WO2023026072A1

WO2023026072A1 - Process for producing a raw material of basidiomycete fungal mycelium and residual constituents of an agricultural side stream

Info

Publication number: WO2023026072A1
Application number: PCT/IB2021/057703
Authority: WO
Inventors: Alexander Stephan
Original assignee: Science & Life Consulting Gbr
Priority date: 2021-08-23
Filing date: 2021-08-23
Publication date: 2023-03-02

Abstract

The invention aims to create a process for providing a virtually allergen-free raw material as fermentation product of an allergenic, protein-rich agricultural side stream for virtually allergen-free food or feed and/or a virtually allergen-free food or feed preparation for humans and animals formed of a predominant fraction of basidiomycete fungal mycelium, and also the virtually allergen-free raw material as such. This is achieved by expanding upon known methods for fermentation of agricultural side streams with basidiomycetes through a separate preselection of at least one suitable basidiomycete by means of propagation samples on agar plates with fractions of the agricultural side stream containing allergenic proteins. In the further preculture step for propagating sufficient amounts of the selected basidiomycete, the ergosterol content of the basidiomycete mycelium is analysed. In addition, the amount of the allergenic protein in the agricultural side stream to be fermented is determined before the inoculation thereof. During the fermentation the ergosterol content of the fermentate is determined at least once. The raw material accrued (fungal mycelium + residual materials) is lastly analysed in the laboratory for residual amounts of allergenic proteins. The raw material consists of a predominant fraction of a fungal mycelium of a basidiomycete and a relatively small fraction of residual materials from the agricultural side stream, where the content of ergosterol is at least 11 mg/kg, based on the dry mass.

Description

Process for producing a raw material from basidiomycete fungal mycelium and residual components of an agricultural by-product

The invention relates to a method for producing a raw material from basidiomycetes mycelium and residual components of an agricultural by-product for food or feed and/or food or feed preparations including fermentation mixtures, functional mixtures and flavor mixtures, for humans or animals and the raw material as such.

I. Field of application

The aim of the invention is to provide large quantities of an almost allergen-free raw material consisting of basidiomycetes fungal mycelium and residual components from widely available agricultural side-streams containing allergenic vegetable and/or animal proteins by fermentation. This almost allergen-free raw material can be used for food or feed and/or food or feed preparations, including fermentation mixes, functional mixes and flavor mixes, for humans or animals.

II. Technical background

The present invention deals with in the food and feed industry in large quantities accruing the agricultural by-products such as wheat draff or by-products from soybean production.

These agricultural by-products are usually rich in carbohydrates and roughage and are therefore attractive for further use.

For some time now, agricultural side-streams have been analyzed for possible further use in terms of their ingredients, in order to then subject them to fermentation processes, whereby the side-streams in fermented form are converted into raw materials, e.g. for the food or animal feed industry, resulting in new products.

Known fermentation aids are usually microorganisms selected from bacteria, yeasts or molds, but pillar fungi (Basidiomycetes) are also used as fermentation aids.

Attractive agricultural by-products are also those that contain a particularly large amount of plant and/or animal proteins, since protein-rich raw materials have been used in various foods or feedstuffs that make an essential contribution to human or animal nutrition, particularly in the last decade.

However, all the proteins contained therein (e.g. milk protein, soy protein or wheat protein) can trigger an allergic reaction after ingestion by animals and/or humans, i.e. in the case of a food allergy an immunoglobulin E (IgE ) response, which in turn can lead to an allergic cascade in the organism.

The food or animal feed industry reacts to this on the one hand by declaring the main allergens according to European legislation.

In addition, protein-rich fractions are increasingly being used as starting fractions, which do not fall under the obligation to declare, but which also have a negative effect in the event of increased consumption. tive IgE response, such as peas, chickpeas, field beans, potatoes or mung beans.

The present invention deals exclusively with those protein-rich agricultural by-products that contain, in particular, allergenic proteins.

Raw materials for food or feed and/or food or feed preparations that have been produced by fermentation of selected, particularly carbohydrate-rich agricultural by-products using basidiomycetes are already known, and with such fungal mycelia of some basidiomycetes cultivated in the fermenter, dried and pulverized, has been experimented with for some time as a basic ingredient in meat-free products, but according to the current state of the art, fermentation is always carried out with the sole aim of improving the techno-functional properties of the end product, for example its water solubility, as shown e.g. in DE102016110653A1 has been. This document is seen as the closest prior art.

