F1ELD OF THE INVENTION

The present invention relates to a food product comprising fungal my celium. Fungal mycelium is used as a binding agent to bind food particles of dif ferent origin into mycelium network. Using fungal mycelium for binding of non animal based food particles, a meat-like structure to the resultant product can be provided.

BACKGROUND OF THE INVENTION

Use of plant-based protein in human and animal nutrition has in creased in recent years. Especially, vegetarian alternatives to meat, having a meat-like structure, are of great interest. However, use of plant proteins in food applications generally involves challenges regarding their technological, nutri tional and sensory properties. As a consequence they act poorly as structural agents, for example in making appealing food structure or meat-like fibrous struc ture.

lt is known to use mycelium of edible of filamentous fungi as meat- substitutes in food products. Quorn is a mycoprotein product made of fungal my celium of Fusarium venenatum, and it consists almost completely of mycelium. Tempeh is traditional lndonesian food and made by fermenting soy beans with Rhizopus oligosporus. The fungus uses soy beans as a substrate and grows around the beans making solid food.

There are also other type of meat alternatives available in the market, such as soy based (tofu and Oumph), wheat based (seitan), oats based (Pulled Oats) and faba bean based (Harkis) products.

WO 02/090527 discloses an aqueous formulation comprising edible fungi for use in foodstuffs as a fat mimetic. Fusarium species is cultivated in aque ous media in conditions where mycelium forms small particles (100-200 gm in diameter), and these particles are mixed with food.

US 4212947 discloses a method for obtaining fungal mycelium from the genus Polyporus grown in submerged cultivation. The mycelium can be adapted as a food product or as an additive in food products.

WO 2010/0086647 A1 discloses feed or food compositions comprising fungal material. The fungal material in the compositions enhances survival and/or support growth of normal, healthy animals, and modulate the microbial populations in the digestive tract. Conventional binding agents used in food products include egg white, gelatin, starch and wheat gluten. Egg white and gelatin are not suitable, for exam ple, in vegan diets continuing to increase in popularity.

There is a need for sustainable vegan binding agents which are suita ble in the industrial scale production of vegetarian food products with appealing sensory properties, such as structure and texture.

BR1EF DESCRIPTION OF THE INVENTION

ln an aspect, the invention provides a food product comprising non toxic edible fungal mycelium as a binding agent for food particles. Fungal myceli um itself is vegan, and it is rich in protein and in fiber. The fungal mycelium has a network structure in which the food particles are bound. The food particles can be derived from a plant or an animal source.

ln another aspect, the invention provides use of fungal mycelium as a binding agent for food particles.

ln a further aspect, the invention provides a method for the production of a food product, comprising the steps of:

- providing a fungal strain,

- providing food particles,

- cultivating the fungal strain in a liquid culture media in a fermenter to provide fungal mycelium,

- inactivating the fungal mycelium,

- separating the fungal mycelium from the culture media,

- optionally drying the inactivated fungal mycelium,

- mixing the inactivated fungal mycelium with the food particles to provide a food mixture,

- optionally subjecting the food mixture to heat treatment at a temper ature of about 70°C to about 250°C to provide a food product.

The invention provides vegetarian or vegan food products which are completely free from ingredients derived from an animal source. These food products include bakery products, especially gluten-free bakery products, protein bars, snacks. The food product of the invention can have a meat-like structure.

The food product of the invention can also be used as an ingredient in food products, such as a milk substitute in yogurts, smoothies and table spread.

The invention also provides a suitable alternative food product for in dividuals who cannot tolerate wheat gluten or egg white. The invention further provides an economic and ecological production of new food products in which food main streams and side streams and residues from food processes are used as a raw material in the food production. Since fun gal mycelium itself has a structure modifying properties, new food products from a wide variety of raw materials can be produced without food additives.

BR1EF DESCRIPTION OF THE DRAW1NGS

Figure 1 shows appearance of food products produced from various food particles with and without Roligosporus mycelium as a binding agent.

Figure 2 shows hardness values from texture profile analysis of food products produced from various food particles with and without Roligosporus mycelium as a binding agent.

Figure 3 shows hardness values of texture profile analysis of food products from rice protein and Roligosporus mycelium as a binding agent, and varying mycelium contents.

