Mycelial biomass, mycelial biomass production process, use of mycelial biomass in food production, mycelial biomass-based food products and production process thereof

Abstract

The present invention relates to mycelial biomass and a process for producing same; the present invention also relates to mycelial biomass-based food products with high nutritional properties and low concentrations of additional ingredients and a process for producing same.

Description

[Technical Field]

[0001] The present invention relates to mycelial biomass.

[0002] The present invention also relates to mycelial biomass-based food products with high nutritional properties and low concentrations of additional ingredients.

[0003] The invention also relates to a process for producing mycelial biomass and a process for obtaining a food product. [Background technology]

[0004] In recent years, there has been an increasing interest in the environment, sustainability and the search for healthier, more natural diets, including the reduction or elimination of animal protein consumption.

[0005] Studies predict that the world's population will grow to 9.8 billion by 2050, which will require an increase in the range, diversity, sustainability and economy of food production.

[0006] Currently, attachment to plant or vegetative plants is becoming increasingly stronger, resulting in a significant increase in plant and vegetative populations, and with it an increasing need for product substitutes to replace animal proteins, i.e., products derived from the plant kingdom and fungi.

[0007] Over the past two years, the domestic market for alternative protein-based products ("plant-based" foods) has grown dramatically, particularly in the plant-based chilled meat category.

[0008] The majority of plant-based foods are substitutes for ground meat products of any animal species, such as sausages, hamburgers, nuggets, etc. Data compiled by the U.S. Agricultural Marketing Service (AMS) shows that only 36% of animal protein meat products are ground meat, with the remaining products consisting of whole products such as steaks / steaks.

[0009] Since 1960, processed soy protein has served as a substitute for ground meat products. The same techniques used to obtain processed soy protein have been used to obtain other processed plant proteins, such as wheat and pea.

[0010] Fungal-derived protein sources, also known as mycoproteins, are an affordable alternative that is attracting attention from large and small businesses.

[0011] Although mycoprotein contains less protein than animal meat, it has a higher amount than plant-derived mycoprotein, and its composition includes a large amount of fiber, providing an average of about 6 grams of fiber per 100 grams of food. It is worth noting that fungal protein has high nutritional quality, is low in sodium, sugar, and fat, and is rich in essential amino acids, vitamins B12 and B9, calcium, phosphorus, magnesium, and zinc. In addition, compared to beef and chicken production, it produces a smaller carbon footprint and consumes less water.

[0012] The fungal kingdom has an estimated diversity of 1.5 million species, of which 117 are traditionally used as food.

[0013] The production of fungal / mycelial biomass for use in the food supply can be divided into two categories: (a) mushroom production, which has been practiced for thousands of years, which requires cultivation cycles long enough to produce fruiting bodies and is limited in shape and final size; and (b) (I) liquid culture (introduced in 1980 as Quorn™), which results in a fungal cell paste without fiber alignment and therefore requires processing to create a cohesive and acceptable texture, or (II) production of mycoprotein by solid culture, which facilitates the achievement of edible fungal / mycelial biomass with a cohesive structure and texture. Both methods of obtaining mycoprotein can provide a unique nutritional profile, as well as a sensory and texture suitable for meat substitutes.

[0014] Furthermore, unlike the large animal protein (animal meat) industry, the production of fungal proteins (mycoproteins) requires very little space and allows for a variety of textures to be obtained from different types of meat, which is not possible with plant-based proteins (soy, wheat, and pea).

[0015] Therefore, faced with this promising scenario, large and small companies are beginning to consider using fungi to produce alternative foods to meat, which, in addition to being characterized as a vegan option, would contribute to halving the deforestation caused by cattle fattening.

[0016] To obtain meat substitutes composed of fungal mycelium, fungi must be grown through a process commonly known as fermentation. This fermentation is somewhat similar to the fermentation used to make beer, using sugars and other nutrients in the process. Furthermore, the filamentous nature of fungi triggers a process that produces fibrous strands called hyphae, which, when processed in certain ways, resemble the texture of animal meat. The fungi most commonly used to produce fungal proteins are Fusarium venenatum, Fusarium flavolapis, Neurospora intermedia, and Neurospora glabra.

[0017] Faced with this scenario, some researchers are conducting research to improve the process of cultivating fungi, to obtain mycelia with desirable structure and texture, to improve the process for obtaining fungal mycelium-based foods, to scale up and reduce the costs, as well as to work with other species of filamentous fungi to find new alternatives to use in the production of mycelia for use in the food industry.

