Method of making a fermented plant-based food product and products therof

Abstract

The invention presented herein describes a fermented plant-based food product comprising a matrix of substrate interconnected by at least a filamentous fungus having a geometrical shape with one dimension longer than the other two orthogonal dimensions, with a length being the longest direction of the fermented plant-based food and the two other length are at least 3 cm each and preferably at least 7 cm each, the length is at least 3 cm and preferably at least 7 cm, wherein a cross section parallel or perpendicular to the longest direction of the fermented plant-based product presents a whiteness of 15% or more and preferably 20% or more.

Description

[0001] Method of making a fermented plant-based food product and products thereof

[0002] TECHNICAL FIELD

[0003] The present application relates to a method of producing a fermented plant-based food product by assembling and binding at least one substrate by the growth of a filamentous fungus. Thus, a fungus-containing food product is formed.

[0004] BACKGROUND

[0005] Current meat consumption is depleting natural resources while fuelling climate change. The current world-wide meat consumption is unsustainable and therefore meat alternatives must be developed to counteract the ever-growing consumption of meat and at least partially replace the consumption of conventional meat.

[0006] Various approaches are currently used to produce plant-based meat. The recurring problem is to produce plant-based meat that simulates meat-based product in terms most of sensorial aspects such as color, texture, and taste. A first category of plant-based food is produced by extrusion and the products are made from vegetables proteins such as soy proteins or pea proteins. Those products may have a fibrous texture mimicking meat product, but the taste and the color are not close to the meat products.

[0007] Another category of products is fermented products. Tempeh is a well-known vegan product which is usually made from soybeans fermented and bound by a filamentous fungus, normally Rhizopus oligosporus. In recent times, the term Tempeh is used more broadly, including the fermentation and binding of grains or food processing by-products in addition to soybeans. One of the disadvantages of currently available tempeh products is that they lack taste, texture, and shape if compared to real meat and / or meat-derived foods. The thickness of the tempeh is limited to several centimetres, typically tempeh is typically comprised between 2 and 6 cm thick, limited due to the heat generated by the fermentation, which can be particularly problematic as the thickness of the pieces increases. Another disadvantage is that the product is not very juicy, nor very positively flavored per se, requiring heavy marinades and usually deep frying as the most common cooking procedure.

[0008] Most recent inventions combine the two approaches by fermenting several pieces of extrudate and using a filamentous fungus to form a larger plant-based product (see, e.g. EP4082355, EP3923741 and EP3968776). Certain inventions utilize wet extrudates fermented with filamentous fungi (see, e.g. EP2835058). However, a significant challenge arises from the dense nature of wet extrudates such as poor heat dissipation, which is necessary for optimal fungal growth, especially when the extrudates exceed a couple of millimeters in size. Aggregating multiple wet extrudates exacerbates the issue, necessitating puncturing or creating holes in the substrate to facilitate fungal growth (see, e.g. WO2023094619). Nonetheless, this approach often results in poor fermentation within the wet extrudates. Other applications use a textured substrate that can be made of several pieces of textured vegetable protein (TVP) (see, e.g., W02023068701) which are bound together using at least one fungus. One of the limiting parameters in the production of mycelium-based foods is the mycelial density of fungus within the substrate. Growth of the fungus can be limited by available nutrients, lack of oxygen, material properties of substrate such as hardness, and / or heat released as the fungus grows which actively ferments the substrate. Competing bacterial and fungal contamination could also jeopardize the myceliation process. Poor fungal development within a substrate leads to low mycelial density which decreases fibrousness, limits liquid and fat absorption and therefore juiciness and texture complexity during mastication, reduces the binding strength between the different parts of the substrate, increases the population of unwanted microbial species as the fungi also act as a protective culture; the color may deviate from meat products and / or an off-flavor profiles can appear due to competing microbial contamination and unfermented compounds and / or antinutrients present in the vegetable protein fraction of the substrate. To limit this poor fungal development, it is common practice limiting the thickness of the final product to 2 or 3 cm thick, resulting in food products that are not as complex as meat products. Another disadvantage is that the pieces to ferment are rather small and to have good aeration for all the small pieces a large fermenter room is needed.

[0009] In view of the above, there is a need for a fermentation process that leads to plant-based meat alternative with increased thickness, more complex texture and taste and a better fermentation quality.

[0010] SUMMARY OF THE INVENTION

[0011] A fermented plant-based food product and a method described herein aim to address these challenges by achieving robust filamentous fungus growth both externally and internally within a scaffolding substrate larger than 3 centimetres. To accomplish this, the scaffolding substrate is constructed using a method that creates a complex structure, enhancing heat dissipation and oxygenation during fermentation. Consequently, the resulting fermented plant-based food product closely resembles whole cuts of meat. Wholesale cuts or whole cuts, in culinary terms, refer to minimally divided sections of meat, often containing only 1 to 3 cuts or remaining undivided. They are utilized for specific culinary purposes, such as slicing muscles into smaller parts suiting the respective application (retail cuts). Whole cuts exhibit irregular patterns and distinct fibrosity, typically comprising two phases: one composed of muscle and the other of interstitial tissue. The scaffolding substrate may mimic muscles and the filamentous fungus mimic interstitial tissues.

[0012] To achieve the desired characteristics, an assembly of a plurality of substrates, which are preferably be combined and / or deformed to form the scaffolding substrate. These substrates should allow to incorporate enough liquids providing the necessary water activity and nutrients for fermentation by the filamentous fungus. Essentially, the substrates serve as a framework or support structure for the filamentous fungus to grow. The method can generally include providing a plurality of substrates, such as a textured non-animal (vegetable, fungal and / or other) protein substrates, which may or may not be subjected to various pre-processing steps used to help growing the mycelium of the filamentous fungus. The filamentous fungus, thanks to the mycelia, binds the plurality of substrates together. The problem of overheating and hypoxia at the center of the scaffolding substrate is solved by the specific assembling step and the different embodiments used to maintain the plurality of substrates during the incubation while adding voids inside of the scaffolding substrate. Voids can be filled by the mycelium of the filamentous fungus thus creating a more complex fermented plantbased food product. The specific assembly method creating voids between substrates increase the quality of the fungal growth for large scaffolding substrates. According to a first aspect, the invention relate to a fermented plant-based food product comprising a matrix of substrate interconnected by at least a filamentous fungus having a geometrical shape with one dimension longer than the other two orthogonal dimensions, with a length being the longest direction of the fermented plant-based food and the two other length are at least 3 cm each and preferably at least 7 cm each, the length is at least 3 cm and preferably at least 7 cm, wherein a cross section parallel or perpendicular to the longest direction of the fermented plant-based product presents a whiteness of 15% or more and preferably 20% or more.

[0013] Such high whiteness in this large fermented plant-based food product allow to closely mimic the texture of meat food products, such as whole cuts. This means the fermented plant-based food product closely resembles the natural structure and textural properties (juiciness, fibrosity, bite) of meat, visually and in its mouthfeel.

[0014] The juiciness results in the capacity of absorbing liquids, said fermented plant-based product can absorb at least 17% of its own weight, preferably 20% and preferably 25% of liquids, the liquids preferably being water, oil or a mixture thereof, preferably wherein the mixture comprising 40 wt% to 70 wt% of water, more preferably 45 wt% to 65 wt% of water, the rest being oil.

[0015] The shape of the fermented plant-food product can be a sausage with a diameter that is at least 3 cm and preferably at least 7 cm, the diameter measured at the middle of the longest direction.

[0016] According to a second aspect, the invention relate to a method to produce the fermented plantbased food product, the method comprising: a. providing a plurality of substrates with a first density, b. inoculating the substrates with at least one filamentous fungus, c. assembling the substrates to form a scaffolding substrate to reach a second density, d. fermenting said scaffolding substrate to allow the at least one filamentous fungus to grow inside, outside, or inside and outside said scaffolding substrate and / or substrates, wherein the substrates are mechanically deformed, bent and / or folded and stacked on top of each other during step c. and the second density is 10 % to 20 % higher than the first density.

