Hydration of fermented foods

Abstract

The present disclosure describes methods for preparing a hydrated fermented substrate comprising a) providing one or more edible ingredients; b) subjecting the one or more edible ingredients to one or more preparation steps to form a fermentation substrate; c) contacting the fermentation substrate with one or more microorganisms under conditions allowing the microorganisms to colonize and 5 proliferate on / in the fermentation substrate and to produce a fermented substrate; d) contacting the fermented substrate with a hydrating composition under conditions allowing for fermented substrate to absorb the hydrating composition to produce the hydrated fermented substrate, and e) isolating the hydrated fermented substrate. 10 * * *

Description

Hydration of fermented foodsTechnical Field

[0001] The present application concerns methods for rehydrating fermented edible products and the rehydrated edible products resulting therefrom, which optionally from the rehydration are improved in one or more quality properties of flavour, aroma, texture, cooking experience, visual appearance, nutritional and / or dietary value, spoilage time, and / or carbon foot-print.Background

[0002] The practice of food fermentation and its diverse techniques can be traced back to ancient times and is currently prevalent across various food cultures worldwide. One only need to consider the examples of beer, cheese, and yoghurt to get an idea about the magnitude of the use fermentation in the food and beverage industry. However, there is now an increased demand for foods based on vegetable sources. Increasingly, consumers prioritize food options that align with climate and ecological sustainability, specifically emphasizing the utilization of raw materials that would otherwise be classified as waste streams or currently unfit for human consumption or food production. There exists a demand to create high-quality food options based on plant sources, while simultaneously delivering the desired textures, flavours, nutrients, and other characteristics reminiscent of meat. Additionally, it is preferable to utilize vegetable raw materials sourced from agricultural or industrial waste streams (upcycling) either partially or entirely. The existing technologies used for meat alternatives are inadequate or unappealing due to their reliance on artificial additives and isolated proteins to achieve meat-like textures and flavours. PCT / EP2023 / 069428 (unpublished) describes how raw materials and pretreatment of such raw materials to prepare a substrate for a solid state fermentation can be selected to improve the quality parameters of the fermented food product made. However, it has been found that the fermentation of the substrate also alters the matrix of the substrate, and the fermentation process reduces or consumes moisture potentially leading to an undesirably dry food product.Summary

[0003] The rehydration methods and rehydrated edible products described herein provide improvements that address the drawbacks and limitations of background approaches. Rehydration of fermented food product can be used to optimize the quality parameters for the product including theflavour, aroma, texture, cooking experience, visual appearance, nutritional and / or dietary value, spoilage time, and / or carbon footprint, and can further enable the creation of meat-like properties while simultaneously delivering superior health and environmental benefits.

[0004] Accordingly, in a first aspect the present disclosure provides a method for preparing a hydrated fermented substrate comprising: a) providing one or more edible ingredients; b) subjecting the one or more edible ingredients to one or more preparation steps to form a fermentation substrate; c) contacting the fermentation substrate with one or more microorganisms under conditions allowing the microorganisms to colonize and proliferate on / in the fermentation substrate and to produce a fermented substrate; d) contacting the fermented substrate with a hydrating composition under conditions allowing for fermented substrate to absorb the hydrating composition to produce the hydrated fermented substrate and e) isolating the hydrated fermented substrate.

[0005] In a further aspect the present disclosure provides a hydrated fermented substrate obtained by the method described herein.Description of drawings and figures

[0006] The figures included herein are illustrative and simplified for clarity. They merely show details that are essential to the understanding of the invention, while other details may have been left out. The figures and drawings included herein depict the content described in this document.Figure 1 shows a flow chart of the process of making a fermented substrate.Figure 2 shows fermented substrate cut into 5x5cm cubes.Figure 3 shows obtained values of t / q in terms of time show a linear tendency.Figure 4 shows the amount of liquid absorbed per gram of composite during a specified duration of submersion.Figure 5 shows obtained values of t / q in terms of time show a linear tendency. A linear tendency of the data is observed, indicating that the obtained data fits into the proposed model.Figure 6 shows absorbed liquid per gram of composite when the sample is submerged in a specific amount of time.Figure 7 shows a linear trend in the data, suggesting that the collected data aligns with the proposedmodel.Figure 8 shows the amount of liquid absorbed per gram of composite during a specified duration of submersion.Figure 9 shows a representative picture of the control colour profile.Figure 10 shows a representative picture that shows the reference colour profileFigure 11 shows a comparison of normalized RGB colours.Figure 12 shows a visualization of obtained sample colours.Figure 13 shows the final product protein content when the fermented substrate is hydrated with nitrogen free hydrating solution.Figure 14 shows the relation between the protein content of the final rehydrated fermented substrate in relation to the absorbed hydrating solution. Each line represents a different concentration of bakers' yeast supplement in the hydrate (w / w).Figure 15 shows the relation between the protein content of the final rehydrated fermented substrate in relation to the absorbed hydrating solution. Each line represents a different concentration of hydrolyzed peas supplement in the hydrate (w / w).Figure 16 shows a symmetric plot of the principal coordinate analysis (PCoA) of the association between sensory attributes and samples. The blue triangle shows the common attributes among the samples.Figure 17 shows a typical plot of instrumental texture profile analysis (Centre for Industrial Rheology. Available at: Texture Analysis And Texture Profile Analysis - Rheology Lab).Figure 18 and 19 shows the effect of absorption density (%) on hardness (N) and chewiness (N) of the fermented composite.Figure 20 and 21 shows the effect of absorption density (%) on the resilience and cohesiveness of the fermented composite.Figure 22 shows a representative diagram of the hydration by submersionFigure 23 shows the logarithmic part of the kinetic PFO Langmuir model is presented versus the time. Figure 24 shows the time over the absorption density (t / q) of the data are plotted versus t, indicating a linear tendency.Figure 25 shows the selected predicted absorption data by the selected model and the experimental data.Figure 26 shows a representative diagram of the hydration by absorption of a fixed volume. Where t is time, FA is the free (non-absorbed) hydrating solution, W is the weight of the matrix, AA is theabsorbed hydrating solution and WO is the initial weight of the matrix.Figure 27 shows the absorbed liquid over the time when using absorption of a fixed hydrating solution volume.Figure 28 shows the submersion-like performance during the first seconds in a fixed volume hydration causing a higher absorption rate during the first seconds.Figure 29 shows the effect of temperature in an injection hydration process.Incorporation by reference

[0007] All publications, specifically, patents, and patent applications referred to herein are incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. In the event of a conflict between a term herein and a term in an incorporated reference, the term herein prevails and controls.Detailed DescriptionThe features and advantages of the present invention are readily apparent to a person skilled in the art by the below detailed description of embodiments and examples of the invention with reference to the figures and drawings included herein.Definitions

[0008] The term "substrate" as used herein refers to any composition or matrix of ingredients, and "fermented substrate" refers to such composition or matrix of ingredients to which a microorganism has been added an allowed to proliferate metabolising the ingredients.

[0009] The terms "hydrate", "marinade", "hydrating composition" and "hydrating solution" and similar expressions are used herein interchangeable and refers to any mixture or solution used for adding moisture and optionally oils and / or flavours to a fermented substrate.

[0010] The terms "hydration", "rehydration" and "marination" as used herein interchangeable refers to the process of applying a hydrate to a fermented substrate to allow the fermented substrate to absorb moisture and optionally oils and / or flavours to a fermented substrate.

[0011] Percentages given herein as "%wt" refers to weight percent. Weight percents eg. for edible ingredients are in some embodiments calculated as the percentage of a substance in the natural form of the ingredient, such as raw unpeeled beets, while weight percents in other embodiments are calculated as the percentage of a substance in an ingredient which is processed such as by peeling,shredding, drying etc.

[0012] The term "comprise" and "include" as used throughout the specification and the accompanying items as well as variations such as "comprises", "comprising", "includes" and "including" are to be interpreted inclusively. These words are intended to convey the possible inclusion of other elements or integers not specifically recited, where the context allows.

[0013] The articles "a" and "an" are used herein refer to one or to more than one (i.e. to one or at least one) of the grammatical object of the article. By way of example, "an element" may mean one element or more than one element.

[0014] Terms like "preferably", "commonly", "particularly", and "typically" are not utilized herein to limit the scope of the itemed invention or to imply that certain features are critical, essential, or even important to the structure or function of the itemed invention. Rather, these terms are merely intended to highlight alternative or additional features that can or cannot be utilized in a particular embodiment of the present invention.

[0015] Where a numerical limit or range is stated herein, the endpoints are included. Also, all values and sub ranges within a numerical limit or range are specifically included as if explicitly written out.

[0016] The term "and / or" as used herein is intended to represent an inclusive "or". The wording X and / or Y is meant to mean both X or Y and X and Y. Further the wording X, Y and / or Z is intended to mean X, Y and Z alone or any combination of X, Y, and Z.

[0017] For clarity, for compositions including multiple ranges in percent of compounds or ingredients, the sum of all compounds or ingredients in the composition equals 100 percent and the sum of ranged compounds or ingredients does not exceed 100 percent. As an example, a composition comprising 20- 40% A, 10-30% B, and 20-40% C, the sum of A, B, and C does not exceed 100%, while where the sum of A, B, and C is below 100%, the remainder can be other ingredients adding up to 100%.

[0018] All methods described herein can be performed in any suitable order of steps unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0019] Unless specifically defined herein, all technical and scientific terms used have the same meaning as commonly understood by a skilled person in the field of fermented foods.

[0020] All methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, with suitable methods and materials being describedherein. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, shall prevail. Further, the materials, methods, and examples are illustrative only and are not intended to be limiting, unless otherwise specified.

[0021] The first aspect provided for herein concerns a method for preparing a hydrated fermented substrate comprising: a) providing one or more edible ingredients; b) subjecting the one or more edible ingredients to one or more preparation steps to form a fermentation substrate; c) contacting the fermentation substrate with one or more microorganisms under conditions allowing the microorganisms to colonize and proliferate on / in the fermentation substrate and to produce a fermented substrate; d) contacting the fermented substrate with a hydrating composition under conditions allowing for fermented substrate to absorb the hydrating composition to produce the hydrated fermented substrate and e) isolating the hydrated fermented substrate.

[0022] One objective with the hydration of the fermented substrate is to improve one or more quality parameters or properties such as flavour, aroma, texture, cooking experience, visual appearance, nutritional and / or value, spoilage time, and / or carbon footprint

[0023] While a significant objective of the method for hydrating the fermented substrate is to transfer moisture to the fermented substrate thereby replacing moisture consumed in the fermentation process or adding further moisture to the fermented substrate, the main component of the hydrating composition is water. However, the hydrating composition may also suitably comprise further component adding functionality to the fermented substrate and contributing to the improvement of quality parameters. Accordingly in some embodiments hydrating composition comprises water and optionally one or more edible dissolved, dispersed or emulsified components selected from a salt, a protein, a carbohydrates, a surfactant or emulsifier, a colourant, a preservative, an oil or a fat, a vitamin, an amino acid, a mineral, a sweetener, a spice, an acidifier, a dietary fiber, a prebiotic, and / or a probiotic.

[0024] Salts not only add to the flavour of the fermented substrate, but it also binds moisture in the fermented substrate and in some cases preserves the fermented substrate from fouling. A preferred salt is sodium chloride, and in special embodiments the hydrating composition comprises between 0,001 g / L to 50 g / L of sodium chloride, such as between 0,001 g / L to 25 g / L, such as between 0,001g / L to 10 g / L, such as between 0,001 g / L to 5 g / L, such as between 0,001 g / L to 1 g / L, such as between 0,001 g / L to 0,5 g / L, such as between 0,01 g / L to 0,25 g / L, such as between 0,01 g / L to 0,1 g / L, such as between 0,025 g / L to 0,05 g / L .

[0025] Adding proteins and / or carbohydrates to the fermented substrate via the hydrating composition improves the nutritional value of the fermented substrate and also add to flavour, aroma and texture, so in some embodiments the hydrating composition comprises protein and / or carbohydrates. The protein and / or the carbohydrate is preferably added to the hydrating composition extracts of biomass and / or vegetables, which also contains other useful components adding to the nutritional or dietary value or to the flavour and / or aroma. The biomass is in some embodiments a fungal biomass, such as a yeast biomass, optionally a baker's or brewer's yeast - in particular yeasts of the genus Saccharomyces, such as the species S. cerevisiae. The biomass is suitably inactivated so as not to introduce further fermentation of the substrate. Yeast can suitably and gently be inactivated by hydrolysis using sous-vide cooking for 20-30 hours, preferably about 24 hours, at elevated temperature such as 50°C to 70°, preferably 56°C to 60°C.

[0026] Where the protein and / or carbohydrate source is a vegetable, legumes are preferred, such as beans. In preferred embodiments the bean is a P. sativa pea. When the source of proteins and / or carbohydrate is a vegetable, it is preferred to prepare the extract by cooking mechanically shredded or split vegetables, such as split peas, and adding the cooking water to the hydrating composition. The vegetable is suitably cooked with water in a vegetable-to-water ratio of 1:4 to 1:1, preferably 1:2, at 100% steam for 1- to 40 minutes, ideally 30 minutes and then straining the cooking liquid before adding it to the hydrating composition.

[0027] In further embodiments the hydrating composition has a protein concentration which is higher than the protein concentration of the fermented substrate. This can be important where the hydrating composition may wash out protein of the fermented substrate, because the protein concentration in the fermented substrate is higher than the concentration in the hydrating composition. In other embodiments the hydrating composition has a protein content which is at least 5% higher, such as at least 10% higher, such as at least 20% higher, such as at least 30% higher, such as at least 40% higher, such as at least 50% higher, such as at least 60% higher, such as at least 70% higher, such as at least 80% higher, such as at least 100% higher, such as at least 150% higher, such as at least 200% higher, such as at least 500% higher than the protein concentration of the fermented substrate. Alternatively or additionally, the hydrating composition can have a protein content of more than 10 wt%, such as more than 12 wt%, such as more than 15 wt%, such as more than 20 wt%, such as more than 25 wt%.

[0028] The hydrating composition can also advantageously be used to apply a desired colour to the fermented substrate. Such colours are preferably natural colours including but not limited toBeta-Carotenes - which can be obtained from carrots, sweet potatoes, and other vegetables, and which gives a yellow or orange colour to foods.Anthocyanins - which can be obtained from fruits like berries, grapes, and red cabbage and which provide red, purple, or blue hues.Chlorophyll - which can be obtained from green leafy vegetables like spinach and parsley, and which provides a green colour.

[0029] The colours may in some embodiments be provided in the form of extracts, powders or juices of plants such as beets (provides a vibrant red or pink hue); Turmeric (provides a yellow colour); saffron (provides a yellow-orange colour); paprika (provides red, orange, or brown colouring); Annatto (extracted from the seeds of the achiote tree and providing a yellow to orange colour); spinach (provides a green colour); red cabbage (provides shades of purple and blue); spirulina (provides blue or green colours); carmine (cochineal extract obtained from the crushed bodies of female cochineal insects, providing a red colour); and / or safflower (Carthamus, providing a red or yellow colour). Accordingly, in some embodiments the colourant is an extract from carrot, pumpkin, red beet, beet root, barley malt, carthamus, radish, and or apple. Preferably the colourant is an extract selected from a) carrot, pumpkin and apple, b) red beet, barley malt and Carthamus, c) radish and carrot, d) reed beet and barley malt, e) pumpkin and apple, f) beetroot and carrot, and / or g) red beet. Particularly, attractive colourant is a colourant extract from a) carrot, pumpkin and apple, and / or b) red beet, which provides the hydrated fermented substrate with the most meat-like appearance.

[0030] In further embodiments the hydrating composition comprises a surfactant and / or emulsifier. Surfactants are useful for improving the wetting capacity of the hydrating composition (the ability of the hydrating composition to penetrate the matrix of the fermented substrate). Emulsifying surfactants are useful for supplementing non-polar compounds, colourants, preservatives, prebiotics and the like to the fermented substrate. Suitable surfactants / emulsifiers include those selected from lecithin, monoglycerides, diglycerides, polysorbates (optionally Polysorbate 80), sorbitan esters (optionally sorbitan monostearate), propylene glycol esters (optionally propylene glycol monostearate), glycerol esters (optionally glycerol monostearate), acetylated monoglycerides, sucrose esters (optionally sucrose stearate), polyglycerol esters (optionally polyglycerol polyricinoleate - PGPR), and or lactic acid esters of mono- and diglycerides (LACTEM).

