following composition for producing component A: 0.45 wt % C, 0.026 wt % N, 1.24 wt % O and 0.326 wt % P, wherein the nitrogen source is CSL (10 g / l) and the carbon source is dextrose.

[0500] Medium B (synthetic medium) comprises the following composition: 0.41 wt% C, 0.02 wt% N, 1.2 wt% O and 0.326 wt% P for the production of component B, wherein the amino acids are selected from arginine (0.62 g / l) and the vitamins are derived from 1 mg / L D-biotin, 1 mg / L folic acid, 1 mg / L nicotinamide, 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 glucose.

[0501] Medium C (natural complex medium) comprises a lees extract extracted using steam pretreatment at 170°C for less than 5 minutes, in particular 3 minutes, followed by nutrient recovery by washing in water, wherein the final protein content of the extract used for fermentation is about 10 g / l, the total glutamic acid concentration is 566 mg / L, the total aspartic acid concentration is 216 mg / L, and the medium of C comprises the following composition: 0.772 wt% C, 0.059 wt% N, 1.6 wt% O and 0.326 wt% P. The spent grains have a particle size distribution, wherein 2-4 mm constitutes the highest range of the distribution, accounting for about 35 wt% of the total particle size distribution present. The carbon-nitrogen ratio of the extract is about 11 (extract composition based on CHNO analysis: 1 wt% C, 0.093 wt% N, 0.81 wt% O and 0.12 wt% H).

[0502] 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), zinc (0.05 g / l).

[0503] Mycelium production is divided into two different strains. Seed system and specific main fermentation in respective culture medium. Seed system is divided into three different steps. First, mycelium is cultured on a culture dish on PDA culture medium. Then, culture at 28 ℃ for 7 days in one of culture medium A or B or C. Then the whole culture is cultured at ambient temperature. For each corresponding main fermentation, a culture from seed system is used and aerobically cultured for about 5 to 6 days in a fermentation system cultured in a fermenter until the carbon source is completely consumed. In all three cases (A, B and C), fermentation is carried out aerobically at ambient temperature (25 ℃) and a pH of 4-5. In these embodiments, the pH of the fermentation medium is about 5. In fermentation, it is also common to control dissolved oxygen. Then the ingredients are harvested and prepared for further processing after washing with acidic water of pH 3.5. It should be understood that unless the contrary is clearly stated, the culture medium composition used herein (e.g., the specific composition of culture medium A, B and C) corresponds to the culture medium composition at the beginning of fermentation. As described above, it will be apparent to the skilled person that over a period of time the composition of the culture medium may change and, for example, certain components (eg, carbon source) may be at least partially depleted.

[0504] Freeze drying method of components A, B and C

[0505] Freeze drying was carried out in a "Christ Alpha 1-2" freeze dryer connected to an external vacuum pump, where each sample (A, B or C) was freeze dried at -50°C, with a vacuum of 6 mbar applied directly. All samples were pre-frozen at -80°C. The samples were freeze dried for a duration of 72 hours.

[0506] BET surface by gas adsorption (according to DIN-ISO9277, DIN66131 respectively)

[0507] In the gas adsorption process (according to DIN-ISO 9277 and DIN 66131, respectively), the specific surface area of ​​solid substances is determined by default by nitrogen adsorption at 77.4 K using the BET method. Sample preparation is carried out at the respective specified temperature. At the same time, the sample surface must not change. In the statistical volume method, a specific amount of measuring gas is metered onto a sample arranged in a vacuum. The determination of the amount of adsorbed gas is based on the gas equation and the pressure measurement in a volume calibration system. In the dynamic method, the determination of the amount of adsorbed gas is carried out by a thermal conductivity sensor that detects the change in the gas composition in the N2 / He mixture. The molecular weight in the monolayer on the surface of the solid substance is calculated from the measurement readings (adsorption volume Va versus relative pressure p / p0). The evaluation is carried out in the relative pressure range mentioned in the general validity range of the BET method of p / p0=0.05-0.3. For the determination of very small surfaces, krypton adsorption (at 77.4K) is used. Because the expected surface area is very low, the sample cell has been completely filled with sample (about 0.5-0,6 g / sample - see attached data sheet) and krypton has been used, because it is a suitable adsorbent for measuring low surface areas. Prior to analysis, the samples were degassed under vacuum at room temperature for about 65 hours. Analyses were performed on a Quantachrome Quadrasorb.

[0508] Mercury porosimetry (according to DIN-ISO 15901-1, DIN66133)

[0509] Pore ​​analysis by mercury porosimetry was performed using Quantachrome Poremaster-60GT.

[0510] The method is based on the Washburn-Equation, which describes the relationship between pore size and the pressure exerted on a non-wetting liquid such as mercury. With the Poremaster-60GT, the filling of the penetrometer is carried out in a horizontal position before the analysis: due to this fact, hydrostatic mercury pressure on the sample is also avoided as the pore filling is not detected. The resulting intrusion curves are plotted against pressure, respectively pore size. Because the measurement is started at low pressure, the large pores will be filled first, so that in the standard case the pore size decreases from the left to the right of the diagram. The method upper limit is about 1 mm pore size, which means that the detection of larger pores is not the goal of this experiment.

[0511] Free base, nucleosides and 5′NMP nucleotides

[0512] The freeze-dried fraction was dissolved in 25 mg / mL water, heated at 100°C for 10 minutes and homogenized. For protein precipitation, TCA was added to a final concentration of 5% and incubated at -20°C for 16 hours. 5 μL of the deproteinized extract was then injected and analyzed by ion-pair HPLC.

[0513] RNA Quantification Methods

[0514] For RNA quantification, nuclease was added to the dissolved sample (25 mg / mL) and incubated at 60°C for 16 hours to convert RNA to 5'NMP. For protein precipitation, TCA was added to a final concentration of 5% and incubated at -20°C for 16 hours. 5 μL of the deproteinized extract was then injected and analyzed by ion pair HPLC.

[0515] The known flavoring agents in the form of 5' nucleotides (5'NMP) were measured in g / kg, including 5'-inosine monophosphate (IMP), 5'-guanosine monophosphate (GMP) and 5'-adenosine monophosphate (AMP). Ingredient A showed a good abundance of 3.56 g / kg of 5'NMP, while B showed 1.78 g / kg. However, the use of spent grain extract resulted in a 10-fold increase in ingredient C compared to C, resulting in 10.43 g / kg of 5'NMP. In addition, ingredient C showed the highest concentration of 3.56 g / kg of uridine monophosphate (UMP), compared to 1.26 g / kg and 0.71 g / kg for A and B, respectively.

[0516] In the absence of any additional ribonucleic acid (RNA) reduction steps during the production of the ingredients by additional heating or pH treatment, the methods of the present invention result in ingredients with inherent RNA levels below 2 wt%, specifically 1.88 wt% for A, 1.65 wt% for B and 2 wt% for C in one specific embodiment.

[0517] Table 1: 5'-nucleotide and RNA content of mycelial components A, B and C

[0518]

[0519]

[0520] Amino Acid Quantitation Methods and Data

[0521] Amino acid analysis was performed by acid hydrolysis of the samples followed by HPAEC-IPAD. Tryptophan was not detected using this method because tryptophan undergoes degradation during acid hydrolysis of the samples.

[0522] All ingredients A, B and C exhibit at least about 20 wt% umami amino acids, more specifically A at 21.48 wt%, B at 19.5 wt%, and C at 22.64 wt%.

[0523] All ingredients A, B and C show a BCAA content of at least about 20 wt. %, more specifically 21.73 wt. % for A, 23.1 wt. % for B and 19.6 wt. % for C.

[0524] All ingredients A, B and C exhibit an essential amino acid content of at least about 40 wt %, more specifically 40.81 wt % for A, 40.52 wt % for B and 39.61 wt % for C.

[0525] 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 C (163.43 mg / g).

[0526] Table 2: Amino acid content of mycelium components A, B and C

[0527]

[0528]

[0529] Biomass characterization and EUC concentration

[0530] Mycelial component A was grown on a medium containing a carbon-nitrogen ratio (medium) of about 17. Mycelial component B was grown on a medium containing a carbon-nitrogen ratio (medium) of about 20. Mycelial component B was grown on a medium containing a carbon-nitrogen ratio (medium) of about 13.

[0531] The insoluble fiber content increases from about 23 wt % to about 34 wt % to about 54 wt % from C to A to B, while the protein content decreases from about 60 wt % to about 39 wt % to about 32 wt % from C to A to B. This demonstrates the relationship between protein and fiber discussed previously, which are inversely proportional and can be controlled by changing the fermentation medium composition.

[0532] The fat content of ingredient A is about 3% by weight. The fat content of ingredient A is about 2% by weight.

[0533] The fat content of ingredient C is about 7% by weight. The fat contains saturated fatty acids, monosaturated fatty acids, polyunsaturated fatty acids and trans fatty acids, as shown in the table below. The saturated fatty acid content of mushroom mycelium is the highest (A and B up to 1.5g / 100g, C up to 2g), while C is rich in polyunsaturated fatty acids (3.63g / 100g). The ingredients contain the least amount of trans fatty acids (up to 0.02-0.03g / 100g). It is worth noting that the content of ω-6 fatty acids (linoleic acid) is 0.77% by weight for A, 0.46% by weight for B and 3.5% by weight for C. All ingredients show a calculated carbohydrate content of less than 0.1% by weight. B is the most abundant in all glucans, with a value of 31% by weight. Mycelial ingredient A has a total glucan value of about 26% by weight. Mycelial ingredient C has a total glucan value of about 17% by weight.

[0534] Table 3: Composition and nutritional characteristics of mycelial components A, B and C

[0535]

[0536]

[0537] The EUCs of the mycelial components were calculated to be 302% for A, 34% for B, and 2892% for C.