Figure 4 shows appearance of food products from rice protein and Roligosporus mycelium using lyophilized (A) and pasteurized (B) mycelium as a binding agent.

Figure 5 shows hardness values from texture profile analysis of food products produced from rice protein isolate with and without Roligosporus myce lium as a binding agent. Comparison of fresh and dry mycelium.

Figure 6 shows hardness values from texture profile analysis of food products vegetable mixtures and Roligosporus mycelium as a binding agent, and varying mycelium contents.

Figure 7 shows appearance of extruded food products from faba pro tein and Roligosporus mycelium.

DETA1LED DESCRIPTION OF THE INVENTION

The invention provides a food product comprising non-toxic edible fungal mycelium as a binding agent for food particles ln an embodiment, the food product contains about 1 wt-% to about 30 wt-% of the fungal mycelium, on dry matter basis of fungal mycelium ln another embodiment, the amount is about 5 wt-% to about 20 wt-%. ln a further embodiment, the amount is about 10 wt-% to about 15 wt-%. ln an embodiment, the food product is protein-rich vegetarian food product free from ingredients from animal source.

The fungal mycelium has a branched, fibrous network structure and binds food particles into its network forming a solid compact structure ln an em- bodiment, the food product has a meat-like structure.

The fungal mycelium is produced separately before it is combined with food particles. The mycelium can be obtained from any non-toxic edible filamen tous fungi including macrofungi and moulds ln an embodiment, the filamentous fungi is Rhizopus oligosporus.

The cultivation of fungal mycelium is performed in a fermenter in a conventional manner known to a skilled person ln an embodiment, fungal myce lium of edible filamentous fungi is produced as a liquid cultivation using edible cultivation media components. Fungal mycelium is cultivated in the optimal con ditions characteristics of each fungi. Typically, 10-20 g mycelium (dry weight)/L of the cultivation media is obtained. Cultivation time in a fermenter takes usually several days depending on the fungal strain.

ln an embodiment, the fungal mycelium obtained from liquid cultiva tion is used directly in the production of a food product.

The fungal mycelium typically has a high protein content and is a good source of protein in the food product. The fungal mycelium is also a good source of fibres and beta glucan that are good for digestion. The fibre content of the my celium is typically in the range of 1% to 6% by weight.

The food particles can be derived from a plant or an animal source and can be provided in various forms. The food particles can also be protein and fibre fractions of plant and animal-based raw material, or plant cells. The particles can be for example pieces of meat from bovine, pork or poultry. The plant-based par ticles can be derived from any plant suitable for human or animal nutrition in cluding, but is not limited to, cereals such as wheat, oats, rye, barley, corn and mil let, rice, vegetables, nuts, fruits and berries ln the case of plant-based food parti cles, the particles are typically in the form of flakes, grains, strips, crush etc. ln an embodiment, the food particles are selected from vegetables, a vegetable protein fraction, a vegetable protein isolate, cereals and a mixture of these.

The plant-based food particles can also be derived from side streams or residues obtained from various food processes, such as brewer’s spent grains, wheat bran, and berry press cake. Side streams and residues with low value can thus be converted to higher-value food products instead of composting or provid ing as a feedstuff.

Food particles can also be a mixture of particles obtained from various sources. The taste and texture of the food product can be modified by selecting different food particles. The food product of the invention comprising fungal mycelium and food particles exhibits a solid, compact structure ln an embodiment, the appear ance of the food product is similar to meat. The fibrous structure, typical of meat, of the food product can be increased by using extrusion. The fungal mycelium provides appealing soft mouthfeel to the meat-like food product.

ln another aspect, the invention provides use of fungal mycelium as a binding agent for food particles ln an embodiment, fungal mycelium is used in an amount of about 1 wt-% to about 30 wt-%, on dry matter basis of fungal myceli um. ln another embodiment, fungal mycelium is used in an amount is about 5 wt- % to about 20 wt-%. ln a further embodiment, fungal mycelium is used in an amount is about 10 wt-% to about 15 wt-%.