[0031] In further embodiments the hydrating composition comprises a preservative suitable for preventing or retarding fouling of the fermented substrate. Sodium Chloride in various forms is atraditional preservative that helps inhibit the growth of bacteria and molds. Sugar (sucrose) acts as a preservative by reducing water activity and inhibiting the growth of microorganisms. Acetic acid in vinegar helps preserve food by creating an acidic environment that inhibits the growth of bacteria. Citric acid is used as a preservative to enhance acidity and prevent spoilage. Ascorbic acid (Vitamin C) is used to prevent the oxidation helping to maintain colour and freshness. Tocopherols (Vitamin E) are used to prevent the oxidation of fats and oils, helping to extend the shelf life of products containing these ingredients. Rosemary extract, rich in natural antioxidants, is used to prevent lipid oxidation and maintain product quality. Natamycin is used to inhibit the growth of molds. Sorbic acid and potassium sorbate are used as preservatives in a variety of food products. Benzoic acid and sodium benzoate are used against yeast and mold. Calcium propionate is used to inhibit the growth of mold and bacteria. BHA (butylated hydroxyanisole) and BHT (butylated hydroxytoluene) are synthetic antioxidants used to prevent the oxidation of fats and oils in various foods. Accordingly, in some embodiments preferred preservatives include those selected from sodium chloride, sugars, vinegars, citric acid, ascorbic acid (vitamin C), tocopherols (vitamin E), rosemary extract, natamycin, sorbic acid or its salts, benzoic acid or its salts (optionally sodium benzoate), calcium propionate, BHA (butylated hydroxyanisole), and / or BHT (butylated hydroxytoluene).

[0032] In further embodiments the hydrating composition comprises an oil or a fat. Oils / fats are useful for adding nutritional value, flavour and texture to the fermented substrate. In some embodiments preferred oils / fats includes those selected from olive oil, canola oil, soybean oil, corn oil, sunflower oil, coconut oil, peanut oil, sesame oil, avocado oil, grapeseed oil, rape seed oil, butter, lard, shortening, coconut butter, palm oil, walnut oil, and / or flaxseed oil. In some embodiments the addition of an oil / fat to the hydrating composition is combined with addition of a surfactant or emulsifier.

[0033] In some embodiments the hydrating composition is an oil / fat-water emulsion. Adding oil to the substrate in the form of an emulsion provide for and even distribution of the oil and moisture in the substate greatly benefiting the organoleptic properties, including taste, texture, mouthfeel and softness / bite. In some embodiments the oil / fat-water emulsion comprise 2,5 %wt to 10 %wt of oil or fat for optimal result. The oil / fat is preferably a vegetable oil as mentioned above, optionally Sunflower oil. In further embodiments the oil / fat-water emulsion comprises salt and or spices, in particular peppers like black pepper, for further improvement of the organoleptic properties.

[0034] In further embodiments the hydrating composition comprises one or more added vitamins. Vitamins are useful for increasing the nutritional value of the fermented substrate. In some embodiments preferred vitamins are those selected from vitamin A (Retinol, Retinal, Retinoic Acid),vitamin Bl (Thiamine), vitamin B2 (Riboflavin), vitamin B3 (Niacin), vitamin B5 (Pantothenic Acid), vitamin B6 (Pyridoxine), vitamin B7 (Biotin), vitamin B9 (Folate, Folic Acid), vitamin B12 (Cobalamin), vitamin C (Ascorbic Acid), vitamin D (Cholecalciferol, Ergocalciferol), vitamin E (Tocopherols, Tocotrienols), vitamin K (Phylloquinone, Menaquinones), and / or vitamin P (Bioflavonoids).

[0035] In further embodiments the hydrating composition comprises added amino acids. Amino acids are useful for increasing the nutritional value of the fermented substrate and for some amino acids the aroma and flavour. In some embodiments preferred amino acids are those selected from essential amino acids (Lysine, Leucine, Isoleucine, Valine, Methionine, Phenylalanine, Threonine, and / or Tryptophan), non-Essential amino acids (Alanine, Arginine, Asparagine, Aspartic Acid, Cysteine, Glutamic Acid, Glycine, Proline, Serine, and / or Tyrosine), conditional amino acids (Arginine, and / or Cysteine), and / or semi-essential amino acids (Histidine).

[0036] In further embodiments the hydrating composition comprises added minerals. Minerals are useful for increasing the nutritional value of the fermented substrate. In some embodiments preferred minerals are those selected from a salt of Calcium, Phosphorus, Potassium, Sodium, Magnesium, Iron, Zinc, Copper, Manganese, Selenium, Iodine, Fluoride, Chromium, Molybdenum, and / or Cobalt.

[0037] In further embodiments the hydrating composition comprises added sweetener. Sweeteners are useful for improving the flavour of the hydrated fermented substrate. A preferred sweetener is a natural sweetener, preferably selected from sugar (sucrose), fructose, glucose, glucose syrup, stevia, honey, maple syrup, agave nectar, coconut sugar, date sugar, monk fruit sweetener, molasses, rice syrup, and / or yacon syrup. In other embodiments the sweetener is an artificial sweetener selected from aspartame, xylitol, saccharin, sucralose, acesulfame potassium, steviol glycosides, neotame, cyclamate, and / or advantame.

[0038] In further embodiments the hydrating composition comprises added spices. Spices are useful for the flavour of the fermented substrate. In some embodiments preferred spices are those selected from thyme, basil, garlic, onion, black pepper, green peppers, chili peppers, cinnamon, cumin, coriander, paprika, turmeric, ginger, nutmeg, cloves, cardamom, bay leaves, oregano, rosemary, and / or sage.

[0039] In further embodiments the hydrating composition comprises an acidifier. Acidifiers are used for improving flavour and preservation of the fermented substrate. In some embodiments preferred acidifiers are those selected from vinegar, acetic acid, and / or citric acid.

[0040] In further embodiments the hydrating composition comprises added dietary fibers / prebiotics. Dietary fibers / prebiotics are useful for maintains gut health and a well-functioning digestion. In some embodiments preferred fibers / prebiotics are those selected from betaglucan, pectin, inulin,Fructooligosaccharides (FOS), Galactooligosaccharides (GOS), Resistant Starch, Arabinoxylan, Lactulose, Konjac Glucomannan, and / or Polydextrose.

[0041] The pH of the hydrating composition was found to highly influence the absorption profile of the hydrating composition - and the amount of absorbed hydrating composition influences the texture and the organoleptic properties of the hydrated fermented substrate including appearance, flavour, and mouthfeel. Acidic or alkaline pH values improved the absorption profile while neutral pH was less useful. Accordingly in some embodiments the pH of hydrating composition is acidic or alkaline. Alternatively or additionally the pH of hydrating composition is preferably between pH 5 and pH 6, such as between pH 5,1 and pH 5,3.

[0042] Also, the temperature of the hydrating composition was found to highly influence the absorption profile of the hydrating composition, where temperatures between 5°C to 45°, particularly 10°C to 40°C, more particularly 15°C to 35°C, such as between 20°C to 30°C, was found the produce the best absorption profile.

[0043] In further embodiments of the method the hydrating composition is filtered to remove microorganisms prior to contacting the fermented substrate with the hydrating composition. Contamination of the fermented substrate with microorganisms from the hydrating composition accelerates fouling of the fermented substrate and reduce the shelf-life of the product. It was found that using a filter pore size of 0.45 pm removed microbial contamination for the hydrating composition without negatively affecting the taste properties of the hydrating composition. Accordingly, in some embodiments a pore size of 0.45 pm or below can be applied, in particular pore sizes between 0,22 pm and 0,45 pm. Such filters are commercially available.

[0044] The fermented substrate comprises one or more edible ingredients, which can be sourced from any suitable source. It is particularly useful when sourcing edible ingredients from industrial or agricultural waste streams which are not usually used or even suitable for human consumption due to lack of appropriate properties. One example of such edible ingredients is Brewers Spent Grain (BSG, typically barley or wheat), which is the cereal residue remaining after the cereal has been used for brewing of for example beer or other beverages. BSG contains, among other things, leftover starch, sugars, and fibres not spent during the brewing process. However, BSG is not very attractive as a food ingredient for humans. Therefore, prior to the availability of the method provided for herein, BSG has typically been used for animal fodder or incinerated for heat.

[0045] The choices of edible ingredients and the microbial fermentation, whether separately or together, all influence the properties of the edible product in particular flavour, aroma, texture, cooking experience, visual appearance, nutritional and / or dietary value, spoilage time, and / or carbonfootprint. Accordingly in further embodiments, the edible ingredients can comprise one or more of: a) a dry component comprising i) starch providing nutrient to the one or more microorganism and ii) fibres providing nutritional and / or dietary value to the edible product, the dry component being capable of absorbing moisture from other edible ingredients; b) a high-moisture and high starch component providing for the formation of gelatinized starch, binding other edible ingredients together and providing nutrient to the one or more microorganisms, as well as texture to the edible product; c) a component providing minerals and vitamins for the one or more microorganisms as well as a natural colour to the edible product; d) a high-protein and low moisture component providing flavour and firmness and structure to the texture of the edible product; or e) a high starch component providing sustained nutrients for the one or more microorganism and providing softness to the texture of the edible product.

[0046] The choice ingredients can be based on the ingredient's individual nutritional components, and their combined qualities as a growth medium for the fungal culture used.

[0047] In some embodiments, the one or more edible ingredients comprise at least 2, such as at least 3, such as at least 4, such as at least 5 components selected from the dry component, the high- moisture and high-starch component, the component providing minerals and vitamins, the high- protein and low-moisture component, and / or the high-starch component. In a special embodiment, the one or more edible ingredients comprise all 5 ingredients selected from the dry component, the high-moisture and high-starch component, the component providing minerals and vitamins, the high- protein and low-moisture component, or the high-starch component.

[0048] In more specific embodiments, the edible ingredients are selected from cereal grains, root vegetables, and legumes or a combination or extract thereof have proved to be particularly useful.

[0049] The cereal grain is very useful as the dry component, in particular cereals selected from maize (corn), rice, wheat, barley, sorghum, millet, oats, triticale, rye, and / or fonio. In some embodiments the cereal grain is spent grain from the fermentation of alcoholic beverages, brewers spent grain (BSG). More specifically the use of oats as cereal grain is particularly attractive. The one or more edible ingredients preferably comprise 5 %wt to 60 %wt of cereal grain, such as 5 %wt to 10 %wt, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt, optionally 19 %wt to 21 %wt of cereal grain, particularly oats.

[0050] The root vegetable is very useful as the high-moisture and high starch component and / or thecomponent providing minerals and vitamins. The root vegetable can in particular be a modified plant stem vegetable, a root-like stem vegetable, or a true root vegetable.

[0051] Modified plant stem vegetables include Corms (bulbo-tuber / bulbotuber), Rhizomes orTubers. Corm is suitably selected from Amorphophallus konjac (konjac), Colocasia esculenta (taro), Eleocharis dulcis (Chinese water chestnut), Ensete spp. (enset), Nymphaea spp. (waterlily), Pteridium esculentum, Sagittaria spp. (arrowhead or wapatoo), Typha spp., Xanthosoma spp. (malanga, cocoyam, tannia, yautia and other names), and / or Colocasia antiquorum (eddoe or Japanese potato). A rhizome is suitably selected from Curcuma longa (turmeric), Panax ginseng (ginseng), Arthropodium spp. (rengarenga, vanilla lily, and others), Canna spp. (canna), Cordyline fruticosa (ti), Maranta arundinacea (arrowroot), Nelumbo nucifera (lotus root), Typha spp. (cattail or bulrush), and / or Zingiber officinale (ginger, galangal). Tuber is suitably selected from Apios americana (hog potato or groundnut), Cyperus esculentus (tigernut or chufa), Dioscorea spp. (yams, ube), Dioscorea polystachya (Chinese yam or white name), Helianthus tuberosus (Jerusalem artichoke or sunchoke), Hemerocallis spp. (daylily), Lathyrus tuberosus (earthnut pea), Oxalis tuberosa (oca or New Zealand yam), Plectranthus edulis and P. esculentus (kembili, dazo, and others), Solanum tuberosum (potato), Stachys affinis (Chinese artichoke or crosne), Tropaeolum tuberosum (mashua or anu), and / or Ullucus tuberosus (ulluku). In a preferred embodiment the Tuber is of the genus Solanum, in particular Tuber of the species Solanum tuberosum (potato), such as the Solanum tuberosum red skin potato.

[0052] The root-like stem vegetable is suitably Zamia integrifolia (Florida arrowroot).

[0053] The true root vegetable is in some embodiments selected from taproot or tuberous root. The tuberous root vegetable can be selected from Amorphophallus galbra (yellow lily yam), Conopodium majus (pignut or earthnut), Dioscorea polystachya (nagaimo, Chinese yam, Korean yam, mountain yam), Hornstedtia scottiana (native ginger), Ipomoea batatas (sweet potato), Ipomoea costata (desert yam), Manihot esculenta (cassava or yuca or manioc), Mirabilis expansa (mauka or chago), Psoralea esculenta (breadroot, tipsin, or prairie turnip), and / or Smallanthus sonchifolius (yacon). The taproot vegetable can be selected from Arracacia xanthorrhiza (arracacha), Beta vulgaris (beet and mangelwurzel), Brassica spp. (kohlrabi, rutabaga, and turnip), Bunium persicum (black cumin), Burdock (Arctium, family Asteraceae), Carrot (Daucus carota subsp. sativus), Celeriac (Apium graveolens rapaceum), Daikon - the large East Asian white radish (Raphanus sativus var. longipinnatus), Dandelion (Taraxacum) spp., and / or Lepidium meyenii (maca). Preferably the taproot vegetable is Beta vulgaris, optionally subspecies vulgaris, in particular B. vulgaris var. conditiva (red beet).

[0054] In some embodiments of the described method the one or more edible ingredients comprise5 %wt to 60 %wt root vegetable, such as 5 %wt to 10 %wt, such as 1 0%wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt, 20 %wt to 60 %wt of a root vegetable, optionally 38 %wt to 42 %wt. In some embodiments, the one or more edible ingredients comprise 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of Solanum tuberosum. In additional or alternative embodiments, the one or more edible ingredients can comprise optionally 15 %wt to 2 5%wt, optionally 19 %wt to 21 %wt of B. vulgaris var. conditiva. In still further embodiments the one or more edible ingredients comprise 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of Solanum tuberosum, and 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of B. vulgaris var. conditiva, the sum of potato, and red beet not exceeding 100 percent.

[0055] The legume is very useful as the high-protein and low moisture component and / or the high starch component providing sustained nutrients for the one or more microorganisms, providing softness to the texture of the edible product. Useful legumes include and can be selected from the genera of Phaseolus, Pisum, Vigna, Cajanus, Lens, Cicer, Vicia, Arachis, Glycine, Macrotyloma, Mucuna, Lupinus, Ceratonia, Canavalia, Cyamopsis, Lablab, Psophocarpus, Clitoria, Lathyrus, Trifolium, Medicago, Melilotus, and / or Tamarindus.