[0018] As can be seen in the state of the art, document PI7800368 describes a process for texturing mycelial fungal masses by defined steps and subsequent freezing.

[0019] WO 2021092051 describes improved mycelia suitable for use as food products. U.S. Patent Application Publication No. 20200305486 describes a food product comprising filamentous fungi of the genus Fusarium, agar, and particles of an edible hydrocolloid, and WO 2020061502 describes a method for producing a protein food product based on filamentous fungal mycelia.

[0020] The teachings of US Patent Application Publication No. 20190373934 disclose a system for growing fungal mycelium and an edible meat substitute product containing a defined weight percentage of filamentous mycelial mass.

[0021] Therefore, focusing on this new need and seeking improvements related to the latest technology, including the development of edible product alternatives that can replace animal protein, the present researchers developed mycelium biomass to produce food products with high nutritional properties. In addition to developing a simple process, it is easy to scale up and has low production costs.

[0022] It is important to emphasize that in the state of the art there is no equivalent solution to that presented by the present invention, which combines the technological differences to obtain a "clean label" product, a simpler production process, economic advantages and environmental conservation. [Prior art documents] [Patent documents]

[0023] [Patent Document 1] International Publication No. 2021092051 [Patent Document 2] US Patent Application Publication No. 20200305486 [Patent Document 3] International Publication No. 2020061502 [Patent Document 4] US Patent Application Publication No. 20190373934 Summary of the Invention

[0024] The main object of the present invention is to provide a mycelial biomass with high nutritional properties and a process for its production.

[0025] In a second aspect, the present invention aims at using mycelial biomass as the basis for the production of food products, which mycelial biomass is the object of the present invention, making it possible to obtain food products that are substitutes for animal meat, with a high protein index, low caloric value and low fat content, with more than 95% of the product corresponding to the mycelial biomass.

[0026] Mycelial biomass-based food products and processes for their production remain the object of the present invention. [Brief explanation of the drawings]

[0027] [Figure 1] 1 shows a food product containing mycelial biomass obtained by a cultivation process using culture medium 1. [Figure 2] 1 shows a food product containing mycelial biomass obtained by a cultivation process using culture medium 3. [Figure 3] 1 shows hybrid burger patties obtained by the process of the present invention, 50:50 beef and mycelium biomass (left) and 100% beef (right). [Figure 4] A visual comparison between the products is shown: 3 - 100% beef burger patty, 2 - product from Figure 3 with no added natural coloring, 1 - product from Figure 3 with 1% added natural beet coloring. DETAILED DESCRIPTION OF THE INVENTION

[0028] The main objective of the present invention is the realization of a mycelial biomass with high nutritional properties and its use as a basis for the production of food products. The mycelial biomass object of the present invention can be pressed in a single step, resulting in obtaining a cut that mimics chicken breast, without the need for cutting the pressed material and then a new pressing, as described in the state of the art.

[0029] The present invention is also directed to the process, the process for obtaining mycelial biomass and the process for preparing food products.

[0030] Furthermore, the mycelial biomass object of the present invention is the basis for the production of food products that are substitutes for animal meat, with a high protein index and a low caloric and fat value.

[0031] Ultimately, the present invention aims to provide a mycelium biomass-based food product and a process for its production.

[0032] Biomass for the purposes of the present invention is composed of filamentous fungal mass without the presence of chemical additives. The biomass is composed of 100% filamentous fungal mass and does not contain added chemical additives.

[0033] If there are no chemical additives present in the mycelial biomass, the biomass can be classified as "clean label" and the food can be obtained as "clean label".

[0034] According to the present invention, a "clean label" product is one that is produced with fewer ingredients and is free of chemical additives such as dyes, preservatives, stabilizers, etc.

[0035] According to the present invention, the filamentous fungi used to obtain the biomass are Pleurotus ostreatus, Ganoderma lucidum, Rhizopus oligosporus, Rhizopus microsporus var. oligosporus, Rhizopus oryzae, and preferentially the fungus Rhizopus oligosporus.

[0036] The biomass production process that is also the object of the present invention is simple, fast to implement and easy to scale up, in addition to having low production costs.

[0037] Mycelial biomass is obtained from a process that includes the steps of cultivating filamentous fungi, filtering, rinsing, and inactivating the fungi.