The invention also provides a method of using wherein fungal myceli um is mixed with food particles.

ln a further aspect, the invention provides a method for the production of a food product comprising non-toxic edible fungal mycelium, comprising the steps of:

- providing a fungal strain,

- providing food particles,

- cultivating the fungal strain in a liquid culture media in a fermenter to provide fungal mycelium,

- inactivating the fungal mycelium,

- separating the fungal mycelium from the culture media,

- optionally drying the inactivated fungal mycelium,

- mixing the inactivated fungal mycelium with the food particles to provide a food mixture,

- optionally subjecting the food mixture to heat treatment at a temper ature of about 70°C to about 250°C to provide a food product.

ln an embodiment, the fungal mycelium separated from the culture media and inactivated is mixed with food particles without drying. The moisture content of the separated mycelium is typically in the range of 80% to 97%.

ln another embodiment, the separated fungal mycelium is dried to powder form. The drying is performed, e.g., by freeze drying, ring-drying or spray drying. Dry mycelium as such is not able to entangle the food particles in the my celium directly but must be rehydrated to a moisture content of at least 50% be fore it is mixed with food particles ln an embodiment, the dry matter content of the dried fungal mycelium is adjusted close to that of the fungal mycelium ob- tained from cultivation.

The activity of the fungal mycelium of the final food product is stopped in order to prevent fungus from using food particles as nutrient for its growth lnactivation of the fungal mycelium also prevents expression of potentially harm ful secondary metabolites during possible growth ln an embodiment, the fungal mycelium is inactivated in a fermenter before the mycelium is separated from the culture media and before the mycelium is mixed with food particles ln another embodiment, the fungal mycelium is inactivated after it has been separated from the culture media lnactivation of the mycelium can be performed by heat treat ment, such as pasteurization, ultra high temperature treatment (e.g. at 135°C for 1 sec, or at 140-150°C for 2 sec), high pressure treatment, or chemical treatments (e.g. with alkali, acids or ethanol). The inactivation can also be performed by au toclaving the fungal culture (e.g. at 120°C for 20 min) ln an embodiment, the fun gal mycelium is inactivated after the separation by autoclaving at 120°C for 20 min. Drying of the fungal mycelium partially inactivates the mycelium.

The inactivated mycelium is gently mixed with food particles in order not to break down the network structure of the mycelium lf desired, excess water is removed, e.g., by filtration.

ln an embodiment, the food mixture of food particles and mycelium is baked in the oven, or frying on the stove, for example. On baking, the structure of the food product is also desirably stabilized. The interaction time between myce lium and food particles is minimized to prevent fungus from using food particles as nutrient for its growth.

ln an embodiment, the invention provides a method for the production of a food product comprising non-toxic edible fungal mycelium, comprising the steps of:

- providing a fungal strain,

- providing food particles,

- cultivating the fungal strain in a liquid culture media in a fermenter to provide fungal mycelium,

- inactivating of the fungal mycelium in the fermentation by heat treatment,

- separating the inactivated fungal mycelium from the culture media,

- drying the inactivated fungal mycelium,

- mixing the inactivated fungal mycelium with the food particles to provide a food mixture, - optionally subjecting the food mixture to heat treatment at a temper ature of about 70°C to about 250°C to provide a food product.

ln another embodiment, the invention provides a method for the pro duction of a food product comprising non-toxic edible fungal mycelium, compris ing the steps of:

- providing a fungal strain,

- providing food particles,

- cultivating the fungal strain in a liquid culture media in a fermenter to provide fungal mycelium,

- separating the fungal mycelium from the culture media,

- inactivating the separated fungal mycelium by autoclaving,

- drying the inactivated fungal mycelium,

- mixing the inactivated fungal mycelium with the food particles to provide a food mixture,

- optionally subjecting the food mixture to heat treatment at a temper ature of about 70°C to about 250°C to provide a food product.

The amount of the fungal mycelium in the final food product is from about 1 wt-% to 30 about wt-%, on dry matter basis of fungal mycelium, of the weight of the food product ln an embodiment, the amount is about 5 wt-% to about 20 wt-%. ln another embodiment, the amount is about 10 wt-% to about 15 wt-%. The fungal mycelium typically has a high protein content and is a good source of protein in the food product.

The following examples are presented for further illustration of the in vention without limiting the invention thereto.