[0056] In particular embodiments thePhaseolus bean is selected from the species P. vulgaris (Kidney Bean, Pinto Bean, Navy Bean Haricot Bean, Black Beans, Borlotti Beans), P. lunatus (Lima Bean), P. coccineus (Runner Bean, Flat Bean), and / or P. acutifolius (Tepary Bean));Pisum bean is selected from P. sativum peas (Green Peas, White Peas, Yellow Peas, Field Peas, Snow Peas, Snap Peas);Vigna bean is selected from V. radiata (Mung Bean), V. mungo (Urad), V. unguiculata (Cowpea, Yardlong bean, Black-eyed Pea), V. aconitifolia (Moth bean), and / or V. angularis (Adzuki bean); Cajanus bean is selected from C. cajan (Pigeon Pea);Lens bean is selected from L. culinaris (Lentil, Red Lentil, Green Lentil, Puy Lentil);Cicer bean is selected from C. arietinum (Chickpea, Garbanzo Bean);Vicia bean is selected from V. faba (Fava Bean, Broad Bean), V. ervilia (Bitter vetch), and / or V. sativa (common vetch);Arachis bean is selected from A. hypogaea (peanut);Glycine bean is selected from G. max (soybean);Macrotyloma bean is selected from M. uniflorum (Horsegram);Mucuna bean is selected from M. pruriens (velvet bean);Lupinus bean is selected from L. albus (white lupin, sweet lupin), L. mutabilis (Tarwi / Andean Lupin), L. hirsutus and / or L. angustifolius;Ceratonia the bean is selected from C. siliqua (Carob bean);Canavalia bean is selected from C. gladiate (Sword bean), and / or C. ensiformis (Jack bean);Cyamopsis bean is selected from C. tetragonoloba (Guar bean);Lablab bean is selected from L. purpureus (Hyacinth Bean, lablab bean);Psophocarpus bean is selected from P. tetranoglobulus (winged bean);Clitoria bean is selected from C. ternatea (butterfly pea);Lathyrus bean is selected from L. sativus (grass pea) and / or L. tuberosus (tuberous pea); Trifolium bean is selected from T. repens (white Clover), and / or T. pratense (red clover); Medicago bean is selected from M. sativa (alfalfa);Melilotus bean is selected from M. officinalis (sweet clover); and / or Tamarindus bean is selected from T. indica (tamarind);

[0057] The one or more edible ingredients can suitably comprise 5% wt to 60% wt of legumes, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt, optionally 38 %wt to 42 %wt. Additionally or alternatively, the legume can comprise 5 %wt to 60 %wt of protein, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt. Additionally or alternatively the legume can comprise 5%wt to 60 %wt of starch, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt. The one or more edible ingredients can also comprise at least two legumes, of which at least one legume contains a high level of protein, while at least one other legume contains high levels of starch. More specifically the high-protein level legume can comprise 5 %wt to 60 %wt of protein, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt, whereas the high starch level legume can comprise 5 %wt to 60 %wt of protein, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt. In desired embodiments the legume comprises Pisum beans and / or Lupinus beans, particularly P. sativum peas and L. albus lupin seeds, more particularly yellow split P. sativum peas andsweet L. albus lupin seeds. In further useful embodiments the one or more edible ingredients comprise 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of yellow split P. sativum peas. Alternatively, the one or more edible ingredients comprise 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of sweet L. albus lupin seeds. Further, the one or more edible ingredients comprise both 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of yellow split P. sativum peas and 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of sweet L. albus lupin seeds.

[0058] In a preferred embodiment the one or more edible ingredients comprise oats (Avena sativa), potato (Solanum tuberosum), red beet (B. vulgaris var. conditiva), P. sativum peas and L. albus lupin seeds more specifically 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt each of oats, potato, red beet, P. sativum peas and L. albus lupin seeds, the sum of oats, potato, red beet, peas and lupin seeds not exceeding 100 percent.

[0059] For further improvement of properties, the edible ingredients further can also comprise plant extract, such as extracts of oats, optionally aqueous extracts also known as oats milk. In addition, or alternatively the extract may be an almon extract, optionally an aqueous extract of almonds also known as almonds milk. Such aqueous extract can suitably be leftovers from actual production of plant extracts, such as the press cake from such extract production.

[0060] Prior to fermentation, the edible ingredients are advantageously pre-processed in a manner improving the conditions for subsequent handling and / or fermentation. As disclosed herein, it has been found that the preprocessing of ingredients significantly influences the effect of fermentation and the quality parameters of the fermented substrate. In some embodiments, the preprocessing is mechanical and includes chopping, grinding, and / or shredding, while in other embodiments the preprocessing is thermal or chemical, such as heating, microwaving, boiling and / or steaming, especially prior to forming the fermentation substrate.

[0061] In some embodiments cereals are milled or ground to a particle size having a maximum diagonal / diameter of 0,5mm to 10mm, such as 0,5mm to 1mm, such as 1mm to 2mm, such as 2mm to 3mm, such as 3 to 4mm, such as 4mm to 5mm, such as 5mm to 6 mm, such as 6mm to 7mm, such as 7mm to 8mm, such as 8mm to 9mm, such as 9mm to 10mm, optionally to an average particle size of 3,5mm. In particular, where the cereal is oats and / or BSG, the oats and / or BSG, before being formed into the fermentation substrate, undergo pre-processing by milling them into particles ranging between 2 and 5 mm in size, optionally between 3 and 4 mm, and optionally to an average particle size of 3,5 mm.

[0062] In other embodiments the one or more edible ingredients are pre-processed, particularly priorto forming the fermentation substrate, by heat, for example by microwaving, boiling, or steaming, to gelatinize among others starch components. For the fermented substrate described herein steaming is particularly useful, as it provides some water for gelatinization while still controlling the amount of water in the fermentation substrate prior to fermentation. Steaming leads to a more precise cooking, while e.g. boiling would remove nutrients and increase the water content in the fermentation substrate.

[0063] More specifically, tubers such as potatoes or other high-starch containing ingredients can be advantageously pre-processed through heat treatment to expose starch granules for the fermenting microorganism and to gelatinize the starch fully or partially through methods such as microwaving, boiling, or steaming. Where the ingredients include both cereals and root vegetables, particularly oats and potatoes, these can also be co-ground prior to forming the fermentation substrate, optionally to a degree where no visible chunks of the root vegetable can be observed. In some embodiments, in particular where the cereal is oats and the root vegetable is potato, the edible ingredients include coground cereal and root vegetable, preferably in a ratio of 0.5:1 to 1:0,5 of cereakroot vegetable, optionally 1:1. In some embodiments cereakroot vegetable ratio is oats:potato. In the grinding or cogrinding of cereals and root vegetables, in some embodiments these ingredients are milled or ground to a particle size having a maximum diagonal / diameter of 0,5mm to 10mm, such as 0,5mm to 1mm, such as 1mm to 2mm, such as 2mm to 3mm, such as 3 to 4mm, such as 4mm to 5mm, such as 5mm to 6 mm, such as 6mm to 7mm, such as 7mm to 8mm, such as 8mm to 9mm, such as 9mm to 10mm, optionally to an average particle size of 3,5mm.

[0064] In still further embodiments of pre-processing where the root vegetable is a taproot vegetable such as Beta vulgaris, optionally subspecies vulgaris, in particular B. vulgaris var. conditiva (red beet), the tap root is pre-processed by i) boiling and / or steaming the tap root and ii) grinding the tap root prior to forming the fermentation substrate. In this embodiment the taproot is suitably ground to an average particle size in its longest diagonal / diameter as described, supra, preferably between 1,7mm to 2,8mm.

[0065] In the method described herein the one or more edible ingredients may comprise P. sativum peas, and wherein the P. sativum peas, prior to forming the fermentation substrate, is pre-processed by i) drying and splitting the P. sativum peas, ii) microwaving, boiling, or steaming the P. sativum peas and iii) grinding the P. sativum peas to an average particle size as described, supra, preferably between 1,7-2, 8 mm.

[0066] Where the edible ingredients include legumes, these are preferably also pre-processed, prior to forming the fermentation substrate to both gelatinize and open the ingredient components to themicroorganism and to remove undesired components such as toxic and / or bitter tasting compounds, such as alkaloids. This pre-processing suitably includes soaking the legume is an aqueous liquid to extract undesired compounds to microwave, boil or steam the legume and to grind the legume. For example, where the legume is beans / seeds of Lupinus, such as from L. albus (white lupin, sweet lupin), L. mutabilis (Tarwi / Andean Lupin), L. hirsutus and / or L. angustifolius, in particular from L. albus; these can advantageously be soaked in fresh or salted water to wholly or partially extract toxic or bitter tasting alkaloids and ground to an average particle size in the longest diagonal / diameter as described, supra, preferably between 1,7 mm to 2,8 mm.

[0067] In a further embodiment the fermentation substrate is prepared from red beet, potato, oatmeal, split peas, and lupin beans in an even mix of 20 %wt. Red beets and potatoes are preferably peeled, cut, and steamed, oatmeal is preferably ground to a flour, split peas are preferably were boiled, and lupins are preferably soaked for removing undesirable components (such as at least for 6 hours) before boiling. Red beets, potatoes, and lupins are preferably blended separately to increase the surface area. The potatoes are preferably blended with the oatmeal flour in a 1:1 ratio (W / W) to decrease stickiness.

[0068] In preferred embodiments the method described herein may also include a step of forming the edible ingredients, whether pre-processed or not, into the fermentation substrate. This step suitably includes operations selected from further mechanical, ultrasound and / or thermal treatment optionally as well as addition processing aids or other functional ingredients.

[0069] Mechanical steps include mixing of ingredients, further grinding of the mixtures, and / or shaping the mixtures into a desired shape suitable for inoculation by the microorganism and for the fermentation process. Thermal treatment can also include additional steps such as microwaving, boiling, or steaming of the ingredient mixtures, including procedures to sanitize the formed substrate, such as pasteurization.

[0070] In some embodiment the fermentation substrate forming step includes mixing the ingredients, whether pre-processed or not, and optionally further grinding them to an average particle size in the longest diagonal / diameter of between 50 pm to 500 pm, such as between 50 pm to 100 pm, such as between 100 pm to 150 pm, such as between 150 pm to 200 pm, such as between 200 pm to 250 pm, such as between 250 pm to 300 pm, such as between 300 pm to 350 pm, such as between 350 pm to 400 pm, such as between 400 pm to 450 pm, such as between 450 pm to 500 pm,. In other embodiments the fermentation substrate forming step include grinding ingredients to an average particle size in the longest diagonal / diameter of between 0,5 to 10mm, such as 0,5 to 1 mm, such as 1 to 2 mm, such as 2 to 3 mm, such as 3 to 4 mm, such as 4 to 5 mm, such as 5 to 6 mm,such as 6 to 7 mm, such as 7 to 8 mm, such as 8 to 9 mm, such as 9 to 10 mm. Optionally the fermentation substrate forming step includes pasteurizing the mixture before and / or after grinding. This approach is particularly useful when the ingredients include oat, potato, red beet, P. sativum peas and L. albus lupin beans.

[0071] Pasteurization of the fermentation substrate prior to inoculation and fermentation is important to avoid contamination of the fermentation substrate with microorganisms which could negatively impact on the fermentation. Pasteurization of substrates can typically be accomplished by steaming at 120°C for 30 minutes.

[0072] In important embodiments, the water content of the fermentation substrate is also adjusted prior to fermentation where the best fermentation performance can be achieved with a moisture content of between 25 %wt to 50 %wt, such as between 25 %wt to 30 %wt, such as between 30 %wt to 35 %wt, such as between 35 %wt to 36 %wt, such as between 36 %wt to 38 %wt, such as between 38 %wt to 39 %wt, such as between 39 %wt to 41 %wt, such as between 41 %wt to 43 %wt, such as between 43 %wt to 45 %wt, such as between 45 %wt to 50 %wt.

[0073] Other important embodiments include those in which the fermentation substrate is divided and formed into a number of discrete portions with a desired size and / or shape, and then placed into a container optimized for the subsequent fermentation and / or final use of the fermented substrate. Preferred portion sizes are between 25g to 3000g, such from 25g to 50g, such from 50g to 100g, such from 100g to 200g, such from 200g to 300g, such from 300g to 400g, such from 400g to 500g, such from 500g to 750g, such from 750g to 1000g, such from 1000g to 1500g, such from 1500g to 2000g, such from 2000g to 2500g, such from 2500g to 3000g. The portions are formed into desired shapes, such as granules, balls, blocks, sheets, or patties, and then placed into containers that are useful for maintaining both the shape and the fermentation process. Such containers may be of open or closed design for optimal fermentation results and / or handling, and they may be made from wood, plastic, metal, or composites thereof.

[0074] The method described herein may also include the step of contacting the fermentation substrate with one or more microorganisms under conditions allowing the microorganisms to colonize and proliferate on / in the fermentation substrate. The contacting includes any suitable way of introducing or inoculating the microorganisms onto or into the fermentation substrate, include the mixing into the fermentation substrate or spraying or dusting onto a surface of the fermentation substrate of the microorganism. The microorganism can be added to the fermentation substrate as a lyophilized powder or as a pre-inoculation cell suspension, preferably in amounts of 0.25g to 2g of microorganism per kilogram substrate. In some embodiments the fermentation substrate isinoculated with between 0,01 g to 0,05 g koji spores per kg substrate, such as between 0,02 g to 0,04 g, such as about 0.033g koji spores per kg. substrate.

[0075] The microorganism used in the method described herein is preferably a GRAS (Generally Recognized as Safe) strain, which is particularly useful in the sense that less safety studies are required for the fermented edible product to be market approved. Further, in some embodiments the microorganism is a fungus, such as a filamentous fungus or mould, optionally of the genus Aspergillus, optionally of the species Aspergillus oryzae or Aspergillus sojae. Other preferred moulds include those of the genus Rhizopus optionally of the species Rhizopus oligosporus. Useful subvariants of A. oryzae include A. Oryzae "sweet koji" and A. Oryzae "protein koji".

[0076] The fermentation substrate inoculated with the microorganism(s) is allowed to ferment under conditions allowing the microorganisms to colonize and proliferate on / in the fermentation substrate preferably during a predetermined time interval, temperature, and humidity to achieve optimal improvement of the properties. In preferred embodiments the fermentation time is selected from between 20 hours to 90 hours, such as between 20 hours to 30 hours, such as between 30 hours to 40 hours, such as between 40 hours to 50 hours, such as between 50 hours to 60 hours, such as between 60 hours to 70 hours, such as between 70 hours to 80 hours, such as between 80 hours to 90 hours. In preferred embodiments the fermentation temperature is selected from between 10°C to 70°C, such as from 10°C to 19°C, such as from 19°C to 20°C, such as from 20°C to 25°C, such as from 25°C to 30°C, such as from 30°C to 31°C, such as from 31°C to 35°C, such as from 35°C to 40°C, such as from 40°C to 50°C, such as from 50°C to 60°C, such as from 60°C to 70°C. To stop fermentation, the fermentation substrate and microorganism are suitably cooled, preferably to below 10 °C, and subsequently pasteurized at 90 °C for 3 minutes to heat inactivate any remaining microbial activity.

[0077] During fermentation further moisture can be supplied, to wholly or partially maintain a moisture level optimal for the fermentation. However, the fermentation process may still spend moisture, and the fermented substrate may require rehydration to maintain attractive properties. Accordingly, in one embodiment the fermented substrate is rehydrated by the adding water to the product to reach a moisture content between 20 %wt to 90 %wt, such as 20 %wt to 30 %wt, such as 30 %wt to 40 %wt, such as 40 %wt to 50 %wt, such as 50 %wt to 60 %wt, such as 60 %wt to 70 %wt, such as 70 %wt to 80 %wt, such as 80 %wt to 90 %wt.

[0078] The method described herein improves or enriches the fermented substrate in one or more properties selected from flavour, aroma, texture, cooking experience, visual appearance, nutritional and / or dietary value, spoilage time, and / or carbon footprint compared to a parent un-hydrated product. Such properties are, if not directly quantifiable by analytical methods, usually assessed anddetermined by one or more panels of seasoned and experienced food scientists and / or chefs.

[0079] Improved flavour properties include enhancements in selected properties such as sourness, sweetness, bitterness, saltiness, umami, or a combination thereof, compared to the unfermented parent product. Improved aroma properties include improvements or enhancements in the aromas of raw ingredients, characterized by a strong vegetal and bean aroma, while fermented substrates exhibit milder aromas of toasted cereal, sweet porridge, stone fruit, and mushroom.

[0080] Improved texture properties include improvements in properties selected from firmness, softness, cohesiveness, juiciness, chewiness, sandiness, or a combination thereof compared to a parent unfermented substrate. Improved cooking properties include improvements in (a) browning properties of the fermented edible product (Maillard reaction) when roasting or frying the fermented edible product compared to roasting or frying a parent unfermented substrate, and / or (b) structure and integrity of the fermented substrate by the microorganism biomass binding together the edible ingredients compared to a parent unfermented substrate. Improved visual properties include improvement in colour of the fermented substrate, compared to the colour of a parent unfermented substrate. Improved nutritional properties include improvements in content and / or distribution of proteins, lipids, carbohydrates, fibres, vitamins, minerals, amino acids, or a combination thereof compared to a parent unfermented substrate. Still further improved spoilage time includes improvement in the shelf-life of the fermented substrate. This applies to both ambient and cooled conditions, as well as to packaged and unwrapped conditions, resulting from the microorganisms' control and competition effects on other microbial fauna in the fermented substrate. Improved carbon footprint follows and results not only from the improved or extended spoilage time, but also from the inclusion of ingredients sourced from industrial waste streams. This reduces or lowers the need and / or demand for new (virgin) agricultural produce, thereby decreasing and / or reducing both fossil fuel consumption during crop cultivation and the utilization of land areas.