[0017] The incubation and fermentation of the scaffolding substrate by the filamentous fungus enhances the flavour profile, the texture, the colour and / or the juiciness of the fermented plant-based food product.

[0018] During the step c. the assembly, the substrates are deformed and bent and / or folded increasing their density then stacked on top of each other, due to the deformation of the substrate voids are created between the substrate and the density of the scaffolding substrat is increase by 10% to 20%. The voids created between the substrat enhance the fungal growth.

[0019] Preferably, the substrates are each porous, fibrous, anisotropic, foamed, have a lamellar structure and / or an extrudate and comprise non-animal proteins or are each made from non-animal proteins such as textured vegetable protein. The filamentous fungus having a better growth in such substrates.

[0020] At least one, and preferably more or all of the substrates have a shape of a sheet, wherein the sheet extends along a first direction and a second direction, which is preferably orthogonal to the first direction, the sheet having a thickness along a third direction perpendicular to the first and second directions and the thickness is preferably between 1 mm and 10 cm, preferably between 5 mm and 3 cm.

[0021] The voids created by this method can be measured by taking a cross section parallel or perpendicular to the longest direction of the scaffolding substrate, a ratio of an area occupied by voids between the substrates to a total area of the cross section is between 0.1 and 0.4 and preferably between 0.1 and 0.3.

[0022] To creates and / or increase the voids any technics can be used, the substrates can each be bent and / or folded into a V-shape and / or a conical shape, preferably wherein the V-shape or the conical shape forms an internal angle of between 20° and 160°, preferably between 40° and 140°, and more preferably between 60° and 120°.

[0023] A line linking the tips of the V-shapes can be substantially colinear to a direction of assembly, the direction of assembly preferably being a direction along which the substrates are stacked to form the scaffolding substrate.

[0024] Alternatively, for each V-shape, an axis passing through the tip of the respective V-shape and bisecting an internal angle is defined, wherein a majority of the axes, preferably 75% of the axes, extend at an angle to the direction of assembly of between 0° and 45° and preferably between 0° and 25°.

[0025] The step b) can be carried out before, during or after the step c). The inoculation can be done after the scaffolding substrate has been created, the fungal growth inside the scaffolding substrate is still possible thanks to the voids created by the assembly.

[0026] The filamentous fungus can be Rhizopus.

[0027] The substrates can be inoculated with at least one filamentous fungus and at least one other microbial species such as a fungus, a bacterium or a yeast.

[0028] Since the plurality of substrate are bend and / or stacked it is possible that the scaffolding substrate doesn't stable therefore the scaffolding substrate can be held in place during step d).

[0029] To ensure an optimal growth of the filamentous fungi, the scaffolding substrate may be held in place during step d) by a container having apertures to allow air to flow therethrough.

[0030] After the incubation, the fermented plant-based food product can undergo a post-processing stage that adds significant versatility to the fermented plant-based food product. This optional postprocessing step involves various techniques and treatments to produce a diverse range of products or retail products. The fermented plant-based food product can then be processed with a plurality of means such as deactivating the fungal growth and / or the ones used in the meat industry. The post process steps can comprise cutting or marinating the fermented plant-based food product.

[0031] BRIEF DESCRIPTION OF THE FIGURES In Fig. 1 is represented a flow diagram illustrating a method for producing a fermented plant-based food product according to various embodiments described herein.

[0032] In Fig. 2 is represented a standard assembly according to prior art.

[0033] In Fig. 3 is represented an assembly method according to an embodiment.

[0034] In Fig. 4 is represented an assembly method according to another embodiment.

[0035] In Fig. 5 is represented an assembly method according to another embodiment.

[0036] In Fig. 6 is represented an assembly method according to another embodiment.

[0037] Fig. 7 is a picture of two fermented products, on top a fermented plant-based food product according to one embodiment and at the bottom a standard food product produced by a conventional method.

[0038] In Fig. 8 the relative oxygen concentration in a fermented plant-based food product according to one embodiment, in a standard food product according to prior art and in a substrate is represented.

[0039] In Fig. 9 is represented the whiteness of the standard food product on the left and the fermented plant-based food product according to one embodiment on the right.

[0040] DETAILED DESCRIPTION

[0041] The present invention describes a fermented plant-based food product made exclusively from nonslaughtered products. Said fermented plant-based food product comprises extruded plant proteins and at least a filamentous fungus.

[0042] FIG. 1 presents a method 100 for preparing a fermented plant-based food product, comprising steps of providing 110 a plurality of substrates, inoculating 120 said substrates with at least one filamentous fungus, assembling 130 said substrates to form a larger scaffolding substrate and incubating 140 the scaffolding substrate, allowing the filamentous fungus to grow inside, outside, or inside and outside said scaffolding substrate to form a fermented plant-based food product.

[0043] In step of providing 110 any substrates suitable for food preparations can be used. Preferably, substrates are plant-based, more preferably vegetable protein substrates. The substrates may additionally contain (poly)saccharides, fibers, mineral salts and / or amino acids necessary for the growth of the filamentous fungus.

[0044] The choice of the properties of substrates such as their size and thickness can also influence the chewing, juiciness, texture, and mouthfeel of the fermented plant-based food product. Thus, by modulating the size of substrate, it is possible to create a wide variety of textures and mouthfeel sensations for fermented plant-based food products.

[0045] Substrates used in this invention can be chunks or sheets. Preferably mostly of the substrates are sheets; in other words, they are elongated in two directions compared to a third direction which is smaller than the two others. Preferably, they are characterized by their elongated width and length, with a relatively uniform thickness. The aspect ratio of the sheets which is defined as a ratio of the width to the length of a respective sheet is between 1:1 and 1:10 and preferably between 1:1 and 1:6 and more preferably between 1:1 and 1:3. The thickness of the substrate should be between 1 mm and 10 cm, preferably between 5 mm and 3 cm.

[0046] Preferably, substrates comprise one or more proteins derived from plant-based sources, including pulses or legumes, such as soy, pea, fava, beans, oilseeds such as rapeseeds or sunflower seeds, grains such as wheat or oat grains, or any other sources. Substrates may comprise a mixture of proteins from different sources, or a single protein not blended with proteins from other sources. For example, substrates are composed of a blend of soy protein and pea protein. Alternatively, substrates are composed of a blend of rice protein and pea protein. Alternatively, substrates are composed of a blend of at least 2 plant-based proteins, preferably wherein at least one is soy protein or pea protein. Alternatively, substrates are composed of soy protein and no proteins from other sources, and further comprise starch. Alternatively, substrates are composed of pea protein without proteins from other sources, and further comprise starch. Preferably, substrates comprise other non-slaughtered ingredients originating from single cell organisms, cellular agriculture, algae, plants, or fungi.

[0047] In one embodiment substrates are be formed by any texturization process; it can be high moisture extrusion cooking (HMEC) or shear cell (SC) processing, where proteins are molten under high temperature, pressurized at moisture contents of 15-80% and subsequently cooled under shear, resulting in the formation of a solidified fibrous structure.

[0048] In one embodiment, substrates are formed by freeze structuring, where the substrates are frozen in order to creates ice crystals in the substrate to form a fibrous substrate.

[0049] In one embodiment, substrates are formed by fiber spinning, where a solution comprising proteins are loaded into a spinning apparatus, which comprise a reservoir and a nozzle (also call a spinneret). The solution is then forced through the nozzle under high pressure. The nozzle typically has multiple holes to form thin filaments creating the fibers of the substrates.

[0050] In one embodiment, substrates mainly comprise starch and carbohydrates.