In DE102016110653A1 specifically carbohydrate-rich agricultural side streams were selected as starting fractions for further fermentation, which according to the invention described there were not fermented using special bacteria, but using various available pillar fungi (Basidiomycetes) of the genus fungi, who surprisingly split the carbohydrates present in the agricultural by-product in such a way that the end product is much easier to digest.

The end product described in DE 102016110653A1, a raw material from basidiomycetes mycelium and residues of the carbohydrate-rich agricultural by-product, for the substitution of vegetable proteins in the food and feed sector, was then considered to be inventive in Regarding very good techno-functional properties revealed due to good solubility, good water binding capacity and also a good oil-binding capacity of the disclosed raw material, which above all has a significant influence on sensory parameters of the subsequent end product, such as a vegan sausage based on mushroom mycelium.

The research focus of this closest prior art was therefore primarily to develop a raw material containing mushroom mycelium, which has various techno-functional properties favorable for further processing, in order to be able to be used as a substitute for vegetable proteins, e.g. in vegan sausages. i.e. the raw material was examined, for example, at which temperature the mushroom protein denatured, how well the raw material combined with other water- or fat-containing components of the sausage, what color and taste the raw material had, etc.

However, at the time DE 102016110653A1 was published, the raw material was not examined to determine how high its content of allergenic proteins was, since it had to be expected that the fermentation product, i.e. the raw material obtained from fungal mycelium and residues of the agricultural by-product , still has a large number of allergenic proteins from the original fraction.

The problem that has not yet been solved remained that many agricultural by-products not only contain carbohydrates in particular, but also contain large amounts of allergenic proteins (epitopes) of plant or animal origin, which means that even after a fermentation process according to the disclosure from DE 102016110653A1 End product (raw material) containing mushroom mycelium can still trigger allergic reactions in animals and humans.

That certain basidiomycetes are suitable, in known fermentation processes of agricultural by-products, an almost complete conversion of allergenic or potentially allergenic epitopes of the plant and/or animal proteins in the protein-rich fractions used into fermentation products from fungal mycelium that practically no longer trigger allergies and converting residues was not previously known and is a surprising discovery of further investigations on the raw materials obtained by the fermentation process from DE 102016110653A1, which led to further inventive modifications of the fermentation process using basidiomycetes and is the basis of the present patent application.

III. object of the invention

It is therefore the object of the present invention to provide a process for providing an almost allergen-free raw material as a fermentation product for almost allergen-free food or feed and/or an almost allergen-free food or feed preparation for humans and animals from a predominant proportion of basidiomycete to create mushroom mycelium.

Likewise, a fermentation product is to be created as a raw material for further use as such, in which in the manufacturing process, depending on the starting fraction, which is a widespread, particularly allergenic, protein-rich agricultural by-product, through the fermentation with specific, particularly suitable and selected Basidiomycetes of the genus fungi, cultivated submersed or emersed, the number of epitopes of one or more proteins in the fraction used is minimized or converted.

The finished end product as a raw material is therefore a mixture of a predominant proportion of a protein-rich fungal mycelium of at least one specific basidiomycete and a smaller proportion of residues from the agricultural by-products used in the fermentation process.

In this raw material according to the invention, the amount of remaining allergenic proteins of the starting fraction should be so low that it can no longer be detected using an ELISA test and the human or animal organism no longer produces a perceptible IgE response. IV. Solution of the task

This problem is solved by an extension of the well-known fermentation process of agricultural by-products with basidiomycetes, which includes the following new steps:

The method according to the invention can be applied to agricultural by-products which contain quantities of allergenic proteins of plant or animal origin that are problematic for further use. As a new, essential process step, the process step of a separate selection of at least one suitable basidiomycete is first introduced. In order to select and check the growth behavior of the possible, suitable basidiomycetes in the agricultural side-stream to be fermented, according to the invention a first pre-culture is set up, in which agar plates are used whose agar contains malt extract contains 1.5 - 3 percent by weight of the agricultural side stream containing allergenic proteins, whereby the warm liquid agar is also adjusted to pH 7 with sodium hydroxide solution or hydrochloric acid (20 g agar-agar for microbiology; 20 g agricultural side stream calculated on the dry matter and 960g distilled water).

Various pieces or mycelia of selected basidiomycetes are then applied to these special agar plates of the first preculture (placed in the middle with about 1 cm ² ) and cultivated in a manner customary for the person skilled in the art for preferably four to five days at preferably 24.degree , usually after a few days, to check the growth behavior of the selected fungal mycelia of various basidiomycetes on the nutrient medium containing the agricultural by-product.