[0081] In the method provided herein the contacting of the fermented substrate with the hydrating composition preferably comprises partial or full submersion of the fermented substrate in the hydration composition. The fermented substrate is preferably partially or fully submersed in the hydration composition for a selected time span to allow the fermented substrate to absorb the amount of moisture and optionally other functional ingredients in the hydrating composition which provides the desired improvement of quality properties of the hydrated fermented substrate. An optimal time span has been identified to between 1 to 40 seconds. In further embodiments, the fermented substrate is contacted with the hydrating composition at conditions allowing the fermented substrate to absorb more than 50% of the hydration composition, such as more than 60%,such as more than 70%, such as more than 80%, such as more than 90%, such as more than 95%, such as more than 98% of the hydration composition. In other embodiments, the fermented substrate is contacted with the hydrating composition at conditions allowing the fermented substrate to absorb between 50% to 100%, such as between 60% to 90%, such as between 70% to 80%, such as between 75% to 80% of the weight of the un-hydrated fermented substrate. The weight of hydration composition absorbed per weight of fermented substrate is also referred to as the absorption density of the fermented substrate. Accordingly, in some embodiments for achieving optimal texture the amount of hydrating composition is 0,5 gram to 1 gram, such as between 0,6 gram to 90 gram, such as between 0,7 gram to 80 gram, such as between 0,75 gram to 0,8 gram per gram of fermented substrate. In some preferred embodiments the fermented substrate can also be rehydrated by injecting the hydration composition into the fermented substrate, for example by inserting hollow tubes (such as needles) into the fermented substrate and pumping the hydration composition into the fermented substrate through the tubes. Tubes / needles having an inner diameter between 0,5 mm to 5 mm, such as between 1 mm to 3 mm, such as about 2mm are preferred.

[0082] The method provided herein may also further after treatment including vacuum packaging and heat treatment of the hydrated fermented substrate, to ensure to maintain the improved quality parameters. The heat treatment ideally comprises heating the vacuum packaged hydrated fermented substrate to a temperature of at least 92°C for at least 5 minutes.

[0083] The methods described herein produce unique hydrated fermented substrates with improved properties. Additionally, as a separate aspect described herein, a hydrated fermented substrate is provided, which is produced using the methods disclosed herein. The hydrated fermented substrate preferably comprises one or more filamentous fungi or the mycelium thereof and 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt each of cereal, root vegetable, and legume, optionally 10 %wt to 30 %wt, optionally 1 5%wt to 25 %wt, optionally 19 %wt to 21 %wt each of oats, potato, red beet, P. sativum peas and L. albus lupin seeds. In a further embodiment the hydrated fermented substrate comprises more than 5 %wt of dietary fibres, such as between 5 %wt to 20 %wt, such as 5 %wt to 10 %wt, such as 10 %wt to 15 %wt, such as 15 %wt to 20%wt.

[0084] The hydrated fermented substrate is typically a food, a pet food, or an animal feed composition, particularly a food composition, since for foods for human consumption there are particularly high requirements for flavour, aroma, texture, cooking experience, visual appearance, nutritional and / or dietary value, spoilage time, and / or carbon footprint.ExamplesThe following non-limiting examples illustrate the methods and products of this disclosure.Example 1 - Impact of pH on absorption capacity of fermented substrate

[0085] The kinetic absorption curve of a fermented plant matrix substrate was assessed as a function of the pH of the hydration liquid. The water absorption curve was established by immersing standardsized samples at various time intervals and plotting the relative absorbed liquid against the duration of submersion.

[0086] A pre-fermentation substrate was prepared which consisted of a mixture of 4 main ingredients: Cooked beet roots (5 mm of average particle size), cooked potato oat and flour (0.5mm average particle size), grinded cooked split peas (1.93 mm of average particle) and grinded cooked lupines (5 mm of average particle). These ingredients were mixed in quantities of each of the ingredients according to table 1 below, heat treated (92°C, 6 min), cooled to 35°C and then inoculated with 2.7xl09CFU of Aspergillus oryzae conidial spores per kilogram of substrate.Table 1.

[0087] Subsequently, the inoculated substrate was divided into molds and left to incubate for 40 hours at 31°C and relative humidity of 75%. Upon completion of the fermentation process, a composite material consisting of mycelium and plant matrix (fermented substrate) with a thickness of 2 cm was obtained.

[0088] 5cm x 5cm samples of fermented substrate was cut out of the fermented substrate (see figure 2) and submerged into an aqueous hydrating solutions having different pH values for a specific amount of time. Four sets of experiments were conducted, each involving a distinct solution of demineralized water with a corresponding pH (5,6,7 and 8). The pH of the absorbate was adjusted using sodium bicarbonate and / or citric acid to achieve the desired value. Subsequently, the samples were fully immersed in the solutions for specific durations ranging from 0 to 40 seconds at 20°C.

[0089] The weight of samples before and after the submersion was recorded, and the amount ofabsorbed water per initial fermented substrate weight was calculated. Each experiment was performed in triplicate.

[0090] The results in each of the set of experiments were fitted into an empirical kinetic pseudosecond order Langmuir model [1]:Where KE2 is the apparent rate constant (s-1) of the model, qeis the absorption density at equilibrium (g of absorbate / g of absorbent) and q is the absorption density (g of absorbate / g of absorbent) at any time, t (seconds).

[0091] The kinetic absorption performance of the fermented substrate was compared at the different evaluated pH conditions.Results / conclusion

[0092] To ensure that the experimental kinetic absorption behaviour could be represented by the empirical kinetic pseudo-second order Langmuir model, plots of time versus absorption density over time were generated (see figure 3a and 3b) showing a linear tendency of the data, indicating that the obtained data fits into the proposed model.

[0093] Minimal variations were observed in the absorption curve when altering the pH of the absorbent. Based on our findings, it can be concluded that the recommended pH for the hydration of the studied fermented composite application falls within the acidic range of 5-6.

[0094] The resulting data revealed a linear trend, suggesting that the collected data aligns with the proposed model. Table 2 below provides the calculated values for Ifeand qe2.Table 2

[0095] The kinetic absorption capacity of the hydrated fermented substrate samples under the various pH conditions are shown in figure 4. As illustrated, specifically, for a pH value of 7, theabsorption occurred at a slower rate over time compared to all the other measured pH values (5, 6, and 8), all of which exhibited higher absorptions. The higher absorptions at pH 5,6 and 8 suggest that these pH levels create a more favourable environment resulting in a faster absorption rate compared to the pH 7. The best absorption rates were observed within the acidic range and the optimal pH for absorption was within the acidic range of 5-6 in terms of absorption rate and capacity. There was a high content of fiber (chitin, b-glucan) in the fermented substrate, and it is contemplated that due to the structure and solubility properties of these fibers they absorbed water faster in acidic pH, because at acidic conditions, the protonation of functional groups on the fiber molecules could occur leading to changes in the electrostatic interactions and hydrogen bonding within the fiber structure. As a result, the fibers may become more water-soluble or water-accessible in an acidic environment. Additionally, the protonation of functional groups can alter the overall charge distribution on the fiber molecules, affecting their interactions with water molecules. This change in charge distribution may enhance the hygroscopic properties of the fibers, leading to increased water absorption.

[0096] Example 2 - Impact of salt concentration on absorption capacity of fermented substrate

[0097] The kinetic absorption curve of a fermented plant matrix substrate was assessed as a function of the salt content of the hydration liquid. The water absorption curve was established by immersing standard-sized samples at various time intervals and plotting the relative absorbed liquid against the duration of submersion.

[0098] Samples of 5cm x 5cm of fermented substrate were prepared as described in example 1. Four sets of experiments were conducted, each involving a distinct solution of demineralized water with a corresponding concentration of sodium chloride (0, 0.1, 0.25 and 0.5 g / mL). Subsequently, the samples were fully immersed in the solutions for specific durations ranging from 0 to 40 seconds at 20°C. The weight of samples before and after the submersion was recorded, and the amount of absorbed water per initial fermented substrate weight was calculated. Each experiment was performed in triplicate.

[0099] The results in each of the set of experiments were fitted into an empirical kinetic pseudosecond order Langmuir model [1]:Where KE2 is the apparent rate constant (s-1) of the model, qeis the absorption density at equilibrium(g of absorbate / g of absorbent) and q is the absorption density (g of absorbate / g of absorbent) at any time, t (seconds).

[0100] The kinetic absorption performance of the fermented substrate was compared at the different evaluated salt concentrations.Results / conclusion

[0101] To ensure that the experimental kinetic absorption behaviour could be represented by the empirical kinetic pseudo-second order Langmuir model, plots of time versus absorption density over time were generated (see figure 5a and 5b) showing a linear tendency of the data, indicating that the obtained data fits into the proposed model.

[0102] The resulting data revealed a linear trend, suggesting that the collected data aligns with the proposed model. Table 3 below provides the calculated values for KE?and qe2.Table 3

[0103] The kinetic absorption capacity of the hydrated fermented substrate samples under the various salt concentrations of the rehydrating liquid are shown in figure 6. As illustrated, differences in salt concentration of the rehydrating liquid had an effect in the kinetic absorption rate of the fermented substrate. Higher salt concentrations lowered the velocity of absorption and decreased the equilibrium absorption density in comparision with the triplicates executed at 0.025 g / mL. These data indicates that a rehydrating solution with 0.025g / mL salt will be absorbed two times faster than a solution with O.lg / mL. This emphasizes the importance of having control with the salt concentration in rehyrating solution. From the results it can be observed that salt concentrations between 0.025g / mL and 0.05 g / mL appear to be an optimal range.Example 3 - Impact of temperature on absorption capacity of fermented substrate

[0104] The kinetic absorption curve of a fermented plant matrix substrate was assessed as a function of the temperature of the hydration liquid. The water absorption curve was established by immersing standard-sized samples at various time intervals and plotting the relative absorbed liquid against the duration of submersion.

[0105] Samples of 5cm x 5cm of fermented substrate were prepared as described in example 1. Four sets of experiments were conducted, each involving a distinct solution of demineralized water with corresponding temperatures of 10°C, 20°C, 30°C and 40°C. Subsequently, the samples were fully immersed in the solutions for specific durations ranging from 0 to 40 seconds at 20°C. The weight of samples before and after the submersion was recorded, and the amount of absorbed water per initial fermented substrate weight was calculated. Each experiment was performed in triplicate.

[0106] The results in each of the set of experiments were fitted into an empirical kinetic pseudosecond order Langmuir model [1]:Where KE2 is the apparent rate constant (s-1) of the model, qeis the absorption density at equilibrium (g of absorbate / g of absorbent) and q is the absorption density (g of absorbate / g of absorbent) at any time, t (seconds). The kinetic absorption performance of the fermented substrate was compared at the different evaluated temperatures.Results / conclusion

[0107] To ensure that the experimental kinetic absorption behaviour could be represented by the empirical kinetic pseudo-second order Langmuir model, plots of time versus absorption density over time were generated (see figure 7a and 7b) showing a linear tendency of the data, indicating that the obtained data fits into the proposed model.

[0108] The resulting data revealed a linear trend, suggesting that the collected data aligns with the proposed model. Table 4 below provides the calculated values for KE?and qe2.Table 4

[0109] The kinetic absorption capacity of the hydrated fermented substrate samples under the various temperatures of the rehydrating liquid are shown in figure 8. As illustrated, the temperature of the rehydrating liquid discernibly influences the kinetic absorption capacity of the fermented substrate. The rehydration kinetics shows an initial steep increase in water absorption followed by a decrease in the rehydration rate. This asymptomatic behaviour is attributed to the diminishing driving force for water transfer as rehydration progresses and the system approaches equilibrium. The rehydration temperature notably influences the amount of absorbed water. A temperature of between 20°C to 30°C appears to be optimal for the absorption capacity of the fermented substrate. Conversely, at lower temperatures (10°C), there is a reduction in absorption capacity, while at higher temperatures (40°C), absorption takes place more rapidly, reaching a plateau around 20 seconds, following a pattern similar to that observed for 20°C and 30 °C. However, as there is no significant increase in water uptake at higher temperatures it presents an advantage to avoid heating the fermented substrate during rehydration as the water uptake equilibrium at 20°C to 30°C was reached within approximately 15 seconds.Example 4 - Application of colourant to fermented substrate from hydrating solution

[0110] In this investigation of supplying colour to a fermented substrate via a rehydrating solution, seven different colourants were assessed to identify the colourants providing the most attractive appearance of the rehydrated fermented substrate.

[0111] Samples of 5cm x 5cm of fermented substrate were prepared as described in example 1.

[0112] A rehydrating stock solution of distilled water containing 2,81wt% inactivated Saccharomyces biomass (commercially available from Lallemand or White Lab®) and 2,81 wt% NaCI was prepared and adjusted pH to 5.2. Subsamples were prepared by mixing commercially available colourants with stock solutions according to table 5.Table 5

[0113] Following the preparation of the coloured rehydrating liquid, the samples of fermented substrate was submerged in the rehydration liquid until a total of 0.76 grams of rehydration liquid was absorbed per gram of fermented substrate for each sample. A total of 360 grams of rehydrated fermented substrate was produced for each colourant. Each of the samples were then vacuum packed and heat treated to reach a temperature of 92°C for 6 mins. Afterwards, the samples were cooled to 5°C. A commercially available meat "beef-like" analogue reference sample was further included.

[0114] The colour of the samples was evaluated from 32 MP photos of the samples taken under standard white lighting. The average colour profile and standard deviation of the colours were determined by measuring the Red-Green-Blue (RGB) values of 10 random spots in each photo using Adobe Photoshop software. The average RGB colour and standard deviation were measured for each sample, and the colour profiles of each of the samples were compared with the control and the reference sample.Results / Conclusions:

[0115] For establishing comparison point, the colour profile of the control sample (without colourant) was defined. A photo of the control sample and its RGB colour profile is shown in figure 9 and table 6 respectively. Then the colour profile of a desired target (reference sample) was established. A photo of the reference sample and its RGB colour profile is shown in figure 10 and table 6 respectively.Table 6The colour profile of each colorant sample was recording from their photos as shown in table 7.Table 7.

[0116] To mathematically compare the colours the saturation was equalized of all the data by normalizing the data. The equation used for normalizing the obtained colour was:Normalized Red = 100Normalized Green = 100Normalized Blue = 100After normalizing the data, the vectorial distance from the colorant samples to the reference sample was calculated. The results are shown in Table 8.Table 8.

[0117] The normalized data was then plotted into a ternary diagram as shown in figure 11. Photos of all samples are shown in figure 12.

[0118] Sample with colorant FR-02 exhibited an appearance most similar to the reference sample, followed by the EO sample as the second most similar. An additional internal survey involving nine participants yielded results consistent with those obtained through RGB analysis. These findings suggest that the incorporation of colourants into the rehydrating liquid is a technically attractive way of altering the appearance of the rehydrated fermented substrate and provide it a colour similar to the a "beef-like" meat analogue.Example 5 - Application of nutrients from a hydrating solution to fermented substrate

[0119] In this investigation of supplying additional nutrients to a fermented substrate via a rehydrating solution, different sources of protein and / or other nutrients were assessed to identify suitable concentrations of the protein and / or other nutrients in the rehydrating liquid.

[0120] Samples of 5cm x 5cm of fermented substrate were prepared as described in example 1. Water content, protein content and starch and sugars content in the fermented substrate was determined. The water content of the fermented substrate was determined by drying a sample of the fermented substrate at 70°C for 20 hours. The water content of the sample was calculated as follows: g Initial sample (o) — Dry sample (o)Water content (—) = - g Initial sample (g)

[0121] The protein content of the fermented substrate was determined by the Dumas method (AOAC 922.15). In this method the 200 to 300 mg of sample were incinerated at 850°C in an oxygen stream. The resulting water and carbon dioxide were absorbed, and the nitrogen content was measured by its thermal conductivity. A calibration method with a known nitrogen content such as ethylene diamine tetraacetic acid (EDTA) was used as described in Nollet, L. M., & Toldra, F. (Eds.). (2008). Handbook of processed meats and poultry analysis. CRC Press. The protein content was calculated from the total quantity of nitrogen by the factor 6.25.