[0051] In one embodiment, substrates are solid, gelled or partly gelled at a first temperature and liquified at a second temperature. The first and the second temperatures can be adjusted to assemble the scaffolding substrate. In one embodiment, the first temperature is 0°C and the second temperature is 25°C or 30DC or 35°C or more. Alternatively, substrates can liquefy at a temperature above 0°C but the time to completely liquefy said substrates is long so that said substrates can be assembled to form the scaffolding substrate. In other words, the substrates could slowly liquefy during this assembly phase and / or during the inoculation and / or during the incubation and / or after. These substrates can melt during the growth of the filamentous fungus and can completely disappear in the fermented plant-based food product. The gelled substrates can be made of water, fat, oil, gelatin agar-agar or hydrocolloids or a combination thereof.

[0052] In another embodiment, substrates can be made by any kind of edible textured vegetable protein product or a combination thereof.

[0053] In another embodiment, substrates is fermented by microbial species and preferably by a filamentous fungus. In another embodiment, substrates comprise a fungus and preferably a filamentous fungus.

[0054] In a preferred embodiment the substrates comprise a porous substrate such as a foamed substrate or textured vegetable protein (TVP) substrate. TVP substrates are formed using an extrusion process in which a pressurized molten protein mixture exits the extruder; the sudden drop in pressure causes rapid expansion into a spongy structure. This spongy structure provides pores or channels allowing filamentous fungus to grow both on the surface but also inside the substrate and receive sufficient oxygen and nutrients. The TVP substrates may be dried after extrusion leading to a lower water content and higher solid (dry matter) and protein content or may be only partly dried or not dried prior to further processing the TVP substrate.

[0055] Substrates, in particular TVP substrates, typically comprise more than 30% wt of protein in the dry matter, preferably at least 40 wt% of protein in the dry matter, more preferably at least 50 wt% of protein in the dry matter.

[0056] TVP substrates can be rehydrated prior to incubation which decreases the relative weight percentage of protein in the overall substrate by adding water. After hydration, the density of such TVP substrates is typically comprised between 0.4 g.cnr3and 0.9 g.cm’3.

[0057] Preferably, substrates comprise one or more proteins derived from plant-based sources, including pulses or legumes, such as soy, pea, fava, beans, oilseeds such as rapeseed or sunflower seeds, grains such as wheat or oat grains, or any other sources. Substrates may comprise a mixture of proteins from different sources, or a single protein not blended with protein from other sources. For example, substrates are composed of a blend of soy protein and pea protein. Alternatively, substrates are composed of a blend of rice protein and pea protein. Alternatively, substrates are composed of a blend of at least 2 plant-based proteins, preferably wherein at least one is soy protein or pea protein. Alternatively, substrates are composed of soy meal or flours. Alternatively, substrates are composed of vegetable flours. Alternatively, substrates are composed of soy protein and no proteins from other sources and comprises both protein and starch. Alternatively, substrates are composed of pea protein and no protein from other sources and comprise both protein and starch. Preferably, substrates comprise other non-slaughtered ingredients originating from single cell organisms, cellular agriculture, algae, plants, or fungi.

[0058] In one embodiment, various kinds of substrates are provided. In one embodiment, a first kind substrate with a high concentration of protein and a second kind of substrate with a high concentration of starch are provided. Alternatively, a first kind of substrate with soy protein and a second kind of substrate with pea protein are provided. In another embodiment, a first kind substrate with a high concentration of protein and a second kind of substrate which is a gelled substrate are provided.

[0059] The substrates can be prepared prior to inoculation 120. During the preparation, the substrates can be hydrated with an aqueous solution and treated to reduce the number of and / or eradicate other microbial species present inside and / or outside of the substrates that may constitute a source of contamination. The water activity of the substrates is adjusted to the conditions desired for microbial growth, preferably the water activity is between 0.8 and 1.0, more preferably between 0.9 and 1.0, even more preferably between 0.96 and 1. In some embodiments the aqueous solution is composed of water. The ratio between the mass of substrates before preparation and the mass of the aqueous solution (e.g., water) can be from 1:0.5 to 1:4 and more preferably between 1:1 and 1:3.

[0060] In other embodiments the aqueous solution can comprises water but also color, flavors, any compounds useful to grow the filamentous fungus such as nutrients, vitamins, and / or any compounds that stop the growth and / or eradicate other non-wanted microbial species.

[0061] The hydration provides a growth condition for the filamentous fungus to develop and grow into the substrates. It is also the case for non-wanted microbial species. To reduce the number of microbial species present or inhibit the growth of unwanted microbial species, substrates can be pasteurized, sterilized, ozonated, treated by antibacterial compounds and / or treated by one or several acids. For example, the substrates may be sterilized in an autoclave at up to 121°C and 1.5 psi. Another example is acidification, in which the surface of the substrates is treated by an acid, preferably a food-grade acid, more preferably hydrochloric acid, hydrogen peroxide, lactic acid, acetic acid, malic acid, citric acid, or succinic acid, preferably to reach a pH of below 6, more preferably below pH 5, most preferably between pH 4.4 and 4.6 at the surface. Even more preferably, the pH on the surface is adjusted to ensure growth of the filamentous fungus used and to reduce growth of other microorganisms.

[0062] In another embodiment the substrates and the aqueous solution are treated separately to then be brought into contact and thus hydrate the substrates.

[0063] The plurality of substrates is then inoculated 120 with at least one filamentous fungus. The inoculation can be carried out in any way and consequently additional subsequent steps may be performed to obtain a good development of filamentous fungus inside and / or around the substrates.

[0064] Inoculation 120 can be done with an inoculum. Inoculum contains an amount of colony forming units of the filamentous fungus that can be spores, active vegetative hyphae, and active parts of fungal tissue, and it may include as well dormant hyphae and parts of fungal tissue, e.g., dehydrated or lyophilized hyphae and parts of fungal tissue. Colony-forming units refer to a number of microbial cells (bacteria, spores, active vegetative hyphae, and active parts of fungal tissue etc.) that are viable and able to multiply for example via binary fission under controlled conditions. The inoculum is introduced into and / or onto a larger volume of the substrates to initiate growth of the filamentous fungus on the substrates. The inoculum can take different forms, such as spores, mycelial fragments, or liquid or solid culture suspensions, and may contain the vegetative and reproductive structures of the fungus and a spawning substrate. Spawning substrate can be grains, vermiculite, sawdust or other appropriate powdered or granulated substrate that ensures a large amount of colony forming units once the inoculum has been mixed with the final substrates.

[0065] The number of colonies forming units in the inoculum and its composition can vary depending on the desired outcome and the characteristics of the filamentous fungus being used. In some embodiments, the inoculum is directly introduced into and / or on the substrates to deliver filamentous fungus both on the surface and / or inside. In other embodiments, the filamentous fungus and / or the spores are in an aqueous suspension and the inoculum is introduced into and on the substrates to deliver filamentous fungus both on the surface but also inside. Said fungi are preferably selected from the group consisting of ascomycetes, basidiomycetes, deuteromycetes, oomycetes, and / or zygomycetes, in particular edible species belonging to the genus Rhizopus, Aspergillus, Penicillium, Ganoderma, Sporidiobolus, Mucor or Pleurotus. More specifically, the species Rhizopus oligosporus, Rhizopus delemar, Rhizopus oryzae, Rhizopus microsporus, Aspergillus oryzae, Aspergillus luchuensis, Aspergillus sojae, Penicillium nalgiovense, Penicillium camemberti, Penicillium roqueforti, Penicillium salami, Penicillium olsonii, Ganoderma lucidum, Mucor indicus, Actinomucor repens, Actinomucor taiwanensis, Mucor circinelloides, Mucor hiemalis, Mucor racemosus, Pleurotus ostreatus, Pleurotus eryngii, or a combination thereof.

[0066] In some embodiments the substrates are co-inoculated with at least two microbial species comprising at least one filamentous fungus. The second microbial species can be a fungus, a yeast, or a bacterium.

