WO2020074782A1 - Food product comprising fungal mycelium material - Google Patents

Food product comprising fungal mycelium material Download PDF

Info

Publication number
WO2020074782A1
WO2020074782A1 PCT/FI2019/050725 FI2019050725W WO2020074782A1 WO 2020074782 A1 WO2020074782 A1 WO 2020074782A1 FI 2019050725 W FI2019050725 W FI 2019050725W WO 2020074782 A1 WO2020074782 A1 WO 2020074782A1
Authority
WO
WIPO (PCT)
Prior art keywords
mycelium
food
fungal
fungal mycelium
food product
Prior art date
Application number
PCT/FI2019/050725
Other languages
French (fr)
Inventor
Veera Virtanen
Arja PAANANEN
Geza SZILVAY
Leevi VIRTANEN
Anni NISOV
Anniina SUHONEN
Original Assignee
Teknologian Tutkimuskeskus Vtt Oy
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Teknologian Tutkimuskeskus Vtt Oy filed Critical Teknologian Tutkimuskeskus Vtt Oy
Publication of WO2020074782A1 publication Critical patent/WO2020074782A1/en

Links

Classifications

    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/008Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from microorganisms
    • AHUMAN NECESSITIES
    • A21BAKING; EDIBLE DOUGHS
    • A21DTREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D13/00Finished or partly finished bakery products
    • A21D13/04Products made from materials other than rye or wheat flour
    • A21D13/045Products made from materials other than rye or wheat flour from leguminous plants
    • AHUMAN NECESSITIES
    • A21BAKING; EDIBLE DOUGHS
    • A21DTREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D13/00Finished or partly finished bakery products
    • A21D13/04Products made from materials other than rye or wheat flour
    • A21D13/047Products made from materials other than rye or wheat flour from cereals other than rye or wheat, e.g. rice
    • AHUMAN NECESSITIES
    • A21BAKING; EDIBLE DOUGHS
    • A21DTREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D13/00Finished or partly finished bakery products
    • A21D13/06Products with modified nutritive value, e.g. with modified starch content
    • A21D13/064Products with modified nutritive value, e.g. with modified starch content with modified protein content
    • A21D13/066Gluten-free products
    • AHUMAN NECESSITIES
    • A21BAKING; EDIBLE DOUGHS
    • A21DTREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D2/00Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/24Organic nitrogen compounds
    • A21D2/26Proteins
    • A21D2/267Microbial proteins
    • AHUMAN NECESSITIES
    • A21BAKING; EDIBLE DOUGHS
    • A21DTREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D2/00Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/36Vegetable material
    • A21D2/362Leguminous plants
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/20Proteins from microorganisms or unicellular algae
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/22Working-up of proteins for foodstuffs by texturising
    • A23J3/225Texturised simulated foods with high protein content
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/22Working-up of proteins for foodstuffs by texturising
    • A23J3/225Texturised simulated foods with high protein content
    • A23J3/227Meat-like textured foods
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/50Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/40Meat products; Meat meal; Preparation or treatment thereof containing additives
    • A23L13/42Additives other than enzymes or microorganisms in meat products or meat meals
    • A23L13/424Addition of non-meat animal protein material, e.g. blood, egg, dairy products, fish; Proteins from microorganisms, yeasts or fungi
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/40Meat products; Meat meal; Preparation or treatment thereof containing additives
    • A23L13/45Addition of, or treatment with, microorganisms
    • A23L13/46Addition of, or fermentation with fungi, e.g. yeasts; Enrichment with dried biomass other than starter cultures
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065Microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00Edible extracts or preparations of fungi; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10Cereal-derived products
    • A23L7/104Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms

Definitions

  • 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.
  • a meat-like structure to the resultant product can be provided.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • the invention provides use of fungal mycelium as a binding agent for food particles.
  • the invention provides a method for the production of a food product, comprising the steps of:
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • the invention provides a method for the production of a food product comprising non-toxic edible fungal mycelium, comprising the steps of:
  • 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%.
  • 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.
  • 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.
  • the food mixture of food particles and mycelium is baked in the oven, or frying on the stove, for example.
  • 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.
  • the invention provides a method for the production of a food product comprising non-toxic edible fungal mycelium, comprising the steps of:
  • the invention provides a method for the pro duction of a food product comprising non-toxic edible fungal mycelium, compris ing the steps of:
  • 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.
  • Rhizopus oligosporus strain from commercial tempeh starter 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).
  • growth medium pH 6.2
  • malt extract Maltax 10, Senson Ltd.
  • yeast extract BD Bacto, US
  • 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).
  • Rhizopus oligosporus strain from commercial tempeh starter 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).
  • 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.
  • TPA Texture Analyser
  • 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.
  • FIG. 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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-
  • 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.
  • 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.
  • 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.
  • 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).
  • 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.
  • 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.
  • TPA Texture Analyser
  • 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.
  • 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.
  • 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.
  • 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).
  • Gluten-free bread with mycelium as a binding agent was produced.
  • the mycelium was produced as in Example 8.
  • Breads with 1% (dry matter basis) fresh mycelium and 5% dried mycelium were baked. Control breads were baked without mycelium.