[0538] In a separate study, the fruiting bodies of Pleurotus pulmonarius were grown on three types of forestry waste (pine, poplar and honeysuckle), showing that the EUC values ​​of the fruiting bodies of Pleurotus pulmonarius were between 72.31% and 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 ​​of the fruiting bodies of the same fungal strains. Taking component C as an example, the difference of component C is 25-40 times higher than the reported range of the fruiting bodies of Pleurotus pulmonarius.

[0539] It is also known that the highest value of EUC of mushroom fruiting bodies is reported to be 4465% for (Straw mushroom), which is 1.54 times lower than that of ingredient C.

[0540] In addition, EUC concentrations were measured in beef, chicken, real peas, and soybeans (for the exact products used for comparison, see Figure 5 ) to compare them with mycelial fractions A, B, C, where the EUCs found were 78.77%, 134.7%, 65%, and 1%, respectively.

[0541] Total Sugar Quantification Method

[0542] About 300mg of the ingredients are added to the pressure tube. 3mL72% H2SO4 is added by an automatic titrator, and the weight of the acid added is recorded. The sample is thoroughly mixed with the acid using a glass rod. The test tube is transferred to a water bath maintained at 30°C. These steps are repeated for duplicate samples. The glass rod used for each pressure tube is stirred every 10 minutes so that the acid reaches all parts of the sample and complete hydrolysis occurs. Exactly after it is placed in a water bath for 1 hour, the pressure tube is taken out and placed on a balance and 84mL of water is added (record the weight of the added water). This water is used to remove any acid / sample on the rod from the rod at this time. The cap is screwed on the tube and the tube is inverted several times to ensure the thorough mixing of the acid. Two sugar recovery solution (SRS) pressure tubes are prepared to monitor the sugar loss associated with the second stage hydrolysis. This includes the following steps: (a) 348 microliters of 72% H2SO4 are added to a test tube containing a solution containing a known weight (about 10g) of a sugar standard. The standard should have a sugar composition similar to that expected for the sample being analyzed. The acid and sugar solution are thoroughly mixed. (b) The sugar-acid mixture is transferred to a pressure tube and then sealed. All SRS and sample pressure tubes are placed in an autoclave running at 121 degrees Celsius for 60 minutes. After the temperature in the autoclave drops to less than 80°C, the test tubes are taken out and left in the laboratory (sealed) until they reach room temperature. The hydrolyzate is then filtered (using vacuum suction) through a filter crucible of known weight, and the resulting filtrate is stored. Any remaining solids are washed out of the tube using deionized water until all acid-insoluble residues remain on the filter crucible. After the acid hydrolysis step, the hydrolyzate (filtrate from vacuum filtration) is diluted 5 times using a water-based fucose solution. Fucose is an internal standard for the chromatographic analysis of the hydrolyzate. After this dilution, the sample is immediately placed on the chromatographic system. The equipment used is a NIR spectrophotometer (FOSS XDSNIR) and an ion chromatography (ICS-3000).

[0543] Sugars in water extract data

[0544] The components were hydrolyzed into sugar (glucan, xylan, arabinan, galactan, mannan, rhamnan) content using acid. However, this hydrolysis was performed on the original sample without removing extractables (e.g. ethanol or water-soluble components). All data are expressed as a percentage of the total dry mass.

[0545] Table 4: Sugar profiles of mycelial components A, B and C

[0546]

[0547] As a percentage of the total sugar content, the most abundant sugar was 88% for dextran in A, 92.5% for dextran in B, and 78.6% for dextran in C (here dextran is based on all glucose, including glucose derived from other polysaccharides, such as heteropolysaccharides and / or exopolysaccharides).

[0548] Sugars in one or more water extracts shown in the above table, which contain sugars extracted from fresh mycelial components by water. Water extraction was performed in an 11 ml stainless steel extraction cell using ASE 200 (accelerated solvent extractor), using deionized water at 100°C and 1500 psi (heating time 5 minutes, static time: 7 minutes, flushing volume: 150%, purge time: 180 seconds, static cycle: 3). The total sugars in the water extracts of components A, B and C were 6.25%, 7% and 3.26%, respectively, wherein, for A, the sugars in the water extract (expressed as % of total extract sugars) included disaccharides (trehalose 20.1%), hexoses (glucose 22%, fructose 0.5%, mannose 0.1%, galactose 1.4%), pentoses (arabinose 0.1%) and sugar alcohols (mannitol 55%, sorbitol 0.7%). For B, the sugars in the water extract expressed by the % of sugars in the water extract include disaccharides (trehalose 19.2%), hexoses (glucose 43.3%, fructose 0.4%, galactose 0.1%), pentoses (arabinose 0.1%), and sugar alcohols (mannitol 35.7%, sorbitol 1%). For C, the sugars in the water extract expressed by the % of sugars in the water extract include disaccharides (trehalose 46%), hexoses (glucose 16%, galactose 0.1%), pentoses (arabinose 0.03% and xylose 0.15%), and sugar alcohols (mannitol 22.7%, arabitol 13.5%, xylitol 1%, sorbitol 0.64%).

[0549] Elemental analysis, ash content and gross calorific value determination methods

[0550] The final (element) analysis of biomass will provide the mass concentration of the main elements (carbon, oxygen, hydrogen, nitrogen and sulfur) in the sample. According to the procedure outlined in European standard EN 15104:2011 ("Solid biofuels-Determination of total contentof carbon, hydrogen and nitrogen-Instrumental methods"), the carbon, hydrogen, nitrogen and sulfur content of the sample are measured. Elementar Vario MACRO Cube elemental analyzer is used. Oxygen content is calculated by difference according to the following formula: oxygen (%)=100-carbon (% dry basis)-hydrogen (% dry basis)-nitrogen (% dry basis)-sulfur (% dry basis)-ash (% dry basis). In order to perform this calculation, the ash content of the sample is also measured in a Nabertherm L-240H1SN muffle furnace and heated to 575°C. The sample is then weighed to calculate the ash content. As outlined in EN 14918:2009, higher heating value (HHV, commonly referred to as gross heating value) is directly determined using an oxygen bomb calorimeter. The lower heating value (LHV, often referred to as net heating value) is calculated based on the HHV and the elemental composition of the sample.

[0551] Elemental analysis, ash content and gross calorific value data

[0552] 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.

[0553] The ash contents of components A, B and C are 9.39%, 9.73% and 8.40%.

[0554] Elemental analysis of these components C, H, N, O, S analysis results in:

[0555] For A: 44.45% (carbon C), 5.95% (H), 6.25% (N), 0.32% (S), 33.53% (O), 0.0815% (chlorine);

[0556] Carbon to nitrogen ratio: 7.11.

[0557] For B: 43.92% (carbon C), 5.83% (H), 4.62% (N), 0.26% (S), 35.64% (O), 0.0788% (chlorine);

[0558] Carbon to nitrogen ratio: 9.5.

[0559] For C: 46.90% (carbon C), 6.26% (H), 9.57% (N), 0.42% (S), 28.45% (O), 0.0703% (chlorine);

[0560] Carbon to nitrogen ratio: 4.9.

[0561] The lower nitrogen content in sample B indicates a sample rich in fiber content but with lower protein content, while sample C (based on spent brewers grains) has the highest nitrogen content, indicating a high protein content with lower fiber content. Sample A falls in between the two. This suggests that protein and fiber content can be adjusted by adjusting the synthetic culture medium composition or by using a nutrient side stream with a known elemental composition, allowing for the production of fiber-rich or protein-rich specialty foods.

[0562] The higher heating value (HHV, often referred to as gross calorific 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 calorific value) was calculated based on the HHV and the elemental composition of the sample. 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

[0563] Other determination methods

[0564] Chitin analysis

[0565] For the determination of chitin content, the method described in the following paper was used (https: / / doi.org / 10.1155 / 2020 / 5084036). This ensured the maximum recovery of glucosamine. The glucosamine content was estimated by ion chromatography. Chitosan was used as an experimental control.

[0566] Analysis of Ergothioneine

[0567] In order to quantify ergothioneine, about 1.25g sample was weighed into a 25mL volumetric flask. The volumetric flask was filled to the mark with cold 70% ethanol, 10mM dithiothreitol was added thereto and ultrasonicated for 15 minutes. The sample was then centrifuged at 4000rpm for 20 minutes to separate insoluble matter. 2mL of the supernatant obtained was evaporated to dryness. The residue was resuspended in 1mL water and centrifuged at 10,000rpm for 1 minute. Finally, the supernatant was filtered through a 0.45-μm filter and further diluted with water before HPLC injection.

[0568] The HPLC analysis of thioneine is carried out on the Prominence system (Shimadzu, Duisburg, Germany) equipped with LC-20AD high performance liquid chromatography (HPLC) pump, SIL-20AC HT automatic sample injector, SPD-M20A diode array detector (DAD), CBM-20A communication bus module and LabSolutions MultiLC Data System Manager. EC 250 / 4Nucleosil 100-5C18 post (Macherey-Nagel, Düren, Germany) with matching pre-column is used. The injection volume is 10 μL. The mobile phase used is 3% acetonitrile and 0.1% acetic acid aqueous solution. Separation 15min is carried out at an isocratic elution and ambient temperature of 0.7mL / min at flow velocity, and detection is carried out at 254nm. External calibration is carried out to quantify using thioneine (5 μg mL-1 to 100 μg mL-1) dissolved in water.