Example 1. Cultivation of Rhizopus oligosporus strain

Rhizopus oligosporus strain from commercial tempeh starter (Raprima Tempeh starter, Indonesia) was cultured in growth medium (pH 6.2) containing 2% (w/v) malt extract (Maltax 10, Senson Ltd.), 0.3% (w/v) yeast extract (BD Bacto, US), and 0.5% wheat peptone (Solabia, France). A 200 ml aliquot of the medium was inoculated using 6xl0 ⁷ spores in 500 ml Erlenmayer flasks (2L cul tivation in total), followed by cultivation at 30°C with agitation at 150 rpm for 2 days. The amount of mycelium after 2 days cultivation was 6.7 g/L (dry weight). The produced fungal mycelium was then concentrated by vacuum filtration using an 11 cm diameter GF/C filter (Whatman, UK) and a Buchner funnel to about 50% of the volume. The final concentration of the mycelium was 14 g/L (dry matter). Example 2. Cultivation of Rhizopus oligosporus strain

Rhizopus oligosporus strain from commercial tempeh starter (Raprima Tempeh starter, Indonesia) was cultured in growth medium (pH 6.2) containing 2% (w/v) malt extract (Maltax 10, Senson Ltd.), 0.3% (w/v) yeast extract (BD Bacto, US), and 0.5% (w/v) wheat peptone (Solabia, France). A 200 ml aliquot of the medium was inoculated using 6xl0 ⁷ spores in 500 ml Erlenmayer flasks (2L cultivation in total), followed by cultivation at 30°C with agitation at 150 rpm for 19 h. The amount of mycelium after 2 days cultivation was 5.7 g/L (dry weight). The produced fungal mycelium was then concentrated by vacuum filtration using an 11 cm diameter GF/C filter (Whatman, UK) and a Buchner funnel to about 50% of the volume. The final concentration of the mycelium was 15.7 g/L (dry weight).

Example 3. Food product with Rhizopus oligosporus mycelium

Three vegetarian foods products were produced using fresh R. oli gosporus mycelium ln one food product, rice protein isolate was used as food par ticles. ln another product, grated carrot-cabbage mixture containing 50% of car rot and 50% of cabbage was used as food particles ln a third product, oat bran was used as food particles.The mycelium was produced as shown in Example 2. The concentration of Roligosporus mycelium was 15.7 g/L (dry weight) lt was mixed with vegetable raw material so as to provide a mixture of 6.7 g in total mass and containing 15 wt-% of mycelium on dry matter basis. Mixing was con ducted thoroughly with spoon. After mixing, the mixture was filtered in a Buchner funnel (5.5 cm in diameter) until no liquid was separating or to same final mass. The food products were then baked in an oven (150°C) for 30 minutes.

Control samples from each vegetable raw material above were pre pared analogously except that fungal mycelium was not added (total mass 6.7 g).

Figure 1 shows how mixture of fungal mycelium and rice protein iso late makes a uniform and compact food sample while the control samples without mycelium is strong and brittle. The clear difference is also observable with the carrot-cabbage sample and oat bran sample compared to the corresponding con trol samples. The control samples do not hold together, while samples containing mycelium had more uniform structure.

The texture of the food samples was instrumentally measured with a Texture Analyser (TA.XTPlus, Stable Micro Systems) using a texture profile analy sis (TPA) test that emulates the mouthfeel. During the TPA test the food sample was compressed twice with a cylindrical probe (diameter 20 mm) to 20% of the sample height. The crosshead speed was 1.0 mm/s. The maximum force during the first compression cycle was recorded as hardness.

Figure 2 shows the results from the TPA test where hardness values upon compression are presented. The results show that the food products con taining mycelium were softy and springy and had a mouthfeel closer to that of a meat patty. For example, rice protein patty was very hard and brittle without fun gal mycelium. Carrot-cabbage mixture did not hold together after baking without fungal mycelium. The TPA result of a meat patty is shown for comparison lt is clearly seen how the hardness value of the food sample containing fungal myceli um approaches to the hardness value of a meat patty.