[0067] In one embodiment a filamentous fungus of the genus Rhizopus and preferably Rhizopus oligosporus or Rhizopus delemar and a second microbial species, which is a yeast of the genus Saccharomyces, Schizosaccharamyces, Zygosaccharomyces, Torulaspora, Candida, Kluyveromyces, Pichia, Hansenula, Yarrowia, Schwanniomyces, Arxula Brettanomyces, Dekkera, Debaryomyces, Sporidiobolus, Geotrichum, and Xanthophyllomyces, are inoculated simultaneously or successively. The inoculation of the filamentous fungus and the yeast can be performed by spreading colony forming units on the substrates. For this embodiment, the colony forming units are spores for the filamentous fungus species and yeast for the yeast. A spore concentration of the filamentous fungus species, in terms of spores per kg of the substrates, ranges from 1 x 104to 1 x 1012, preferably from 1 x 10sto 1 x IO10, and more preferably from 1 x 108to 1 x 109, and a yeast concentration is 1 x 104to 1 x 1012, preferably from 1 x 10sto 1 x 1011, and more preferably from 1 x 107to 1 x 1010for the yeast in term of yeast per kg of the substrates.

[0068] In one embodiment a filamentous fungus of the genus Rhizopus and preferably Rhizopus oligosporus or Rhizopus delemar and a second microbial species of a bacterium of the genera Lactilplantobacillus, Limosilactobacillus, Levilactobacillus, Lentilactobacillus, Ligilactobacillus, Lactobacillus, Lacticaseibacillus, Pediococcus, Weissella, Lactococcus, Streptococcus, Leuconostoc, Enterococcus, Oenococcus, Staphylococcus, Bifidobacterium, or Bacillus are inoculated simultaneously or successively. The inoculation of the filamentous fungus and / or the bacteria can be performed by spreading colony forming units on the substrates. For this embodiment, the colony forming units are spores for the filamentous fungus species and bacteria for the bacteria. A spore concentration of the filamentous fungus species, in terms of spores per kg of the substrates, ranges from 1 x 104to 1 x 1012, preferably from 1 x 106to 1 x IO10, and more preferably from 1 x 108to 1 x 109and a bacteria concentration of ranges from 1 x 104to 1 x 1013, preferably from 1 x 106to 1 x

[0069] 1012, and more preferably from 1 x 107to 1 x IO10in term of bacteria per kg of the substrates.

[0070] In one embodiment a first filamentous fungus of the genus Penicillium and preferably Penicillium nalgiovense and a second filamentous fungus of the genus Rhizopus and preferably Rhizopus oligosporus or Rhizopus delemar are inoculated simultaneously or successively. The inoculation of the first and / or second filamentous fungus can be performed by spreading colony forming units on the substrates. For this embodiment, the colony forming units are spores of the respective filamentous fungus species. A first spore concentration of the first filamentous fungus species, in terms of spores per kg of the substrates, ranges from 1 x 104to 1 x 1012, preferably from 1 x 106to 1 x IO10, and more preferably from 1 x 108to 1 x 109, and a second spore concentration of the second filamentous fungus species, in terms of spores per kg of the substrates, ranges from 1 x 104to 1 x

[0071] 1013, preferably 1 x 105to 1 x IO10, and more preferably 1 x 107to 1 x 108. Preferably the ratio of the first spore concentration to the second spore concentration ranges from 0.01 to 1000, preferably 0.1 to 500, and more preferably 50 to 150.

[0072] In one embodiment the substrates are co-inoculated with at least three microbial species comprising at least one filamentous fungus and at least one bacterium. In one embodiment a first filamentous fungus of the genus Penicillium and preferably Penicillium nalgiovense and a second filamentous fungus of the genus Rhizopus and preferably Rhizopus oligosporus or Rhizopus delemar and a bacterium of the genera Lactilplantobacillus, Limosilactobacillus, Levilactobacillus, Lentilactobacillus, Ligilactobacillus, Lactobacillus, Lacticaseibacillus, Pediococcus, Weissella, Lactococcus, Streptococcus, Leuconostoc, Enterococcus, Oenococcus, Staphylococcus, Bifidobacterium, or Bacillus are inoculated simultaneously or successively. The inoculation of the first and / or second filamentous fungus and / or the bacteria can be performed by spreading colony forming units on the substrates. For this embodiment, the colony forming units are spores of the respective filamentous fungus species. A first spore concentration of the first filamentous fungus species, in terms of spores per kg of the substrates, ranges from 1 x 104to 1 x 1012, preferably from 1 x 106to 1 x 1010, and more preferably from 1 x 108to 1 x 109; a second spore concentration of the second filamentous fungus species, in terms of spores per kg of the substrates, ranges from 1 x 104to 1 x 1013, preferably 1 x 105to 1 x 1010, and more preferably 1 x 107to 1 x 10s; and a bacteria concentration ranges from 1 x 104to 1 x 1013, preferably from 1 x 106to 1 x 1012, and more preferably from 1 x 107to 1 x 1010in term of bacteria per kg of the substrates. Preferably the ratio of the first spore concentration to the second spore concentration ranges from 0.01 to 1000, preferably 0.1 to 500, and more preferably 50 to 150.

[0073] In one embodiment the substrates are co-inoculated with more microbial species.

[0074] During step 130, a scaffolding substrate is created. This assembly phase aims to generate a scaffolding substrate with ample internal voids capable of being filled by the filamentous fungus. The assembly process may involve the mechanical deformation of substrates to generate voids between substrates as they are assembled together.

[0075] In one embodiment, these substrates are sheets or carpets. Simply stacking them in parallel (prior art assembly method) may be insufficient to facilitate growth of the filamentous fungus due to insufficient oxygenation and inadequate heat dissipation. Additionally, using such a method may result in a fermented plant-based food product that appears uniform and unnatural, as illustrated in Fig. 2. To address these issues, the substrates preferably undergo mechanical deformation and / or be stacked in a manner that creates a scaffolding substrate with increased void volume between substrates and at the same time slightly increase the density of the substrate. A transversal cut parallel or perpendicular to the longest direction of a scaffolding substrate should present a ratio of the area of voids to the area of substrates between 0.1 and 0.4 and preferably between 0.1 and 0.3. Also, millimetric voids may be visible by naked eyes as presented in Fig. 3. The presence of these voids enhances the airflow within the scaffolding substrate, promoting improved aeration. This, in turn, improve oxygenation of the filamentous fungus and enhances heat dissipation, ultimately leading to enhanced growth of the fungus in the substrates and in the scaffolding substrate.

[0076] In one embodiment, a scaffolding substrate is formed using a continuous or semi-continuous substrate. Continuous and semi-continuous substrates refer to extruded materials with a length at least 10 times, preferably 100 times longer than the smaller dimension of the extrudate. For cylindrical extrudates, cutting along at least one direction passing through the radius of the extrudate creates a carpet-like structure. The longest dimension aligns with the extrusion direction. This continuously extruded substrate is then employed to construct the scaffolding substrate. To form the scaffolding substrate, any technique that allows to have between 10 % and 40% of voids in the scaffolding substrate can be used. Preferably, the density of the scaffolding substrate is higher than the substrate's density by 10%, even though the presence of 10% to 40 % of voids between the substrates. To achieve this, several techniques can be employed, such as rolling the continuous or semi-continuous substrate along an axis perpendicular to the extrusion. Alternatively, rolling can be executed along any axis other than parallel to the extrusion.

[0077] For semi-continuous substrate it is possible to roll it along any axis in the plane made by the substrate.

[0078] Another approach involves pushing the continuous or semi-continuous substrate against a stationary surface orthogonal to the extrusion direction. This mechanical action folds the continuous or semi-continuous substrate in a wave-shape. By modulating the strength, it is possible to finely tune the wavelength and therefore the density of the scaffolding substrate.

[0079] Another technique applicable to cylindrical substrates or plane substrates shaped into cylindrical shapes involves compressing the cylindrical substrates in multiple directions parallel to the diameter of the cylinder. This compression can occur simultaneously, resulting in the formation of waves or wrinkles on the cylindrical substrates. Alternatively, the compression of the cylindrical substrates can be performed sequentially.