[0038] For the purposes of the present invention, for biomass production, the filamentous fungus is selected from the group consisting of Pleurotus ostreatus, Ganoderma lucidum, Rhizopus oligosporus, Rhizopus microsporus var. oligosporus, Rhizopus oryzae, but is not limited to these, and is preferentially the fungus Rhizopus oligosporus.

[0039] To obtain fungal mycelial biomass, it is necessary to cultivate the fungus through a specific process that allows the filamentous nature of the fungus to produce fibrous bundles that allow the formation of a biomass with characteristics similar to the texture of animal flesh.

[0040] To obtain mycelial biomass, the following steps are carried out: (a) Growth of fungi in solid media; (b) pre-inoculation; (c) fermentation in bioreactors; (d) filtration of filamentous fungal mass; (e) Biomass washing / rinsing; and (f) Fungal inactivation is necessary.

[0041] For the solid medium growth stage, the fungus is cultured in a slant agar tube on BDA medium (potato dextrose agar) for 4 to 10 days in an incubator at 25 to 39°C, preferably 30 to 35°C. The fungus used is a filamentous fungus selected from the group consisting of Pleurotus ostreatus, Ganoderma lucidum, Rhizopus oligosporus, Rhizopus microsporus var. oligosporus, Rhizopus oryzae, Neurospora intermedia, and Neurospora glabra, with Rhizopus oligosporus being preferred.

[0042] After growth on solid medium, in the pre-inoculation step, spores are suspended in sterile distilled water and inoculated at a concentration of 105 spores per mL of liquid medium. For this step, 500 mL Erlenmeyer flasks are used. The culture is maintained in an incubator with orbital agitation at 130 rpm at a temperature of 30-38°C, preferably 35°C, for 15-24 hours.

[0043] In this step, a defined liquid culture medium is used that may contain (i) a carbon source: mono-, di-, tri- and polysaccharides, preferably corn starch, (ii) a nitrogen source: yeast extract, malt extract, hydrolyzed or non-hydrolyzed cereal bran, ammonia and its salts, preferably brewer's residual yeast, and (iii) micronutrients as sources of magnesium, phosphorus and potassium, preferably magnesium sulfate, monobasic potassium phosphate and / or mixtures thereof, and that is responsible for the growth of fungi with the desired characteristics to obtain the mycelial biomass that is the object of the present invention. The liquid culture medium of primary interest is composed of a mixture of the following: Culture medium 1 contains corn starch, ammonium nitrate, magnesium sulfate, monobasic potassium phosphate, and yeast extract. Culture medium 2 contains corn starch, malt extract, yeast extract and magnesium sulfate. Culture medium 3 contains corn starch, malt extract, yeast extract and magnesium sulfate. Culture medium 4 contains brewer's waste yeast and corn starch.

[0044] Preferably, the liquid culture medium is as follows: Culture medium 1 contains 20-60 g / L cornstarch, 0.125 g / L ammonium nitrate, 0.2 g / L magnesium sulfate, 0.05 g / L monobasic potassium phosphate, and 5-20 g / L yeast extract. 50 g / L cornstarch and 12 g / L yeast extract are preferred. Culture medium 2 contains 20-60 g / L corn starch, 2-25 g / L malt extract, 5-20 g / L yeast extract, and 0.2 g / L magnesium sulfate. 50 g / L corn starch, 2 g / L malt extract, and 12 g / L yeast extract are preferred. Culture medium 3 contains 20-60 g / L corn starch, 2-25 g / L malt extract, 5-20 g / L yeast extract, and 0.2 g / L magnesium sulfate. 50 g / L corn starch, 25 g / L malt extract, and 4 g / L yeast extract are preferred. Culture medium 4 contains 50% (v / v) residual brewer's yeast and 20 g / L corn starch.

[0045] The process using culture medium 1 or 2 yields about 12 grams of dry mycelium per liter, using culture medium 3 yields about 11 grams of dry mycelium per liter, and using culture medium 4 yields about 14 grams of dry mycelium per liter.