Example 4. Food product with Rhizopus oligosporus mycelium

Vegetarian food product was produced from rice protein isolate using fresh fungal mycelium produced as described in Example 2. The concentration of Roligosopus mycelium was 15.7 g/L (dry weight) lt was mixed with vegetable raw material so as to provide a mixture of 6.7 g in total mass and containing 5, 10, 15 and 25 wt-% of mycelium on dry matter basis. Mixing was conducted thor oughly with spoon. After mixing, the mixture was filtered in a Buchner funnel (5 cm in diameter) until no liquid was separating or to same final mass. The food products were then baked in an oven (150°C) for 30 minutes.

Control sample from rice protein isolate was prepared analogously ex cept that fungal mycelium was not added (total mass 6.7 g).

The mixture of fungal mycelium and rice protein isolate makes a uni form and compact food sample even with the smallest mycelium content (5%) while the control sample without mycelium is hard and brittle. The TPA results presented in Figure 3 show the trend how hardness of the rice protein samples decreases with increasing mycelium content.

Example 5. Food product with Rhizopus oligosporus mycelium

Concentrated fungal mycelium from Example 1 was lyophilized for 2 days (Hetosicc, CD52) and stored in a desiccator before use. Vegetarian food product was produced from rice protein isolate and the lyophilized fungal myce lium. 0.23 g of the dry mycelium was rewetted with 30 ml water for 30 min. lt was mixed with vegetable raw material so as to provide a mixture of 2.3 g in total mass and containing 10 wt-% of mycelium on dry matter basis. Mixing was con ducted thoroughly with spoon. After mixing, the mixture was filtered in a Buchner funnel (5.5 cm in diameter) until no liquid was separating. The food products were then baked in an oven (150°C) for 30 minutes.

Example 6. Food product with Rhizopus oligosporus mycelium

Concentrated mycelium from Example 1 was pasteurized at 80°C for 20 min). Some shrinkage of the mycelium was observed during the heat treat ment.

Vegetarian food product was produced from rice protein isolate and the pasteurized fungal mycelium. The concentration of Roligosopus mycelium was 14 g/L (dry weight) lt was mixed with vegetable raw material so as to pro vide a mixture of 6.7 g in total mass and containing 25 wt-% of mycelium on dry matter basis. Mixing was conducted thoroughly with spoon. After mixing, the mix ture was filtered in a Buchner funnel (5.5 cm in diameter) until no liquid was sep arating. The food products were then baked in an oven (150°C) for 30 minutes. Control sample from rice protein isolate was prepared analogously except that fungal mycelium was not added (total mass 6.7 g).

Figure 4 shows appearance of food products produced in Examples 5 and 6. The food product "A" is prepared in Example 5, and the food product "B" is prepared in Example 6. ln both products the mycelium bound rice protein parti cles providing a compact structure.

Example 7. Cultivation of Rhizopus oligosporus strain

The R. microsporus var. oligosporus strain (VTT D-82192/ ATCC 22959) (later referred to as R. oligosporus) was selected for the production of fungal mycelium in a 20 L bioreactor. The medium for pre-culture and bioreactor cultivations was identical in composition (20 g/L glucose, VWR Chemicals; 10 g/L yeast peptone, X-Seed ^® Peptone, Barentz ApS, Denmark; 6 g/L yeast extract, X- Seed ^® Cell Kat, Barentz ApS, Denmark) except that 1 mL/L of antifoam agent (Clerol FBA 3107) was added into the bioreactor medium to prevent foam for mation. The pH was adjusted to 5.0 with hydrochloric acid in the pre-culture me dium and with 15% phosphoric acid in the production medium. The media were autoclaved at 121°C for 15 min.