In addition, at the end of the growth phase of the first preculture, each agar plate is sniffed and the basidiomycete which has developed a bad odor and which is perceptible is sorted out. If a certain basidiomycete has developed particularly well on the special agar of this first preculture, which also contains the agricultural sidestream to be fermented later and has shown strong mycelia growth, it is selected as suitable and used as an inoculum in the subsequent fermentation process to become.

For the process according to the invention, numerous first pre-cultures have already been carried out on seven common agricultural by-products with the following results:

Agricultural side streams:

Malt germ protein extracted malt with a high husk content

Residual stream from soy protein production

Solubles from pea starch production

Residual stream from mung bean protein manufacture

chickpea flour

rice protein

The following fungi (basidiomycetes) were tested:

1. LED lentinula edodes

2. POS Pleurotus ostreatus

3. PSS Pleurotus stramineus salmoneus (Pleurotus djamor)

4. MCO Marasminus cohortalis

5. PSA Pleurotus sapidus

6. FVE Flammulina velutipes

7. PPO Pleurotus pulmonarius

8. PER Pleurotus eryngii

9. PDY Pleurotus dryines

10. WCO Wolfoporia cocos

11. BAD Bjerkandera adusta

12.CLG Clitocybe lignatilis

13.CPE Clitopilus passeckerianus 14.CHE Cyathus helenae

15.CAP Ganoderma applanatum

16. HCN Hypholoma capnoides

17. IBN Ischnoderma benzoinum

18.KMT Kuehneromyces mutabilis

19.LSQ Lentinus squarrosulus

20.MSO Marasmius scorodonius

21.NVT Nidula niveo- tomentosa

22.POT Panellus serotinus

23. PCY Phanerochaete chrysosporium

24.PCI Pleurotus citrinopileatus

25.POU Pleurotus eous

26.PML Polyporus melanopus

27.PCE Postia caesia

28.PYS Pycnoporus sanguineus

29.TSV Trametes suaveolens

30.TMV Trametes versicolor

31.TYC Tyromyces chioneus

32.EPI Fomitopsis pinicola

According to the first pre-cultivation trials, the following basidiomycetes have proven to be particularly suitable for the seven agricultural side-streams mentioned, which occur widely and occur in large quantities (compare Table 1):

Malt germ protein: MCO, PPO, PER, PML Extracted malt with high husk content:

MCO, PPO, PER, NVT, POT, PML, TYC

Residual stream from soy protein production:

POS, PSS, CPE, MSO, PCI, POU, PYS, TSV

Solubles from pea starch production:

LED, POS, PSS, MCO, CPE, CHE, MSO, PCI, POU, PYS, TSV

Residual stream from mung bean protein production:

LED, POS, PSS, MCO, MSO, PCI, POU, PCE, PYS Chickpea Flour:

POS, PSS, MSO, FVE, CHE, LSQ, PCI, PCE

Rice Protein:

POS, PSS, FVE, CPE, CHE, LSQ, PCI, TYC

After this new process step according to the invention, the first preculture for the selection of suitable basidiomycetes for the subsequent fermentation of the existing agricultural sidestream, according to the invention, as a second preculture process, further fungal mycelium of the at least one selected basidiomycetes is first grown on standard agar plates with malt extract Cultivated using known culture techniques to obtain a larger quantity of mushroom mycelium for the fermentation process, i.e. for approx. 4 - 5 days at 24°C.

The further mycelium propagation of the second preculture process is then continued in liquid standard nutrient medium made from agar and malt extract, since the inoculum for the subsequent submerse cultivation as well as for an emerse cultivation is always introduced into the fermenter as a liquid inoculum for the agricultural bypass stream to be fermented.

During this second preculture process for growing sufficient amounts of the inoculum on malt agar plates or in malt agar medium, the ergosterol content of the at least one selected basidiomycete mycelium is determined in the laboratory using known digestion methods as a further process step before the start of fermentation according to the invention. and extraction methods are determined in order to obtain a reference value. This determination is advantageously carried out on mycelium which was cultivated on the standard agar-malt plate. Depending on the type of basidiomycete, the mushroom mycelium contains its own specific amount of ergosterol. In the test series of the present invention, the ergosterol content was determined as a reference content according to method no. 81 from 2015 of the International Fruit and Vegetable Juice Association.