[0080] In one embodiment, the plurality of substrates is assembled 130 to form a scaffolding substrate. The scaffolding substrate comprising several substrates that can be chunks and / or sheets, the number thereof being from 2 to 500 and preferably 2 to 100.

[0081] Preferably, the majority of substrates should be in sheet form. The sheet extends along a first direction and a second direction, which is preferably orthogonal to the first direction, the sheet having a thickness is in a range of 0.1 cm to 10 cm, preferably 0.5 cm to 3 cm along a third direction perpendicular to the first and second directions. Sheets are folded during the creation of the scaffolding substrate to add voids inside of the scaffolding substrate. The scaffolding substrate is formed by mechanically deforming the substrates, either before or simultaneously stacking them to create a larger scaffolding substrate. When mechanical deformation is applied to the substrate, it bends, taking a V-shape form or U-shape form. V-shape substrates are defined by a tip and two sides extending therefrom. Consequently, substrates are stacked on top of each other, and the scaffolding substrate exhibits a preferred orientation, although lacking clear parallelism among the different substrates, as depicted in Fig. 4. These substrates form V-shaped layers due to the mechanical deformation form an internal angle formed between the sides of the V-shape of between 20° and 160°, preferably between 40° and 140°, and more preferably between 60° and 120°, wherein the internal angle is determined in a cross section parallel to the longest direction of the scaffolding substrate. V-shaped substrates exhibit the average angles 0, measured on at least 10 V-shaped substrates, ranging between 10° and 80°, preferably between 20° and 70° and preferably between 30° and 60°. The tips of the V-shape substrates may point in a direction substantially parallel to the stacking direction (sometimes also called "direction of assembly" herein), as presented in Fig. 4. Alternatively, the axis passing through the tip of the respective V-shape and bisecting the internal angle of the V-shape is at an angle a to the direction of assembly of between 0 and 50° and preferably between 0° and 20° relative to the stacking direction. In one embodiment the stacking of substrates is carried out in an alternative manner: defining a V- shape substrate by a tip and two sides extending therefrom, the tip of the first and the second sides of a first V-shape substrate is offset from the central axis of the scaffolding substrate, and the tip of the first and the second sides of a second V-shape substrate is offset from said central axis, preferably offset to an opposite side with respect to the first V-shape substrate. This pattern consisting of 2 substrates having an offset between them may be repeated in the stacking direction, as to form a pinecone-like assembly, the axis passing through the tip of the respective V-shape and bisecting the internal angle of the V-shape is at an angle a to the direction of assembly of between 10 and 90° and preferably between 30° and 60° relative to the stacking direction, as illustrated in Fig. 5.

[0082] Alternatively, the substrate can be folded in half to create a V-shaped substrate, which is then stacked on top of each other alternately, with the V formed by the substrate facing one direction while the next substrate faces a substantially opposite direction. The axis passing through the tip of the respective V-shape and bisecting the internal angle of the V-shape is at an angle to the direction of assembly of between 45 and 135° and preferably between 75° and 115° as illustrated in Fig. 6.

[0083] These assembly methods allow for the creation of voids between substrates and at the same time increase the density of the scaffolding substrate. A transversal cut parallel or perpendicular to the longest direction of a scaffolding substrate should present a ratio of the area of voids to the area of substrates between 0.1 and 0.4 and preferably between 0.1 and 0.3.

[0084] When substrates are stacked atop one another in a vertical or horizontal arrangement, they can form a rectangular or a cylindrical shape or any other shape. Each substrate contributing to the overall structure of the scaffolding substrate. As more substrates are added, the scaffolding substrate grows in length while substantially maintaining the same cross-section. The scaffolding substrate can have a shape similar to a sausage.

[0085] These assembly methods involving multiple mechanically folded substrates generate inherent voids between substrates that facilitate the growth of filamentous fungus mycelium within them. The size of these voids can be finely adjusted by applying pressure during substrate assembly. High pressure results in smaller voids at least 10% of the total area, and a higher density of the scaffolding substrate, whereas low pressure yields larger voids at most 40% and a lower density of the scaffolding substrate.

[0086] Alternatively, the arrangement of these substrates is completely random, the different and / or irregular shapes of the substrates are sufficient to have between 10 % and 40 % and preferably between 10 % and 30 % of the area of voids compare to the area of substrates when taking a transversal cut parallel or perpendicular to the longest direction of a scaffolding substrate.

[0087] In one embodiment, substrates with irregular shapes and / or that cannot be deformed are assembled with low pressure, wherein each substrate is deposited on top of each other with minimal applied pressure. This assembly method ensures that substrates maintain their individual integrity and structural integrity is preserved throughout the scaffolding substrate.

[0088] Alternatively, pressure can be applied to the substrates to form a scaffolding substrate. Pressure may be applied manually or by means of machines. By applying a sufficient pressure, the substrates can be hold together prior to incubation and this can help to maintain the scaffolding substrate in the desired shape. Pressure can be exerted perpendicularly onto the surface of the substrates to create a denser scaffolding substrate. Pressure can be applied to bend the substrates.

[0089] Alternatively, deformation can be applied to all the substrates at the same time after being assembled to form a scaffolding substrate. Deformation can be applied by pressure for example. For example a pressure orthogonal to the direction of assembly can be applied to bend the substrates.

[0090] In some embodiments at least two substrates are stacked as to form a small pillar and then rolled into one cylinder. Once the substrates are stacked, they are rolled together into a single cylinder using any conventional rolling apparatus or method. The rolling process may be performed manually or automatically.

[0091] In some embodiments, substrates are deposited on top of each other with an offset before rolling them into a cylinder. The offset may be achieved by shifting each successive substrates slightly relative to the previous one, creating an offset arrangement. The offset arrangement ensures that the edges of the substrates do not align perfectly.

[0092] This assembly process can create a self-supporting scaffolding substrate. The resulting scaffolding substrate can vary in size and length depending on the number of substrates used and the desired dimensions of the scaffolding substrate.

[0093] In some embodiments the plurality of substrates is maintained during the assembly 130 and during the incubation 140. In some embodiments the scaffolding substrate is hold in place by a container during incubation 140 to maintain the plurality of substrates together at a distance sufficient to have the filamentous fungus to bind the plurality of substrates to form a fermented plant-based food product. This step should be adapted to have sufficient oxygenation of the filamentous fungus to have a growth inside and outside of the scaffolding substrate. The container used should then have sufficient aperture to have an oxygen flow of at least 0.02 m3 / h and preferably 0.05 m3 / h per kilogram of scaffolding substrate. The container should also allow sufficient thermal conductivity either by the aperture or by having a sufficient high thermal conductivity to remove heat, created by the growth of the filamentous fungus, from the scaffolding substrate during incubation. The container may include, but is not limited to, a wrap, a net, a tray, a mold, a casing, plastic bags with holes, skewers, string passing through the scaffolding substrate or any other devices with holes to have air passing through. The container may be in any suitable material such as metallic, plastic, vegetable or any combination of it. In some embodiments the container can be sacrificial. Sacrificial means that the container is fermented by the filamentous fungus and become part of the fermented product. Such sacrificial containers can be made by vegetable materials such as rice paper, banana leaf, corn husk, jute fiber, bamboo or a combination of it. Alternatively, the container can be non- sacrificial and made in metal or non-fermentable plastic or a combination of it.

[0094] In some embodiments the container is used to bend the substrates during the incubation step to add a more complex structure to the scaffolding substrate. In some embodiments the substrates are folded in a direction orthogonal to the smallest dimension of the substrate to form a V-shape substrate. Alternatively, substrates can be folded in several directions orthogonal to the smallest dimension of the substrate to form a conical substrate. Alternatively, the inoculation step can be performed after the assembly has been done. In other words, the substrates can be assembled to form the scaffolding substrate 130 and then the scaffolding substrate is inoculated 120 with the method describe at the step 120.