[0122] The starch and sugar content of the fermented substrate was determined by SGS Analytics Sweden AB in Linkbping, Sweden, using methods known in the art.

[0123] The nutritional composition of the fermented substrate prior to rehydration is shown in table 9Table 9.

[0124] It was observed that prior to rehydration the fermented substrate had a protein concentration of 13.3% of the entire weight of the matrix.

[0125] Two sources of supplemental protein were tested. These supplemental proteins were tested by preparing a solution of demineralized water with a specific amount of the studied protein at 25°C. Afterwards, the solution was absorbed by the fermented matrix by submersion. The protein sources are described in table 10.Table 10.Results / Conclusions

[0126] The protein content in the rehydrated fermented substrate was reduced when rehydrated with pure water. However the rehydration is necessary for improving the mouthfeel parameters of the fermented substrate. Based on previous investigations in the relation of the absorbed liquid and texture of the product, an optimal amount of rehydration liquid absorbed by the fermented substrate was between 0.75 to 0.80 grams of absorbed rehydration liquid per gram of fermented substrate. However, in a meat analogue like the fermented substrate, protein content is an important parameter to control. In table 11 the protein contents of reference market products are shown:Table 11

[0127] Wet weight (WW) and dry weight (DW) protein content comparison of mycoprotein (Quorn) and meat.

[0128] It was found that where the fermented substrate is rehydrated with water without a protein supplement, the total protein content of the final product decreased in proportion to the absorbed water (see figure 13). By considering an absorption density of 0.75 g / g, the final protein content of the hydrated product would be 7.6% (w / w).

[0129] To assess the feasibility of using of the rehydration liquid as a carrier of nutrient / protein supplementation, it was important to understand the properties of the selected protein sources. Table 12 shows a comparison of the protein contents and solubility in water.Table 12.Relevant properties of the selected protein sources.The properties detailed in Table 12 were crucial for assessing protein supplementation through the rehydrating liquid. Both the protein content of the source and its water solubility impose constraints on the maximum achievable supplementation through rehydration. If assumed that all dissolved solids were uniformly absorbed as the rehydrating solution in which they are dissolved, the maximum absorbed protein concentration would correspond to the protein content in the rehydrating solution saturated with the protein source.

[0130] Figures 14 and 15 shows the relation between the protein source concentration of the rehydration solution and the final protein content in the rehydrated fermented substrate. It is observed in Figure 14 that the total protein concentration of the rehydrated fermented substrate only increases when the concentration of protein in the rehydration solution is higher than the concentration of protein in the fermented substrate. In the case of the hydrolysed peas supplementation the maximum rehydration solution concentration (defined by the solubility of the fermented peas) is 0.27 grams of fermented pea protein per gram of rehydration solution (equivalent to a rehydration solution protein concentration of 4.5%). Accordingly, it was not possible to increase the protein content of the rehydrated fermented substrate by when using hydrolyzed peas were used as supplementation source.

[0131] Considering the previous studies finding an optimal absorption density between 0,75 g / g to 0,80 g / g the theorical protein content in the rehydrated fermented substrate in relation to the protein concentration in the rehydrating solution which is show in table 13.Table 13.Final product protein content when the matrix absorbs 75% its initial weight of hydrate.BY= Bakers' yeast. HP= Hydrolysed peas. Protein concentration shown in parenthesis in %wt.

[0132] Table 13 shows that the protein supplementation at lower supplement concentrations slightly increases the final protein content of the product. In the case of bakers' yeast, a rehydration solution supplemented with 2.7% protein only increases the final protein content by 0.4%. At the same time, a rehydration solution supplemented with 1.8% of fermented peas seems to increase the protein content of the product by 0.1%.

[0133] When comparing the data of table 13 with the protein content of comparable products (table 11) it seemed that in both cases a supplementation using a rehydrating solution with more than 20% protein were required to achieve a protein content close to mycoprotein. During the use of the two supplement sources in the rehydration solution it was observed that they had different effects also in the organoleptic properties of the product, showing that the supplementation via the rehydrating solution not only affects the final nutrient content of the product but also other properties such as appearance, flavour, and mouthfeel.

[0134] Accordingly, this example not only shows that it is possible to add nutrients via the rehydrating solution, but also opens the possibility of using other additives in the rehydrating solution such as surfactants for improving the wetting capacity of the fermented substrate, emulsifiers for supplementing non-polar compounds, colorants for improving product appearance, preservatives, prebiotics, between other options.Example 6 - Impact of rehydration on fermented substrate flavour.

[0135] In this example the effect of applying a hydration solution containing different amino acid sources on the flavour properties of a fermented substrate was investigated.

[0136] Fermented substrate was prepared as described in example 1 and cut into cubes (2 x 2 cm).

[0137] For the hydration solutions, split peas were cooked with water (ratio 1:2) at 100% steam for30 minutes. The split pea cooking liquid (SPL) was strained. Active brewer's yeast (BRY) from White Lab® was hydrolysed using sous-vide cooking for 24 hours at 58°C. Inactivated fresh baker's yeast (BAY) from Lallemand was mixed with 30 g tap water and occasionally stirred until melted. Then, the mixture was cooked at 120°C for 45 min and additionally cooked at 70°C for 140 min. The resulting dry yeast was blended in a coffee grinder to gain a powder.

[0138] The split pea cooking liquid, brewer's yeast, and baker's yeast solutions were used to formulate three hydration solutions with additional spices. Table 14 shows the ingredient composition of the three hydration solutions. The three hydration solution formulations were based on previous development tests and subsequent evaluation by food science professionals.Table 14Ingredient composition of the hydration solutions.Hydration solutionIngredient (g) BRY BRY + SPL BAY + SPLSPL 75 100BRY 7.5 75BAY 0 10Tap water 100 0 100Salt (NaCI) 2 6 4Garlic powder 0.3 0.2Thyme 0.3 0.2SPL = split pea liquid; BRY = brewer's yeast; BAY = baker's yeast

[0139] The solid fermented substrate cubes were transferred into containers and added the hydration solution and shaken until the hydration solution was absorbed. Approximately 5.9 g hydration solution was absorbed per 9.1 g of fermented substrate. For the testing approximately 15 g samples (9.1 g fermented substrate + 5.9 g hydration solution) were prepared.

[0140] For testing 72 participants were recruited for tasting the samples. The samples were served monadically to the participants following a Williams Latin square design and water was provided to cleanse the palate between samples. The participants were instructed to firstly score their overall liking using a 7-point hedonic scale anchored at l-"dislike very much" and 7-"l ike very much". Next, they were asked to answer check all that apply (CATA) with 18 descriptors related to the sensory characteristics of sauces. The sensory descriptors were determined internally by food industryprofessionals pre the consumer test.

[0141] The participants scoring of the samples was tested by one way ANOVA with post-hoc Tukey's test for multiple comparison of means. Check-all-that-apply data was analysed using Cochran's Q test with post-hoc McNemar-Bonferroni correction to identify significant differences between attributes associated with each sample (Meyners et al. (2013); Existing and new approaches for the analysis of CATA data; Food Quality and Preference; 30(2); 309-319). A principal coordinate analysis (PCoA) was conducted to determine the association between sensory properties, as part of the CATA ballot. All data analyses were performed by using XLSTAT Version 2020.4.01.Results / Conclusions

[0142] The test revealed that there was no statistically significant difference in liking between the three hydration solutions (F (2) = 0.304, p = 0.739). Detailed results are shown in table 15. Yet, the relatively most liked sample seems to be the sample hydrated with the solution containing baker's yeast and split pea liquid (BAY + SPL). This might indicate that the use of spices and their ability to enhance flavours may have been a conditioning factor for the liking of BAY + SPL, since the relatively least liked sample hydrated with brewer's yeast (BRY) does not contain any spices (Dillon, V. M. (2014). Natural Anti-Microbial Systems: Preservative Effects During Storage. Encyclopedia of Food Microbiology: Second Edition, 941-947. The most frequently associated attributes in the CATA test, as shown in table 15, were, in order of occurrence, mushroom flavour (n = 91.0), umami (n = 71.0), salty (n = 47.0), and beef-like (n = 45.0). Subsequently, differences in associated attributes between samples was analysed. Yet, no significant effect of the hydration on the frequency of selected attributes was found (x2 (34) = 24.189, p = 0.894). The fermented substrate might have been dominating the flavour, especially the umami flavour, possibly overweighing any significant differences between the hydration solutions. Umami and mushroom flavour had previously been characterized to be flavour increasing but also greatly contributing to the overall flavour by itself (Maga, J. A. (1983). Flavour potentiators. C R C Critical Reviews in Food Science and Nutrition, 18(3), 231-312. Thereby, the frequently attributed flavours, such as umami, could be credited to the solid fermented substrate being a result of a solid-state fermentation of a plant matrix using the fungus Aspergillus oryzae. The provided substrate included different sources of proteins, carbohydrates, and lipids, which the fungus digests and transforms into smaller and "tastier" molecules such as amino acids, organic acids, esters, sugars, etc. When the hydrated fermented substrate was placed in a frying pan with hot oil, the sugars, present in both the solid fermented substrate and the hydrating solution, were broken down and caramelized, leading to the development of new flavours based on the sugarreduction (Maillard reaction) (Chisti, Y. (2014). Fermentation (Industrial): Basic Considerations. Encyclopedia of Food Microbiology: Second Edition, 751-761; Johnson, A. J. (2016). Artisanal food microbiology. Nature Microbiology, 1, 16039.). However, the lack of significant differences might furthermore also be due to the added concentration of amino acids in the yeasts, which might have been too low to be noticed by participants. Nevertheless, the perceived association of umami is highly present for all samples (33%), indicating that the hydration did not negatively influence the intended flavour. Yet, the BRY + SPL and BAY + SPL samples were slightly more liked by participants, while being characterized with the same umami perception. Hence, it could be discerned that the hydration solutions had increased the kokumi aftertaste sensation, leading to a higher liking of the fermented substrate (Alim, A., Yang, C., Song, H., Liu, Y., Zou, T., Zhang, Y., & Zhang, S. (2019). The behaviour of umami components in thermally treated yeast extract. Food Research International, 120, 534-543.; Liu, J., Song, H., Liu, Y., Li, P., Yao, J., & Xiong, J. (2015). Discovery of kokumi peptide from yeast extract by LC-Q-TOF-MS / MS and sensomics approach. Journal of the Science of Food and Agriculture, 95(15), 3183-3194; Maruyama, Y., Yasuda, R., Kuroda, M., & Eto, Y. (2012). Kokumi substances, enhancers of basic tastes, induce responses in calcium-sensing receptor expressing taste cells. PLoS ONE, 7(4); Miyaki, T., Kawasaki, H., Kuroda, M., Miyamura, N., & Kouda, T. (2015). Effect of a kokumi peptide, y- glutamyl-valyl-glycine, on the sensory characteristics of chicken consomme. Flavour, 4(1); Tang, C. S., Tan, V. W. K., Teo, P. S., & Forde, C. G. (2020). Savoury and kokumi enhancement increase perceived calories and expectations of fullness in equicaloric beef broths. Food Quality and Preference, 83).

[0143] In a last step, a principal coordinate analysis (PCoA) was performed to identify the correlation between hydrating solutions and associated sensory attributes. The results were plotted in a symmetric plot as illustrated in figure 16, with shared attributes highlighted in a blue triangle. These shared attributes again include umami and mushroom flavour, as well as bread / cereal flavour, sweet, beef-like, and yeasty.

[0144] A visual inspection of the PCoA plot suggested a positive correlation of the BRY sample with the first factor of the model. This correlation could be explained by a positive association with the attribute liver flavour and a negative association with the attribute's garlic flavour and salty. Furthermore, the model revealed that the BAY + SPL and BRY + SPL samples were negatively correlated with the first factor explaining the majority of variance (69,9%). The negative correlation might be due to an association with the attributes of garlic flavour, salty, and herbal. It is worth noting that spices such as garlic and thyme were used in the hydrating solution for these samples and both samples contained split pea liquid.Table 15Liking and frequency of CATA attributes associated with the three hydrated samples (n = 72).BRY BRY + SPL BAY + SPL Total count p-valueMean (SD)Liking 5.194 5.278 5.375 - 0.739(0.164) (0.164) (0.164)Check-all-that-apply %3Yeasty 13.9 11.1 11.1 26.0 0.735Beef-like 22.2 19.4 20.8 45.0 0.908Umami 33.3 33.3 33.3 71.0 1.000Sweet 18.1 15.3 15.3 35.0 0.852Herbal111.1 16.7 13.9 30.0 0.580Pork-like 25.0a9.7a16.7a37.0 0.039Chicken-like 9.7 16.7 20.8 34.0 0.152Fruity 2.8 5.6 6.9 11.0 0.529Bitter 19.4 12.5 16.7 35.0 0.405Sour 6.9 6.9 2.8 12.0 0.407Salty 16.7 25.0 23.6 47.0 0.289Vegetal flavour 11.1 16.7 11.1 28.0 0.513Garlic flavour 6.9 12.5 12.5 23.0 0.449Off-flavour 5.6 9.7 11.1 19.0 0.465Bread / cereal flavour 18.1 18.1 16.7 38.0 0.949Liver flavour 15.3 8.3 9.7 24.0 0.291Mushroom flavour 41.7 41.7 43.1 91.0 0.976Alcoholic25.6 5.6 0.0 8.0 0.135SPL = split pea liquid; BRY = brewer's yeast; BAY = baker's yeast1Herbal like thyme, rosemary, etc.2Alcoholic like wine in a stew.3Bold frequencies (%) indicate the max response of the associated CATA words.Note: CATA-data is presented as proportion (%) and total count (n). Means with unlike superscript letters are statistically different at a = 0.05 (post-hoc McNemar correction).

[0145] Moreover, the two samples also differed slightly in their CATA evaluation as represented by the correlation with the second factor explaining 30.4% of variance. Thereby, the BRY + SPL samplewas positively correlated with the second factor (and negatively with the first factor) resulting from an association with flavour attributes such as vegetal or herbal but not pork-like, liver, or bitter. In contrast, the BAY + SPL sample was associated with chicken-like flavour and oppositely related to sour and alcoholic flavour, leading to a n negative correlation with (the first and) second component. These findings indicated that the brewer's yeast in higher concentrations lead to a sour, alcoholic taste as opposed to the baker's yeast. It is well known that the quantitatively largest groups of aroma compounds synthesized by yeasts consist of higher alcohols, fatty acids, and their esters (Suomalainen, H., & Lehtonen, M. (1979). The production of aroma compounds by yeast. Journal of the Institute of Brewing, 85(3), 149-156.). Higher alcohols, also known as fusel alcohols, are responsible for wine-like aromas, while ethanol itself is responsible for the imparting warming, drying, and astringent effects on the palate. As fusel alcohols are the most abundant organoleptic compounds present in beer (Pires, E. J., Teixeira, J. A., Branyik, T., & Vicente, A. A. (2014). Yeast: The soul of beer's aroma— A review of flavour-active esters and higher alcohols produced by the brewing yeast. Applied microbiology and biotechnology, 98, 1937-1949), they resulted in the consumers' association of BRY and BRY + SPL to release a slightly stronger alcoholic flavour. Previous research had determined that yeasts can also significantly contribute to the aroma of fermented foods such as breads (Abbas, C. A. (2006). Production of antioxidants, aromas, colours, flavours, and vitamins by yeasts. Yeasts in Food and Beverages, 285-334. Berlin, Heidelberg: Springer Berlin Heidelberg). The baker's yeast applied in the BAY + SPL sample though alternated the fermented substrate taste more towards fruity and chicken-like notes. Flavour compounds formed based on the addition of baker's yeast have reported to be alcohols, organic acids, carbonyl compounds and browning fermented substrates such as melanin and caramels (Van Dam H. W. (1986) The biotechnology of bakers' yeast: old or new business? In: Blanshard J. M. V., Frazier P. J., Galliard T. (eds) Chemistry and physics of baking. Royal Society of Chemistry, London, 117-131). The associated attributes in the CATA test also provided useful information to understand if there were any taste parameters from the solid fermented substrate which one might want to mask, complement, or avoid in the context of a meat-like taste such as sour, bitter, fruity, sweet, between others.

