Novel food composition and method for production thereof

Abstract

The present disclosure relates to a process for preparing an upcycled material from a dry raw material with a moisture content of 0-30% w / w comprising lignocellulose and derived from spent cereal and / or pseudocereal grain and / or food waste or lignocellulosic biomass or a mixture thereof, the process comprising: a) a pre-treatment comprising the steps of: i) grinding and / or pulverizing the raw material to obtain a powder capable of passing through a sieve having a mesh size of 10-1000 μm and ii) performing autohydrolysis, followed by b) enzymatic hydrolysis of the of powder obtained from step ii), wherein the enzymatic hydrolysis comprises adding an enzyme mixture comprising at least a glucanase and a xylanase, and c) a post-treatment comprising the step of grinding and / or pulverizing the enzymatically hydrolysed mixture obtained from step b) to obtain the upcycled material in the form of a flour-like product.

Description

[0001] NOVEL FOOD COMPOSITION AND METHOD FOR PRODUCTION THEREOF

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to a process of preparing an upcycled material from a dry raw material comprising lignocellulose and typically derived from spent cereal or pseudocereal grain or food waste. The process may further comprise a fermentation step. The invention further relates to an edible product comprising 0.1-100% dry matter by weight flour-like and / or fermented product.

[0004] BACKGROUND OF THE INVENTION

[0005] The global increase of food consumption and its demands are compromising global food security and representing environmental risks. According to the Food and Agriculture Organization of the United Nations (2011), approximately one-third of the food produced for human consumption is lost or wasted globally. This contributes to loss of biodiversity, water pollution, wasteful water use, and increased greenhouse emissions resulting from food production, while hunger and malnutrition are a reality for millions of people. Solving these problems requires implementing strategies throughout the whole food chain. For example, changing to a more sustainable diet on an individual level and upcycling food waste to reduce the amount of food lost or wasted are prominent part of the solution (FAO, 2011; Foley et aL, 2011; Ishangulyyev et aL, 2019).

[0006] Upcycling, i.e., the process of transforming by-products, waste materials, 'useless' or unwanted products into new materials or products so as to create a product of higher quality or value than the original, is one of the cornerstones of a functional 'bio-circular economy'. In Denmark, for example, malting is the main destination of the produced barley. In Europe, the production of beer has been around 36 thousand million litres in the last years (The Brewers of Europe, 2021), which produces approx. 7 million tons per year of brewer's spent grain. Other industrial food or beverage production, such as distilleries or restaurants, also produce side streams of waste products that are currently not upcycled. Additionally, households also produce a huge amount of waste products which either ends up in a landfill or is incinerated. There are however incentives to upcycle waste materials from agricultural, domestic or industrial sources into e.g., biofuels or other value-added upcycled products.

[0007] Brewer's spent grain (BSG) or draff is a food waste that is a by-product of the brewing industry that makes up approx. 85% of brewing waste. BSG is the solid residue left after the separation of the wort during the lautering stage. The majority of BSG is composed of barley malt grain husks in combination with parts of the pericarp and seed coat layers of the barley. Depending on the exact type of beer produced, the BSG may also comprise wheat, maize, rice, oats and other cereal grains. Though the composition of BSG can vary depending on the type of barley used, the way it was grown, and other factors, BSG is usually rich in lignocellulose, i.e., cellulose, hemicelluloses and lignin, proteins, lipids and phenolic compounds. BSG is also naturally high in fibres, making it of great interest as a food additive, replacing low-fibre ingredients.

[0008] BSG has been proposed for its potential in the production of feedstock, fertilizers, enzymes, biologically active substances, spores, lactic acid, xylitol, pullulan, activated carbon, constructions bricks, bioethanol and biomethane, among others. Due to its highly nutritional value and potential health benefits, BSG has also drawn the attention of academic and industrial researchers for human consumption.

[0009] For example, BSG has been added to bakery and snack products to improve shelf life and to increase fibre, protein and lysine content or to frankfurter sausages to reduce the fat content. However, previous studies have reported that the direct incorporation of BSG negatively modifies the product characteristics (texture, colour, aroma, flavour) thus reducing consumer acceptance. Additionally, BSG contains a lot of fibre, which is not easily digestible.

[0010] Various enzymatic treatments have been proposed (e.g., carbohydrases and / or proteinases) to improve the organoleptic properties of the BSG added products. The breakdown of BSG into its constituent components seems to obtain valuable gastronomic elements such as fermentable carbohydrates and amino acids, also improving the rheological properties of products. Enzymatic hydrolysis is a heterogeneous reaction that requires direct physical contact between the enzymes and the target regions of the substrate.