[0569] Ergosterol analysis

[0570] Ergosterol is quantitatively measured by using 7-dehydrocholesterol as an internal standard. For ergosterol analysis, 150-200 mg of each sample is weighed into a derivatization tube and mixed with 50 mg sodium ascorbate and 250 μL 7-dehydrocholesterol stock solution (5 mg mL-1 in 2-butanone). After adding 5 mL of a methanol solution of sodium hydroxide and homogenizing by vortexing, the tube is incubated in an 80 ° C water bath for 1 hour. Oscillate once every 20 minutes. After cooling the sample to room temperature in the dark, each sample is membrane filtered and transferred to a new derivatization tube. Subsequently, three extractions are performed with hexane. The hexane phases are combined in a 15 mL volumetric flask and the flask is filled with hexane. After drying with sodium sulfate, 6 mL is transferred to a new derivatization tube. The solvent is then removed under a stream of nitrogen. The residue is absorbed in 0.5 mL of tetrahydrofuran (THF) and 0.5 mL of N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) and dissolved in an ultrasonic bath. The solution was then heated at 70°C for 2 minutes in a water bath. After each 1 minute, homogenization was performed by vortexing. Silylation was performed overnight. After being transferred to a vial, the sample was analyzed by gas chromatography (GC) and flame ionization detector (FID). A calibration series with each 1mL 7-dehydrocholesterol stock solution and 0.25 to 4.5mL ergosterol stock solution (2mg·mL-1 in 2-butanone) was prepared in a 10mL volumetric flask. 2-Butanone was used to supplement the ad marker, and 1mL was transferred to a derivatization tube and dried under a nitrogen stream. Subsequently, the calibration series was recorded and silylated in THF and MSTFA similar to the sample. The ergosterol content of sample A was determined to be 2.35g / g dry weight, the ergosterol content of sample B was 2.24g / g dry weight, and the ergosterol content of sample C was 5.56g / g dry weight.

[0571] Further analysis

[0572] The protein content was determined according to DIN EN ISO 16634-1, 2009-07, (N*6.25) DUMAS. The fat content was determined according to Weibull-Stoldt. The fiber content was determined according to the enzymatic-gravimetric method ASU L 00.00-18. The carbohydrate content was determined via HPLC according to SOP M2569.

[0573] Vitamins Vitamin content was determined using 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 α / β glucan determination. Thermogravimetric analysis (TGA) was performed in a TA Instruments Q500 TGA unit that allows monitoring of sample weight loss up to 1000°C under nitrogen atmosphere (temperature ramp used: equilibration at 35°C, isothermal for 2 minutes, equilibration at 105°C, isothermal for 5 minutes, ramp to 900°C at 20°C / min).

[0574] Brands of commodities purchased for EUC calculation: beef and chicken (METZGERFRISCH, Rinder-SupppenFleisch and -Innenfilets), Sunat and Rapunzel.

[0575] Density measurement

[0576] The density of the mycelial component samples was measured according to the Archimedean principle using an analytical density balance KERN EMB-V (KERN SOHN GmbH, Balingen, Germany) and YDB-01 Set (KERN SOHN GmbH, Balingen, Germany). The test was carried out at room temperature ***21°C and 50-60% relative humidity and was repeated three times. The kit provides a weighing plate, a platform, a beaker and an immersion basket for the sinking and floating of solid substances. The sample was weighed in air and distilled water (or other suitable liquids with a density below 1 g / cm3) and the density was calculated directly in g / cm3.

[0577] In one example, the density of the composition was found to be 1.012 g / cm3. The density of the mycelial composition ranges from 0.1-2.5 g / cm3, most preferably from 0.7-1.2 g / cm3.

[0578] Table 5: Density of mycelial components (SD = standard deviation)

[0579]

[0580]

[0581] Shear Force Methodology

[0582] Shear force was generated using a TA.XTplus100 texture analyzer (Stable Micro Systems, Surrey, United Kingdom). Tests were performed using 50 kg and 100 kg load cells. Samples were analyzed at ambient temperature (≈21° C.). Due to the heterogeneity of the biomass, 5 g of the sample was sheared, compressed and extruded into blocks using a Miniature Kramer Shear / Ottowa Cell (HDP / MKS5). The results are given as the average value of the force (N) required to cut through the sample. In addition to the force, the area under the curve (i.e., shear work) (N·s) was also calculated.

[0583] Table 6: Shear forces of mycelial components

[0584] Test ID Maximum force (N) Area (N·s) Sample 1 43.019 281.352 Sample 2 44.097 300.464 Sample 3 40.733 284.220 average 42.616 288.679 SD 1.718 10.307 The coef of the variant. 4.031 3.570

[0585] Water Holding Capacity (WHC) and Released Water (RW)

[0586] Water holding capacity (WHC) describes the ability of a material to retain water during processing. Among the many procedures used to measure WHC, in this study, a method modified from van der Sman et al. and Liu et al. was used to determine WHC. (Van der Sman, R. G. M.; Paudel, E.; Voda, A. Khalloufi, S. Hydration properties of plant foods explained by the 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 the water holding capacity of grass carp surimi gels by ozone water washing. Food Science and Technology 2021, 150, 111919.)

[0587] After separation of the biomass and supernatant, 3 g of wet biomass was placed in a 50 ml centrifuge tube consisting of cotton balls and a filter paper layer at the bottom. The sample was centrifuged at 4500 rpm for 10 minutes at 23 ° C. During the centrifugation, water was removed from the mycelial particles through the filter and collected in the lower part. After centrifugation, the biomass was carefully removed and weighed. The falcon tube with cotton and filter paper was also weighed for comparison of the results. WHC (%) was calculated according to the following equation:

[0588]

[0589] The released water was measured in triplicate according to the study of Seon-Tea Joo (Joo, S. Determination of water-holding capacity released water method using optimal load). The released water was measured in triplicate. Korean journal for food science of animal resources 2018, 38 (4), 823.). About 1.0 g of the drained biomass was weighed and carefully placed on two thin plastic films (PP5) and filter paper (MN 615, 12.5 cm in diameter) that had been dried in advance. A metal round plate was used as a weight and a 100 g weight was applied for 5 minutes. After accurately taking out the compressed biomass sample, the wet filter paper and two plastic films were weighed. RW (%) (%):

[0590]

[0591] Table 7: WHC and RW of mycelial components

[0592] Test ID WHC(%) RW(%) Sample 1 42.00 49.00 Sample 2 52.32 43.73 average 47.16 46.37 SD 7.30 3.73

[0593] Piercing test and spreadability of dairy analogs (cream cheese)

[0594] Puncture test

[0595] The measurements were performed using 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 puncture tests were performed using a cylindrical plexiglass probe (P / 20P) with a diameter of 20 mm. The probe pierced the cheese sample 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. The 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 that the probe fully penetrated the coextrudate. The force (N) versus time (s) data were recorded and some mechanical parameters were determined using index software (Stable Micro Systems, Surrey, UK). The maximum positive force (N), maximum negative force (N), positive area (N·s), and negative area (N·s) parameters were determined. The maximum force is highly correlated with the hardness of the cheese samples (Journal of Texture Studies, 32:41-55).

[0596] Cream cheese spreadability

[0597] Spreadability testing was performed using a TA.XTplus100 texture analyzer (Stable Micro Systems, Surrey, United Kingdom). Testing was performed using a 5kg load cell. Samples were analyzed at 16°C to minimize differences in solid fat content. The TTC spreading device (HDP / SR) includes a male 90° conical probe and five precisely matched female plexiglass conical product holders. Fill the cream cheese sample with a spatula and then smooth the surface. Before testing the sample, the sample holder was stored at the above temperature. During the test, the degree of spreading was calculated when the product was forced to flow outward at 45° between the male and female conical surfaces. The force-time curve was recorded (below), and the force (N) at the maximum penetration depth was taken as the sample hardness. The area under the positive curve (N·s) represents the total amount of force required to carry out the shear process, and it is considered to be a good instrumental measurement of spreadability in cream cheese and other spreadable products. The smaller the value of this area, the easier it is to spread. The force (N) of the largest negative peak indicates sample viscosity, and the largest negative area (N·s) indicates the work of adhesion (Bayarri, S.; Carbonell, I.; Costell, E. Viscoelasticity and texture of spreadable cheeses with different fat contents at refrigerated and room temperature. Journal of Dairy Science 2012, 95(12), pp. 6926-6936.).

[0598] Comparison between two mycelium-based cream cheeses

[0599] Sample 1 is an example of a dairy analog comprising mycelial component A, 8.0% pea starch and 2.5% corn starch. Sample 2 is an example of a dairy analog comprising mycelial component A, about 3.0% wheat starch and 2.5% corn starch. Spreadability was measured and puncture tests were performed. It was observed that the lower the starch content, the softer the texture became, and therefore a lower maximum force (i.e., the maximum force required for diffusion (N), i.e., the lowest positive area (Ns) and the lowest puncture force (N)) was required to spread, and these values ​​increased with increasing starch content. But this generally depends on the type of starch and the combination of starches used.

[0600] Table 8: Spreadability data for mycelium-based cream cheese

[0601]

[0602] Table 9: Piercing data for mycelium-based cream cheese

[0603]

[0604]

[0605] Cutting strength method

[0606] 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 cooling temperature (≈7°C). The meat balls were cut into 80% of their initial height using a blade. The results are given as the average value of the maximum force (N) required to cut through the sample. In addition to the force, the area under the curve (N·s) is also calculated. The force-time curve is recorded and the maximum force is taken as the maximum cutting strength. The area under the positive curve (Ns) represents the total amount of force.

[0607] Table 10: Cutting strength of soft mycelium based meat analogs

[0608]

[0609]

[0610] Texture Profile Analysis (TPA) Method

[0611] TPA was performed using a TA.XTplus100 texture analyzer (Stable Micro System, Surrey, United Kingdom). The test was performed using a 50 kg load cell. The samples were analyzed at a cooling temperature (≈7° C.). Unfried meat balls cooked at about 85° C. for 5 to 10 minutes were compressed twice in a row by 60% of their initial height at a test speed of 1 mm / sec. Therefore, a compression plate (SMS P / 75) was used. The results are given as the average value of the maximum force (N) required to cut through the sample.