Ergosterol is a building material of cell membranes that occurs primarily in fungi and is suitable for detecting fungal growth in fermentation processes. It is known, for example, to detect mold infestation of tomato juices by determining the ergosterol content in tomato juices. In this case, it is used to detect the percentage of basidiomycetes in the end product. Before the start of fermentation, in a further inventive process step, the agricultural by-product stream containing allergenic proteins is then analyzed in a laboratory according to known methods in order to determine the amount of the allergenic-reaction-triggering proteins therein before the start of fermentation. This amount of allergenic proteins is said to decrease significantly during fermentation with increasing growth of fungal mycelium in the agricultural side-stream.

For example, the determination of the content of soy allergens in an agricultural side stream containing soy allergens before the start of the fermentation process was carried out in our own test series according to IFP 002822 (ELISA): 2020-01 (a) ELISAFast® Soy with a limit of quantification of 1 mg/kg carried out.

For example, the determination of wheat allergen in an agricultural side stream containing wheat allergen before the start of the fermentation process was carried out in our own test series according to IFP 002822 (ELISA): 2020-07(a) once with R-Biopharm "RIDASCREEN® Gliadin competitive" (Art. R7021, competitive ELISA) , detection limit gluten 4.6 mg/kg, limit of quantification 10 mg/kg with a limit of quantification of 10 mg/kg and carried out once with ELISAFast® gluten (sandwich ELISA based on R5 antibodies by Prof. Mendez) detection/quantification limit 3 mg/kg to detect all epitopes that cause a detectable IgE response in humans and animals.

Now the actual fermentation process begins. For this purpose, according to the invention, a specific amount of the agricultural by-product, liquid or solid, is transferred to a fermenter of a suitable size and inoculated with the at least one selected basidiomycete mycelium as a liquid inoculum.

The fermentation of the agricultural sidestream used by basidiomycetes can be carried out either as a submerged cultivation of the fungal mycelium in a liquid environment or as an emerald cultivation of the fungal mycelium in a merely moist environment. The fermentation of the agricultural by-product now proceeds, submerged or emersed, in the manner known for both processes and already disclosed under suitable temperature conditions over a normal period of time, generally 4 - 6 days at 24°C.

With both cultivation methods, submersed or emersed, an allergen-reduced, protein-containing basidiomycete mycelium grows in the fermenter, which also contains ergosterol.

During the fermentation process, the ergosterol content of the fermentate is also measured at least once according to known laboratory methods as a further, new and inventive process step. Determining the ergosterol content in the respective fraction during the fermentation process can be used to demonstrate increasing fungal growth with a simultaneous decrease in the amount of the specific allergenic proteins in the respective fraction. It is advantageous to carry out these measurements at least once to be carried out after 1 to 6 days.

By analyzing the parameters "amount of allergen" and "ergosterol content" during the fermentation or after the end of the fermentation, it can be seen directly whether the fungus has grown sufficiently and whether the allergen has been broken down during the fermentation.

This interplay is illustrated below using two tables (Table 2 and Table 3) from our own test series.

Table 2: Presentation of the allergen soy, ergosterol and protein

PSS SN = Pleurotus stramineus salmoneus cultivated on soya side stream

SN = soy sidestream

Table 3: Presentation of the allergen wheat as well as ergosterol and protein

PER WN= Pleurotus eryngii cultivated on wheat draff

PER MN= Pleurotus eryngii cultivated on malt germ protein

It was found that by fermenting a certain agricultural by-product containing allergenic proteins with specific basidiomycetes of the fungi, submersed or emersed genus, tailored to the starting fraction, Fourth, the amount of allergenic epitopes in the fraction is minimized or degraded in such a way that after the end of the fermentation, when the raw material produced from it is consumed, there is no longer any noticeable allergic reaction in the consumer (human or animal) (acting according to the so-called VITAL concept of the Federal Institute for Risk Assessment, Berlin).

The allergenic proteins originally present in the agricultural side-stream used are transformed by the specially selected basidiomycetes used and/or integrated into their own fungal protein matrix and lose an allergenic potential of at least 40% and at most 99.9%.

After completion of the fermentation process, the method according to the invention finally leads to a specific quantity of a raw material, which consists of a predominant proportion of a fungal mycelium suitable for the food and animal feed industry of at least one basidiomycete and a smaller proportion of residues from the original jump faction.

A further process step according to the invention now includes a laboratory analysis of the raw material obtained by means of ELISA tests, whereby in the context of all experiments that led to the basis for the process according to the invention, the raw material as a whole had almost no traceable contained more allergenic proteins from the agricultural by-products used.