[0011] Various enzymes, for example, endoxylanases, p-xylosidases, feruloyl esterases, acetyl esterases, glucuronidases, glucuronoyl esterases and a-L-arabinofuranosidases have been proposed to complete the degradation of hemicellulose to its monomer constituents. Enzymatic hydrolysis of cellulose is a reaction that can be carried out by cellulase enzymes, such as cellulases, exoglucanase, endoglucanase, and p-glucosidase.

[0012] US8658407B2 describes a composition for the generation of ethanol (fuel from a lignocellulosic feedstock. This publication describes a solid-state fermentation product followed by a cooking and saccharification process in a conventional distillery cooking system.

[0013] US20220195475 Al discloses a method for hydrolysing Brewer's spent grain, the product of which may be directly incorporated into food compositions, supplements or food products. However, the processing and the nutritional value of the food product is to be further optimised.

[0014] OBJECT OF THE INVENTION

[0015] It is an object of embodiments of the invention to provide a process for preparing an upcycled material from a dry raw material comprising lignocellulose and derived from spent cereal, pseudocereal or food waste to provide a nutritious edible product which may directly be incorporated into human food or may be used as a substrate for microorganisms to further process and enhance the nutritional value of the spent cereal or food waste.

[0016] SUMMARY OF THE INVENTION

[0017] It has been found by the present inventor(s) that by processing a dry raw material comprising lignocellulose according to the claims, a nutritious, edible product may be obtained that can directly be consumed by humans and / or can be added to recipes to replace other, traditionally used materials, such as flour. The edible product exhibits an unprecedent degree of utilisation - where priori art product produced from BSG have typically been used as substitute ingredients in foods and feeds to constitute a relatively low percentage of an edible end product, the product obtained can constitute very high percentages of edible products.

[0018] So, in a first aspect the present disclosure relates to a process for preparing an upcycled material from a dry raw material with a moisture content of 0-30% w / w comprising lignocellulose and derived from spent cereal and / or pseudocereal grain and / or food waste or lignocellulosic biomass or a mixture thereof, the process comprising: a) a pre-treatment comprising the steps of: i) grinding and / or pulverizing the raw material to obtain a powder capable of passing through a sieve having a mesh size of 10-1000 pm and ii) performing autohydrolysis, followed by b) enzymatic hydrolysis of the of powder obtained from step ii), wherein the enzymatic hydrolysis comprises adding an amount of an enzyme mixture comprising at least a glucanase and a xylanase, preferably so as to add a controlled amount of enzyme(s), and c) a post-treatment comprising the step of grinding and / or pulverizing the enzymatically hydrolysed mixture obtained from step b) to obtain the upcycled material in the form of a flour-like product.

[0019] In a further aspect the disclosure relates to a process for preparing a beverage comprising fermentation by microorganisms of a carbohydrate-containing substrate, which comprises or consists of a flour-like product obtainable by the process described herein.

[0020] In further aspect the disclosure relates to a chocolate-like product comprising 20-45% matter by weight flour-like product obtained at the end of step c) and / or fermented product obtained at the end of step d), 10-25% by weight sugar, 25-50% by weight butter and 0.1-1.2% by weight lecithin.

[0021] In an additional aspect the disclosure relates to a chocolate-like product comprising 10-25% by weight sugar, 25-50% by weight fats, 0.1-1.2% by weight lecithin and 0-0.01% by weight theobromine.

[0022] In a further aspect the disclosure relates to an edible product comprising 0.1-100% dry matter by weight flour-like product obtained at the end of step c) and / or the fermented product obtained at the end of step d).

[0023] In a further aspect the disclosure relates to a flour-like product obtainable by the process defined herein, for use in cosmetics, dietary supplements and / or nutraceuticals

[0024] In a further aspect the disclosure relates to a non-alcoholic beverage prepared using a flour-like product as defined herein, wherein the beverage comprises less than 0.2 g / lOOg maltose.

[0025] In a further aspect the disclosure relates to a fermented beverage, wherein the fermentation broth at the end of the fermentation comprises less than 0.2g / 100g maltose and less than 1% ABV of ethanol, and wherein the microorganism used in the fermentation is of the genus Pichia spp., such as Pichia kluyveri.

[0026] BRIEF DESCRIPTION OF FIGURES

[0027] Fig. 1. Particle size distribution of enzymatically treated (E-BSG12 and EBSG24) and untreated (NE-BSG12 and NE-BSG24) samples following 12 hrs or 24 hrs of conching.