[0612] Record the force-time curve and take the maximum force (N) at the first peak as the sample hardness. The area under the first negative peak (Ns) represents the adhesion of the sample. The height recovery represents the elasticity of the sample (%), 2 nd Divide the area by 1 st The area (%) indicates the cohesiveness of the sample. The value of stickiness (N) is obtained by multiplying hardness (N) and cohesiveness (%), while chewiness (N) is the product of stickiness (N) and elasticity (%).

[0613] Table 11: TPA data for soft mycelium based meat analogs

[0614]

[0615] The recipes for mycelium-based cream cheese were prepared by three different methods:

[0616] By acidification of ingredients

[0617] 10-50 wt% of Pleurotus pulmonaria mycelium (edible fibrous mycelium) is homogenized with about 55 wt% of drinking water, about 0-7 wt% of cashew nuts (vegetable fat component), 10-40 wt% of coconut oil (vegetable fat component), up to 2 wt% of salt (sodium chloride source), up to 2 wt% of yeast flakes, up to 5 wt% of sucrose (vegetable sugar source), up to 5 wt% of lemon juice (natural vegetable acid source), and up to 5 wt% of citric acid.

[0618] The homogenized slurry is heated at a temperature of 75-95° C. for a maximum of 60 seconds under constant mixing. In order to coagulate the homogenized slurry, a previously prepared coagulant solution is applied once a temperature of 25° C. to 40° C. has been reached during this previously defined heating step. The coagulant solution comprises up to 15% by weight of each of: - water, wheat starch and hydrolyzed corn starch.

[0619] The resulting slurry is quickly removed from the heat source and the mixture is subsequently allowed to settle to room temperature (21°C). Once at room temperature, the coagulated slurry is stored in a dark and cooled environment, preferably between 4°C and 7°C.

[0620] Acidification by microbial fermentation

[0621] 10-50 wt% of Pleurotus pulmonaria mycelia (edible fibrous mycelia) are homogenized with about 55 wt% of drinking water, about 0-7 wt% of cashew nuts (vegetable fat component), 10-40 wt% of coconut oil (vegetable fat component), up to 2 wt% of salt (sodium chloride source), up to 2 wt% of yeast flakes, and up to 5 wt% of sucrose (vegetable sugar source).

[0622] The homogenized slurry is heated at a temperature of 75-95° C. for a maximum of 60 seconds under constant mixing. In order to coagulate the homogenized slurry, a previously prepared coagulant solution is applied once a temperature of 25° C. to 40° C. has been reached during this previously defined heating step. The coagulant solution comprises up to 15% by weight of each of: - water, wheat starch and hydrolyzed corn starch.

[0623] The resulting slurry is quickly removed from the heat source, and the mixture is subsequently allowed to settle to room temperature (21°C). Once the obtained mixture reaches a temperature of 40°C or less, 0.1 g to 0.25 g of acid-producing bacteria, particularly lactic acid bacteria, is added to induce microbial acidification. In order to provide a hospital environment for 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 can be placed at a temperature of 20 to 45°C for 1 to 6 hours. After the microbial fermentation is complete, the coagulated slurry is stored in a dark and cold environment, preferably between 4°C and 7°C.

[0624] Cream cheese based on two fungal strains

[0625] The same instructions as in Example 1 and Example 2 were followed, except that a portion of the edible fibrous mycelium of Pleurotus pulmonatus was replaced with the edible fibrous mycelium of Morchella rubra. Assuming the amounts listed above, 60-90% by weight of the total mycelium was derived from the Pleurotus pulmonatus mycelium, and 10-40% by weight of the total mycelium component was derived from the mycelium of Morchella rubra (edible fibrous mycelium).

[0626] Formulation preparation of vegetarian mycelium-based meat analog compositions :

[0627] 50-95% by weight of Pleurotus pulmonarius mycelia (edible fibrous mycelia) is mixed, and 1-40% by weight of the following ingredients are added: rapeseed oil, salt, egg white, gluten. Optionally, up to 1 to 40% by weight of each of the following ingredients are added: methylcellulose, hydrocolloid, texturized vegetable protein, starch-based ingredients, and flavor components. The obtained dough is then shaped by a molding machine and / or an extruder or a combination thereof to have a final product as a meat ball or sausage or an extruded product.

[0628] Formulation preparation of vegetative mycelium-based meat analog compositions :

[0629] 50-95% by weight of Pleurotus pulmonarius mycelia (edible fibrous mycelia) is mixed, and rapeseed oil, salt, and gluten are added at 1-40% by weight or less. The missing egg white is calculated by adding additional dry mycelia, and the equivalent ratio is 1 to 1.4 respectively. Optionally, up to 1 to 40% by weight of each of the following ingredients is added: methylcellulose, hydrocolloid, texturized vegetable protein, starch-based ingredients, and flavor components. The obtained dough is then formed via a forming machine and / or an extruder or a combination thereof to have a final product as a meat ball or sausage or an extruded product.

[0630] Other examples and / or embodiments of the invention are disclosed in the following numbered items.

[0631] 1. Edible mycelium component A derived from submerged fermentation, having an elemental composition of mycelium with a C:N ratio between 6 and 8.

[0632] 2. Edible mycelium component B derived from submerged fermentation, having an elemental composition of mycelium with a C:N ratio of between 8 and 12.

[0633] 3. Edible mycelial component C derived from submerged fermentation, having an elemental composition of the mycelial C:N ratio between 2 and 6.

[0634] 4. The edible mycelial component of item 1, wherein the inherent RNA level is less than 1.88% by weight on a dry basis, without further processing steps to reduce the RNA.

[0635] 5. The edible mycelial component of item 2, wherein the inherent RNA level is less than 1.65% by weight on a dry basis, without further processing steps to reduce the RNA.

[0636] 6. The edible mycelial component of items 1-3, wherein the inherent RNA level is at most 2% by weight on a dry basis, without further processing steps to reduce the RNA.

[0637] 7. The edible mycelium component of item 1, wherein the ergothioneine content of the mycelium is in the range of 70-100 mg / kg.

[0638] 8. The edible mycelium ingredient according to item 2, wherein the mycelium has an ergothioneine content of 110-150 mg / kg.

[0639] 9. The edible mycelium component according to item 3, wherein the mycelium has an ergothioneine content of 380-455 mg / kg.

[0640] 10. The edible mycelium component of item 1-3, wherein the 5'-inosine monophosphate (IMP) is 0 g / kg.

[0641] 11. The edible mycelium component of item 1, wherein the content of umami 5' nucleotides (5'NMP) containing guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is at most 6 g / kg.

[0642] 12. The edible mycelium component of item 2, wherein the content of umami 5' nucleotides (5'NMP) containing guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is at most 3 g / kg.

[0643] 13. The edible mycelium component of item 3, wherein the content of umami 5' nucleotides (5'NMP) containing guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is at most 20 g / kg.

[0644] 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% after enzymatic processing of AMP into IMP.

[0645] 15. The edible mycelium component of item 3, wherein the concentration of uridine monophosphate (UMP) is 3.56 g / kg.

[0646] 16. The edible mycelium component of any one of items 1-3, wherein the amount of branched chain amino acids (BCAAs) is at least about 19% by weight of the total amount of amino acids present.

[0647] 17. The edible mycelium component of any one of items 1-3, wherein the amount of umami amino acids is at least about 19% by weight of the total amount of amino acids present.

[0648] 18. The edible mycelium component of items 1-3, wherein the amount of essential amino acids is at least about 40% by weight of the total amount of amino acids present.

[0649] 19. The edible mycelium component according to item 1, wherein the amount of BCAA and the amount of umami amino acids are 40-100 mg / g respectively.

[0650] 20. The edible mycelium component according to item 2, wherein the amount of BCAA and the amount of umami amino acids are 20-80 mg / g respectively.

[0651] 21. The edible mycelium component of item 3, wherein the amount of BCAA is 50-100 mg / g, and the amount of umami amino acids is 70-100 mg / g.

[0652] 22. The edible mycelium component of item 1, having an EUC concentration of at least 500%.

[0653] 23. The edible mycelium component of item 2, having an EUC concentration of at least 400%, preferably about 460%.

[0654] 24. The edible mycelium component of item 3, having an EUC concentration of at least 500%, more preferably at least 5000%, most preferably at least 10000%.

[0655] 25. The edible mycelium component of item 1-2, in N 2 It has a thermal stability of up to 220°C under atmosphere.

[0656] 26. The edible mycelium component as described in item 3, wherein 2 It has thermal stability up to 190°C in atmosphere.

[0657] 27. The edible mycelium component according to any one of items 1 to 3, wherein the calorific value ranges from 300 to 600 Kcal / 100 g.

[0658] 28. The edible mycelium component of item 1, having an insoluble fiber content of 30-60% by weight, preferably 30-40% by weight.

[0659] 29. The edible mycelium component of item 2, having an insoluble fiber content of 40-60% by weight, preferably 40-50% by weight.

[0660] 30. The edible mycelium component according to item 3, having an insoluble fiber content of 10-40% by weight, preferably 20-30% by weight.

[0661] 31. The edible mycelium component of item 1, having a protein content of 30-50% by weight, preferably 30-40% by weight.

[0662] 32. The edible mycelium component according to item 2, having a protein content of 30-50% by weight, preferably 30-40% by weight.

[0663] 33. The edible mycelium component of item 3, having a protein content of 30-65% by weight, preferably 45-65% by weight.

[0664] 34. The edible mycelium component of item 1-2, having an ergosterol content of 2-4 mg / g.

[0665] 35. The edible mycelium component of item 3, having an ergosterol content of 4-7 mg / g.

[0666] 36. The edible mycelium component of any one of items 1 to 3, having a carbohydrate content of at most 1%.

[0667] 37. The edible mycelium component according to any one of items 1 to 3, wherein the uronic acid content is 0.1-5% by weight.

[0668] 38. The edible mycelium component according to any one of items 1 to 3, wherein the chitin content ranges from 6 to 11% by weight.