[0146] These findings suggested that the fermented substrate has a dominant influence on the flavour profile, resulting in the lack of significant differences between hydration solutions. The umami flavour is most likely to be credited to the plant matrix of the fermented substrate itself, being constant over the different hydration solutions. A slightly higher liking and alternation of associated sensory attributes resulted from the addition of hydration solutions containing spices and split pea liquid and possibly the occurrence of a kokumi sensation possibly being more present in thosesamples.

[0147] In addition, the attributes identified in the CATA test offered valuable insights into potential taste parameters in the solid fermented substrate that may need to be either masked, complemented, or avoided to achieve a meat-like taste profile. These parameters encompass characteristics such as sourness, bitterness, fruitiness, sweetness, and others.

[0148] Finally, the baker's yeast and brewer's yeast both in combination with split pea liquid altered the sensory properties of a fungi-fermented plant substrate. Thereby, the baker's yeast produced a more chicken-like and fruity flavour with very little sour and alcoholic taste and was thus the superior alternative to apply as a hydration solution. The baker's yeast was in addition to the positive alteration of sensory properties also the most economically viable yeast.Example 7 - Impact of rehydration of fermented substrate on texture.

[0149] In this example the effect of rehydration of a fermented substrate on the texture parameters was investigated.

[0150] Fermented substrate was prepared as described in example 1. Subsequently, the fermented substrate was re-hydrated by submersion in a demineralized water solution to achieve absorption densities of 50%, 70%, 75%, 80% and 100%, respectively, where the absorption density was the amount of liquid absorbed by the matrix (g) divided by the initial weight of the same matrix (g). Prior to further testing, the samples were kept at room temperature for approximately 2 hours.

[0151] Samples of fermented substrate were cut out with a cylindrical cutter (30 mm) and placed on a Stable Micro Systems Texture Analyzer measuring platform. Six sets of experiments were conducted. A double compression cycle test was performed (45% strain) with an aluminium cylinder probe P / 50 (Stable Micro Systems, 50 mm). A time of 5 seconds was allowed to elapse between the two compression cycles. Force-time deformation curves were obtained with 5 g load cell applied at a crosshead speed of 1 mm / s. The texture profile parameters (figure 17) were calculated using Stable Micro Systems' standard TPA macros (included with the Exponent software) and determined as follows: (1) Hardness was defined as the peak force required in the first compression cycle (45% compression). (2) Cohesiveness was determined from the area of work during the second compression divided by the area of work during the first compression. (3) Chewiness was taken from the peak stress and the areas and time of the first and second compression. (4) Resilience was measured by dividing the upstroke area by the downstroke area of the first compression cycle (9,12). Six replicates of each sample were measured.(1) Hardness (A?) = Peak force of the first compression (Fi)( '2) ' Cohesiveness =(~ad++be'))Results / Conclusions

[0152] The observed adverse correlation of hardness and chewiness with the re-hydration concentration (shown in figure 18 and 19) could be attributed to several factors associated with the hydration process and the interplay between the fermented substrate and absorbed liquid. When fermented substrate underwent rehydration, it absorbed moisture from the hydrating solution leading to an increase in its overall moisture content. This contributed to more tender consistency, as water acts as a plasticizer, diminishing the stiffness and thereby contributing to the softening of the fermented substrate. This property is due to presence of starch and proteins. During rehydration, these components absorb water and undergo swelling (Ergezer, H., Akcan, T., & Serdaroglu, M. (2014). The effects of potato puree and bread-crumbs on some quality characteristics of low-fat meatballs. Han-gug Chugsan Sigpum Hag-hoeji, 34(5), 561-569 and Kurt, S., & Kilincceker, O. (2012). The Effects of cereal and legume flours on the quality characteristics of beef patties. Kafkas Universitesi Veteriner Fakultesi Dergisi, 18(5), 725-730), while p-glucans with their high water-holding capacity (Berggren, S. (2018). Water holding capacity and viscosity of ingredients from oats: the effect of b-glucan and starch content, particle size, pH and temperature and Chema Borchani, Fabienne Fonteyn, Guilhem Jamin, Jacqueline Destain, Luc Willems, Michel Paquot, Christophe Blecker & Philippe Thonart (2016). Structural Characterization, Technological Functionality, and Physiological Aspects of Fungal p-D-glucans: A Review, Critical Reviews in Food Science and Nutrition, 56:10, 1746-1752), also play a role.

[0153] Numerous studies have delved into this phenomenon, particularly in ingredients rich in B- glucans, such as cassava (Onodu, Bona & Culas, Richard & Nwose, Ezekiel. (2018). Facts about dietary fibre in cassava: Implication for diabetes' medical nutrition therapy. Integrative Food, Nutrition and Metabolism), demonstrating a substantial enhancement in the texture of meat patties, leading to reduced hardness and chewiness (W.Y. Akwetey, C.L. Knipe, Sensory attributes and texture profile of beef burgers with gari, Meat Science, Volume 92, Issue 4, 2012, Pages 745-748). This observation aligns with previous findings that highlight an inverse correlation between total moisture content and both hardness and chewiness (Mabrouki Sabah, Alberto Brugiapagli, Sara Glori, Sonia Tassone,Salvatore Barbera (2023) Texture profile analysis of homogenized meat and plant-based patties. International Journal of Food Properties. 26:2, 2757-2771). The hardness and chewiness values obtained at 75% and 80% absorption density closely resemble those reported in comparable studies on products like beef patties and plant-based sausages (Mabrouki et al (2023) and Paredes, J., Cortizo- Lacalle, D., Imaz, A.M. et al. Application of texture analysis methods for the characterization of cultured meat. Sci Rep 12, 3898 (2022). Accordingly, an absorption density within that range was considered optimal to meet the specific requirements.

[0154] No discernible changes were observed when examining various concentrations of hydrating solution concerning resilience and cohesiveness (figure 20 and 21). Given that resilience is linked to the plastic deformation of the material, these findings suggested that none of the investigated samples exhibited a permanent deformation after the initial compression, and that all of them recovered their shape to similar extent. Cohesiveness offers insights into how well a food retains its form between first and second chew and from the results, it was concluded that the samples demonstrated similar structural integrity.

[0155] The values obtained signified a moderate level of cohesiveness, indicating that the fermented substrate was neither extremely soft and crumbly, nor excessively firm and rigid. While the obtained cohesiveness values were within the range of 63-67 %, Malbrouki et al. reported cohesiveness measurements of 50-55 % in their study where they compared textural characteristics of meat patties and plant-based analogues. The marginally lower values they observed may be linked to the elevated fat content in the patties relative to our composite, a factor that exhibits a negative correlation with this parameter (Mabrouki et al 2023).

[0156] Table 16 below provides values obtained for 4 main parameters analyzed:Table 16All values were the mean ± standard error of six replicates.

[0157] Accordingly, the analysis of the 5 samples with different concentrations showed that these samples exhibit similar texture characteristics for parameters such as cohesiveness and resilience, while parameters such as hardness and chewiness showed adverse relationship with the re-hydration concentration. In conclusion, it was found that an absorption density within the range of 75-80% is well-suited for the fermented substrate.Example 8 - Impact of process for rehydrating fermented substrate on moisture uptake.

[0158] In this example the effect of three different modes of rehydrating a fermented substate was investigated.

[0159] Fermented substrate was prepared as described in example 1 and a hydrating solution of deionized water with 0.025 g / g of sodium chloride and 0.021 g / g of powder of inactivated Saccharomyces biomass was prepared.Hydration by submersion

[0160] Samples of 5cm x 5cm x 2cm of fermented composite prepared. The samples were totally submerged into the hydrating solution in specific amounts of times from 0 to 40 seconds at 20°C. Sample weights before and after the submersion were registered and the amount of absorbed hydrating solution per initial fermented substrate weight was calculated. Four replicates of all the experiments were made.

[0161] The obtained data of the absorption density (absorbed liquid / initial weight) versus the submersion time was analyzed. For this, the obtained data was fitted into two models. First the results were fitted into a kinetic pseudo-first order Langmuir model (PFO) [1]:Second the results were fitted into an empirical kinetic pseudo-second order Langmuir model (PSO) [1]:Where KEi and KE2are the apparent rate constants (s1) of the models, qeis the absorption density at equilibrium (g of hydrating solution / g of absorbent) and q is the absorption density (g of hydrating solution / g of absorbent) at any time, t (seconds). The model's suitability was determined based onthe best fit. In addition, the standard deviation between replicates was used to assess the reliability of the unit operation.Hydration by absorption of fixed volume

[0162] Circular samples of 10.5cm diameter and 2cm thickness of fermented substrate were prepared for these experiments. The samples were placed into an empty cylindrical container with 11.5 cm of internal diameter. A volume of hydrating solution equivalent to 0.75 times the weight of fermented substrate was poured into the container with the sample at 20°C. The samples were kept into the container with the hydrating solution during specific number of times between 0 to 80 seconds. After that, the sample was separated from the non-absorbed hydrating solution.

[0163] The samples' weights before and after the hydration were registered and the amount of absorbed hydrating solution per initial fermented substrate weight was calculated. Three replicates of all the experiments were made. Observations such as absorption time, weight standard deviations and kinetic absorption curve were evaluated for mode of re-hydration.Hydration by injectionCircular samples of 10.5cm diameter and 2cm thickness of fermented substrate were prepared for these experiments. For the injection a JBT Schroder IMAX Injector (Model: 350C) was used. In this mode of rehydration the samples were positioned on a conveyor belt. Here, the samples were perforated two times by a set of 2mm needles (0.45 needles / cm2). During the perforation, the needles were expelling hydrating solution at 1.9 bar at 45 injections per minute. Temperature was a variable parameter in this test, with values between 28°C to 68°C. Triplicates of the studied hydrating solution injection temperatures were executed.Results / ConclusionsHydration by submersion

[0164] Considering that in a manufacturing process the desired absorbed volume must be around 75% of the initial weight of the matrix. Factors such as the time of submersion (ts) and hydrating solution temperature (Thydratin solution) must be controlled in this process. Because of the volume of the hydrating solution in the submersion tank is significantly high in comparison of the absorbent mass, the volume of hydrating solution (Vhydratin solution) can be considered as infinite. absorbate00ts= constantabsorbate = constant

[0165] Figure 22 shows the obtained absorption densities versus the submersion time are shown. It was observed that the distribution of the absorption values over the time seem to follow a normal kinetic absorption behaviour. That indicates that it was possible to fit the information into a kinetic model such as the Langmuir kinetic models (Islam, M. A., Chowdhury, M. A., Mozumder, M. S. I., & Uddin, M. T. (2021). Langmuir adsorption kinetics in liquid media: Interface reaction model. ACS omega, 6(22), 14481-14492). Accordingly, in order to understand the performance of the absorption of the fermented substrate, the data was fitted first in into a kinetic pseudo-first order Langmuir model (PFO) (Chen, H. (2013). Modern solid-state fermentation. Netherlands: Springer):

[0166] By considering that the value KEI corresponds to a constant rate of the model. It is presumed that if the PFO model represents the absorption behaviour of the studied system, the data would fit in a linear function when the logarithmic part of the model is plotted versus the submersion time. In addition, it would be expected that in this case, the intersection of the linear function would be close to the origin.

[0167] Figure 23 shows that the experimental data did not fit totally in a kinetic pseudo first order Langmuir model. Because of that, the results were fitted into an empirical kinetic pseudo-second order Langmuir model (PSO) [1]:

[0168] By considering that the values of qeand kE2 are the absorption capacity and the rate constants it can be assumed that the PSO model would properly represent the absorption behaviour of the obtained data if the values of t / q versus the time follow a linear trend.

[0169] Figure 24 shows that when plotting t / q versus t there is an evident linear tendency of the data indicating that the kinetic PSO Langmuir model fits correctly into the obtained data. By following the structure of the model, the slope of the linear regression would be equal to l / qeand the constant of the regression corresponds to l / (qe2x kE2). Based on that information the absorption capacity of the matrix and the constant rate of the PSO Langmuir model are shown in table 17.Table 17.Absorption parameters of the studied hydrating solution absorbed by the fermented matrix for a kinetic PSO Langmuir model.

[0170] The model could also be represented graphically as shown in figure 25 showing a comparison of the predicted data by the Langmuir kinetic pseudo-second order model and the data obtained experimentally is observed. When comparing the predicted versus the experimental data it is observed that the model effectively represents the data. The absorption capacity of the model of 98% was calculated meaning that that is the maximum amount of liquid that the matrix could absorb under the studied conditions in comparison to the initial weight of the same matrix.

[0171] Based on prior studies, an absorption density ranging from 75% to 80% appeared to be optimal forthe intended application of the fermented substrate as a meat analogue. This implied that, in the case of submersion hydration, the absorption time of the fermented substrate should fall between 7 to 10 seconds. However, concurrently, a notable variability in experimental results is observed within the 5 to 10-second range, exhibiting variations up to 10% among samples at the same experimental point. This high variability was attributed to factors such as the intrinsic heterogeneity of the contact surface, density variations among samples, the presence of air bubbles during sample submersion, and potential human errors during execution. It is evident that the control of submersion hydration is highly contingent on external variables, emphasizing the need for precise control in preceding unit operations to meet quality criteria.Hydration by absorption of a fixed volume.

[0172] Hydration by absorption of a fixed volume consisted of placing a circular fermented-matrix sample into a container with a slightly higher diameter. After that, an amount of hydrating solution equal to 0.76 times the initial weight of the sample was poured into the container. The absorption process is shown in figure 26.One of the main advantages of this mode of rehydration is that the final absorbed volume of the sample is limited by the amount of liquid that is poured into the container. That means that the final weight in an infinite amount of time should be equal to the initial weight of the sample (W0) plus the added free hydrating solution (FA) if the weight of the hydrating solution is not more than the absorption capacity of the fermented substrate (that based in the last analysis is around 98% of theinitial weight but varies depending on hydrating solution and matrix condition). Because on the current experimentations the added free hydrating solution corresponded to 76% of the weight of the fermented substrate, it can be assumed that W=W0+0.76*W0. The obtained results can be represented graphically as showed in figure 27. Figure 27 shows that the maximum absorption density was observed at all the samples after 45 seconds. Relatively high standard deviations are observed from the seconds five to 44. These higher standard deviations are attributed to differences in surface, presence of air bubbles, shape and density of the samples. However, high variability seems to be overcome at higher time ranges. It is relevant to mention that from the seconds 0 to 5 the fastest absorption rate was observed. This absorption rate is possible comparable to the observed during the first seconds of hydration by submersion (see figure 28). It is highly possible that during the first seconds the fermented matrix is totally submerged. Possibly, after the 2ndsecond the amount of free hydrating solution is low enough for having the entire matrix submerged causing that the solid / liquid contact area is reduced, hence the absorption rate is reduced.

[0173] The obtained data showed that absorption by fixed hydrating solution volume could be used as an effective way for the hydration unit operation of the studied fermented substrate. One of the main advantages of fixed volume hydration is that in larger hydration times, the maximum absorption density (that is limited by the amount of added hydrating solution) is reached. Within the studied conditions it seems that a hydration time of 56 seconds was enough for having all the samples totally rehydrated.Hydration by injection of hydrating solution

[0174] The hydration by injection comprised in the penetration of the fermented substrate by needles having a hollow tube that allowed the liquid to enter and be absorbed by the penetrated fermented substrate. The needle created channels within the matrix, allowing the liquid to enter. This process took advantage of the pressure difference between the injected liquid and the fermented substrate. At the same time the process was limited by factors such as the absorption capacity of the injected material, the liquid pressure, viscosity of the liquid, temperature, design of the needle, diffusivity of the liquid inside the solid, among others.

[0175] A preliminary test where the hydrating solution was injected to the fermented substrate by using a 2.5 mm needle was executed. In that test, the injection pressure was increased gradually until observing that the fermented substrate was not able to hold more liquid from one injection point. Because of the tools used it was not possible to determine the maximum tolerable pressure for the experiment mentioned. But the experiment was taken as a reference to understand the referenceliquid absorption capacity and reference liquid distribution area when the fermented substrate was injected from one central point.