[0028] DETAILED DISCLOSURE OF THE INVENTION

[0029] Definitions

[0030] The term "upcycled material" refers to a product obtained by the process of transforming lignocellulose containing by-products, waste materials (such as food waste - food that may otherwise be destined to be thrown out), products that are conventionally considered to be "useless" for human use or in other ways unwanted, or products destined for animal feed, into new materials or products, which have a higher quality or environmental value than the byproduct or waste material it originates from. In the context of present application, this term is typically used in connection with materials originating from the food or beverage industry. However, the waste may also be originated from e.g., food waste (household food waste that currently is incinerated or used for biomass conversion) or leftovers from agricultural or industrial sources. Food waste is any food, including inedible parts of food, removed from the food supply chain to be recovered or disposed (including composted, crops ploughed in / not harvested, anaerobic digestion, bio-energy production, co-generation, incineration, disposal to sewer, landfill or discarded to sea). As such waste product also includes materials which have been used in industrial production which is otherwise destined for further side stream use as e.g., animal feed, fertilizer or biofuel, but which side stream can be redirected back to products suitable for human use or human consumption i.e., upcycling of such materials for human use or human consumption. Examples of such side stream usages are e.g., the use of brewers spent grain (BSG) or rapeseed cakes in animal feed. Such side stream usages may e.g., be redirected back to human consumption using the upcycling processes as disclosed herein.

[0031] The term "dry raw material" refers to a raw material comprising lignocellulose, which is dry or is dried prior to the first processing step. Dry refers in this context to a moisture content of 0-30%. For example, Brewer's spent grain (BSG) may be obtained from industrial sources and may be wet due to processing. Therefore, prior to upcycling BSG or other source materials, the raw material is dried using any conventional methods known in the art to obtain a dry material, such as by dehydration.

[0032] The term "spent grain cereal" refers to cereals, i.e., any plant of the grass Poaceae) familily cultivated for the edible components of its grain, which are "spent", i.e., are considered to be food waste or by-products of a food manufacturing process. For example, BSG is a food waste, which is a by-product of the brewing industry and is obtained as a mostly solid residue after wort production in the brewing process. The spent grain cereal may originate from any suitable raw material source.

[0033] The term "pseudocereal grain" refers to grains that do not originate from grasses and which are used in much the same way as cereals. Pseudocereals typically have good nutritional profiles with high levels of amino acids, essential fatty acids, minerals and some vitamins. Spent pseudocereals may also originate from conventional food manufacturing processes, such as the beverage industry. The pseudocereal grain may also be spent pseudocereal grain.

[0034] The term "food waste" typically refers to food or components from food raw materials that are not eaten or consumed by humans but is destined to be thrown away.

[0035] The term "lignocellulose" refers to plant dry matter (or biomass), which is composed of two kinds of carbohydrate polymers: cellulose and hemicellulose and an aromatic-rich polymer called lignin. Each three components have different properties and chemical behaviour; therefore, the processing of lignocellulose remains a challenge. The term "pre-treatment" refers to a treatment conducted prior to the enzymatic hydrolysis step.

[0036] The pre-treatment may comprise several steps that may be conducted in any suitable order.

[0037] The terms "grinding and / or pulverizing" refer to a processing step in a suitable device, such as a mill, a structure, a machine or a kitchen appliance, that breaks solid materials into smaller pieces by grinding, crushing or cutting. The mill may be powered by hand, wind, water or electricity. Any conventionally known device may be used to grind the material to obtain smaller particles.

[0038] The term "autohydrolysis" refers to a hydrolysis process in which the hydrolysis of molecules, such as e.g., protein or oligosaccharides is facilitated by water. In most instances, the autohydrolysis is catalysed by heat, such as by autoclaving the watery mixture to catalyse the autohydrolysis of the molecules.

[0039] The term "enzymatic hydrolysis" refers to a process, in which enzymes facilitate the cleavage of bonds in molecules with the addition of water. As such, enzymatic hydrolysis may be obtained from addition of purified hydrolytic enzymes or it may be obtained from culturing with microorganisms expressing hydrolytic enzymes. Preferably, the present disclosure relates to the addition of controlled amounts purified hydrolytic enzymes, as compared to the uncontrolled amount of hydrolytic enzymes obtained when culturing with microorganisms expressing hydrolytic enzymes. The use of controlled amounts also enables that balanced amount of the individual enzyme species can be added in order to optimize the enzymatic activity exerted.

[0040] The term "post-treatment" typically refers to a processing step that follows after the enzymatic hydrolysis step. The post -treatment may comprise several steps that may be conducted in any suitable order.

[0041] The term "dehydration step" or "dehydrating" refers to a processing step during which moisture is removed from the product or material and so the product or material is dried (dehydrated or desiccated). The dehydration may be by any conventional drying methods, such as freeze-drying, irradiation, by electrical dehydrators (conventional household appliances), spray-drying, microwave-vacuum drying, by using an oven or by sun-or wind-drying. Preferably, any dehydration step is followed by a grinding and / or milling step as during dehydration, the particles tend to coalesce into a single block. To break this up into smaller particles, the dehydrated material, such as the raw material or the product is preferably grinded and / or milled to obtain a powder.