[0046] After this period, the culture is transferred to a bioreactor for the fermentation stage. A pre-inoculum suspension is prepared at a ratio of 10% pre-inoculum to 90% sterile fresh culture medium for addition to the reactor. The suspension is transferred to the bioreactor, and aeration is adjusted. To obtain the desired biomass, aeration is adjusted to 0.5 to 3 VVM. The bioreactor used contains a set of flat-blade air intakes near the air inlet and an elephant-ear downflow valve at the top to optimize oxygen transfer and mixing time in the system. The pre-inoculum suspension is maintained in the bioreactor for 24 to 48 hours without the need for pH control throughout the process. The lack of pH control makes the process easier and less expensive.

[0047] Alternatively, pH control can be performed if this is of interest.

[0048] In the filtration step, the biomass produced in the above step is separated from the fermentation medium by simple filtration using a filter material with a porosity of 5 to 17 mesh, preferably a filter material with a porosity of 14 mesh.

[0049] The formed biomass is rinsed / washed under running water until the pH is neutralized. This step aims to remove all residues from the medium used in the process.

[0050] Furthermore, with the aim of inactivating the fungi, reducing the RNA content and extending the shelf life of the product, the mycelial biomass undergoes a heating step at 70°C for 2-45 minutes, preferably 10-15 minutes, preferably 15 minutes, after which the mycelial biomass is ready for pressing, freezing and / or use in the production of food products.

[0051] The fungal mycelium biomass formed is rich in fiber, has a high protein content and a very low energy value and fat index compared to similar products of plant origin (known as "plant-based").

[0052] According to the invention, the mycelial biomass obtained at the end of the pressing process has at least 1% to 10%, i.e., 1 g to 10 g of fiber for every 100 g portion. Animal proteins do not have fiber in their composition.

[0053] Fungal mycelium biomass has 60-150 kcal, preferentially 78 kcal and less than 4 g of fat per 80 g portion of biomass. This represents a very low energy value and very low fat content compared to similar plant-based products or traditional chicken. Similar plant-based products on the market have approximately 150 kcal, essentially double the 78 kcal biomass, while traditional chicken breast has approximately 126 kcal.

[0054] The market leading plant-based product has 10.8g of fat per 80g portion of product, while traditional chicken has 3.5g of fat per 80g portion of product, i.e. the plant-based product has 10 times more fat compared to the mycelial biomass of the fungus that is the object of the present invention, while traditional chicken has 3.5 times more fat.

[0055] In terms of protein content, the biomass of interest in this invention has a high protein content relative to the calories the product delivers to the consumer. In the same example above, biomass with a moisture content of 78% has an average of 10.5 g of protein per 78 kcal, i.e., a protein to calories ratio of 13.5%. Market-leading plant-based products have 11 g of protein per 150 kcal, i.e., a protein to calories ratio of 7%.

[0056] The mycelial biomass that forms the basis of the food product is composed, on average, of: aspartic acid 0.70%; glutamic acid 0.86%; serine 0.30%; glycine 0.40%; histidine 0.22%; taurine <0.01 (LQ); arginine 0.42%; threonine 0.33%; alanine 0.46%; proline 0.24%; tyrosine 0.29%; valine 0.44%; methionine 0.10%; cystine 0.07%; isoleucine 0.38%; leucine 0.53%; phenylalanine 0.32%; lysine 0.74%; hydroxyproline <0.01 (LQ); 6.81% total amino acids and 7.00 g / 100 g carbohydrates. The energy value is 78.12 kcal / 100g; moisture and volatile matter 78.11%; crude protein 10.46%; ether extract 0.92%; dietary fiber 2.09g / 100g; insoluble dietary fiber 2.09g / 100g; soluble dietary fiber <0.10g / 100g; mineral matter 1.42%. This gives the biomass high nutritional properties.

[0057] Furthermore, the following mycotoxins: total aflatoxins (B1+B2+G1+G2) and ochratoxin; and heavy metals: arsenic; cadmium; lead and mercury were not detected.

[0058] The mycelial biomass is the basis for the production of a food product with high nutritional properties and a visual appearance and texture similar to chicken meat. The product can be flavored.

[0059] To obtain mycelium biomass-based food, the following steps are carried out: (a) Preparation of mycelial biomass (b) Pressurization (c) Addition of additional ingredients (d) Heating and (e) Freezing is necessary.

[0060] In the mycelial biomass preparation step, biomass is obtained as described above.

[0061] After preparation, the obtained biomass is placed in a mold having a desired shape (for example, chicken breast) with perforations, and pressurized to 10 to 100 psi, preferably 20 psi.