[0095] During incubation 140, the scaffolding substrate is incubated. The filamentous fungus inoculated during the inoculation 120 grows and develops into and on the scaffolding substrate. To facilitate the growth of the filamentous fungus, this step is carried out under controlled parameters, such as temperature, humidity, oxygen concentration and / or time. Preferably, the incubation conditions are adjusted to avoid formation of unpleasant off-flavours, such as resulting from formation of ammonia, organic acid accumulation, and / or before sporulation.

[0096] Incubation 140 can be carried out in any way and consequently additional subsequent steps may be performed to obtain a good development of the filamentous fungus inside and / or around the scaffolding substrate. Incubation 140 can be, for example, liquid fermentation or more preferably, solid-state fermentation (SSF). The fermentation conditions and time may be adjusted to the filamentous fungus, the available nutrients, the composition of the scaffolding substrate and to the desired result. Preferably, the substrates are incubated at a temperature between 4 and 70°C, in particular between 10°C and 50°C, more preferably between 14°C and 40°C and more preferably between 22°C and 38DC.

[0097] In one embodiment, the incubation 140 is terminated prior to spore formation.

[0098] Fungal growth can be paused, for example by decreasing the temperature below temperature conditions required for growth of the filamentous fungus by any means such as cooling, freezing and / or by decreasing the water activity to below the water activity required for growth of the filamentous fungus by any means such as drying the fermented plant-based food product. Alternatively, fungal growth may be paused by decreasing oxygen concentration to below the critical level required by the respective filamentous fungus to grow. Preferably, the growth of the filamentous fungus is terminated by killing the filamentous fungus by heating the fermented plantbased food product to above 60°C, preferably above 71°C for at least 1 minute and more preferably more than 30 minutes, whereas the temperature is measured in the centre of the product.

[0099] Alternatively, fungal growth is not interrupted. Instead, the food product is stored in the fridge or freezer until consumption and is preferably consumed prior to sporulation or spoilage.

[0100] In one embodiment, the scaffolding substrate is inverted during incubation. Regular rotation of the scaffolding substrate during incubation ensures that all sides of the scaffolding substrate are equally in contact with airflow, thereby evening out oxygenation and heat removal. As a result, the scaffolding substrate develops more uniform fungal growth.

[0101] The fermented plant-based food product preferably comprises at least two different phases; preferably, the two different phases can be distinguished by human eye, more preferably a first phase comprising the substrates and a second phase comprising the filamentous fungus, preferably, wherein a ratio of the second phase of the fermented plant-based food product, which is define as a ratio of an area occupied by the second phase in a cross section of the fermented plant-based food product to a total area of said cross section, ranges from 0.15 to 0.4, the second phase being measured by a whiteness measurement described in detail later in the examples. The second phase made principally by the mycelium of the filamentous fungus is more lipophilic than the first phase made of substrates.

[0102] Once the fermented plant-based food product is formed, according to method 100 described above, further processing may be performed to improve taste, preservation and / or transportation as well as to change size and shape. In particular, the fermented plant-based food product may be marinated, spiced, smoked, cured, dehydrated, pressed, infused, soaked, tumbled, injected with liquids such as water, marinade and / or oil, steamed, boiled, cut, rolled, pressed, aged, fermented assembled with other products such as fat, or post-processed in any other way as typically done with animal meat products. Aging processes can be used to develop flavors and textures. During aging, enzymes and microbes present in the fermented plant-based food product continue to break down proteins and / or fats, leading to changes in texture and / or flavor. The length and conditions of aging can vary depending on the desired characteristics of the fermented plant-based food product. The post process can be used to replicate the shape and / or color of the meat product it is supposed to replace to facilitate easy recognition and identification by consumers.

[0103] In particular, the fermented plant-based food product is cut to closely resemble the size, thickness, and overall form of the meat product it is imitating. For example, if the imitated meat product is a traditional meat retail cut, the fermented plant-based food product will have a shape that is similar in size and thickness to the traditional meat retail cut such as ribeye, chicken breast, pork belly, ham, duck breast, pastrami, T-bone, rump, loin, porterhouse, filet, brisket, entrecote, tuna steak, coppa, pork chop, sirloin, steak, pancetta, prosciutto, bacon, salmon steak, lardons, jowl, lomo, guanciale, flank or any other retail cuts. On the contrary, by cutting the fermented plant-based food product to smaller pieces such as chunks it is possible to imitate burger and patty. Alternatively, by fermenting the fermented plant-based food product a second time it is possible to reassemble processed fermented plant-based food product such as chorizo, salchichon, pepperoni, fuet, salami, soppressata, bresaola, salami. For this second fermentation typical fermentation process of said processed fermented plant-based food product can be used, for example using Penicillium nalgiovense and / or Penecillium salamii to imitate salami.

[0104] The fermented plant-based food product also considers any unique features of the meat product being imitated. For instance, if the meat product has a distinctive texture or pattern on its surface and / or inside, the fermented plant-based food product can be designed to replicate that texture and pattern as closely as possible.

[0105] In another embodiment, the fermented plant-based food product can be shaped using various methods to achieve a desired shape, texture, and appearance. One such method is vacuum shaping, which involves placing the fermented plant-based food product in a mold and applying vacuum pressure to shape the product.

[0106] Another method of shaping the fermented plant-based food product is mechanical shaping, which involves using mechanical force to shape the product. This can be accomplished through a press, extruder, or similar device. Mechanical shaping can be particularly useful for shaping fermented plant-based food products that require a high degree of consistency and uniformity in their shape and size.

[0107] It should be noted that the post-processing methods are not limited to the methods described herein, and that other methods and variations of the above methods may be used as well. The shaping method used will depend on various factors such as the type of fermented plant-based food product, the desired shape and texture, and the manufacturing process used to produce the fermented plant-based food product.

[0108] In one embodiment, the fermented plant-based food product is more nutritious than the edible substrate, preferably wherein the proteins in the edible substrate are more digestible than prior to fermentation. Fermentation reduces antinutritional factors to enhance bioavailability of micronutrients. Some non-limiting possible examples are higher antioxidant activity, higher total phenolic content, increased soluble proteins, higher ellagic acid (anti-carcinogenic), increased content of L-DOPA and / or decrease in cholesterol.

[0109] The fermented plant-based food product is designed to provide a satisfying and juicy experience for consumers, similar to that of a traditional meat product, for example by releasing juices when bitten or cut, further enhancing the overall juiciness experience.

[0110] The fermented plant-based food product produced using method 100 can be compared with a standard food product produced without the assembly method 130. The standard food product follows the exact same process as the method 100, except for the assembly method, which differs. In the standard food product assembly, the TVP substrates are stacked substantially parallel to each other to form the scaffolding substrate. The fermented plant-based food product produced by the method 100 is at the top of Fig. 7 and the standard food product is at the bottom of Fig. 7.

[0111] The difference of assembly between the assembly method described herein and the previous art assembly method can be determined by measuring the oxygen diffusivity in both food products and in the substrate. The measure of the diffusivity is done by an oxygen meter (GREISINGER G1690) that has been calibrated with fresh air to 20.9% (initial oxygen concentration). After 25 h of fermentation, a whole is drilled into the center of the products with the same diameter as the sensor to create an airtight insertion. To eliminate the effect of metabolic activity in the three products, all products were covered by a plastic foil such that the fungi will use substantially all the oxygen inside the products indicated by a remaining oxygen level of below 2%. Once the oxygen depletion is complete, i.e. oxygen below 2% and stationary, the plastic foils are removed and the increase in oxygen level is measured. In the substrate, the time required to recover the initial oxygen concentration is more than 12 minutes. In the standard food product, the time required to recover the initial oxygen concentration is more than 20 minutes. As expected, the densely standard food product exhibits a poor oxygen diffusivity. For the fermented plant-based food product, the time to retrieve the initial concentration is about 7 minutes as presented in Fig. 8.