Abstract

It is disclosed a food product comprising non-toxic edible fungal mycelium as a binding agent for food particles. It is also disclosed use of fungal mycelium as a binding agent for food particles. It is further disclosed 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 temperature of about 70°C to about 250°C to provide a food product.

Description

FOOD PRODUCT COMPRISING FUNGAL MYCELIUM MATERIAL
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 6xl07 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 6xl07 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 (107 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 (107 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
Figure imgf000015_0001
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.

Claims

CLA1MS
1. A food product comprising non-toxic edible fungal mycelium as a binding agent for food particles, wherein the food product contains about 1 wt-% to about 30 wt-% of the fungal mycelium, on dry matter basis of fungal mycelium.
2. The food product of claim 1, wherein the food product contains about 5 wt-% to about 20 wt-% of the fungal mycelium, specifically about 10 wt- % to about 15 wt-%.
3. The food product of claim 1 or 2, wherein the fungal mycelium is Rhizopus oligosporus.
4. 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.
5. The method of claim 4, wherein the inactivation of the fungal myce lium is performed before or after the separation from culture media.
6. The method of claim 4 or 5, wherein the inactivation is performed by heat treatment, high pressure treatment or chemical treatment.
7. The method of any one of claims 4 to 6, 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.
8. The method of any one of claims 4 to 6, 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,
- 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.
9. The method of any one of claims 4 to 8, wherein the food particles are selected from plant-based food particles, animal-based food particles or a mixture thereof.
10. The method of any one of claims 4 to 9, wherein the fungal myceli um is added in an amount of about 1 wt-% to about 30 wt-% on dry matter basis, specifically about 5 wt-% to about 20 wt-%, more specifically about 10 wt-% to about 15 wt-%, based on the weight of the food product.
11. Use of fungal mycelium in an amount of about 1 wt-% to about 30 wt-%, on dry matter basis of fungal mycelium, as a binding agent for food parti cles.
12. The use of claim 11, wherein the amount of fungal mycelium is about 5 wt-% to about 20 wt-%, specifically about 10 wt-% to about 15 wt-%.
13. The use of claim 11 or 12, wherein the fungal mycelium is Rhizopus oligosporus.
PCT/FI2019/050725 2018-10-11 2019-10-10 Food product comprising fungal mycelium material WO2020074782A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FI20185851 2018-10-11
FI20185851 2018-10-11

Publications (1)

Publication Number Publication Date
WO2020074782A1 true WO2020074782A1 (en) 2020-04-16

Family

ID=68296511

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FI2019/050725 WO2020074782A1 (en) 2018-10-11 2019-10-10 Food product comprising fungal mycelium material

Country Status (1)

Country Link
WO (1) WO2020074782A1 (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112641042A (en) * 2021-01-20 2021-04-13 李红光 Steamed bun for preventing senile dementia
GB2592103A (en) * 2019-11-29 2021-08-18 Marlow Foods Ltd Foodstuffs
EP3942937A1 (en) * 2020-07-03 2022-01-26 Mycorena AB A food product comprising a pure fungi biomass
WO2022091089A1 (en) * 2020-10-28 2022-05-05 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Food products comprising fungal mycelium, process for their preparation and uses thereof
SE2150532A1 (en) * 2021-04-27 2022-10-28 Mycorena Ab A dry food product comprising fungi biomass and methods for manufacturing a dried fungi biomass food product

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3885048A (en) * 1971-02-08 1975-05-20 James J Liggett Method for preparing simulated meat, fish and dairy products
US4212947A (en) 1975-05-22 1980-07-15 Dso "Hranmash" Method for obtaining mycelium from the genus Polyporus
WO1996021362A1 (en) * 1995-01-12 1996-07-18 Zeneca Limited Texturised foodstuffs from gelled edible fungus and hydrocolloid mixtures
EP0986960A1 (en) * 1998-09-15 2000-03-22 Dsm N.V. Mucorales fungi for use in preparation of textured products for foodstuffs
WO2002090527A1 (en) 2001-05-04 2002-11-14 Marlow Foods Limited Edible fungi
WO2010086647A2 (en) 2009-01-29 2010-08-05 Ontology-Partners Ltd. Data processing in a distributed computing environment
WO2013087558A1 (en) * 2011-12-12 2013-06-20 Nestec S.A. Vegetable-based minced meat alternative
WO2016033241A1 (en) * 2014-08-26 2016-03-03 Mycotechnology, Inc. Methods for the production and use of mycelial liquid tissue culture
WO2016120594A1 (en) * 2015-01-27 2016-08-04 Marlow Foods Limited Edible fungi
GB2551738A (en) * 2016-06-28 2018-01-03 Marlow Foods Ltd Foodstuff
WO2018211243A1 (en) * 2017-05-16 2018-11-22 Marlow Foods Limited Foodstuffs