The pre-cultures for the bioreactor cultivation were grown in sterile 500 mL Erlenmeyer flasks containing 170 mL of the medium. The flasks were in oculated with 1% (v/v) freshly prepared spore suspension (10 ⁷ spores/mL) and incubated under 150 rpm shaking at +30°C for 16.5 h. A 20-L bioreactor (B. Braun Biostat C20-2) was inoculated with 10% (v/v) of the pre-culture. The total initial volume, including the inoculum, was 17 L. The cultivation was carried out at +30°C for 48 h with 8.5 -10 L/min of aeration and stirring speed of 300 to 800 rpm to ensure adequate air supply. The pH was controlled at five by adding 2 M sodium hydroxide. After 15 h of cultivation, 55% (w/v) glucose solution was fed into the reactor was started at the rate of 19 - 25 g/h-

After 48 h of cultivation, the fungal culture was autoclaved (121°C, 20 min) in order to inactivate biomass. Subsequently, the mycelium was separated from the medium by straining. Dry weight of the mycelium was determined at the end of fermentation by filtering a small portion of the culture through pre weighted filter (GF/B, Diameter 47mm, 100 circles, CAT No. 1821-047), followed by drying in an oven at 103°C to constant weight and re-weighing of the filter. The amount of mycelium after 48 h cultivation was 12.7 g/L (dry weight). The collect ed mycelium was freeze-dried and stored in moisture tight bags at -20°C.

Example 8. Cultivation of Rhizopus oligosporus strain

The R. microsporus var. oligosporus strain (VTT D-82192/ ATCC 22959) (later referred to as R. oligosporus) was selected for the production of fungal mycelium in a 200 L bioreactor (lnfors HT Techfors 300L). The medium for pre-culture and bioreactor cultivation was identical in composition (20 g/L glu cose, VWR Chemicals; 10 g/L yeast peptone, X-Seed ^® Peptone, Barentz ApS, Den mark; 6 g/L yeast extract, X-Seed ^® Cell Kat, Barentz ApS, Denmark) except that 4 mL/L of antifoam agent (Sunflower oil) was added into the bioreactor medium to prevent the foam formation. The initial pH was adjusted to 5.0 with hydrochloric acid in the pre-culture medium and with 15% phosphoric acid solution in the bio reactor medium. The media were autoclaved at 121°C for 15 min.

For the preparation of the pre-culture, sterile 500 mL Erlenmeyer flasks containing 200 mL of the medium (2 L cultivation in total) were inoculated with 1% (v/v) spore suspension (10 ⁷ spores/mL). The flasks were incubated at +30°C with agitation at 150 rpm for 14.5 h.

The starting volume of the 200 L bioreactor cultivation was adjusted to 190 L and the medium was inoculated with 1% (v/v) of the pre-culture. The cultivation was carried out at +30°C for 40 h at maximum stirring speed (400 rpm). Aeration was increased from an initial value 064 L/min to towards the end of batch fermentation. The pH was maintained at five by adding 2 M sodium hy- droxide. After glucose depletion, 20% (w/v) glucose solution was fed into the bio reactor at the average rate of 950g/h.

After 40h, the bioreactor cultivation was autoclaved (121°C, 20 min) in order to inactivate the biomass. The mycelium was separated from the culture media by straining. Afterwards, the collected mycelium was freeze-dried and stored in moisture tight bags at -20°C. The amount of mycelium after 40 h cultiva tion was 10.5 g/L (dry weight).

Example 9. Testing protocol for vegetable patties

To ensure the uniformity of the vegetable patties, a protocol for the preparation and testing of the samples was created. The vegetable patties were prepared by using rice protein isolate as a food particle and fresh or freeze-dried R. oligosporus mycelium as a binding agent. The dry matter content of the freeze- dried mycelium powder was 95.88%, for fresh mycelium 9.44% and for rice pro tein isolate it was 97%. The mycelium content of 5% (of the total dry matter con tent) were used in the vegetable patties.

The freeze-dried mycelium was rehydrated for >10 min before mixing with the food particles in order to match the moisture content of fresh mycelium. Raw materials were mixed and water (approximately 1:1) was added to obtain decent moisture content for the dough. After that, the dough was placed in metal molds and the surface of each sample was smoothed. The patties were baked at 150°C for 30 min. Control samples were prepared as above except mycelium was not added. Water was mixed to the dough to obtain similar moisture content as it was in the mycelium containing patties.

The texture of the food samples was instrumentally measured with a Texture Analyser (TA.XTPlus, Stable Micro Systems) using a texture profile analy sis (TPA) test that emulates the mouthfeel. During the TPA test the food sample was compressed twice with a cylindrical probe (diameter 20 mm) to 20% of the sample height. The crosshead speed was 1.0 mm/s. The maximum force during the first compression cycle was recorded as hardness.