[0669] 39. The edible mycelium component of any one of items 1 to 3, wherein the β-glucan content is at least 80% of the total glucans.

[0670] 40. The edible mycelium component according to item 1, wherein the total glucan content is 20-35% by weight.

[0671] 41. The edible mycelium component according to item 2, wherein the total glucan content is 25-50% by weight.

[0672] 42. The edible mycelium component according to item 3, wherein the total glucan content is 10-20% by weight.

[0673] 43. The edible mycelium component of any one of items 1 to 3, wherein at least 96% by weight of their polyunsaturated fatty acids comprise linoleic acid.

[0674] 44. The edible mycelium component of item 3, having 2-5% by weight of ω-6 fatty acid (linoleic acid).

[0675] 45. The edible mycelium component of items 1-2, having a fat content of up to 3% by weight.

[0676] 46. ​​The edible mycelium component of item 3, having an enriched fat content, wherein the fat content is up to 8% by weight.

[0677] 47. The edible mycelium component according to any one of items 1 to 3, having a total phenolic content (TPC) of 1 to 15 GAE / g and a total flavonoid content (TFC) of 1 to 15 mg QE / g, wherein the flavonoid content of mycelium component A accounts for about 35% to 60% of the total phenolic content, the flavonoid content of mycelium component B accounts for about 70 to 95% of the total phenolic content, and the flavonoid content of mycelium component C accounts for about 50 to 80% of the total phenolic content.

[0678] 48. After freeze drying, the edible mycelium component of item 1 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.

[0679] 49. After freeze drying, the edible mycelium component of item 2 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.

[0680] 50. After freeze drying, the specific pore volume of the edible mycelium component of item 2 is 2.46 cm3 / g, the median pore diameter is 7.1 μm, and the BET surface area is 1.59 m2 / g.

[0681] 51. The edible mycelium component of item 1, when freeze-dried and with a pore size below 1 mm, has 58% of its pore volume corresponding to pore sizes between 1000 and 30 μm, with the most common pore size being in the range of 85 to 185 μm, and 42% of its pore volume corresponding to pore sizes between 30 and 2 μm, with the most common pore size peak being equal to 16 μm, which is the most common pore size in the range of 1000 to 2 μm.

[0682] 52. The edible mycelium component of item 2, when freeze-dried and having a pore size less than 1 mm, has 81.5% of its pore volume corresponding to pore sizes between 1000 and 30 μm, with the most common pore size being 147

[0683] The pore size range is 2.1 μm, which is also the most common pore size in the range of 1000-2 μm, and 18.5% of the pore volume corresponds to pore sizes between 30-2 μm, with the most common pore size peak equal to 15 μm.

[0684] 53. The edible mycelium component of item 3, when freeze-dried and with a pore size below 1 mm, has 20% of its pore volume corresponding to pore sizes between 1000 and 20 μm, and 80% of its pore volume corresponding to pore sizes between 20 and 2 μm, with the most common pore size peak being equal to 5.5 μm.

[0685] 54. The edible mycelium component of any one of items 1 to 3, wherein the mycelium has a shear force of at least 15 N and a water holding capacity of 20-90% and a water release of 25-70%.

[0686] 55. A method for producing the mycelium component of any one of items 1-3 by submerged fermentation, wherein the fermentation medium contains 5-60 g / L carbon source, 0.1-60 g / L nitrogen source, 0.01-15 g / L minerals and 0.01-50 mg / L vitamins.

[0687] 56. The method of item 55, wherein the production of component A of item 1 comprises the step of culturing at least one fungal species in a defined medium, wherein the defined medium comprises a plurality of amino acids and vitamin sources, minerals based on a complex nitrogen source and at least one carbon source, wherein the carbon-nitrogen ratio in the medium ranges between 14 and 19.

[0688] 57. The method of item 55, wherein the production of component B of item 2 comprises the step of culturing at least one fungal species in a basic synthetic medium comprising an amino acid and a vitamin, wherein the carbon-to-nitrogen ratio in the medium is between 16 and 23.

[0689] 58. The method of claim 55, wherein the production of ingredient C of claim 3 comprises the step of culturing at least one fungal species in a natural synthetic culture medium comprising a vinasse C5-sugar extract, wherein the carbon-to-nitrogen ratio in the culture medium ranges between 10 and 25, and wherein at least 35% by weight of the extracted vinasse ranges between 2 and 4 mm.

[0690] 59. Preferably, the edible fibrous mycelium mass is obtained from at least one fungal strain selected from the group consisting of Basidiomycota, Ascomycota, Pezizomycota, Agaricaceae, Pezizomycota, Agaricaceae, Coleomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morelaceae, Truffleaceae, Pleurotaceae, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus eryngii, Trichodermaceae, Omphalaceae, Pyriculariaceae, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericiumaceae, Polyporaceae, Cordyceps, Auriculariaceae, and Bolognese.

[0691] 60. The method of any one of items 55-58, 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.

[0692] 61. The method described in any one of items 55-58, further comprising the step of recovering the supernatant from the culture medium.

[0693] 62. The method of item 61, wherein the recovery comprises the step of crystallizing or precipitating the obtained supernatant.

[0694] 63. A method for producing a soft or hard meat analog composition comprising at least one mycelium component according to any one of items 1-3, the method comprising the method for producing the corresponding mycelium component according to any one of items 55-58, and further comprising the step of preparing such a meat analog composition by mixing at least one mycelium component according to any one of items 1-3 from at least one fungal strain with a composition comprising at least one protein-rich component, at least one vegetable lipid-rich component and optionally at least one constituent component.

[0695] 64. A method for producing a dairy analog composition comprising at least one mycelial component of any one of items 1-3 from at least one fungal strain, the method comprising the method for producing the corresponding mycelial component of any one of items 55-58, and further comprising the step of preparing such a dairy analog composition by the following steps: (1) forming a slurry comprising at least one mycelial component of any one of items 1-3 and a composition comprising at least one protein-rich ingredient, at least one plant-based lipid-rich ingredient and at least one constituent ingredient, and (2) mixing the slurry with at least one constituent ingredient, in particular a texturizing agent or a thickening agent or a carbohydrate-rich ingredient.

[0696] 65. An edible non-animal dairy substitute product according to any one of items 1 to 5, which also contains up to 25% by weight, preferably up to 25% by weight, and most preferably in the range of 1% to 5% by weight of an edible plant or algae-based fat component, a fat component derived from fungi or yeast.

[0697] 66. A method according to any one of items 64 to 64, wherein the edible plant-based fat component is selected from coconut oil, sunflower oil, rapeseed oil, palm oil, cottonseed oil, olive oil, canola oil, algal oil or oil derived from oleaginous yeast.

[0698] 67. The edible product of item 64, wherein the at least one protein-rich ingredient or the at least one constituent ingredient is selected from agar, egg white, yeast flakes, edible starch, guar gum, locust bean gum, wheat gluten, a sugar source, cellulose or its derivatives, lemon juice, colorants and / or edible flavors, such as salt, extracts and / or spices.

[0699] 68. A meat analogue obtained as described in item 63, comprising mycelial mass in the range of 1 wt% to 99 wt% and water in the range of up to 99 wt%, wherein the edible fibrous mycelium has an insoluble fiber content of 40 to 50 wt%, preferably about 45% w / w.

[0700] 69. A dairy analog obtained according to item 64, comprising in the range of 1 wt% to 99 wt% mycelium mass and in the range of up to 99 wt% water, wherein the insoluble fiber content in the edible fibrous mycelium is at least about 40 wt%.

[0701] 70. The edible non-animal dairy alternative product of claim 64, comprising within the range of 50% to 95% by weight of the fibrous mycelial mass.

[0702] 71. The edible non-animal dairy alternative product of claim 63, comprising said fibrous mycelial mass in the range of 1 wt% to 50 wt%.

[0703] 72. Products according to item 63 or 64, characterised in that these products can have a soft or non-soft / harder texture.

[0704] 73. The food of item 63, characterized by a soft texture, preferably having a hardness of 10-55N, an elasticity of 35-85%, a cohesion of 15-70%, a stickiness of 1-40N, a chewiness of 0.3-35N, a cutting strength of 1-25N and an adhesion of 0N.s to -0.3Ns.

[0705] 74. The food of item 63, characterized by a non-soft or relatively hard texture, preferably having a hardness of 30-100N, an elasticity of 20-70%, a cohesiveness of 20-85%, a stickiness of 6-85N, a chewiness of 1-60N, and a cutting strength of 1-50N.

[0706] 75. The food 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 viscosity of -15 to -100 N and a piercing force of 1 to 30 N.s.

[0707] 76. The food according to item 64, characterized by a non-soft or relatively hard texture, preferably having a hardness of 20 to 100 N, a spreadability of 1 to 20 N.s, a viscosity of -1 to -14 N and a puncture force of 40 to 100 N.s.

[0708] Other embodiments of the present invention are disclosed in the following numbered paragraphs:

[0709] 1. Edible mycelial component C derived from submerged fermentation, having an elemental composition of the mycelial with a C:N ratio between 2 and 6.

[0710] 2. The edible mycelium component of item 1

[0711] wherein the inherent RNA content is at most 4 wt.-%, preferably at most 2 wt.-% (on a dry basis), without further processing steps to reduce the RNA, and / or wherein the content of umami 5'-nucleotides (5'NMP) containing guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is at most 40 g / kg, preferably at most 20 g / kg, and / or

[0712] wherein 5'-inosine monophosphate (IMP) is 0 g / kg, and / or

[0713] wherein the umami taste from the 5'-nucleotide can be further enhanced by at least 40% after enzymatic treatment (preferably with 5'-adenylate deaminase) to convert AMP into IMP, and / or

[0714] wherein the amount of branched chain amino acids (BCAA) is at least about 19% by weight of the total amount of amino acids present, and / or

[0715] wherein the amount of essential amino acids is at least about 40% by weight of the total amount of amino acids present, and / or

[0716] The amount of BCAA is between 50 and 150 mg / g, and the amount of umami amino acids is between 70 and 100 mg / g.