This practically allergen-free raw material obtained according to the invention can be used as food or feed or as a food or feed preparation of the food after fermentation of the agricultural by-product used has been completed in large fermenters with a large capacity and, if necessary, subsequent drying and/or pulverization of the raw material - and animal feed industry are provided in large quantities, as it is extremely rich in non-allergenic mushroom proteins, contains, among other things, roughage, minerals and trace elements and some fat.

The method according to the invention thus includes, in an inventive way, the selection of particularly suitable basidiomycetes for special, protein-rich agricultural side streams in a first preculture process, since not every possible basidiomycete in the fermentation process has a suitable raw material in sufficient quantity with the required properties in every agricultural side stream. property, namely a raw material that contains a predominant proportion of a special, non-allergenic fungal mycelium of at least one specific basidiomycete, which incorporates the previously allergenic proteins in the agricultural by-product in a proportion of at least 40% to a maximum of 99.9% into its own fungal protein matrix has.

According to the invention, the content of the non-allergenic mycelium in the raw material (mixture) for an almost allergen-free food or feed or for an almost allergen-free food or feed preparation is 50.01 to 99.9% by weight, based on the total weight of the raw material quantity. This gives users the opportunity to provide a raw material for an almost allergen-free food or feed preparation for humans or animals, depending on the specific requirements of this food or feed preparation.

It is also provided that the raw material according to the invention, which is almost allergen-free and consists of a predominant proportion of mushroom mycelium, can be used in a fermentation mixture and/or functional mixture and/or flavor mixture. These can then in turn be used in a food or feed preparation, e.g. B. for further fermentation of a food preparation and/or for fermentation or stabilization of the food preparation using suitable additional microorganisms or for seasoning, for example, a vegan sausage mass. The raw material according to the invention, which is almost allergen-free, can also be used as animal feed either individually or in mixtures with other non-allergenic proteins. It is used to feed animals and makes a major contribution to animal health, muscle building or as a substitute for animal meal. The raw material according to the invention can be used as feed directly or in mixtures in a proportion of 0.01 to 99.5% by weight, in particular from 5 to 90% by weight, with other feed ingredients such as fibers, vitamins, various carbohydrates or fats are used.

Furthermore, the almost allergen-free raw material according to the invention can be used in all known foodstuffs that are defined in the guidelines for bread and biscuits and/or the guidelines for fine baked goods and potato products. It is used here as a well-tolerated source of protein, e.g. as a substitute for gluten. The raw material according to the invention can also be used in various concentrations in pasta according to the guiding principles for pasta.

The raw material according to the invention can also be used as an alternative ingredient in all currently known meat and sausage products according to the guiding principles for meat and sausage products. It serves as a non-allergenic source of protein to substitute a certain proportion of meat protein.

With various mixtures of other, non-allergenic vegetable proteins, additives according to European Regulation 1333/2008 of the European Parliament and of the Council, hydrocolloids, starches, sugars, spices, spice extracts, flavorings according to European Regulation 1334/2008 of the EU European Parliament and Council, minerals and fats, any type of vegan or vegetarian products can be produced by adding the raw material according to the invention. The method according to the invention in the form of the two possible fermentation processes, submersed or emersed, are explained in more detail below:

Description of the submerged cultivation of basidiomycetes according to the invention:

The submerged cultivation of basidiomycetes can be carried out with various agricultural by-products containing allergenic proteins, such as brewer’s grains, soy by-products, protein-rich fractions from juice production such as carrot pomace and/or apple pomace, or shells or press cakes from oil production such as sunflower seed shells and/or sunflower seed pomace or All other protein-containing agricultural side streams that are in liquid form for fermentation. If the agricultural side-stream is in the form of a solid, it is liquefied using distilled, sterilized water before the start of the fermentation process.

After the selection of at least one suitable basidiomycete in a first preculture process, a second preculture process follows for the submerse cultivation, in which a sufficient amount of the desired basidiomycete mycelium from the first preculture for the submerse cultivation is first placed on standard agar plates with malt extract , then grown in liquid medium.

The nutrient media for the standard plates or the agar for the liquid medium of the second preculture usually consist of a mixture of 15.0 g/L agar-agar and 20 g/L malt extract (MAE) that are heated at 121 °C for 20 min autoclaved and then poured into sterile Petri dishes.