[0042] The term "flour-like product" refers to a product obtained at the end of the process according to the first aspect. The flour-like product may be a wet flour-like product, i.e., might resemble the look and texture of wet flour. The flour-like product may also be a dry product, which resembles the look and texture of conventional flour products with no water or other solvents added. Typically, the flour-like product is a wet flour-like product, which may be dehydrated to obtain a dry flour-like product.

[0043] The term "chocolate-like product" refers to a product that resembles chocolate in terms of taste, texture, consistency etc. however it is not made solely using "traditional" chocolate components, such as roasted and ground cacao seed kernels. The chocolate-like product may comprise cacao derivatives or the like, however it is understood that these are not the main constituents of the chocolate-like product and are typically viewed as additives to further enhance the properties of the chocolate-like product. In embodiments, the chocolate-like product contains an amount of theobromine below 100 mg / 100 g product, such as below 50 mg / 100 g, 40 mg / 100 g, 30 mg / 100 g, 20mg / 100g, 10 mg / 100 g product, preferably, below 5 mg / 100 g product. In embodiments, the chocolate-like product contains no more than trace amounts of theobromine. Furthermore, the chocolate-like product should have a snap comparable to the snap of an actual chocolate product, either evaluated by a 3-point measurement or evaluated by the sensory perception. Additionally it is preferable that the sensory perception of the chocolate-lie product is comparable to that of actual chocolate products, such analysis may be conducted as described in e.g., Example 3, 4 or 14. Especially Example 14 suggests how the particle size of the flour like product can be correlated with the sensory perception and mouth-feel of the chocolate-like product.

[0044] As used herein the term "snap" refers to the point at which a product, such as a chocolate bar, snaps from an applied pressure according to the standard ISO 178. The snap is generally measured in MPa. The snap of a chocolate-like product as disclosed herein should preferably be in the range of 1-500 MPa, preferably in the range of 5-100 MPa, more preferably 5-40 MPa, for products that are 70-100 mm long, 30-60 mm wide and 5-15 mm thick, preferably about 95 mm long, 40 mm wide and 10 mm thick, and tested according to the Three Point Test of the ISO 178 standard. In example, the snap may be evaluated as follows, where the snap is measured at room temperature 20-25°C where the chocolate bars (or chocolate-like bars) are placed on two supports (making two points of contact), and a force is applied to the center of the bar (making the 3rd point of contact in the 3-point bending test). The flexural strength of the bar is essentially the highest stress that the material experiences during its moment of rupture (failure) and can be calculated from the following equation (formula 1):

[0045] 1.5P L

[0046] Formula 1: a = - -

[0047] (w-ty2where o is the flexural strength (MPa), P is the applied force (N), L is the span length (mm), w is the width of the bar (mm), and t is the thickness of the bar (mm)). Sample texture is analyzed with a TA-XT2 Texture Analyzer (Texture Technologies Corp., Scarsdale, NY).

[0048] Ranges of Flexural strength for chocolate are normally 5-100 MPa. The "snap" of a chocolate bar or a chocolate-like bar may also be perceived as a sensory perception of the snap-point of the bar. Such a sensory perception of the snap may be evaluated by a group of people testing the snap and reporting on the sensory perception of the snap on a similarity scale, such as that described in Example 3, where the scale was 1-9 points was used with 1 being 'extremely dislike' and 9 being 'extremely like'.

[0049] The term "non-alcoholic beverage" or "non-alcoholic beer" refers to beverages or beer with 0- 0.5% alcohol by volume (ABV).

[0050] The term "edible" refers to an item that is safe for humans to eat, i.e., is fit for human consumption and is non-poisonous.

[0051] Specific embodiments of the invention

[0052] The present disclosure relates to a process for preparing an upcycled material from a dry raw material comprising lignocellulose and derived from spent cereal and / or pseudocereal grain and / or food waste or lignocellulosic biomass or a mixture thereof, the process comprising: a) a pretreatment comprising the steps of: i) grinding and / or pulverizing the raw material to obtain a powder capable of passing through a sieve having a mesh size of 10-1000 pm and ii) performing autohydrolysis, followed by b) enzymatic hydrolysis of the of powder obtained from step ii), wherein the enzymatic hydrolysis comprises adding an enzyme mixture comprising at least a glucanase and a xylanase, and c) a post-treatment comprising the steps of grinding and / or pulverizing the enzymatically hydrolysed mixture obtained from step b) to obtain a flour-like product. Preferably, the moisture content of the upcycled material is in the range of 0-30% w / w.

[0053] As is shown in the examples, the pre-treated raw material that was passed through the sieve had a darker colour and toasted aromas, while the non-sifted fibre had a lighter colour and a straw-like aroma. Accordingly, the pre-treatment may for example be used to select for aroma(s), texture, and appearance of the lignocellulose comprising raw material, which is beneficial for several applications of the produced product.