[0062] In the additional ingredient addition step, the pressed biomass is immersed in a solution containing flavoring, chickpea flour, and gum arabic until the product absorbs 10% of its weight from the solution. The final concentration of the product can contain 0-4% gum acacia, 0.5-4% chickpea flour, and 0.05-0.7% flavoring.

[0063] Finally, the resulting product is heated to 70° C. for 15 minutes and frozen in a conventional freezer at −12° C. and −15° C. This step can be performed as a pre-freezing or pre-pressing step.

[0064] The resulting food product mimics a whole cut of chicken breast, has a neutral color similar to that of chicken breast, and has more than 50% horizontally aligned hyphal fibers, as shown in Figures 1 and 2. The product has a texture of 35-40 N WBSF (Warner-Bratzler shear force).

[0065] Furthermore, the food product of the present invention comprises 90-99% mycelial biomass by weight of the total product and 1-10% additional compounds by weight of the total product. More preferably, the food product comprises 95% or more mycelial biomass by weight of the total product, preferably mycelial biomass of the fungus Rhizopus oligosporus, and 5% or less other ingredients by weight of the total product.

[0066] According to the present invention, the additional compounds are understood to be acacia gum, starch, chickpea flour, flavorings, bamboo fiber, oat fiber, xanthan gum, coconut oil, corn oil or any other vegetable oil with a neutral flavor, olive oil, and / or mixtures thereof. Such a mixture of compounds is present in a proportion of 5% or less of the total weight of the product. A preferred additional component is a mixture of acacia gum, chickpea flour, flavorings, and / or mixtures thereof. The mixture contains 0-4% acacia gum, 0.5-4% chickpea flour, and 0.05-0.7% flavoring. Preferably, the mixture contains 2% acacia gum, 1.5% chickpea flour, and 0.2% flavoring. The mixture used can be in the form of a solution or suspension.

[0067] The resulting food has a moisture content of 65 to 90% and can be directly frozen and stored at -12 to -15°C in a standard freezer.

[0068] While preferred implementations of the present invention have been described, it is to be understood that the scope of the present invention encompasses other possible variations of the inventive concepts described and is limited only by the content of the appended claims, including possible equivalents.

[0069] Example 1: Fungal Cultivation Process The fungus Rhizopus oligosporus is inoculated onto solid BDA medium "potato dextrose agar" in a slant agar tube and incubated in an incubator at 30°C for 7 days. The grown spores are then resuspended in sterile distilled water. For the preparation of the preinoculum, approximately 105 spores / mL are transferred to 200 mL of liquid medium in an Erlenmeyer flask and maintained at 35°C. The liquid medium used consists of liquefied cornstarch (50 g / L); malt extract (2 g / L); yeast extract (12 g / L); and MgSO4.7h2O (0.2 g / L). The preinoculum is incubated in an orbital incubator with shaking at 130 rpm at 35°C for 24 hours. The culture is transferred to a bioreactor for the fermentation stage with 10% preinoculum and 90% fresh culture medium. To obtain the desired biomass, aeration is set at 0.75 VVM until the dissolved oxygen in the medium reaches 50% saturation, then aeration is set at 1.5 MVV using a set of flat blades near the air inlet and elephant ear blades. The culture is maintained in the bioreactor for 44 hours, and the pH is maintained at 5.5.

[0070] Example 2: Process for obtaining biomass The mycelial biomass obtained in Example 1 is filtered through a 14 mesh filter. The biomass is rinsed under running water and the residue from the growth medium is discarded.

[0071] The mycelial biomass can be heated to 70°C for 10 minutes and pressed and frozen or used in the production of food products as described in Example 3.

[0072] Example 3: Process for obtaining food The mycelial biomass obtained in Example 2 is immersed in a solution containing acacia gum (2%), chickpea flour (1.5%) and flavorings (0.2%). The biomass is left in contact with the solution for 15 minutes.

[0073] The mycelium mass is then transferred to a chicken breast-shaped stainless steel mold with perforations on the bottom and sides. The dough is then pressed until the final moisture content of the product is 80%. The final product has aligned fibers and a texture of 35-40N (WBSF). The sample is then heated to 70°C for at least 10 minutes and then frozen in a standard freezer.