These MAE agar plates for the second preculture are each inoculated with an approx. 1 cm ² piece of agar covered with mycelium from a basidiomycete selected in the first preculture step, sealed with Parafilm and placed in an incubator at 24°C for one to several days cultivated. The to approx. 80% overgrown plates are stored at 4°C and regularly inoculated using the same method. This cultivation serves as a stick for submerged cultivation.

According to the invention, the ergosterol content of the cultivated mushroom mycelium is now determined by laboratory analyzes in order to obtain a reference value for the analyte ergosterol for further investigations in the later fermentate from the submerged cultivation.

The final cultivation of the selected basidiomycetes mycelium culture for the subsequent fermentation process in the submersed cultivation process for the production of even larger amounts of the raw material according to the invention (process of the second preculture) takes place in a liquid malt agar medium, since the inoculum for submersed cultivation must be provided in liquid form. For this purpose, the fungal mycelium that has grown on the malt agar plates is finally transferred, for example, to a sterile Erlenmeyer flask containing autoclaved, liquid malt agar medium.

This entire second, liquid pre-culture is then used to inoculate the main culture, the agricultural by-product.

Filling a fermenter as follows has proven to be advantageous as a recipe for submerged cultivation according to the method according to the invention:

A 0.5 to 150,000 L fermenter is inoculated with 5-35% dry or liquid agricultural sidestream (calculated on dry matter) with 40 - 90% water and 5 - 25% liquid inoculum to a maximum nominal amount of 55 to 90 % filled.

For fermentation on an industrial scale, the necessary amounts of all components are adjusted accordingly.

Fermentation then preferably takes place in the dark for 4 to 19 days (d) in the fermenter or, depending on the production size, also in an incubator shaker at 24 °C in a closed system with an agitator speed of 150 - 250 rpm and an air dosage of 0.1 to 3 volumes per fermenter volume.

During the submerged fermentation process, the ergosterol content of the fermentate is measured at least once in order to be able to estimate the growth of the fungal mycelium and to determine whether the amount of allergenic proteins in the agricultural by-product is continuously decreasing.

After completion of the fermentation process, the fermentate is separated by filtration (solid/liquid), with the liquid supernatant of the fermentate being discarded. The fermentation product obtained (the raw material from mushroom mycelium and residues) is then processed into a powder with the help of commercially available drying technologies such as lyophilization, roller drying or spray drying, which at least has a water content of between 0.1 and 15%, but preferably in the range of 0.5 to 13% and particularly well in the range 0.75 to 11%. The drier the raw material is, the more stable it remains for further use.

Description of the emeral cultivation of basidiomycetes according to the

Invention:

The agricultural side-stream present as dry matter is weighed into the fermenter for emerald cultivation. The particle size is advantageously between 0.01-1 cm. The substrate moisture is then adjusted to preferably 65% by adding distilled, sterilized water.

Subsequently, the liquid inoculum suspension produced from the second preculture step according to the invention of the at least one basidiomycete selected in the first preculture and cultivated in larger amounts in the second preculture is introduced and mixed with the substrate mix well mixed. This is then indeed moist, so that the fungus grows on the surface of the substrate mixture, but not as liquid as in submerged cultivation, where the fungus grows in a liquid.

Filling a fermenter as follows has proven to be advantageous as a recipe for an emerald cultivation according to the method according to the invention:

A solids fermenter with an agitator and an atomizing device is filled with at least 25 - 85% (calculated on the nominal volume) dry agricultural bypass flow (calculated on dry matter) with an advantageous particle size of 0.1 - 1 cm and by adding distilled, sterile - adjusted water to a substrate moisture content of 50 - 80%, preferably 65%. Then, advantageously, 10% liquid inoculum is filled into the fermenter via an atomizing device and everything is stirred together.

Thereafter, stationary fermentation takes place at a preferred temperature of 32° C. and a preferred pH value of 3.5 to 6.9. The fermentation material advantageously has an average bulk density of 0.1 g DM/mL and is advantageously gassed axially with sterile water-saturated air during the fermentation in order to obtain a preferred atmospheric humidity of 88% in the solids fermenter, since Mushroom mycelium grows better with increased humidity than with dry air. The course of the emer cultivation can be followed visually and by the CCk exhaust gas content.

According to the invention, the ergosterol content in the fermentate is determined at least once. The fermentation is ended after preferably about 120 hours. If necessary, however, non-fermented agricultural side stream can be added again after a fermentation time of at least 8 hours and at most 30 hours.