[0112] An optical analysis of the myceliation can be achieved by measuring the mycelium visible on a transversal cut of the fermented plant-based food products. This measurement correlates with the contrast offered by the whiteness of healthy fungal mycelium against a darker background corresponding with the non-myceliated scaffolding substrate. Fig. 9 represents 3 standard food products on the left and 3 fermented plant-based food products on the right. The percentage of white pixels in selected areas is analyzed and the myceliation levels within the different food products is compared. Images of the transversal cuts are converted from RGB into grey scale images according to the Imaged definition (( R+G+B) / 3, 8-bit, 0 to 255 pixel values). The grey scale images are processed with a default B&W threshold of 199 into binary images using a Make binary command. After obtaining the binary image, the area of interest in the image i.e. the area of the transversal cut is selected. The percentage of white pixels in the selected area is then calculated as: number of pixels with value > 199 in the selected area % whiteness = total number of pixels in the selected area

[0113] The standard food product presents a percentage of white pixels equal to 10% + / - 2% whereas the fermented plant-based food product from method 100 presents a percentage of white pixels equal to 22% + / - 2%. The method 100 allow for a better growth of the filamentous fungus inside of the product. By changing the assembly method of the scaffolding substrate, it is possible to finely tune the percentage of whiteness between 15% and 40%.

[0114] To compare the absorption characteristics of the two food products, cubes are cut from the two food products. Food products are cut into cubes several centimeters wide and weighing about 100 g each. Multiple 100g cubes of each food product are placed in a bag containing an excess of liquid, approximately summing 135g of cube and liquid in total. The liquid fraction was made of 45% of oil and 55% of water. The bag with the cube and the liquid is then vacuum sealed to force the liquid absorption in the cube. After 30 minutes, the cubes are removed from the bag and weight again. Table 1 summarizes the results, and we can observe that the fermented plant-based food product is capable of absorbing more liquid than its counterpart. The results show that the fermented plantbased food product is capable of absorbing the equivalent of 27% + / -2% of its own weight in liquid against 15% + / -3% for the standard food product. By finely tuning the assembly method we can modulate the absorption between 17 % and 45 %.

[0115] Table 1 As seen from the previous results, the fermented plant-based food product presents an increase of mycelium density and a lower density. The fermented plant-based food product presents an increase capacity in liquid absorption and therefore increase the taste, and mouthfeel of the consumer compared to the standard food product. By varying the assembly techniques, for example by applying pressure, changing the thickness of the sheets or the protein sources, it is possible to precisely adjust the texture, firmness, absorption capacity, juiciness and taste of the fermented plant-based food product. This flexibility offers advantages in altering both the juiciness and flavor of the product, influencing the consumer experience. By manipulating the amounts of oil and water, the sensory perception can vary, from resembling chicken to beef or to pork. The sensory experience of the consumer can be gauged by observing the quantity and type of liquid expelled during the pressing of the fermented plant-based food product. Due to the distinct phases in the fermented plant-based food product, the first phase, which is more hydrophilic, retains more water and the second phase retain more oil. The consumer's mouthfeel will be closer to meat products comprising interstitial tissues that comprise more fats and have an oily feeling whereas the muscle fiber will release blood more like water.

[0116] List of all aspects:

[0117] 1. A method to produce a fermented plant-based food product, the method comprising: a. providing at least one substrate(s), b. inoculating the substrates with at least one filamentous fungus, c. assembling the substrates to form a scaffolding substrate, d. fermenting said scaffolding substrate to allow the at least one filamentous fungus to grow inside, outside, or inside and outside said scaffolding substrate and / or said substrates.

[0118] 2. The method of the preceding aspect, wherein the substrates are each porous, fibrous, anisotropic, or foamed, or have a lamellar structure and / or is an extrudate.

[0119] 3. The method of any preceding aspect, wherein the substrates comprise non-animal proteins or are each made from non-animal proteins.

[0120] 4. The method of any preceding aspect, wherein the substrates comprise one or more proteins derived from plant-based sources, including pulses or legumes, such as soy, pea, fava, beans, oilseeds such as rapeseeds or sunflower seeds, grains such as wheat or oat grains, or any other sources and / or any combination thereof.

[0121] 5. The method of any preceding aspect, wherein the substrates comprise soy protein and at least another plant protein.

[0122] 6. The method of any preceding aspect, wherein the substrates comprise pea protein and / or soy protein.

[0123] 7. The method of any preceding aspect, wherein the substrates comprise fungal protein(s) or are each made from fungal protein(s).

[0124] 8. The method of any preceding aspect, wherein the substrates are solid, gelled or partly gelled.

[0125] 9. The method of any preceding aspect, wherein the substrates comprise starch or are each made from starch.

[0126] 10. The method of any preceding aspect, wherein the scaffolding substrate comprises at least 2 different types of substrates.

[0127] 11. The method of the preceding aspect, wherein a first substrate comprises more than 50% wt of protein and a second substrate comprises more than 50% wt of starch.

[0128] 12. The method of any preceding aspect, wherein the substrates comprise a first substrate comprising protein and a second substrate comprising carbohydrate.

[0129] 13. The method of any preceding aspect, wherein at least one, and preferably more or all, of the substrates have the shape of a sheet.

[0130] 14. The method of the preceding aspect, wherein the sheet extends along a first direction and a second direction, which is preferably orthogonal to the first direction, the sheet having a thickness along a third direction perpendicular to the first and second directions.

[0131] 15. The method of the preceding aspect, wherein the thickness is in a range of 1 mm and 10 cm, preferably between 5 mm and 3 cm.

[0132] 16. The method of any of aspects 13-15, wherein the substrate or the substrates are mechanically deformed.

[0133] 17. The method of any of aspects 13-16, wherein step c) comprises stacking the substrates on top of each other.

[0134] 18. The method of any of aspects 13-17, wherein step c) comprises bending and / or folding one or more of the substrates. 19. The method of any of aspects 13-18, wherein step c) comprises stacking the substrates on top of each other in a container having a diameter smaller than the first dimension and / or the second dimension of one or more of the sheets so that the one or more of the sheets are bent and / or folded upon being placed into the container, preferably wherein the container is cylindrical and said diameter is a diameter of a base circle of the container.

[0135] 20. The method of the preceding aspect, further comprising removing the scaffolding substrate from the container.

[0136] 21. The method of any of aspects 13-20, wherein the scaffolding substrate has a pinecone configuration in which preferably the substrates each form a scale of the pinecone.

[0137] 22. The method of any of aspects 13-21, wherein a majority of the substrates, preferably at least 75% or at least 90%, are bent and / or folded into a V-shape and / or a conical shape, preferably wherein the V-shape or the conical shape forms an internal angle of between 20° and 160°, preferably between 40° and 140°, and more preferably between 60° and 120°, preferably wherein the internal angle is determined in a cross section parallel to the longest direction of the scaffolding substrate, preferably wherein the internal angle is formed between the sides of the V- shape.

[0138] 23. The method of the preceding aspect, wherein the tips of the V-shapes point substantially in the direction of assembly, the direction of assembly being a direction along which the substrates are stacked to form the scaffolding substrate.

[0139] 24. The method of the preceding aspect, wherein, for each V-shape, an axis passing through the tip of the respective V-shape and bisecting an internal angle is defined, wherein a majority of the axes, preferably 75% of the axes, extend at an angle to the direction of assembly of between 0° and 45°, preferably of between 0° and 20°, preferably wherein the internal angle is determined in a crosssection extending along the direction of assembly.

[0140] 25. The method of aspect 23, wherein the tip of the V-shapes points substantially orthogonally to the direction of assembly.

[0141] 26. The method of aspect 22 or 25, wherein, for each V-shape, an axis passing through the tip of the respective V-shape and bisecting the internal angle is defined, wherein a majority of the axes, preferably 75% of the axes, extend at an angle to a direction of assembly of between 45 and 135° and preferably of between 75° and 115°, the direction of assembly being a direction along which the substrates are stacked to form the scaffolding substrate.