[0176] Based on the experiments, it was observed that the liquid was able to travel in a radio of 1.15 cm around the injection point and accordingly a needle distribution of more than 0.24 needles / cm2was contemplated to produce better results.

[0177] As mentioned in the methodology, an injection system from the brand JBT Schroder that considers 0.45 needles / cm2was used. Integral fermented substrate, with visually homogeneous internal hydrating solution distribution samples were obtained from all the results. The hydration time for each of the samples was 45 seconds. However, some variations were detected when the temperature of the marinate was modified. The results obtained from varying triplicates at different temperatures of the same hydrating solution are shown in figure 29. Here it is shown that the hydration solution temperature had an effect on the hydration by injection. The results agreed with previous data where an increment in the absorption rate of the fermented substrate was observed when the temperature increased. Also, the results agreed that there was no evident change in the absorption rates between 20°C to 34°C. These results indicated that higher temperatures could increase the hydration rate of the fermented substrate, but at the same time the absorption density would vary at a higher rate. On the other hand, if hydration at lower temperatures is considered, a hydration at a slower rate will be achieved but the absorption density values will be more stable. Overall, the results show that it is possible to obtain an integral rehydrated fermented substrate via injection. In addition, it is possible to control the distribution of the different absorption densities by controlling the temperature. Other parameters such as different pressures, number of injections and needles designs could be used to calibrate this mode to the desired levels.Example 9 Hydration with oil-water mixtures

[0178] The effects of rehydrating a substrate with oil / water mixtures were tested. A range of oil / water compositions, using an exemplary oil and a range of potentially emulsifying ingredients were tested on an exemplary fermented substrates, to identify if rehydrating the substrate using oil / water mixtures would improve organoleptic properties, including taste and texture.Sunflower oil was chosen as the exemplary oil due to its neutral flavour and its general commercial availability. For experiments involving spices in the hydrate, the spices were mixed into the oil for 24 hours and trained before preparing the emulsion.

[0179] Fermented substrate was prepared as described in example 1. Oil / water rehydrating mixtures were prepared by first shearing and mixing non-oil ingredients listed in table 18 into a saline watersolution in a mixer (blender), then adding the sunflower oil slowly into the blender running at high speed (20,000 RPM). Then samples of 5cm x 5cm x 2cm of fermented substrate were prepared and totally submerged into the hydrating mixtures at 20°C until the hydration corresponded to 76.5% of the weight of the sample had been absorbed by the substrate. The samples were then vacuum sealed, and pasteurized, before the organoleptic properties were evaluated by a panel of expert tasters. The results are shown in able 18.Table 18Conclusion

[0180] Results indicated that marinating with emulsified oil improved the taste / texture of the hydrated substrate. High levels of oil provided an attractive softened of the substrate but imparted a strong flavour and unpleasant mouthfeel, while using emulsified oil in amounts between 2.5% - 7.5% or 5% to 10% provided the best qualities.

[0181] The dominant flavour of the oil on the product could be countered by mixing spices or herbs into the oil, thereby masking the oil flavour. Black pepper was found to produce very good results in masking the oil flavour at higher oil levels, as the oil increased the intensity of the black pepper as hydrophobic compounds and oils from the pepper were released into the sunflower oil.

[0182] Marination would water / oil emulsions provides taste, texture and mouthfeel properties suitable for using the hydrated substrate in raw form mimicking a cold cut (processed meat) slice which is served cold and without further cooking.Example 10 Membrane filtration of the hydrate.

[0183] The effect on taste of membrane filtrating hydrate to reduce microorganism load and shelflife was investigated.Hydrate preparationHydrate samples were prepared according to table 19Table 19The individual concentration of each of the ingredients were proportional to the amounts used for sample number 1. Filtration of hydrate solution:

[0184] Subsamples of each of the hydrates were subjected to filtration first through a 10 pm, and then through a 5 pm filter and then through a 0.45 pm filter using a syringe. Then all samples were taken for microbial analysis and tasted comparing to the original sample, strength of thyme, strength of garlic, strength of beet, and salinity. The organoleptic analysis was executed by a panel of taste experts.

[0185] To determine the microbial count by dilution, a series of sample dilutions were prepared (10‘1, 10-2, IO-3). After preparing the dilutions, a constant volume of each dilution was plated on agar medium. The plates were incubated, and the number of colony-forming units (CFUs) per milliliter was counted. The CFU / ml was then calculated based on the number of colonies observed, factoring in the dilution factor for each sample.Table 20 Evaluation of microbial loadScoring of tasteTable 21Table 22 Organoleptic Scores5 = most identical to unfiltered hydrate; 1 = least identical to unfiltered hydrateConclusion

[0186] The filtration of the hydrate through a 0.45um filter did not affect the taste or visual aspect of the hydrate while it was shown the filtration removed the bacterial load. Accordingly, the filtration had a positive effect on the microbiology analysis while not compromising on the flavour of the hydrate and thus in turn a substrate hydrated with the hydrate.* * *

Claims

Claims1. A method for preparing a hydrated fermented substrate comprising: a) providing one or more edible ingredients; b) subjecting the one or more edible ingredients to one or more preparation steps to form a fermentation substrate; c) contacting the fermentation substrate with one or more microorganisms under conditions allowing the microorganisms to colonize and proliferate on / in the fermentation substrate and to produce a fermented substrate; d) contacting the fermented substrate with a hydrating composition under conditions allowing for fermented substrate to absorb the hydrating composition to produce the hydrated fermented substrate and e) isolating the hydrated fermented substrate.

2. The method of claim 1 wherein the hydrating composition comprises water and optionally one or more edible dissolved, dispersed or emulsified components selected from a salt, a protein, a carbohydrates, a surfactant or emulsifier, a colorant, a preservative, an oil or a fat, a vitamin, an amino acid, a mineral, a sweetener, a spice, an acidifier, a dietary fiber, a prebiotic, and / or a probiotic.

3. The method of claim 2 wherein the salt is sodium chloride.

4. The method of claim 3 wherein the hydrating composition comprises between 0,001 g / L to 50 g / L of sodium chloride, such as between 0,001 g / L to 25 g / L, such as between 0,001 g / L to 10 g / L, such as between 0,001 g / L to 5 g / L, such as between 0,001 g / L to 1 g / L, such as between 0,001 g / L to 0,5 g / L, such as between 0,01 g / L to 0,25 g / L, such as between 0,01 g / L to 0,1 g / L, such as between 0,025 g / L to 0,05 g / L.

5. The method of claim 2 wherein the protein and / or the carbohydrate is selected from an extract of biomass and / or legumes.

6. The method of claim 5 wherein the biomass is a fungal biomass, optionally a yeast biomass, optionally a baker's or brewer's yeast.

7. The method of claim 6 wherein the yeast biomass is inactivated yeast, optionally of a a yeast of thegenus Saccharomyces, optionally of the species S. cerevisiae.

8. The method of claim 5 wherein the legume is a legume is a bean.

9. The method of claim 8 wherein the bean is a P. sativa pea.

10. The method of any one of claim 2 to 9 wherein hydrating composition has a protein concentration which is higher than the protein concentration of the fermented substrate.

11. The method of claim 10 wherein the hydrating composition has a protein content which is at least 5% higher, such as at least 10% higher, such as at least 20% higher, such as at least 30% higher, such as at least 40% higher, such as at least 50% higher, such as at least 60% higher, such as at least 70% higher, such as at least 80% higher, such as at least 100% higher, such as at least 150% higher, such as at least 200% higher, such as at least 500% higher than the protein concentration of the fermented substrate.

12. The method of any one of claim 2 to 11 wherein the hydrating composition has a protein content of more than 10 wt%, such as more than 12 wt%, such as more than 15 wt%, such as more than 20 wt%, such as more than 25 wt%.

13. The method of claim 2 wherein the colorant is a colorant extract from carrot, pumpkin, red beet, beet root, barley malt, carthamus, radish, and or apple.

14. The method of claim 13 wherein the colorant is a colorant extract from a) carrot, pumpkin and apple, b) red beet, barley malt and Carthamus, c) radish and carrot, d) reed beet and barley malt, e) pumpkin and apple, f) beetroot and carrot, and / or g) red beet.

15. The method of claim 14 wherein the colorant is a colorant extract from a) carrot, pumpkin and apple, and / or b) red beet.

16. The method of claim 2 wherein the surfactant or emulsifier is selected from lecithin, monoglycerides, diglycerides, polysorbates (optionally Polysorbate 80), sorbitan esters (optionally sorbitan monostearate), propylene glycol esters (optionally propylene glycol monostearate), glycerolesters (optionally glycerol monostearate), acetylated monoglycerides, sucrose esters (optionally sucrose stearate), polyglycerol esters (optionally polyglycerol polyricinoleate - PGPR), and or lactic acid esters of mono- and diglycerides (LACTEM).

17. The method of claim 2 wherein the preservative is selected from sodium chloride, Sugars, vinegars, citric acid, Ascorbic Acid (vitamin C), tocopherols (vitamin E), rosemary extract, natamycin, sorbic acid or its salts, benzoic acid or its salts (optionally sodium benzoate), calcium propionate, BHA (butylated hydroxyanisole), and / or BHT (butylated hydroxytoluene).

18. The method of claim 2 wherein the oil is selected from olive oil, canola oil, soybean oil, corn oil, sunflower oil, coconut oil, peanut oil, sesame oil, avocado oil, grapeseed oil, rape seed oil, butter, lard, shortening, coconut butter, palm oil, walnut oil, and / or flaxseed oil.

19. The method of claim 2 wherein the vitamin is selected from vitamin A (Retinol, Retinal, Retinoic Acid), vitamin Bl (Thiamine), vitamin B2 (Riboflavin), vitamin B3 (Niacin), vitamin B5 (Pantothenic Acid), vitamin B6 (Pyridoxine), vitamin B7 (Biotin), vitamin B9 (Folate, Folic Acid), vitamin B12 (Cobalamin), vitamin C (Ascorbic Acid), vitamin D (Cholecalciferol, Ergocalciferol), vitamin E (Tocopherols, Tocotrienols), vitamin K (Phylloquinone, Menaquinones), and / or vitamin P (Bioflavonoids).

20. The method of claim 2 wherein the amino acid is selected from essential amino acids (Lysine, Leucine, Isoleucine, Valine, Methionine, Phenylalanine, Threonine, and / or Tryptophan), non-Essential amino acids (Alanine, Arginine, Asparagine, Aspartic Acid, Cysteine, Glutamic Acid, Glycine, Proline, Serine, and / or Tyrosine), conditional amino acids (Arginine, and / or Cysteine), and / or semi-essential amino acids (Histidine).

21. The method of claim 2 wherein the mineral is selected from a salt of Calcium, Phosphorus, Potassium, Sodium, Magnesium, Iron, Zinc, Copper, Manganese, Selenium, Iodine, Fluoride, Chromium, Molybdenum, and / or Cobalt.

22. The method of claim 2 wherein the sweetener is a natural sweetener selected from sugar, fructose, glucose, glucose syrup, stevia, honey, maple syrup, agave nectar, coconut sugar, date sugar, monk fruit sweetener, molasses, rice syrup, and / or yacon syrup.

23. The method of claim 2 wherein the sweetener is an artificial sweetener selected from aspartame, xylitol, saccharin, sucralose, acesulfame potassium, steviol glycosides, neotame, cyclamate, and / or advantame.

24. The method of claim 2 wherein the spice is selected from thyme, basil, garlic, onion, black pepper, green peppers, chili peppers, cinnamon, cumin, coriander, paprika, turmeric, ginger, nutmeg, cloves, cardamom, bay leaves, oregano, rosemary, and / or sage.

25. The method of claim 2 wherein the acidifier is selected from vinegar, acetic acid, and / or citric acid.

26. The method of claim 2 wherein the dietary fiber or prebiotic is selected from betaglucan, pectin, Inulin, Fructooligosaccharides (FOS), Galactooligosaccharides (GOS), Resistant Starch, Arabinoxylan, Lactulose, Konjac Glucomannan, and / or Polydextrose.

27. The method of claim 2 wherein the pH of hydrating composition is acidic or alkaline.

28. The method of claim 27 wherein the pH of hydrating composition is acidic, optionally between pH 5 and pH 6, such as between pH 5,1 and pH 5,3.

29. The method of claim 2 wherein the temperature of hydrating composition is between 5°C to 45°, such as 10°C to 40°C, such as 15°C to 35°C, such as between 20°C to 30°C.

30. The method of any preceding claim wherein the fermented substrate comprises one or more edible ingredients selected from: a) a dry component comprising i) starch providing nutrient to the one or more microorganism and ii) fibres providing nutritional and / or dietary value to the edible product, the dry component being capable of absorbing moisture from other edible ingredients; b) a high-moisture and high starch component providing for formation of gelatinized starch binding other edible ingredients together and providing nutrient to the one or more microorganism as well as texture to the edible product; c) a component providing minerals and vitamins for the one or more microorganism as well as a natural colour to the edible product;d) a high-protein and low moisture component providing flavour and firmness and structure to the texture of the edible product; or e) High starch component providing sustained nutrients for the one or more microorganism and providing softness to the texture of the edible product.

31. The method of claim 30 wherein one or more of the edible ingredients are or comprises an industrial waste product, not used for human consumption.

32. The method of any one of claim 30 to 31 wherein the one or more edible ingredients comprise at least 1, such as at least 3, such as at least 4, such as at least 5 components selected from the dry component, the high-moisture and high starch component, the component providing minerals and vitamins, the high-protein and low moisture component, or the high starch component.

33. The method of any one of claim 30 to 32 wherein the one or more edible ingredients comprise 5 ingredients selected from the dry component, the high-moisture and high starch component, the component providing minerals and vitamins, the high-protein and low moisture component, or the high starch component.

34. The method of any one of claim 30 to 33 wherein the one or more edible ingredients are selected from cereal grain, root vegetables, and legumes or a combination or extract thereof.

35. The method of claim 34 wherein the cereal grain is the dry component.

36. The method of any one of claim 34 or 35 wherein the grain is selected from maize (corn), rice, wheat, barley, sorghum, millet, oats, triticale, rye, and / or fonio.

37. The method of any one of claim 34 to 36, wherein the cereal grain is spent grain from fermentation of alcohol (brewers spent grain (BSG).

38. The method of any one of claim 36 to 37, wherein the cereal grain is oats.

39. The method of any one of claim 34 to 38, wherein the one or more edible ingredients comprise ,such as 5 %wt to 10 %wt, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt cereal grain.

40. The method of claim 39, wherein the one or more edible ingredients comprise 15 to 25 %wt, optionally 19 to 21 %wt of oats.

41. The method of claim 34, wherein the root vegetable is the high-moisture and high starch component and / or the component providing minerals and vitamins.

42. The method of claim 34 or 41, wherein the root vegetable a modified plant stem vegetable, a rootlike stem vegetable, or a true root vegetable.

43. The method of claim 42, wherein the modified plant stem vegetable is a Corm (bulbo- tuber / bulbotuber), a Rhizome or a Tuber.

44. The method of claim 43, wherein the Corm is selected from Amorphophallus konjac (konjac), Colocasia esculenta (taro), Eleocharis dulcis (Chinese water chestnut), Ensete spp. (enset), Nymphaea spp. (waterlily), Pteridium esculentum, Sagittaria spp. (arrowhead or wapatoo), Typha spp., Xanthosoma spp. (malanga, cocoyam, tannia, yautia and other names), and / or Colocasia antiquorum (eddoe or Japanese potato).

45. The method of claim 43, wherein the Rhizome is selected from Curcuma longa (turmeric), Panax ginseng (ginseng), Arthropodium spp. (rengarenga, vanilla lily, and others), Canna spp. (canna), Cordyline fruticosa (ti), Maranta arundinacea (arrowroot), Nelumbo nucifera (lotus root), Typha spp. (cattail or bulrush), and / or Zingiber officinale (ginger, galangal).

46. The method of claim 43, wherein the Tuber is selected from Apios americana (hog potato or groundnut), Cyperus esculentus (tigernut or chufa), Dioscorea spp. (yams, ube), Dioscorea polystachya (Chinese yam or white name), Helianthus tuberosus (Jerusalem artichoke or sunchoke), Hemerocallis spp. (daylily), Lathyrus tuberosus (earthnut pea), Oxalis tuberosa (oca or New Zealand yam), Plectranthus edulis and P. esculentus (kembili, dazo, and others), Solanum tuberosum (potato), Stachys affinis (Chinese artichoke or crosne), Tropaeolum tuberosum (mashua or anu), and / or Ullucustuberosus (ulluku).