The fermentation products obtained (a mixture of mushroom mycelium and residues of the agricultural by-products used (raw material)) are then processed into powders, as is the case with the fermentation products from submerged cultivation, with the aid of commercially available drying technologies such as lyophilization, drum drying or spray drying. having a water content between 0.1 to 15%, but more preferably in the range 0.5 to 13% and especially well in the range 0.75 to 11%.

Before the raw material from submerged or emerald cultivation is used in the production of animal feed or foodstuffs, according to the invention it is analyzed in the laboratory using ELISA tests with regard to the residual amount of allergenic proteins from the original fraction and the ergosterol content determined and ultimately only raw materials selected for use in the food and feed industry in which at least 40% to a maximum of 99.9% of the allergenic proteins of plant or animal origin contained in the agricultural by-product were integrated into the fungal protein structure and the content of ergosterol based on the dry matter of the raw material is at least 100 mg/kg.

As a result, powder is produced as the raw material, which, due to the agricultural by-pass flow, can sometimes turn light or sometimes brownish.

Depending on which food application is in focus, various raw materials from various basidiomycetes/agricultural by-products can be used.

The raw materials have a very neutral taste, somewhat reminiscent of nuts or mushroom-like. By drying, a raw material can be produced that is extremely high in protein, contains fiber and some fat. Before the raw material then goes into the production of animal feed or foodstuffs, according to the invention it is analyzed in the laboratory for the residual content of allergenic proteins from the starting fraction and finally only such raw material for use in the foodstuffs and animal feed - selected from the pharmaceutical industry in which at least 40% and at most 99.9% of the allergenic proteins of plant or animal origin contained in the agricultural by-product have been integrated into the fungal protein structure and the ergosterol content, based on the dry matter of the raw material, is at least 100 mg/kg.

The transformation is calculated via the ergosterol content, with the content of the pure fungal mycelium differing among the basidiomycetes. Only the mycelium of the mushroom mycelium (grown on a malt agar plate) is used and the ergosterol content analyzed. This value is equated with 100%. The analyzed ergosterol content of the fermentate is then determined analogously and the concentration of the fungus therein is calculated.

Claims

Claims 1.A method for producing a raw material from Basidiomycetes fungal mycelium and residual components of an agricultural by-product for food or feed and/or food or feed preparations including fermentation onsmixtures, functional mixtures and flavor mixtures, for humans or animals, comprising the Process steps:

- Provision of an agricultural side stream containing allergenic proteins of plant and/or animal origin for fermentation,

- Creation of a first pre-culture of various basidiomycetes on agar plates, in which the malt extract usually contained in the agar is replaced by the agricultural side-stream to be fermented,

- Cultivation of these agar plates from the first preculture for one to seven days at a temperature of 20 - 28 °C,

- quantitative and qualitative evaluation of the mycelia of the various basidiomycetes grown on the agar plates of the preculture,

- selection of at least one basidiomycete for the fermentation of the agricultural sidestream, which was mixed with the agar in the first pre-culture and

- Installation of a second preculture on malt agar plates by applying a piece of mycelium of the basidiomycete species of at least one basidiomycete selected from the first preculture,

- Cultivation of these inoculated malt agar plates for one to seven days at a temperature of 20 - 28 °C to grow a certain amount of mycelium and - laboratory determination of the ergosterol content of the basidiomycete species as a reference value and

- transfer of the mycelium into a liquid malt agar medium to obtain a liquid inoculum,

- laboratory determination of the content of allergenic proteins in the agricultural side-stream as a starting value,

- Transfer of the agricultural side-stream to be fermented into a fermenter in liquid or dry state and, if necessary, adjusting the substrate moisture to a substrate moisture of 50-80%,

- Addition of the liquid inoculum obtained from the second preculture to the liquid agricultural side-stream for submerged cultivation or moist agricultural side-stream for emersed cultivation in the fermenter,

- Process of fermentation, submerged or emersed, in a temperature range of 20 °C to 28 °C for one to 24 days,

- a sample of the fermentate being taken at least once during the fermentation process to determine the ergosterol content in the fermentate in relation to the reference ergosterol content of the at least one basidiomycete used for growth control,

- Removal of the raw material from mushroom mycelium and residues of the agricultural by-product that has grown in the fermenter after completion of the fermentation,

- Drying and pulverization of the raw material and

- Laboratory analysis of the raw material using an ELISA test with regard to the residual amount of allergenic proteins contained in the raw material from the agricultural by-product and

- Laboratory analysis of the raw material to determine the ergosterol content.