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3885048A (en) * 1971-02-08 1975-05-20 James J Liggett Method for preparing simulated meat, fish and dairy products
US4212947A (en) 1975-05-22 1980-07-15 Dso "Hranmash" Method for obtaining mycelium from the genus Polyporus
WO1996021362A1 (en) * 1995-01-12 1996-07-18 Zeneca Limited Texturised foodstuffs from gelled edible fungus and hydrocolloid mixtures
EP0986960A1 (en) * 1998-09-15 2000-03-22 Dsm N.V. Mucorales fungi for use in preparation of textured products for foodstuffs
WO2002090527A1 (en) 2001-05-04 2002-11-14 Marlow Foods Limited Edible fungi
WO2010086647A2 (en) 2009-01-29 2010-08-05 Ontology-Partners Ltd. Data processing in a distributed computing environment
WO2013087558A1 (en) * 2011-12-12 2013-06-20 Nestec S.A. Vegetable-based minced meat alternative
WO2016033241A1 (en) * 2014-08-26 2016-03-03 Mycotechnology, Inc. Methods for the production and use of mycelial liquid tissue culture
WO2016120594A1 (en) * 2015-01-27 2016-08-04 Marlow Foods Limited Edible fungi
GB2551738A (en) * 2016-06-28 2018-01-03 Marlow Foods Ltd Foodstuff
WO2018211243A1 (en) * 2017-05-16 2018-11-22 Marlow Foods Limited Foodstuffs

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2592103A (en) * 2019-11-29 2021-08-18 Marlow Foods Ltd Foodstuffs
EP3942937A1 (en) * 2020-07-03 2022-01-26 Mycorena AB A food product comprising a pure fungi biomass
WO2022091089A1 (en) * 2020-10-28 2022-05-05 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Food products comprising fungal mycelium, process for their preparation and uses thereof
CN112641042A (en) * 2021-01-20 2021-04-13 李红光 Steamed bun for preventing senile dementia
SE2150532A1 (en) * 2021-04-27 2022-10-28 Mycorena Ab A dry food product comprising fungi biomass and methods for manufacturing a dried fungi biomass food product
SE545255C2 (en) * 2021-04-27 2023-06-07 Mycorena Ab A dry food product comprising fungi biomass and methods for manufacturing a dried fungi biomass food product

Similar Documents

Publication Publication Date Title
WO2020074782A1 (en) Food product comprising fungal mycelium material
AU764133B2 (en) Mucorales fungi for use in preparation of foodstuffs
RU2619290C2 (en) Alternative to minced meat, having plant base
RU2360418C2 (en) Bread, containing bread improving agent and its manufacturing method
JP2003526353A (en) Food products containing Mucorales bacteria
US20220000162A1 (en) Food Product Comprising a Pure Fungi Biomass
KR101926741B1 (en) Natural fermented bread using rice
GB2375944A (en) Dough
KR101823087B1 (en) Method for manufacturing breads using lactic acid bacteria
KR102249168B1 (en) Manufacturing method of pastry containing mugwort
KR20160034066A (en) Soy cheese bread roll and a method of manufacturing
KR830001704B1 (en) Manufacturing method of protein containing food
NL2032406B1 (en) Method for the production of a protein matrix composition having a textured structure
RU2737397C1 (en) Method for preliminary activation of pressed bakery yeast
KR101551559B1 (en) Method of preparing bread using lyophilized girasol powder or its hydrolyzates
EP4082355A1 (en) A method and a system for manufacturing a protein-rich biomass comprising edible filamentous fungus
JP6618097B2 (en) Method for producing edible enzyme composition
RU2687375C1 (en) Method of bakery products preparation
RU2267928C2 (en) Method for producing of wheat bread with increased nutrient and biological value
WO2002005657A1 (en) Structured protein food
KR20090056113A (en) Doenjang using pollen and method thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19790576

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19790576

Country of ref document: EP

Kind code of ref document: A1