[0074] Example 4: Sensory test Two blind sensory tests were conducted with seven participants each. Test 1 aimed to evaluate the addition of ingredients to improve the texture of the product. Test 2 aimed to evaluate a mixture of ingredients to improve the texture of the product. For each test, participants tasted the product and were asked to give a score from 1 to 10 regarding how pleasant the product was in terms of: chewiness, hardness, texture, and taste. The means are shown in Tables 1 and 2.

[0075] In experiment 1 (results in Table 1), low (1%) and high (4%) concentrations of biomass + bamboo fiber 90, bamboo fiber 200, and acacia gum were tested. The experiments with bamboo fiber 90 and bamboo fiber 200 resulted in poorer ratings because the product was more brittle and had a dry appearance. The acacia gum experiment resulted in an overall improvement in the scores assigned by participants.

[0076] [Table 1]

[0077] In experiment 2 (results in Table 2), three mixtures of acacia gum, bamboo fiber, chickpea flour and flavorings were used at the following concentrations: ●Mixture A: 2% acacia gum, 0.2% bamboo fiber, 1% chickpea flour, natural or identical chicken, garlic and onion flavor powder, 0.2%. ●Mixture B: Acacia gum 2%, bamboo fiber 0, chickpea flour 1%, natural or identical chicken, garlic and onion flavor powder, 0.2%. ●Mixture C: Acacia gum 2%, bamboo fiber 0.2%, chickpea flour 0, natural or identical chicken, garlic & onion flavor powder, 0.2%.

[0078] The concentration of 2% acacia gum was maintained with or without the addition of chickpea flour (1%) and bamboo fiber 90 (0.2%). It can be observed that the addition of chickpea flour made the product more acceptable, while bamboo fiber, even at a low concentration, resulted in a worse rating. Based on this study, chickpea flour and acacia gum are used in food products.

[0079] [Table 2]

[0080] Example 5: Pilot-scale mycelium production A bioreactor with a working volume of 200 L equipped with an agitation system consisting of Rushton paddles and elephant ear blades was inoculated with 10% (v / v) pre-inoculum as described in Example 1.

[0081] Cultivation in the bioreactor was carried out for 38 hours with pH control at 5.5 and a constant aeration rate. Different aeration rates and mechanical agitation were evaluated. The process yield was compared to the maximum yield obtained at laboratory scale and expressed as a percentage (50% yield means that the conditions produced half of the maximum possible biomass according to the tests performed at laboratory scale).

[0082] [Table 3]

[0083] At pilot scale, it was noted that agitation above 40 rpm resulted in a decrease in biomass production. This must have occurred due to the mechanical stress experienced by the microorganisms. Increasing air injection into the system, especially above 0.75 vvm, had a similar effect.

[0084] Example 6: Inoculation Rate Test Different inoculation rates were evaluated to initiate fermentation in a pilot bioreactor with a working volume of 200 L. Pre-inoculation cultures were prepared as described in Example 1. Test and pilot bioreactors were run in triplicate and under the same aeration rate, pH, and mechanical agitation.

[0085] The pre-inoculum can be made from a spore solution containing 10^5-10^10 spores / mL, preferably 10^8 spores, or from a liquid culture at a ratio of 2-10%, preferably 6%.

[0086] Biomass production on a dry basis was quantified at the end of the fermentation. A 10% inoculation rate produced 12 g / L of mycelium, while a 6% inoculation rate yielded 11.46 g / L and a 2% inoculation rate yielded 8.33 g / L. The possibility to use low inoculation rates is advantageous for scaling the technology. For example, at a 10% inoculation rate, a 200 L culture can inoculate 2,000 L of medium, while a 6% inoculation rate allows for the inoculation of 3,333 L.

[0087] In the second batch of tests, bioreactors containing 200 L of culture medium were inoculated with spore solutions containing 10^5, 10^8, and 10^10 spores. Under these conditions, the fermentation process continued for 50–55 h until total consumption of sugars from the culture medium, which varied in the amount of mycelial biomass produced. Compared to conditions with higher initial spore concentrations, cultures initiated with 10^8 spores resulted in 95% biomass formation, while those initiated with 10^5 spores resulted in 80%.

[0088] Example 7: Production of protein concentrate from dehydrated mycelium The resulting fresh biomass can be processed to remove moisture and obtain a dry protein concentrate. Dehydration can be achieved by a variety of methods, including tray drying, fluidized or spouted bed drying, spray drying, freeze drying, natural (solar), rotary dryers, tunnels, conveyors, cyclones, and screw advances. In this process, the process temperature should preferably be low (preferably 50-60°C), if it is desired to avoid protein hydrolysis, thermal destruction of vitamins, and lipid oxidation, and to maintain the nutritional and sensory properties of the mycelium. Temperatures ranging from room temperature to 100°C can be used.