2.A method for producing a raw material from basidiomycete fungal mycelium and residual components of an agricultural side stream according to claim 1, characterized in that allergenic proteins containing malt germ protein as agricultural side stream and as basidiomycete after the first preculture at least one basidiomycete from the group Marasmius cohortalis, Pleurotus pulmonarius, Pleurotus eryngii and/or Polyporus melanopus is used for the subsequent fermentation of the malt germ protein side stream.

3.A method for producing a raw material from Basidiomycete fungal mycelium and residual components of an agricultural side stream according to claim 1, characterized in that allergenic proteins containing extracted malt with a high proportion of glumes as agricultural side stream and as Basidiomycet after the first preculture at least one Basidiomycete of the group Marasmius cohortalis, Pleurotus pulmonarius, Pleurotus eryngii, Nidula niveotomentosa, Panellus serotinus and/or Polyporus melanopus for the subsequent fermentation of the side stream from extracted malt with a high proportion of glumes.

4.A method for producing a raw material from Basidiomycete fungal mycelium and residual components of an agricultural side stream according to claim 1, characterized in that as an agricultural side stream an allergenic proteins-containing residual stream from the soy protein production and as Basidi- mycet after the first preculture at least one Basidiomycete of the group Pleurotus ostreatus, Pleurotus sta- mineus salmoneus (Pleurotus djamor), Marasmius scorodonius, Pleurotus citrinopileatus, Pleurotus eous, Pycnoporus sanguineus and/or Trametes suaveoiens is used for the subsequent fermentation of the residue stream from soy protein production.

5.A method for the production of a raw material from Basidiomycete fungal mycelium and residual components of an agricultural side stream according to claim 1, characterized in that as an agricultural side stream allergenic proteins containing solubles from pea starch production and as a basidiomycete after the first preculture at least one basidiomycete from the group Lentinula eodes, Pleurotus ostreatus, Pleurotus stamineus salmoneus (Pleurotus djamor), Marasmius cohortalis, Clitopillus passeckerianus,Cyathus helenae, Marasmius scorodonius, Pleurotus citrinopileatus, Pleurotus eous, Pycnoporus sanguineus and/or Trametes suaveoiens for subsequent fermentation of the side stream from Solenubles from pea starch production is used.

6.A method for producing a raw material from Basidiomycete fungal mycelium and residual components of an agricultural side stream according to claim 1, characterized in that as an agricultural side stream an allergenic proteins-containing residual stream from mung bean protein production and as a basidiomycete after the first preculture at least one basidiomycete from the group Lentinula eodes, Pleurotus ostreatus, Pleurotus stamineus salmoneus (Pleurotus djamor), Marasmius cohortalis, Marasmius scorodonius, Pleurotus citrinopileatus, Pleurotus eous, Postia caesia and/or Pycnoporus sanguineus is used for the subsequent fermentation of the side stream from residual stream from mung bean protein production.

7.A method for producing a raw material from Basidiomycetes fungal mycelium and residual components of an agricultural by-product according to claim 1, characterized in that chickpea flour containing allergenic proteins is used as the agricultural by-product and as Basidiomycete after the first preculture at least one Basidiomycete from the group Pleuro- tus ostreatus, Pleurotus stamineus salmoneus (Pleurotus djamor), Marasmius cohortalis, Flammulina velutipes, Cyathus helenae, Lentinus squarrosulus, Pleurotus citrinopileatus and/or Postia caesia is used for the subsequent fermentation of the side stream in the form of chickpea flour.

8.A method for producing a raw material from basidiomycetes fungal mycelium and residual components of an agricultural by-product according to claim 1, characterized in that rice protein containing allergenic proteins as agricultural by-product and as basidiomycete after the first preculture at least one basidiomycete from the group Pleurotus ostreatus , Pleurotus stamineus salmoneus (Pleurotus djamor), Flammulina velutipes, Clitopilus passeckerianus, Cyathus helenae, Lentinus squarrosulus, Pleurotus citrinopileatus and/or Tyromyces chioneus is used for the subsequent fermentation of the rice protein sidestream.

9.Raw material as the end product from the method according to claim 1, characterized in that

- it consists of a majority of 50.01% - 99.9% of a fungal mycelium of at least one basidiomycete and - in relation to the mushroom mycelium contained, there is a lower proportion of residues from an agricultural side-stream containing allergenic proteins prior to fermentation and

- the content of ergosterol related to the dry matter is at least 100 mg/kg.