[0142] 27. The method of any of the preceding aspects, wherein step c) comprises assembling the substrates in a non-random, semi-random, semi-regular or regular manner.

[0143] 28. The method of any preceding aspect, wherein the substrate or substrates comprise or are each made from textured vegetable protein (TVP), preferably wherein the TVP has a water activity preferably between 0.8 and 1.0, more preferably between 0.9 and 1.0, even more preferably between 0.96 and 1.

[0144] 29. The method of any preceding aspect, wherein in a cross section parallel or perpendicular to the longest direction of the scaffolding substrate, a ratio of an area occupied by voids between the substrates to a total area of the cross section is between 0.1 and 0.4 and preferably between 0.1 and 0.3.

[0145] 30. The method of any preceding aspect, wherein the filamentous fungus is Rhizopus. 31. The method of any preceding aspect, wherein substrates are inoculated with at least one filamentous fungus and at least one other microbial species.

[0146] 32. The method of the preceding aspect, wherein the other microbial species is a fungus, a bacterium or a yeast.

[0147] 33. The method of any preceding aspect, wherein the substrates are inoculated with at least two filamentous fungi.

[0148] 34. The method of any preceding aspect, wherein the step b) is carried out before or after the step c).

[0149] 35. The method of any preceding aspect, wherein the scaffolding substrate has the shape of a sausage.

[0150] 36. The method of the preceding aspect, wherein the sausage is at least 3 cm in diameter and preferably at least 7 cm.

[0151] 37. The method of any preceding aspect, wherein the scaffolding substrate is held in place during step d), preferably by skewers or string passing through the scaffolding substrate.

[0152] 38. The method of any preceding aspect, wherein the scaffolding substrate is held in place during step d) by a container, the container preferably being permeable or semi-permeable to oxygen.

[0153] 39. The method of any preceding aspect, wherein the scaffolding substrate is held in place during step d) by a container having apertures to allow air to flow therethrough.

[0154] 40. The method of aspect 38 or 39, wherein the container is rigid, non-rigid or flexible.

[0155] 41. The method of any of aspects 38-40, wherein the container comprises a mold, a net, a membrane, a wrap, a tray, a casing, or a plastic bag with holes.

[0156] 42. The method of any of aspects 38-41, wherein the container is sacrificial.

[0157] 43. The method of any of aspects 38-42, wherein the container is made from vegetables.

[0158] 44. The method of any of aspects 38-43, wherein the container is edible.

[0159] 45. The method of any of aspects 38-44, wherein the container is non-sacrificial and made from metal or plastic.

[0160] 46. The method of any preceding aspect, wherein the scaffolding substrate is rotated during step d).

[0161] 47. The method of any preceding aspect, wherein step d) is carried out under controlled parameters such as temperature, oxygenation and / or humidity.

[0162] 48. The method of any preceding aspect, wherein the fermented scaffolding substrate is post processed.

[0163] 49. The method of the preceding aspect, wherein the post process comprises any of the following steps: marinated, spiced, smoked, cured, dehydrated, pressed, infused, soaked, tumbled, injected with liquids such as water, marinade and / or oil, steamed, boiled, cut, rolled, pressed, aged, fermented and assembled with other products.

[0164] 50. A fermented plant-based food product produced by a method according to any of the preceding aspects, preferably in a shape of a sausage, wherein a diameter of the fermented plant-based food product is at least 3 cm and preferably at least 7 cm.

[0165] 51. The fermented plant-based food product of the preceding aspect, wherein the fermented plantbased food product can absorb at least 17% of its own weight, preferably at least 20% and preferably at least 25% of liquids, the liquids preferably being water, oil or a mixture thereof, preferably wherein the mixture comprising 40 wt% to 70 wt% of water, more preferably 45 wt% to 65 wt% of water, the rest being oil.

[0166] 52. The fermented plant-based food product of the preceding aspect, wherein a cross section of the fermented plant-based food product presents a whiteness of 15% or more and preferably 20% or more. 53. The fermented plant-based food product of any of aspect 50-52, wherein the fermented plantbased food product comprises at least two phases, one being more hydrophilic than the other.

Claims

CLAIMS1. A fermented plant-based food product comprising a matrix of substrate interconnected by at least a filamentous fungus having a geometrical shape with one dimension longer than the other two orthogonal dimensions, with a length being the longest direction of the fermented plant-based food and the two other length are at least 3 cm each and preferably at least 7 cm each, the length is at least 3 cm and preferably at least 7 cm, wherein a cross section parallel or perpendicular to the longest direction of the fermented plantbased product presents a whiteness of 15% or more and preferably 20% or more.

2. The fermented plant-based product according to claim 1, wherein the fermented plant-based product absorbs at least 17% of its own weight, preferably 20% and preferably 25% of liquids, the liquids preferably being water, oil or a mixture thereof, preferably wherein the mixture comprising 40 wt% to 70 wt% of water, more preferably 45 wt% to 65 wt% of water, the rest being oil.

3. The fermented plant-based product according to claim 1 or 2, wherein the shape of the fermented plant-food product is a sausage with a diameter that is at least 3 cm and preferably at least 7 cm, the diameter measured at the middle of the longest direction.

4. A method to produce the fermented plant-based food product according to any of the preceding claims, the method comprising: a. providing a plurality of substrates with a first density, b. inoculating the substrates with at least one filamentous fungus, c. assembling the substrates to form a scaffolding substrate to reach a second density, d. fermenting said scaffolding substrate to allow the at least one filamentous fungus to grow inside, outside, or inside and outside said scaffolding substrate and / or substrates, wherein the substrates are mechanically deformed, bent and / or folded and stacked on top of each other during step c. and the second density is 10 % to 20 % higher than the first density.

5. The method according to claim 4, wherein the substrates are each porous, fibrous, anisotropic, foamed, have a lamellar structure and / or an extrudate and comprise non-animal proteins or are each made from non-animal proteins such as textured vegetable protein.

6. The method according to claim 4 or 5, wherein at least one, and preferably more or all of the substrates have a shape of a sheet, wherein the sheet extends along a first direction and a second direction, which is preferably orthogonal to the first direction, the sheet having a thickness along a third direction perpendicular to the first and second directions and the thickness is preferably between 1 mm and 10 cm, preferably between 5 mm and 3 cm.

7. The method according to any one of claims 4 to 6, wherein in a cross section parallel or perpendicular to the longest direction of the scaffolding substrate, a ratio of an area occupied by voids between the substrates to a total area of the cross section is between 0.1 and 0.4 and preferably between 0.1 and 0.3.

8. The method according to any one of claims 4 to 7, wherein the substrates are each bent and / or folded into a V-shape and / or a conical shape, preferably wherein the V-shape or the conical shape forms an internal angle of between 20° and 160°, preferably between 40° and 140°, and more preferably between 60° and 120°.

9. The method according to claim 8, wherein a line linking the tips of the V-shapes is substantially colinear to a direction of assembly, the direction of assembly preferably being a direction along which the substrates are stacked to form the scaffolding substrate.

10. The method according to claim 8 or 9, wherein, for each V-shape, an axis passing through the tip of the respective V-shape and bisecting an internal angle is defined, wherein a majority of the axes, preferably 75% of the axes, extend at an angle to the direction of assembly of between 0° and 45° and preferably between 0° and 25°.

11. The method according to any one of claims 4 to 10, wherein the step b) is carried out before, during or after the step c).

12. The method according to any one of claims 4 to 11, wherein the filamentous fungus is Rhizopus.

13. The method according to any one of claims 4 to 12, wherein the substrates are inoculated with at least one filamentous fungus and at least one other microbial species such as a fungus, a bacterium or a yeast.

14. The method according to any one of claims 4 to 13, wherein the scaffolding substrate is held in place during step d).

15. The method according to any one of claims 4 to 14, wherein the scaffolding substrate is held in place during step d) by a container having apertures to allow air to flow therethrough.

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