[0717] 3. Edible mycelium ingredients of clause 1 or 2

[0718] The mycelium has an ergothioneine content of 350-800 mg / kg.

[0719] 4. The edible mycelium component of 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.

[0720] 5. The edible mycelium component of 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%.

[0721] 6. The edible mycelium component of any one of clauses 1 to 5

[0722] wherein at least 96% by weight of the polyunsaturated fatty acids comprise linoleic acid, preferably 2-5% by weight of omega-6 fatty acids (linoleic acid), and / or having a high fat content, wherein the fat content is at most 8% by weight, and / or

[0723] The DPPH free radical scavenging activity was associated with 1-15 mg / ml TPC, and

[0724] or wherein the TPC, TFC and polyphenol contents are 1-15 mg GAE / g, 1-15 mg QE / g and 100-1000 mg / kg respectively, wherein the flavonoid content accounts for about 70-95% of the total phenolic content, and wherein catechins and protocatechuic acid each account for about 45-55% of the total polyphenols.

[0725] 7. The edible mycelium component of any one of clauses 1 to 6

[0726] The calorific value range is 300 to 600 Kcal / 100g, and the ingredients are N 2 Thermal stability up to 190°C in an atmosphere, and / or

[0727] wherein the insoluble fiber content is 10-40% by weight, preferably 20-30% by weight, and / or wherein the protein content is 30-65% by weight, preferably 45-65% by weight, and / or wherein the ergosterol content is 4-7 mg / g, and / or wherein the carbohydrate content is at most 5% by weight, and / or

[0728] The content of uronic acid is 0.1-5% by weight.

[0729] and / or wherein the chitin content is 6-11 wt %, and / or wherein the β-glucan content is at least 80 % of the total glucans, preferably wherein the total glucan content is 10-20 wt %.

[0730] 8. The edible mycelium component of any one of clauses 1 to 7

[0731] Among them, when freeze-dried, for pores less than 1 mm, the pore volume is about 15

[0732] to 25%, preferably about 20%, of the pore volume corresponds to pore sizes between 1000 and 20 μm, and about 75 to 85%, preferably about 80%, of the pore volume corresponds to pore sizes between 20 and 2 μm, the most common pore size peak being equal to 5.5 μm, and / or

[0733] The mycelium has a shear force of at least 15 N and a water holding capacity of 20-90% and a water release of 25-70%.

[0734] 9. A method for producing the mycelial component of any of clauses 1 to 8 by submerged fermentation, wherein the fermentation medium comprises 5-60 g / L carbon source, 0.1-60 g / L nitrogen source, 0.01-15 g / L minerals and 0.01-50 mg / L vitamins, wherein the production of component C comprises the step of culturing at least one fungal species in a natural synthetic medium comprising a vinasse C5-sugar extract, wherein the carbon-nitrogen ratio in the medium is in the range between 10 and 25, and wherein at least 35 wt. % of the extracted vinasse is in the range between 2 and 4 mm, preferably wherein the particle size distribution is determined by sieving by an air jet sieving method following DIN 10765 2016-07 after using an automatic sieving tower, i.e. a vibrating sieving method, to remove particles larger than 4 mm.

[0735] 10. The method of clause 9, wherein the at least one fungal species is selected from the group consisting of Basidiomycota, Aspergillus, Pezizomycota, Agaricaceae, Pezizomycota, Agaricaceae, Coleomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morchellaceae, Truffleaceae, Pleurotaceae, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus eryngii, Trichodermaceae, Omphalaceae, Cordyceps, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariales and Bolognese, preferably wherein the at least one fungal strain is selected from the group consisting of Basidiomycota, Aspergillus, Pezizomycota, Agaricaceae, Agaricaceae , preferably wherein the mycelium is obtained from: from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus florida ...

[0736] Optionally, wherein the 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.

[0737] 11. The method of clause 9 or 10, further comprising the step of recovering the supernatant from the culture medium or a portion thereof, optionally wherein said recovery comprises the step of crystallizing or precipitating the supernatant obtained.

[0738] 12. A fungal fermentation medium obtainable according to the method of any one of clauses 9 to 11.

[0739] 13. A method for producing a soft or hard meat analog composition comprising a mycelial component according to any one of clauses 1 to 8 or 12, the method comprising the step of preparing such a meat analog composition by adding the mycelial component to a composition comprising at least one protein-rich component, at least one plant-based lipid-rich component and optionally at least one constituent ingredient.

[0740] 14. A method for producing a dairy analog composition comprising a mycelial component as described in any one of clauses 1 to 8 or 12, the method comprising the steps of preparing such a dairy analog composition by the following steps: (1) forming a slurry comprising the mycelial component and a composition comprising at least one protein-rich ingredient, at least one plant-based lipid-rich ingredient and at least one constituent ingredient, and (2) mixing the slurry with at least one constituent ingredient, particularly a texturizing agent or thickener or a carbohydrate-rich ingredient.

[0741] 15. Edible product obtainable by the method of clause 13 or 14

[0742] Preferably, the food of item 13 is characterized by a soft texture, preferably having a hardness of 10-55N, an elasticity of 35-85%, a cohesion of 15-70%, a stickiness of 1-40N, a chewiness of 0.3-35N, a cutting strength of 1-25N and an adhesion of 0N.s to -0.3Ns, and / or

[0743] Preferably, the food of item 13 is characterized by a non-soft or relatively hard texture,

[0744] Preferably, it has a hardness of 30-100N, an elasticity of 20-70%, a cohesion of 20-85%, a gummy property of 6-85N, a chewiness of 1-60N, a cutting strength of 1-50N, and / or

[0745] Preferably, the food of clause 14 is characterized by a soft texture, preferably having a hardness of 1 to 20 N, a spreadability of 30 to 100 N.s, a viscosity of -15 to -100 N and a puncture force of 1 to 30 N.s, and / or

[0746] Preferably, the food product of clause 14 is characterized by a non-soft or relatively hard texture, preferably having a hardness of 20 to 100 N, a spreadability of 1 to 20 N.s, a viscosity of -1 to -14 N and a puncture force of 40 to 100 N.s.

[0747] Other embodiments of the present invention are disclosed in the following numbered paragraphs:

[0748] 1. The edible mycelial component from submerged fermentation has an elemental composition of mycelial C:N ratio between 6 and 12.

[0749] 2. The edible mycelium component of paragraph 1, which is an edible mycelium component A derived from submerged fermentation in a defined medium, having an elemental composition of the mycelium with a C:N ratio of between 6 and 8.

[0750] 3. The edible mycelium component of paragraph 1, which is an edible mycelium component B derived from submerged fermentation in a synthetic medium, wherein the synthetic medium has an elemental composition of the mycelium with a C:N ratio of between 8 and 12.

[0751] 4. Edible mycelium components of paragraph 2

[0752] wherein the inherent RNA level is at most 2% by weight on a dry basis and there is no further processing step to reduce the RNA, and / or

[0753] wherein the mycelium has an ergothioneine content of 70-270 mg / kg within 5 days, and / or

[0754] wherein the content of umami 5' nucleotides (5'NMP) containing guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is at most 20 g / kg, and / or

[0755] wherein the amount of BCAA and the amount of umami amino acids are 40-100 mg / g respectively, and / or

[0756] Calorific value is 300~600Kcal / 100g, and / or

[0757] having an insoluble fiber content of 30-60% by weight, preferably 30-40% by weight, and /

[0758] or

[0759] having a protein content of 30-50% by weight, preferably 30-40% by weight, and / or

[0760] wherein the total glucan content is 20-35% by weight, and / or

[0761] wherein, when freeze-dried and with a pore size below 1 mm, 55 to 65%, preferably about 58%, of its pore volume corresponds to pore sizes between 1000 and 30 μm, with the most common pore size being in the range of 85-185 μm, and 35 to 45%, preferably about 42%, of its pore volume corresponds to pore sizes between 30 and 2 μm, with the most common pore size peak being equal to 16 μm, which is the most common pore size in the range of 1000-2 μm, and / or

[0762] wherein the flavonoid content accounts for about 35% to 60% of the total phenolic content, and / or

[0763] Among them, catechins and protocatechuic acid each account for about 35-45% of the total polyphenols.

[0764] 5. The edible mycelial component of paragraph 2 or 4, having an EUC concentration of at least 500%.

[0765] 6. Edible mycelium ingredients of paragraph 3

[0766] wherein the inherent RNA level is at most 2% by weight on a dry basis and there is no further processing step to reduce the RNA, and / or

[0767] wherein the mycelium has an ergothioneine content of 100-350 mg / kg, and / or

[0768] wherein the content of umami 5'-nucleotides (5'NMP) containing guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is at most 15 g / kg, and / or

[0769] wherein the amount of BCAA and the amount of umami amino acids are 20-80 mg / g respectively, and / or

[0770] The insoluble fiber content is 40-60% by weight, preferably 40-50% by weight, and / or

[0771] having a protein content of 30-50% by weight, preferably 30-40% by weight, and / or

[0772] wherein the total glucan content is 25-50% by weight, and / or

[0773] wherein, when freeze-dried and with a pore size below 1 mm, 75 to 85%, preferably about 81.5%, of its pore volume corresponds to pore sizes between 1000 and 30 μm, with the most common pore size being in the range of 147 μm, which is also the most common pore size in the range of 1000-2 μm, and 15 to 25%, preferably about 18.5% of the pore volume corresponds to pore sizes between 30-2 μm, with the most common pore size peak being equal to 15 μm, and / or

[0774] wherein the flavonoid content accounts for about 70-95% of the total phenolic content, and / or

[0775] Among them, catechins and protocatechuic acid account for about 25-35% and about 35-45% of the total polyphenols, respectively.