Food products of rice protein isolate with mycelium produced as in Example 7 were tested by this protocol. Comparison of TPA results of fresh and dry mycelium is shown in Figure 5. Rice protein patty was clearly harder and brit tle without fungal mycelium. Example 10. Food product with Rhizopus oligosporus mycelium

Vegetable patties were prepared by using freeze-dried R. oligosporus mycelium produced as in Example 8. Grated carrot-cabbage-onion mixture con taining 35% of carrot, 35% of cabbage and 30% of onion was used as food parti cles. The dry matter content of the dried mycelium powder was 98.01% and for vegetable mixture 10.14%. The mycelium content of the vegetable patties was either 5%, 10% or 15% of the total dry matter content. Reference samples con taining pea protein isolate and soy protein granules instead of mycelium with similar dry matter concentrations were prepared. Control samples from each vegetable raw material above were prepared analogously except that fungal my celium was not added. The mycelium was rehydrated in the ratio of 1:10 for 30 min before mixing with food particles. Reference materials were also soaked in to water in order to match the moisture content of mycelium patties. The raw mate rials were mixed and the mixtures were placed in to the molds. The surface of each sample was smoothed with spatula. The patties were baked at 150°C for 40 min.

The mixture of fungal mycelium and rice protein isolate makes a uni form and compact food sample while the control sample without mycelium is strong and brittle. The clear difference is also observable with the vegetable mix ture sample compared to the corresponding control samples. The control samples do not hold together, while samples containing mycelium had more uniform structure.

The texture of the food samples was as described in Example 9.

Figure 6 shows the results from the TPA test where hardness values upon compression are presented. The results show that the food products con taining where were softy and springy and had a mouthfeel closer to that of a meat patty. Vegetable mixture did not hold together after baking without fungal myce lium. The TPA result of a meat patty is shown for comparison lt is clearly seen how the hardness value of the food sample containing fungal mycelium ap proaches to the hardness value of a meat patty.

Example 11. Food product with Rhizopus oligosporus mycelium

Texturized meat alternatives were produced from a mixture of faba protein (90%) and mycelium (10%) by wet-extrusion with cooling die. Mycelium was produced as in Example 8. Two control sample were prepared with faba pro tein (100%) and mycelium only (100%). The flour feed rate (0.3kg/h) and water feed (300mL/h) were kept constant resulting in extrudate containing 50% water. The extrusions were conducted at varying temperatures from 80 to 150°C. The resulted sample temperature at die, pressure and torque are presented in Table 1. Faba and mycelium controls had weak structure without notable fibrillation at given conditions (Table 1, Figure 7). However, faba and mycelium mixture result ed in structure showing layers on top of the extrudate (Figure 7). The pressure and torque during the extrusion were higher for the faba/mycelium mixture than for the controls showing good water absorption and better structure formation. This indicated that mycelium could be used as a binding and water-absorbing agent in extrudates although mycelium itself does not form fibrous structure.

Table 1. Parameters for extrusion

Example 12. Food product with Rhizopus oligosporus mycelium

Gluten-free bread with mycelium as a binding agent was produced. The mycelium was produced as in Example 8. Basic gluten-free bread recipe, with maize starch and/or faba flour as main components, was chosen as a base. Breads with 1% (dry matter basis) fresh mycelium and 5% dried mycelium were baked. Control breads were baked without mycelium.

Bread with 1% fresh mycelium had better shape compared to the con trol. Control bread was mushroom shaped whereas the mycelium bread had square shape. Texture analysis showed that the bread with mycelium had harder structure. When the breads were teared by hand, the mycelium bread crumbled less.

Bread with 5% mycelium had similar shape to its control bread. How ever, the height and volume were lower. Water evaporation of the mycelium bread was lower during the baking compared to the control suggesting that the mycelium had better water binding capacity. Texture analysis showed that the mycelium bread was harder compared to the control bread. However, the bread with mycelium crumbled less when teared by hand. lt will be obvious to a person skilled in the art that, as the technology advances, the inventive concept can be implemented in various ways. The inven tion and its embodiments are not limited to the examples described above but may vary within the scope of the claims.