[0054] As such, the dry raw material may be derived from a spent cereal or pseudocereal grain selected from the group consisting of millet, maize, sorghum, barley, oats, rice, rye, spelt, teff, triticale, wheat, wild rice, Brewer's spent grain, Distiller's spent grain, amaranth, buckwheat, chia, quinoa, wattle seeds, and acorn.

[0055] The spent cereal grain may be derived from a raw material selected from the group consisting of millet, maize, sorghum, barley, oats, rice, rye, spelt, teff, triticale, wheat and wild rice. The spent cereal grain may be Brewer's spent grain. Further the spent cereal grain may be Distiller's spent grain.

[0056] The spent cereal grain may be obtained from any suitable source, for example, from household waste, industrial kitchens, food processing plants, such as industrial or agricultural processing facilities. The Brewer's spent grain and / or Distiller's spent grain may be obtained from the brewing industry, for example, beer breweries, whiskey distilleries and the like. Spent cereal grains are typically rich in cellulose, hemicelluloses, lignin and proteins and are also naturally high in dietary fibre.

[0057] The raw material may, however, be any food waste selected from the group consisting of Brewer's spent grain, Distiller's spent grain, coffee grounds, okara, wheat bran, rapeseed press cake, palm oil press cake, sunflower press cake, leftover liquorice root, tiger nut, cashew, hemp, coconut, carrot pomace, apple pomace grape pomace, olive pomace, tomato pomace, orange peels, banana peels pineapple core, rice hulls buckwheat hulls, barley husks and hulls, wheat bran oat bran malt rootlets, pumpkin seed, sunflower seed, hazelnut skins, almond skins, sorghum bagasse, coffee husks, cocoa husks, cocoa bean husks, cocoa pod husks, nut pulp leftover from plant-based drinks, such as almond pulp, oat pulps, nut husks, such as hazelnut husks, date seeds, pulses husks, such as chickpea husk, pulses hulls, such as pea hulls, corncob, sugar cane / sugar beet bagasse, fruit skins, vegetable skins, fruit pulp, vegetable pulp, cocoa chaff and coffee chaff and tea leaves.

[0058] In one embodiment, the dry raw material comprising lignocellulose and derived from a spent cereal or pseudocereal grain is selected from the group comprising millet, maize, sorghum, barley, oats, rice, rye, spelt, teff, triticale, wheat, wild rice, Brewer's spent grain, amaranth, buckwheat, chia, quinoa, wattle seeds, and acorn.

[0059] In yet another embodiment, the dry raw material is food waste selected from the group comprising Brewer's spent grain, Distiller's spent grain, coffee grounds, okara, wheat bran, rapeseed press cake, palm oil press cake, sunflower press cake, leftover liquorice root, coffee husks, cocoa husks, nut pulp leftover from plant-based drinks, such as almond pulp, oat pulps, nut husks, such as hazelnut husks, date seeds, pulses husks, such as chickpea husk, pulses hulls, such as pea hulls, corncob, sugar cane / sugar beet bagasse, fruit skins, vegetable skins, fruit pulp, vegetable pulp, cocoa chaff and coffee chaff and tea leaves.

[0060] It is further possible to combine various raw materials in a single batch process. Accordingly, in some embodiments, the raw dry material is a combined raw material from more than one of the raw materials described above. For example, Brewer's spent grain may be mixed with cocoa husks, coffee grounds or any other suitable lignocellulosic waste material prior to the processing. In particular embodiments, the raw dry material is a combination of Brewer's spent grain and cocoa husks. Depending on the intended use, combining various raw materials, such as spent grain cereals and food waste, such as coffee chaffs allow the evolution of different flavours and fragrances, thus resulting in a more complex taste profile and allows for producing more diverse foods and flavours. However, it is also possible to use a single raw material source, e.g., only spent cereal grains in the process.

[0061] It is to be expected that the raw material will have an impact on the tasting notes and general sensory experience of the produced product. As an example, it may be so that products that are produced from e.g., BSG may comprise malty flavours, while cocoa husks may comprise chocolate-like flavours. Nevertheless, the different flavours, in some cases off-flavours, obtained after step c) in the method of the invention may be developed further, modified or removed using fermentation, as also described herein. Accordingly, while the particular tasting notes from the initial flour-like product may contain off-notes, these may be modified using further fermentation. In this context, embodiments of brewing (which include fermentation), which include use of the product obtained from the method of the first aspect of the invention, can advantageously include addition of protease(s), such as endoproteases, in either a mashing step or as part of fermentation as such, so as to e.g. facilitate release of free amino acids. Likewise, p-glucosidase can be added in both these brewing steps. Moreover, both the step of mashing and the step of fermentation may independently benefit form addition of yeast extract / hydrolysate (1-10%), e.g. from Brewers Spent Yeast).