[0776] 7. The edible mycelium component according to any one of paragraphs 1 to 6, wherein the DPPH radical scavenging activity, total phenolic content TPC, total flavonoid content TFC and polyphenol content are between 1 and 15 mg / ml, 1 and 15 mg GAE / g, 1 and 15 mg QE / g and 50 to 900 mg / kg, respectively, relative to TPC.

[0777] 8. The edible mycelium component according to any one of paragraphs 1 to 7, wherein N 2 has a thermal stability of up to 210 to 220°C in a molten-liquid atmosphere, and / or

[0778] wherein 5'-inosine monophosphate (IMP) is 0 g / kg, and / or

[0779] wherein the amount of branched chain amino acids (BCAA) is at least about 19% by weight of the total amount of amino acids present, and / or

[0780] wherein the amount of umami amino acids is at least about 19% by weight of the total amount of amino acids present, and / or

[0781] wherein the amount of essential amino acids is at least about 40% by weight of the total amount of amino acids present, and / or

[0782] Ergosterol - 4 mg / g ergosterol content, and / or

[0783] having a carbohydrate content of at most 5%, and / or

[0784] The uronic acid content is 0.1-5% by weight, and / or

[0785] wherein the chitin content is 6-11% by weight, and / or

[0786] wherein the β-glucan content is at least 80% of the total glucan, and / or

[0787] having a fat content of at most 3% by weight, and / or

[0788] The mycelium has a shear force of at least 15 N and a water holding capacity of 20-90% and a water release of 25-70%.

[0789] 9. A method for producing the mycelial component of any of paragraphs 1 to 8 by submerged fermentation, wherein the fermentation medium comprises 5 to 60 g / L carbon source, 0.1 to 60 g / L nitrogen source, 0.01 to 15 g / L minerals and 0.01 to 50 mg / L vitamins, wherein the carbon-to-nitrogen ratio in the medium ranges from 14 to 23, preferably 14 to 19 for medium A and preferably 16 to 23 for medium B, characterized in that the amount of at least one vitamin or at least one amino acid used to synthesize medium B is characterized in that the ratio of the at least one vitamin or at least one amino acid used to the total amount of all vitamins or all amino acids present in the defined medium A is 0.01 to 10, preferably 0.1 to 10.

[0790] 10. The method of paragraph 9, wherein the at least one fungal species is selected from the group consisting of Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Pezizomycotina, Agaricaceae, Coleomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morchellaceae, Truffleaceae, Pleurotus, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus, Trichodermaceae, Omphalaceae, Cordyceps, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese, preferably wherein The at least one fungal strain is selected from the group consisting of Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Agaricomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morchellaceae, Truffleaceae, Pleurotus eryngii, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Hylomycetes, Omphalaceae, Cordyceps, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese, preferably wherein the mycelium is obtained from Pleurotus pulmonaria and / or Morchella rubrum,

[0791] Optionally, wherein the 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.

[0792] 11. The method of paragraph 9 or 10, further comprising the step of recovering the supernatant from the culture medium or a portion thereof, optionally wherein the recovery comprises the step of crystallizing or precipitating the obtained supernatant.

[0793] 12. A fungal biomass produced according to the method of any one of items 9 to 11.

[0794] 13. A method of producing a soft or hard meat analog composition comprising the mycelial ingredient of any of paragraphs 1 to 8 or 12, the method comprising the step of preparing such a meat analog composition by adding the mycelial ingredient to a composition comprising at least one protein-rich ingredient, at least one plant-based lipid-rich ingredient and optionally at least one constituent ingredient.

[0795] 14. A method for producing a dairy analog composition comprising the mycelial ingredient of any of paragraphs 1 to 8 or 12, the method comprising the steps of preparing such a dairy analog composition by the following steps: (1) forming a slurry comprising the mycelial ingredient and a composition comprising at least one protein-rich ingredient, at least one plant-based lipid-rich ingredient and at least one constituent ingredient, and (2) mixing the slurry with at least one constituent ingredient, particularly a texturizing agent or thickening agent or a carbohydrate-rich ingredient.

[0796] 15. Edible product obtainable by the method of paragraph 13 or 14.

[0797] Preferably, the food of paragraph 13 is characterized by a soft texture, preferably having a hardness of 10-55N, an elasticity of 35-85%, a cohesion of 15-70%, a stickiness of 1-40N, a chewiness of 0.3-35N, a cutting strength of 1-25N and an adhesion of 0N.s to -0.3Ns, and / or

[0798] Preferably, the food of paragraph 13 is characterized by a non-soft or relatively hard texture, preferably having a hardness of 30-100N, an elasticity of 20-70%, a cohesiveness of 20-85%,

[0799] 6-85N adhesiveness, 1-60N chewiness, 1-50N shear strength, and / or

[0800] Preferably, the food of paragraph 14 characterized by a soft texture preferably has a hardness of 1 to 20 N, a spreadability of 30 to 100 N.s, a viscosity of -15 to -100 N and a piercing force of 1 to 30 N.s, and / or

[0801] Preferably, the food product as described in paragraph 14 is characterized by a non-soft or relatively hard texture, preferably having a hardness of 20 to 100 N, a spreadability of 1 to 20 N.s,

[0802] -1 to -14N viscosity and 40 to 100N.s puncture force.

[0803] It will be appreciated that any of these embodiments may be combined with any other definition of the mycelial components A, B or C, their production methods or the corresponding food products as defined above.

Claims

1. An edible mycelial component comprising undifferentiated mycelial biomass having an elemental composition of mycelium having a C:N ratio of 2 to 12, and characterized by an equivalent umami concentration (EUC) of at least 500 monosodium glutamate g MSG / 100g, wherein EUC is defined as: EUC=∑aibi+1218(∑aibi)(∑ajbj), where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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.

2. The edible mycelium component of claim 1, wherein the biomass has an elemental composition of mycelium with a C:N ratio of 2 to 8.

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

4. The edible mycelium component according to any one of claims 1 to 3, Features At least 1000g MSG / 100g EUC.

5. The edible mycelium component according to any one of claims 1 to 4, Features At least 1500g MSG / 100g EUC.

6. The edible mycelium component according to any one of claims 1 to 5, Features At least 2000g MSG / 100g EUC.

7. An edible mycelial component according to any one of claims 1 to 6, wherein the inherent RNA level is at most 4 wt%, preferably at most 2 wt% on a dry basis in the absence of further processing steps for reducing RNA.

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

9. The edible mycelium component according to any one of claims 1 to 8, Features The content of 5'-AMP is 3.5 to 10 g / kg, and / or the content of 5'-GMP is 3.5 to 10 g / kg.

10. The edible mycelium component according to any one of claims 1 to 9, Features The content of 5'-AMP is 4.5 to 7 g / kg, and / or the content of 5'-GMP is 4.5 to 7 g / kg.

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

12. The edible mycelium component according to any one of claims 1 to 11, wherein the umami taste from the 5'-nucleotide is further enhanced by at least 40% after enzymatic treatment, preferably with 5'-adenylate deaminase, to convert 5'-AMP into 5'-IMP.

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

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

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

17. The edible mycelium component of 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. Edible mycelial ingredients according to any one of claims 1 to 17, wherein at least 96% by weight of their polyunsaturated fatty acids comprise linoleic acid, preferably with 2 to 5% by weight enriched in omega-6 fatty acids (linoleic acid), and / or with an enriched fat content, wherein the fat content is at most 8% by weight.

19. The edible mycelial component according to any one of claims 1 to 18, having DPPH radical scavenging activity associated with 1 to 15 mg / ml of TPC.

20. The edible mycelium component according to any one of claims 1 to 19, wherein the TPC, TFC and polyphenol contents are 1 to 15 mg GAE / g, 1 to 15 mg QE / g and 100 to 1000 mg / kg respectively, wherein the flavonoid content accounts for about 70 to 95% of the total phenolic content, and wherein catechins and protocatechuic acid each account for about 45 to 55% of the total polyphenols.

21. The edible mycelium component according to any one of claims 1 to 20, wherein the calorific value is 300 to 600 Kcal / 100 g, wherein the component is N 2 It has a thermal stability up to 190°C under ambient temperature.

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

23. The edible mycelial component according to any one of claims 1 to 22, wherein the mycelial component is characterized by an insoluble fiber content of up to 35% by weight.

24. The edible mycelium component according to any one of claims 1 to 23, wherein the protein content is 30 to 65 wt%, preferably 45 to 65 wt%.

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

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

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

28. An edible mycelium component according to any one of claims 1 to 27, wherein the chitin content is 6 to 11 wt%.

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

30. The edible mycelium component according to any one of claims 1 to 29, in, When freeze-dried, for pore sizes less than 1 mm, about 15 to 25%, preferably about 20%, of the pore volume corresponds to pore sizes of 1000 to 20 μm, and about 75 to 85%, preferably about 80% of the pore volume corresponds to pore sizes of 20-2 μm, with the most common pore size peak being equal to 5.5 μm.

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

32. The edible mycelium component of any one of claims 1 to 31, comprising at least one fungal strain selected from the group consisting of: Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Pezizomycotinae, Agaricaceae, Coleomycetes, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morchellaceae, Truffleaceae, Pleurotaceae, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus eryngii, Trichodermaceae, Omphalaceae, Cordyceps, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae, and Bologneseaceae, Preferably, wherein the at least one fungal strain is selected from the group consisting of: Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Agaricomycetes, Agaricales, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morelaceae, Truffleaceae, Pleurotus, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus, Trichodermaceae, Omphalaceae, Pyriculariaceae, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese.

33. The edible mycelium ingredient of claim 32, wherein the mycelium is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus floridus, Pleurotus aurantii, Pleurotus rosa, Morchella, Morchella black vein, Morchella foetida, and / or Morchella rubrum.

34. An edible mycelium ingredient according to claim 33, wherein the mycelium is obtained from Pleurotus pulmonaria and / or Morchella rubrum.