47. The method of claim 46, wherein the Tuber is of the genus Solanum.

48. The method of claim 47, wherein the Tuber is of the species Solanum tuberosum (potato).

49. The method of claim 48, wherein the Solanum tuberosum is a red skin potato.

50. The method of any one of claim 42 to 49, wherein the root-like stem vegetable is Zamia integrifolia (Florida arrowroot).

51. The method of claim 42 to 49, wherein the true root vegetable is selected from a taproot or a tuberous root.

52. The method of claim 51, wherein the tuberous root vegetable is selected from Amorphophallus galbra (yellow lily yam), Conopodium majus (pignut or earthnut), Dioscorea polystachya (nagaimo, Chinese yam, Korean yam, mountain yam), Hornstedtia scottiana (native ginger), Ipomoea batatas (sweet potato), Ipomoea costata (desert yam), Manihot esculenta (cassava or yuca or manioc), Mirabilis expansa (mauka or chago), Psoralea esculenta (breadroot, tipsin, or prairie turnip), and / or Smallanthus sonchifolius (yacon).

53. The method of claim 51, wherein the taproot vegetable is selected from Arracacia xanthorrhiza (arracacha), Beta vulgaris (beet and mangelwurzel), Brassica spp. (kohlrabi, rutabaga and turnip), Bunium persicum (black cumin), Burdock (Arctium, family Asteraceae), Carrot (Daucus carota subsp. sativus), Celeriac (Apium graveolens rapaceum), Daikon - the large East Asian white radish (Raphanus sativus var. longipinnatus), Dandelion (Taraxacum) spp., and / or Lepidium meyenii (maca).

54. The method of claim 53, wherein the taproot vegetable is Beta vulgaris, optionally subspecies vulgaris.

55. The method of claim 54, wherein the taproot vegetable is B. vulgaris var. conditiva (red beet)56. The method of any one of claim 34 and 42 to 55, wherein the one or more edible ingredientscomprise 5 %wt to 60 %wt root vegetable, such as 5 %wt to 10 %wt, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt, 38 to 42 %wt.

57. The method of claim 56, wherein the one or more edible ingredients comprise 10 to 30 %wt, optionally 15 to 25 %wt, optionally 19 to 21 %wt of Solanum tuberosum.

58. The method of claim 56 wherein the one or more edible ingredients comprise optionally 15 to 25 %wt, optionally 19 to 21 %wt of B. vulgaris var. conditiva.

59. The method of any one of claim 57 to 58, wherein the one or more edible ingredients comprise 10 to 30 %wt, optionally 15 to 25 %wt, optionally 19 to 21 %wt of Solanum tuberosum and 10 to 30 %wt, optionally 15 to 25 %wt, optionally 19 to 21 %wt of B. vulgaris var. conditiva.

60. The method of claim 34, wherein the legume is the high-protein and low moisture component and / or the high starch component providing sustained nutrients for the one or more microorganism and providing softness to the texture of the edible product.

61. The method of any one of claim 34 or 60, wherein the legume is a bean selected from the genus of Phaseolus, Pisum, Vigna, Cajanus, Lens, Cicer, Vicia, Arachis, Glycine, Macrotyloma, Mucuna, Lupinus, Ceratonia, Canavalia, Cyamopsis, Lablab, Psophocarpus, Clitoria, Lathyrus, Trifolium, Medicago, Melilotus, and / or Tamarindus.

62. The method of claim 61 wherein the Phaseolus bean is selected from the species P. vulgaris (Kidney Bean, Pinto Bean, Navy Bean Haricot Bean, Black Beans, Borlotti Beans), P. lunatus (Lima Bean), P. coccineus (Runner Bean, Flat Bean), and / or P. acutifolius (Tepary Bean).

63. The method of claim 61 wherein the Pisum bean is selected from P. sativum peas (Green Peas, White Peas, Yellow Peas, Field Peas, Snow Peas, Snap Peas)64. The method of claim 61 wherein the Vigna bean is selected from V. radiata (Mung Bean), V. mungo (Urad), V. unguiculata (Cowpea, Yardlong bean, Black-eyed Pea), V. aconitifolia (Moth bean), and / or V. angularis (Adzuki bean).

65. The method of claim 61 wherein the Cajanus bean is selected from C. cajan (Pigeon Pea).

66. The method of claim 61 wherein the Lens bean is selected from L. culinaris (Lentil, Red Lentil, Green Lentil, Puy Lentil).

67. The method of claim 61 wherein the Cicer bean is selected from C. arietinum (Chickpea, Garbanzo Bean).

68. The method of claim 61 wherein the Vicia bean is selected from V. faba (Fava Bean, Broad Bean), V. ervilia (Bitter vetch), and / or V. sativa (common vetch).

69. The method of claim 61 wherein the Arachis bean is selected from A. hypogaea (peanut).

70. The method of claim 61 wherein the Glycine bean is selected from G. max (soybean).

71. The method of claim 61 wherein the Macrotyloma bean is selected from M. uniflorum (Horsegram).

72. The method of claim 61 wherein the Mucuna bean is selected from M. pruriens (velvet bean).

73. The method of claim 61 wherein the Lupinus bean is selected from L. albus (white lupin, sweet lupin), L. mutabilis (Tarwi / Andean Lupin), L. hirsutus and / or L. angustifolius.

74. The method of claim 61 wherein Ceratonia the bean is selected from C. siliqua (Carob bean)75. The method of claim 61 wherein the Canavalia bean is selected from C. gladiate (Sword bean), and / or C. ensiformis (Jack bean).

76. The method of claim 61 wherein the Cyamopsis bean is selected from C. tetragonoloba (Guar bean).

77. The method of claim 61 wherein the Lablab bean is selected from L. purpureus (Hyacinth Bean,lablab bean).

78. The method of claim 61 wherein the Psophocarpus bean is selected from P. tetranoglobulus (winged bean).

79. The method of claim 61 wherein the Clitoria bean is selected from C. ternatea (butterfly pea).

80. The method of claim 61 wherein the Lathyrus bean is selected from L. sativus (grass pea) and / or L. tuberosus (tuberous pea).

81. The method of claim 61 wherein the Trifolium bean is selected from T. repens (white Clover), and / or T. pratense (red clover).

82. The method of claim 61 wherein the Medicago bean is selected from M. sativa (alfalfa).

83. The method of claim 61 wherein the Melilotus bean is selected from M. officinalis (sweet clover).

84. The method of claim 61 wherein the Tamarindus bean is selected from T. indica (tamarind).

85. The method of any one of claim to 34 or 60 to 84, wherein the one or more edible ingredients comprise 5 %wt to 60 %wt legumes, such as 10 %wt to 15 %wt, such as 15 %wt to 20 %wt, such as 20 %wt to 25 %wt, such as 25 %wt to 30 %wt, such as 30 %wt to 35 %wt, such as 35 %wt to 40 %wt, such as 45 %wt to 50 %wt, such as 55 %wt to 60 %wt, optionally 38 %wt to 42 %wt.

86. The method of any one of claim 34 or 60 to 85 wherein the one or more edible ingredients comprise a legume which contain more than 25 %wt of protein.

87. The method of any one of claim 34 or 60 to 86 wherein the one or more edible ingredients comprise a legume which contain more than 25 %wt of starch.

88. The method of any one of claim 34 or 60 to 87 wherein the one or more edible ingredients comprise a least two legumes of which at least one legume contains more than 25 %wt of protein and at least one legume contains more than 25 %wt, such as more than 30 %wt, such as between 35 %wtto 45 %wt of starch.

89. The method of claim 61 wherein the one or more edible ingredients comprise Pisum beans and Lupinus beans.

90. The method of claim 89 wherein the one or more edible ingredients comprise P. sativum peas and L. albus lupin seeds.

91. The method of claim 90 wherein the one or more edible ingredients comprise yellow split P. sativum peas and sweet L. albus lupin seeds.

92. The method of claim 61 or 89 to 91 wherein the one or more edible ingredients comprise 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of yellow split P. sativum peas.

93. The method of any one of claim 61 or 89 to 92, wherein the one or more edible ingredients comprise optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of sweet L. albus lupin seeds.

94. The method of any one of claim 61 or 89 to 93 wherein the one or more edible ingredients comprise 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of yellow split P. sativum peas and 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt of sweet L. albus lupin seeds.

95. The method of any one of claim 30 to 94 wherein the one or more edible ingredients comprise oats, Solanum tuberosum, B. vulgaris var. conditiva, P. sativum peas and L. albus lupin seeds.

96. The method of claim 95 wherein the one or more edible ingredients comprise 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt each of oats, potato, red beet, P. sativum peas and L. albus lupin seeds.

97. The method of claim 96 wherein the one or more edible ingredients comprise 20 %wt oats, 20 %wt potato, 20 %wt red beet, 20 %wt P. sativum peas and 20 %wt L. albus lupin seeds.

98. The method of any one of claim 30 to 97 wherein the one or more edible ingredients furthercomprise an aqueous extract of oats (oat milk).

99. The method of any one of claim 30 to 98 wherein the one or more edible ingredients further comprise an aqueous extract of almonds (almond milk).

100. The method of any one of claim 30 to 99 wherein the one or more edible ingredients comprise oats and / or Brewer Spent Grain (BSG) and wherein the oats, or and / or BSG prior to forming the fermentation substrate, are pre-processed by milling into a particle size between 2 and 5 mm, optionally between 3 and 4 mm, optionally to an average particle size of 3,5 mm.

101. The method of any one of claim 30 to 100 wherein the one or more edible ingredients comprise potato and wherein the potato, prior to forming the fermentation substrate, is pre-processed by wholly or partially gelatinizing starch by boiling, microwaving or steaming.

102. The method of any one of claim 100 or 101, wherein the oats and potato, prior to forming the fermentation substrate, is further pre-processed by co-grinding in a ratio of 0.5:1 to 1:0,5 of oats:potato, optionally 1:1, until no visible chunks of potato can be observed.

103. The method of any one of claim 30 to 102 wherein the one or more edible ingredients comprise red beet and wherein the red beet, prior to forming the fermentation substrate, is pre-processed by i) boiling and / or steaming and ii) grinding to an average particle size of between 1,7-2, 8 mm.

104. The method of any one of claim 30 to 103 wherein the one or more edible ingredients comprise P. sativum peas, and wherein the P. sativum peas, prior to forming the fermentation substrate, is pre- processed by i) drying and splitting the P. sativum peas, ii) microwaving, boiling or steaming the P. sativum peas and iii) grinding the P. sativum peas to an average particle size of between 1,7-2, 8 mm.

105. The method of claim any one of 30 to 104 wherein the one or more edible ingredients comprise L. albus lupin seeds, and wherein the lupin seeds, prior to forming the fermentation substrate, is pre- processed by i) soaking the lupin seeds in water to wholly or partially remove toxic or bitter tasting alkaloids, ii) microwaving, boiling or steaming the lupin and iii) grinding the lupin to an average particle size of between 1,7-2, 8 mm.

106. The method of claim any one of 30 to 105 further comprising grinding and mixing the pre- processed one or more edible ingredients into a fermentation substrate and optionally pasteurize the fermentation substrate.

107. The method of any one of claim 30 to 106 wherein the one or more edible ingredients prior to forming the fermentation substrate is ground to an average particle size of between 150 pm to 350 pm, optionally 200 pm to 300 pm, optionally 225 pm to 275 pm, optionally around 250 pm.

108. The method of any one of claim 106 to 107, further comprising adjusting the water content to between 35 to 40 %wt, optionally 36 to 38 %wt in the fermentation substrate.

109. The method of any preceding claim, wherein the one or more microorganisms comprise a GRAS strain.

110. The method of any preceding claim, wherein the one of more microorganisms a fungus.

111. The method of claim 110 wherein the fungus is a filamentous fungus.

112. The method of claim 111 wherein the fungus is a filamentous fungus is of the genus Aspergillus, optionally of the species Aspergillus oryzae and / or Aspergillus sojae.

113. The method of claim 111 wherein the fungus is a filamentous fungus is of the genus Rhizopus optionally of the species Rhizopus oligosporus.

114. The method of any preceding claim, further comprising mixing the one or more microorganisms into the fermentation substrate and allowing the one or more microorganism to colonize and ferment the fermentation substrate at a predetermined time interval, temperature and humidity.

115. The method of claim 114 wherein the fermentation time is between 20 to 90 hours, optionally 30 to 60 hours, optionally 40 to 50 hours.

116. The method of any one of claim 114 or 115 wherein the temperature during incubation is maintained between 20°C and 40°C, optionally between 25°C and 35°C, optionally between 29 °C and31°C.

117. The method of any preceding claim, wherein the hydrated fermented substrate is enriched or improved in one or more properties selected from flavour, aroma, texture, cooking experience, visual appearance, nutritional and / or dietary value, spoilage time, and / or carbon footprint.

118. The method of claim 117 wherein the flavour property is sourness, sweetness, bitterness, saltiness, umami or a combination thereof.

119. The method of claim 117 wherein the enriched or improved texture property is firmness, softness, cohesiveness, juiciness, chewiness, sandiness or a combination thereof.

120. The method of claim 117, wherein the enriched or improved cooking property is improved browning of the fermented edible product (Maillard reaction) when roasting or frying the fermented edible product compared to roasting or frying the edible product prior to fermentation.

121. The method of claim 117, wherein the enriched or improved cooking property is improved structure integrity of the fermented edible product compared to the edible product prior to fermentation, where the microorganism biomass binds together the edible ingredients.

122. The method of claim 117, wherein the enriched or improved visual property is colour.

123. The method of claim 117, wherein the enriched or improved nutritional and / or dietary property is content and / or distribution of protein, lipids, carbohydrates, fibre, vitamins, minerals, amino acids or a combination thereof.

124. The method of any preceding claim wherein the contacting of the fermented substrate with the hydrating composition comprises partially or fully submersing the fermented substrate in the hydration composition.

125. The method of claim 124 wherein the fermented substrate is partially or fully submersed in the hydration composition between 1 to 40 seconds.

126. The method of any preceding claim wherein the amount of fermented substrate and the amount of hydration composition is selected so as to allow the fermentation substrate to absorb more than 50% of the hydration composition, such as more than 60%, such as more than 70%, such as more than 80%, such as more than 90%, such as more than 95%, such as more than 98% of the hydration composition.

127. The method of any preceding claim wherein the hydrated fermented substrate has an absorption density (weight of hydration composition absorbed per weight of fermented substrate) between 50% to 100%, such as between 60% to 90%, such as between 70% to 80%, such as between 75% to 80%.

128. The method of any preceding claim further comprising vacuum packaging and heat treatment of the hydrated fermented substrate.

129. The method of claim 128 wherein the heat treatment comprises heating the vacuum packaged hydrated fermented substrate to a temperature of at least 92°C for at least 5 minutes.

130. The method of any preceding claim wherein the hydrating composition is an oil / fat-water emulsion.

131. The method of claim 130 wherein the oil / fat-water emulsion comprise 2,5 %wt to 10 %wt of oil or fat.

132. The method of any one of claim 130 to 131 wherein the oil / fat is a vegetable oil, optionally Sunflower oil.

133. The method of any one of claim 130 to 132 wherein the oil / fat-water emulsion further comprises salt and or spices.

134. The method of claim 133 wherein the spice is a pepper, optionally black pepper.

135. The method of any preceding claim wherein the hydrating composition is filtered to remove microorganisms prior to contacting the fermented substrate with the hydrating composition.

136. The method of claim 135 wherein the filter has a pore size of 0,45 pm or below.

137. The method of claim 136 wherein the filter has a pore size between 0,22 pm and 0,45 pm.

138. A hydrated fermented substrate obtained by the method of any preceding claim.

139. The hydrated fermented substrate comprising one or more filamentous fungi or the mycelium thereof and 10 %wt to 30 %wt, optionally 15 %wt to 25 %wt, optionally 19 %wt to 21 %wt each of oats, potato, red beet, P. sativum peas and L. albus lupin seeds, the sum of oats, potato, red beet, peas and lupin seeds not exceeding 100 percent.

140. The hydrated fermented substrate of claim 139 further comprising more than 10 %wt of dietary fibres.* * *

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