[0089] Furthermore, the obtained biomass can be subsequently processed in a mill for standardization of the particle size distribution.

[0090] At this stage, 5 kg of fresh mycelial biomass was dried in trays in an oven with forced air circulation at 50 °C for 24 h. Finally, the dried material was processed in a disc mill and analyzed for nutrient composition.

[0091] [Table 4]

[0092] Drying has the added benefit of extending the shelf life of the product and avoids reliance on cold / frozen chains for product storage and transport, but it significantly alters the texture of the product, which does not revert to its original fibrous structure when rehydrated.

[0093] Example 8: Application in meat products Hybrid products were prepared by mixing lean meat with fresh mycelium biomass at 80% moisture content, as shown in Figures 3 and 4, and compared to products containing only meat, with and without dye. Different ratios of beef and mycelium were evaluated. Hamburgers were prepared using 100% beef as a control (comparison) product. In the first test, 100g hamburger patties were prepared by hand and 1% natural beet pigment was added. A sensory test was then conducted with 12 participants (Table 5).

[0094] It was noted that maintaining a beef content of at least 50% resulted in a high product acceptability. Below this content, some loss of chewiness and texture was evident. The poorer flavor perception can be justified by the fact that no flavorings were used in this formulation. Also, the lower score for appearance was related to the color perception, since beet pigments result in a pink product rather than the brownish color of cooked meat, giving an artificial impression.

[0095] [Table 5]

[0096] Example 9: Reducing environmental impact Agriculture and livestock currently account for 70% of water consumption and 25% of greenhouse gas emissions caused by human activities. Animal production stands out because it is directly dependent on agricultural production and brings with it all the environmental impacts that arise during its life cycle, such as the use of fertile land and chemicals (fertilizers, manure, pesticides, etc.). It also contributes significantly to methane emissions, the consumption of water resources, energy consumption, the pollution of water bodies, the generation of liquid and solid waste that is difficult to treat, and the disposal and degradation of soil. Agriculture and livestock are also closely linked to deforestation, as their production is horizontal and the expansion of production areas requires the expansion into new areas.

[0097] Beef, chicken and pigs are currently the most consumed sources of animal protein in the world. In the case of beef, only 40-45% of its weight is meat, of which an average of 25% is protein. A beef cow is slaughtered after 24 months and consumes approximately 15000 calories a day. 2 Considering animals slaughtered at 650 kg (average for 2021), this amounts to just 0.006 kg of protein / day / m 2 The water footprint of beef production is 10.5m per kg of meat. 3 , or 42m per kg of protein 3 It is estimated that...

[0098] The productivity achieved by the process described in this paper allows us to 3 12 g of dry mycelial biomass can be obtained per ml of culture medium. The resulting biomass has a protein content of 48% on a dry basis and is produced in 24 hours. 3 This corresponds to 5.76 kg of protein per day. 3 Considering a bioreactor of approximately 58.5 kg of protein / day / m 2 , i.e., protein productivity nearly 10,000 times higher than that of beef. In terms of water footprint, the inventors estimate that this technology uses at least 10.5 times less water (the calculation could be even lower if considering the use of industrial waste as part of the growth medium).

Claims

1. 1. A mycelial biomass having high nutritional properties, characterized in that it contains a filamentous fungal mass that is free of chemical additives, wherein the filamentous fungus is selected from the group consisting of Pleurotus ostreatus, Ganoderma lucidum, Rhizopus oligosporus, Rhizopus microsporus var. oligosporus, and Rhizopus oryzae.

2. 2. The mycelial biomass of claim 1, wherein the filamentous fungus is Rhizopus oligospourus.

3. 3. Mycelial biomass according to claim 1 or 2, characterized in that it contains 1% to 8% fiber, 60-90 kcal and less than 1 g of fat per 80 g of biomass, with an average of 10.5 g of protein for every 78 kcal.