35. The edible mycelium component of any one of claims 1 to 31, wherein the mycelium is obtained from Sophora sulphurea or Sophora nicotianae.

36. A method for producing an edible mycelium ingredient according to any one of claims 1 to 35 via submerged fermentation, the method comprising the step of 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, 0.01-15 g / L of minerals and 0.01-50 mg / L of vitamins, wherein the medium comprises a spent grain C5-sugar extract, wherein the carbon-to-nitrogen ratio in the medium is 10 to 25, and wherein at least 35% by weight of the extracted spent grain is characterized by a particle size of 2 to 4 mm.

37. The method of claim 36, wherein the spent grain C5-sugar extract is the sole carbon source.

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

39. The method according to any one of claims 36 to 38, wherein the particle size distribution is determined by sieving by air jet sieving in accordance with DIN 10765 2016-07 after using an automatic sieving tower, i.e. a vibrating sieving method, to remove particles larger than 4 mm.

40. A method according to any one of claims 36 to 39, wherein the spent cereal C5-sugar extract has a carbon to nitrogen ratio of 5 to 25, preferably 5 to 18.

41. A method according to any one of claims 36 to 40 wherein the spent cereal C5-sugar extract has a protein content of 7 to 30 g / l protein.

42. The method of any one of claims 36 to 41, wherein the at least one fungal species is selected from the group consisting of: Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Pezizomycotinae, Agaricaceae, Sphaerotheca, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morchellaceae, Truffleaceae, Pleurotaceae, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus eryngii, Trichodermaceae, Omphalaceae, Pleurotus eryngii, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae, and Bolognese, Preferably, wherein the at least one fungal strain is selected from the group consisting of: Basidiomycota, Aspergillus, Pezizomycotina, Agaricaceae, Agaricomycetes, Agaricales, Peziales, Boletales, Cantharellales, Agaricales, Polyporaceae, Russulares, Auriculariales, Hypocreales, Morelaceae, Truffleaceae, Pleurotus, Agaricaceae, Microdermaceae, Cantharellaceae, Odontaceae, Boletaceae, Thinporaceae, Polyporaceae, Strophariaceae, Pleurotus, Trichodermaceae, Omphalaceae, Pyriculariaceae, Schizophyllaceae, Sclerodermaceae, Ganodermaceae, Hydrangeaceae, Hericium, Polyporaceae, Cordyceps, Auriculariaceae and Bolognese.

43. The method of claim 42, wherein the mycelium is obtained from Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus floridus, Pleurotus aurantii, Pleurotus rosa, Morchella, Morchella black vein, Morchella foetida, and / or Morchella rubrum.

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

45. The method of any one of claims 36 to 41, wherein the mycelium is obtained from Sophora solfataricus or Sophora nicotianae.

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 a combination thereof.

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

48. The method of claim 47, wherein the recovery comprises a step of crystallizing or precipitating the resulting supernatant.

49. The process of any one of claims 36 to 48, wherein the pH in the fermentor is maintained between 4.0 and 5.

0.

50. The method according to any one of claims 36 to 49, wherein after 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 component obtainable according to the method of any one of claims 36 to 50.

52. A method for producing a soft or hard meat analog composition comprising a mycelial component according to any one of claims 1 to 35 or 51, the method comprising the step of preparing such a meat analog composition by adding the mycelial component to a composition comprising at least one protein-rich component, at least one vegetable-based lipid-rich component and optionally at least one constituent ingredient.

53. An edible product obtainable by the method according to claim 52.

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

55. A method for producing a dairy analog composition comprising a mycelial component according to any one of claims 1 to 35 or 51, the method comprising the steps of preparing such a dairy analog composition by: (1) forming a slurry comprising the mycelial component with a composition comprising at least one protein-rich ingredient, at least one plant-based lipid-rich ingredient and at least one constituent ingredient, and (2) mixing the slurry with at least one constituent ingredient, in particular a texturizing agent or thickening agent or a carbohydrate-rich ingredient.

56. An edible product obtainable by the method according to claim 55.

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

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

59. An edible mycelium component comprising undifferentiated mycelial biomass having an elemental composition of mycelium with a C:N ratio of 8 to 12, and characterized by an EUC of less than 200 g MSG / 100 g, wherein the EUC Defined as: EUC=∑aibi+1218(∑aibi)(∑ajbj), where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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.

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

61. An edible ingredient according to claim 59 or 60, wherein the ingredient comprises a Pleurotus genus, preferably Pleurotus pulmonaria, or wherein the ingredient comprises Morchella rubrum, or wherein the ingredient comprises Sulphureus sulphureus, or wherein the ingredient comprises Pipe fungus.

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

63. An edible ingredient according to any one of claims 59 to 62, wherein the content of umami 5'-nucleotides (5'NMP) including guanosine 5'-monophosphate (GMP) and adenosine 5'-monophosphate (AMP) is up to 3 g / kg.

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

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

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

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

68. A method for producing an edible mycelium component via submerged fermentation, the edible mycelium component being characterized by an EUC of less than 200 g MSG / 100 g, the method comprising the step of cultivating 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 of 1 to 50, preferably 5 to 50, wherein the fermentation medium provided at the start of the fermentation comprises 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01-15 g / L of minerals and 0.01-50 mg / L of vitamins, and comprises no more than 5 amino acids including arginine and up to 4 other 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. The method of claim 68, wherein the culture medium comprises arginine as the only amino acid.

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

73. An edible mycelial component comprising Pleurotus pulmonaria mycelial biomass having an elemental composition of the mycelium having a C:N ratio of 6 to 8, and characterized by an EUC of 200 to 500 g MSG / 100 g, wherein EUC is defined as: EUC=∑aibi+1218(∑aibi)(∑ajbj), where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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.

74. The edible mycelium component of claim 73, wherein the inherent RNA level is less than 1.88 wt% on a dry basis in the absence of further processing steps to reduce RNA.

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

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

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

78. An edible mycelium component according to any one of claims 73 to 77, wherein the insoluble fibre content is from 25 to 45 wt%, or wherein the insoluble fibre content is from 30 to 60 wt%.

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

80. The edible mycelium component of any one of claims 73 to 79, wherein the total glucan content is 20 to 35 wt%.

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

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

5.

83. An edible product, preferably a meat substitute product or a dairy substitute product, comprising 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of 20 to 60 wt% and an EUC of 200 to 500 g MSG / 100 g, wherein EUC is defined as: EUC=∑aibi+1218(∑aibi)(∑ajbj), where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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. The edible product of claim 83, wherein the mycelial component is characterized by an EUC of about 300 g MSG / 100 g.

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

86. An edible product, preferably a meat substitute product or a dairy substitute product, comprising from 1 to 99 wt% of a mycelial component, wherein the mycelial component is characterized by an insoluble fiber content of from 20 to 60 wt% and an EUC of less than 200 g MSG / 100 g, wherein EUC is defined as: EUC=∑aibi+1218(∑aibi)(∑ajbj), where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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. An edible product according to claim 86, wherein the mycelial component is characterized by an EUC of less than 100 g MSG / 100 g, preferably less than 50 g MSG / 100 g.

88. An edible product according to claim 86 or 87, wherein the mycelial component is characterised by an insoluble fibre content of 30 to 60 wt%, preferably 40 to 60 wt%.

89. An edible product according to any one of claims 83 to 88, wherein the ingredient comprises a Pleurotus genus, preferably Pleurotus pulmonaria.

90. An edible product according to any one of claims 83 to 88, wherein the ingredient comprises Morchella fuchsia.

91. An edible product according to any one of claims 83 to 88, wherein the ingredient comprises Sulphur Fungi.

92. An edible product according to any one of claims 83 to 88, wherein the ingredient comprises Nicotiana tabacum.

93. An edible mycelial component comprising Pleurotus pulmonaria mycelial biomass having an elemental composition of the mycelium having a C:N ratio of 2 to 12, and characterized by an equivalent umami concentration (EUC) of at least 30 monosodium glutamate g MSG / 100 g, wherein EUC is defined as: EUC=∑aibi+1218(∑aibi)(∑ajbj), where EUC is expressed in g MSG / 100 g, ai is the concentration of each umami amino acid Asp or Glu (g / 100 g), aj is the concentration of each umami 5'-nucleotide 5'-IMP, 5'-GMP or 5'-AMP (g / 100 g), bi is the relative umami concentration (RUC) of each umami amino acid 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.

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

95. The edible mycelium component of any one of claims 93 to 94, wherein the protein content is 10 to 65% by weight.

96. The edible mycelium component of any one of claims 93 to 95, wherein the inherent RNA level is at most 2 wt% on a dry basis in the absence of further processing steps to reduce RNA.

97. A method for producing an edible mycelium ingredient according to any one of claims 93 to 96 via submerged fermentation, the method comprising the step of cultivating at least one fungal strain in a fermentation medium, wherein the fermentation medium provided at the start of the fermentation is characterized by a C:N ratio of 1 to 50, preferably 5 to 50, wherein the fermentation medium provided at the start of the fermentation comprises 5 to 60 g / L of a carbon source, 0.1 to 60 g / L of a nitrogen source, 0.01-15 g / L of minerals and 0.01-50 mg / L of vitamins, the method being characterized in that the medium comprises arginine and glutamic acid as the only amino acids.

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

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

100. The edible product according to any one of claims 93 to 99, wherein the at least one fungal strain comprises a Pleurotus genus, preferably Pleurotus pulmonaria, or wherein the at least one fungal strain comprises Morchella rubrum, or wherein the at least one fungal strain comprises Sulphureus sulphureus, or wherein the at least one fungal strain comprises Pipe Fungus.

101. An edible product, preferably a meat substitute product or a dairy substitute product, comprising from 1 to 99 wt% of an edible mycelium component according to any one of claims 1 to 35, wherein the mycelium component is characterized by an insoluble fiber content of from 20 to 60 wt%.

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