[0062] In interesting embodiments of the first aspect of the present invention, BSG (wet or dry) not subjected to enzymatic treatment, is subjected to enzymatic treatment in the mashing or fermentation steps, or both, in a brewing process. That is, step b in the process of the first aspect is carried out as part of the mashing or fermentation steps in a brewing method. Use of 1-10% yeast extract / hydrolysate (eg. From Brewers Spent Yeast)

[0063] For some applications it may be preferred that the dry raw material is cocoa bean husks, cocoa pod husks, or a combination thereof.

[0064] The dry raw material may already be dry when receiving from a suitable source. The term 'dry' refers to a moisture content of between about 0-30%. Alternatively, it is possible to obtain wet raw material, for example, from a distillery process. If wet raw materials are obtained, preferably, the starting material is dehydrated / dried to a moisture content of between 0-30% prior to processing. The dehydration / drying may be by any conventional means. The dehydration can be, for example, between 40-80 °C for 12-48 hours.

[0065] Dehydration of the waste material prior to processing reduces the water activity and inhibits the growth of microorganisms and further, it enables dry grinding and / or milling of the raw material and food waste. Preferably, the material is dehydrated prior to milling / grinding. Enzymatic hydrolysis is used to break down the lignocellulose in the material. During enzymatic hydrolysis, the enzymes and the target regions of the substrate require direct physical contact. The enzymes therefore must be able to diffuse from the bulk aqueous solution to the particle surface, diffuse through the physical barrier present at the surface (lignin) and adsorb on the substrate surface, and then catalyse cellulose and hemicellulose hydrolysis.

[0066] Further, additional steps of enzymatic hydrolysis steps may be used before and / or after enzymatic degradation of lignocellulose in order to degrade e.g., phenolic crosslinking of arabinoxylan, such as that mediated by e.g., ferulic acid. Ferulic acid is a phytochemical i.e., compound found in plants. Ferulic acid has antioxidant properties and is known known for its ability to reduce the presence of free radicals and to boost the effects of other antioxidants, including vitamin A, vitamin C, and vitamin E. Most of the cellular ferulic acid is bound in arabinoxylan fibres, where it, amongst other compounds, mediates crosslinking of the arabinoxylan fibres. Accordingly, the benefits of releasing ferulic acid from the arabinoxylan fibres are two-fold; firstly, the hydrolysis of ferulic acid loosens the arabinoxylan fibres, resulting in e.g., a more efficient subsequent enzymatic hydrolysis of the plant fibres such as e.g., lignocellulose; secondly, the antioxidant properties of ferulic acid makes its release from arabinoxylan fibres favourable for its use e.g., in food products, highly beneficial since it may be used to enhance the shelf-life, protect oxidation labile compounds, or provide health benefits in the individual ingesting the food. Further, an enzymatic treatment of the pre-treated raw material obtained in step a)ii) may be inserted before the enzymatic hydrolysis of step b). As such the additional enzymatic treatment may be done as enzymatic treatment with e.g., a Feruloyl esterase (EC 3.1.1.73) such as e.g., NATE-1624 from Creative Enzymes, NY, USA. Alternatively, the additional enzymatic treatment may be carried out by e.g., lacto-fermentation using a strain expressing a feruloyl esterase, such as e.g., Lactobacillus Fermentum, or Lactobacillus Plantarum. Means for fermenting the pre-treated raw material obtained in step a)ii) by lacto-fermentation are well known in the art. Preferably, the additional enzymatic treatment is conducted before step b) of the process disclosed herein. In addition to a feruloyl esterase, additionally enzymes, such as e.g., xylanases (EC 3.2.1.8) and / or arabinofuranosidases (EC 3.2.1.55) may be added to degrade the arabinoxylan fibres. The current process has the advantage of first pre-treating the raw material comprising lignocellulose to break down the raw material into a powder comprising small particles between about 10-1000 pm and performing autohydrolysis. The sizes refer to the diameter of the particles e.g. as measured by their ability to pass through a sieve or filter having a defined size cutoff. The grinding and / or milling protocol allows decreasing the particle sizes, which in turn eases subsequent steps of the autoclaving and enzymatic treatments. Therefore, in a preferred pre-treatment, the raw material is first grinded and / or pulverized into a powder, which is followed by an autohydrolysis step.

[0067] However, the grinding and / or milling may also be performed after the autoclaving treatment step. Therefore, it is possible to first perform autoclaving on the raw material, followed by the grinding and / or milling steps. Preferably, if the autoclaving step is performed prior to the milling / grinding, the raw material is dehydrated prior to milling / grinding if the moisture content is above about 30%. Generally, the availability of arabinoxylan in the materials (e.g. BSG) which have been treated with the method of the first aspect of the invention, is markedly increased compared to the availability in the untreated materials; the same is true for the availability of cellobiose. Apart from the fact that both substances thereby are made available for further enzymatic degradation as described herein, these two substances are in their own right of value in the product obtained by the method of the first aspect of the invention, since they both are recognized prebiotic substances, meaning that the flour-like product and products comprising the flour-like product will act as beneficial prebiotics.