4. On average, aspartic acid 0.70%, glutamic acid 0.86%, serine 0.30%, glycine 0.40%, histidine 0.22%, taurine <0.01 (LQ); arginine 0.42%, threonine 0.33%, alanine 0.46%, proline 0.24%, tyrosine 0.29%, valine 0.44%, methionine 0.10%, cystine 0.07%, isoleucine 0.38%, leucine 0.53%, phenylalanine 0.32%, lysine 0.74%, hydroxylase 0.82%, hydroxypropyl 1.0 ...

4. The mycelial biomass according to claims 1 to 3, characterized in that it contains <0.01 (LQ)% hydroxyproline; 6.81% total amino acids and 7 g / 100 g carbohydrates; energy value 78.12 kcal / 100 g; moisture and volatile matter 78.11%; crude protein 10.46%; ether extract 0.92%; dietary fiber 2.09 g / 100 g; insoluble dietary fiber 2.09 g / 100 g; soluble dietary fiber <0.10 g / 100 g; and mineral matter 1.42%.

5. (a) Growth of fungi in solid media; (b) Pre-inoculation; (c) fermentation in a bioreactor; (d) filtration of the filamentous fungal mass; (e) Washing / rinsing the dough; (f) fungal inactivation; and (g) Drying The process for producing biomass according to any one of claims 1 to 4, characterized in that it comprises the steps of:

6. 6. A production process according to claim 5, characterized in that in the stage of growth in solid medium, the fungus is inoculated into BDA medium "potato dextrose agar" in agar tubes and kept tilted at a temperature between 25 and 39°C for 4 to 10 days.

7. 6. The production process according to claim 5, wherein the fungus used is a filamentous fungus selected from the group consisting of Pleurotus ostreatus, Ganoderma lucidum, Rhizopus oligosporus, Rhizopus microsporus var. oligosporus, Rhizopus oryzae, Neurospora intermedia, and Neurospora glabra.

8. 6. The production process according to claim 5, characterized in that the liquid medium comprises corn starch, yeast extract, malt extract, brewer's residual yeast, ammonium nitrate, magnesium sulfate, monobasic potassium phosphate and / or mixtures thereof.

9. 6. The production process of claim 5, wherein the fermentation step comprises: preparing a pre-inoculum suspension in a ratio of 2-10% pre-inoculum to 90-98% sterile fresh culture medium; transferring the pre-inoculum suspension to the bioreactor; adjusting the aeration to 0.5-3 VVM; and maintaining the pre-inoculum suspension in the bioreactor for a period of 24-48 hours, without the need for pH control.

10. 6. The production process according to claim 5, characterized in that in the filtration step, the biomass produced in the previous step is separated from the fermentation medium by simple filtration using a filtration material with a porosity of 5 to 17 mesh.

11. 6. Production process according to claim 5, characterized in that in the rinsing / washing stage, the formed biomass is rinsed under running water until the pH is neutralized and all residues of the medium used in the process are removed.

12. 6. The production process according to claim 5, characterized in that in the inactivation step of the fungus, the biomass is heated to 70°C for 10 to 15 minutes.

13. 6. The production process according to claim 5, characterized in that the drying step is carried out in a tray, fluidized or spouted bed, spray dryer, freeze-drying, natural (solar), rotary dryer, tunnel, conveyor, cyclone, advancing screw.

14. Production process according to claims 1 to 13, characterized in that the obtained biomass is pressed and frozen or frozen for subsequent pressing.

15. A product that is a partial or complete substitute for animal protein, characterized in that it contains 40 to 99% of the mycelial biomass according to claims 1 to 4 based on the total weight of the product, 0 to 50% of meat and 1 to 10% of additional compounds based on the total weight of the product.

16. 16. Product according to claim 15, characterized in that said additional compounds are selected from the group consisting of acacia gum, chickpea flour, flavorings, bamboo fiber, oat fiber, xanthan gum, coconut oil, corn oil or any other vegetable oil with a neutral flavor, and / or mixtures thereof.

17. Use of biomass according to claims 1 to 4, characterized in that it is used to prepare food products which are partial or complete substitutes for animal proteins.

18. (a) preparation of mycelial biomass by the process of any one of claims 5 to 14; (b) Pressurization (c) Addition of additional ingredients (d) Heating and (e) Freezing 17. The process for producing a product according to claim 15 or 16, characterized in that it comprises the steps of:

19. 20. The process for producing a product according to claim 18, characterized by the step of adding said additional ingredients, wherein the pressed biomass is immersed in a solution containing flavorings, chickpea flour and gum arabic until the product has absorbed 10% of its weight from the solution.

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