[0068] Typically, the raw material, such as BSG have grain sizes of around 280 to 10000 pm prior to processing. Other suitable raw materials, such as wheat bran typically have a maximum diameter of about 1500 pm such as e.g., 1497 pm prior to processing. Husks typically are in the range of 850 pm (diameter) prior to processing. Other raw materials, such as coffee ground typically have a maximum diameter of around 1000 pm; rapeseed press cake (or general press cake) typically are in the range of 600 pm; cereals are in the range of 10000 pm; pseudocereals are typically up to a maximum diameter of around 215 pm and sugarcane may be in the range of 1715 pm prior to processing. Due to the high variability of grain sizes, if various raw materials are pre-treated together, a uniform grain size may be obtained which facilitates the further processing of the raw material into useful upcycled products.

[0069] The powder or particulate material incapable of passing through a sieve having a mesh size of 10- 1000 pm may be discarded.

[0070] Alternatively, multiple subsequent grinding and / or pulverizing and sieving steps can be carried out in order to reduce waste. Multiple subsequent rounds of grinding and / or pulverizing and sieving allows more or all of the raw material to be broken down to be further processed, thereby reducing waste originating from the upcycled process.

[0071] The mesh size in step a)i) is preferably 15-950 pm, such as 20-900 pm, such as 25-850 pm, such as 30-800 pm, such as 35-750 pm, such as 40-650 pm, such as 50-600 pm, such as 60-550 p, such as 70-500 pm, such as 80-450 pm, such as 90-400 pm, such as about 100-300 pm, more preferably around 15-200 pm, such as 15-180 pm, such as 15-170 pm, such as 15-160 pm, such as 15-150 pm, such as 15-140 pm, such as 15-130 pm, such as 15-120 pm, such as 15-110 pm, such as 15-100 pm, more preferably around 15-90 pm, such as 15-80 pm, such as 15-70 pm, such as 15-60 pm, such as 15-50 pm, such as 15-40 pm, such as 15-30 pm, such as 15-20 pm, such as 15-15 pm, preferably between 15-30 pm , such as 16-29 pm , such as 17-28 pm, preferably around 70 pm.

[0072] The mesh size can be conveniently selected by the skilled person depending on the desired application of the resulting powder. For example, for applications requiring a finer particle size, such as for use as an additive or an ingredient where a smooth texture is required, mesh sizes of around 15-90 pm may be chosen. For other applications, such as coarse flour a larger mesh size may be chosen.

[0073] Autohydrolysis is used to reduce the molecular weight of cellulose and / or hemicellulose and to enhance cellulose and / or hemicellulose solubility, which consequently enhances the accessibility of the enzyme during the later enzymatic hydrolysis step. Autoclaving untangles the 3D network of cellulose, hemicellulose and lignin molecules without compromising the edibility and is a preferred treatment step to other possible methods, such as using strong acid / base treatments.

[0074] Step a)i) of the pre-treatment may further comprise a dehydration step. After the dehydration in step a)i), the moisture content of the raw material is reduced by 45-90% relative to initial moisture content of the raw material. The dehydration reduces the water content and consequently, the water activity of the waste material and inhibits the growth of microorganisms. Preferably, the dehydration is before the grinding and / or milling step to enable dry grinding and / or milling of the raw material and / or food waste.

[0075] If the grinding and / or pulverizing pre-treatment step is performed first, the obtained powder may be mixed with water at a ratio of 1: 1 to 1:6 prior to step a)ii). Otherwise, the raw dry material may be mixed with water at a ratio of 1: 1 to 1:6 prior to performing autohydrolysis.

[0076] Step i) of the pre-treatment comprises the steps of I) milling the dehydrated raw material and II) sifting the milled raw material through a sieve having a mesh size of 10-1000 pm and optionally, III) repeating the steps I) and II) steps at least once, preferably at least twice.

[0077] The pre-treatment step a)ii) comprises autoclaving or other heat or steam treatment or an acidic or alkaline treatment. The acidic treatment may utilize inorganic acids, such as phosphoric acid, nitric acid, hydrochloric acid and sulphuric acid or organic acids, such as acetic acid, citric acid or oxalic acid.

[0078] The ratio between the powder and water during the autoclave step (step a)ii)) is between 1: 1 to 1: 10, such as about 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, and about 1: 10, preferably about 1:3.

[0079] The duration of the autoclave step (step a)ii)) is between 10 minutes and 2 hours, such as about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, and about 120 minutes, preferably about 40 minutes.

[0080] The autoclaving or other heat or steam treatment, or the acidic or alkaline treatment may in some embodiments be performed at elevated pressure; this is known to enhance the effect of the heat treatment. The glucanase is in some embodiments an endo-l,3(4)-beta-glucanase and the xylanase may be an endo-l,4-beta-xylanase; in such embodiments, the enzymatic mixture may further comprise a p-glucosidase.

[0081] In additional embodiments, the enzyme mixture may further comprises one or more proteases, preferably one or more prolyl-specific proteases and / or one or more carboxy-specific protease.

[0082] Suitable and commercially available enzymes may be e.g. : Viscozyme® L and / or Ultraflo® Max from Novozymes, Bagsvaerd, Denmark, or e.g. ROHALASE® (AB enzyme); Axtra® PRIME (IFF).

[0083] A suitable enzyme class may be e.g., Enzyme class: 3.2.1.- and more specifically, hydrolases and glycosylases, i.e., enzymes hydrolysing O- and S-glycosyl compounds.

[0084] The glucanase may be p-l-4(3)-endoglucanase, which attacks regions of low crystallinity in the cellulose fibre creating free chain ends, p-l-4-exoglucanase or cellobiohydrolase degrades the molecule further by removing cellobiose units from the free chain ends, p-glucosidases or cellobiase hydrolyzes cellobiose to produce glucose.

[0085] The enzymatic mixture advantageously hydrolyses lignocellulose, making it suitable for e.g., human consumption. Humans and most mammals cannot readily break down cellulose, thus processing the raw material allows transformation of the lignocellulose into digestible and edible components such as simple sugars and reducing sugars such as glucose, galactose, fructose, lactose, xylose and maltose.

[0086] The enzymes may be added together in a single-step process; however, it is also possible to add the enzymes in a multi-step process, i.e., to first add the glucanase for a first enzymatic treatment step, followed by adding the xylanase in a second enzymatic treatment step or to first add the xylanase for a first enzymatic treatment step, followed by adding the glucanase in a second enzymatic treatment step. When carrying out a multi-step process, the addition of the enzymes may occur in any suitable order.

[0087] Additionally, the enzymatic hydrolysis may in some instances be done by fermentation, wherein the fermenting microorganism used expresses one or more hydrolytic enzymes as described herein. Accordingly, the enzymatic hydrolysis may be either a single step process comprising addition of the one or more enzymes, or it may be a fermentation process comprising addition of one or more microorganisms expressing one or more of the above-mentioned enzymes, such as e.g., Trichoderma reesei, which expresses and secretes a large amount of e.g., cellulolytic enzymes. In most instances the enzymatic hydrolysis as used herein, comprises the addition of a controlled amount of hydrolytic enzymes. The addition of a controlled amount of hydrolytic enzymes is beneficial over the fermentation with a microorganism expressing hydrolytic enzymes, since the addition of the hydrolytic enzymes allows for a more controlled and simple process, which for the current disclosure is beneficial for reproducibility of production of the flour-like product.

[0088] During the enzymatic hydrolysis, the mixture may be under constant agitation, which may be by any conventional means known in the art.

[0089] The enzymatic hydrolysis in step b) is followed by an inactivation of the enzymes which may be by heat or radiation, autoclaving at elevated pressure, or by changing the pH, such as by alkaline or acidic treatment. The inactivation may be by heating the mixture to at least 80°C.

[0090] Typically, 0.1-2% v / w enzyme mixture is added in step b).

[0091] After the enzymatic hydrolysis, the post-treatment step may comprise a, during which the moisture content of the raw material is reduced by 45-90% relative to initial moisture content of the material. The dehydration as a post-treatment step allows the inclusion of the product as an ingredient and allows control the water content for other recipes. Dehydration also inhibits the growth of microorganisms.

[0092] Often step b) is followed by step c) which comprises a post-treatment comprising the step of grinding and / or pulverizing the enzymatically hydrolysed mixture obtained from step b) to obtain a flour-like product. Additionally, step c) of the post-treatment further comprises a dehydration step before the grinding and / or pulverizing step. Typically, after the dehydration step, the moisture content of the enzymatically hydrolysed mixture is reduced to 0-30% w / w.

[0093] Following the enzymatic hydrolysis step, the dry material of the hydrolysed mixture has a tendency to aggregate into larger loosely packed particles. Accordingly, the inventors found that for the product to be useful in foods and a flour-like product, a post treatment comprising grinding and / or pulverizing was needed in order to reduce the particle size into a more suitable range. As is also stated herein, the suitable range of particles sizes is largely dependent on the use of the flour like product. When used in foods the desired size of the particles depends on the particular food. As is also mentioned earlier when used as a chocolate like product, it is preferable that the mean particle size is in the range of 5-20 pm, preferably 8-12 pm. Preferably, less than 10% of the particles are larger than 125um. In other food products, the preferred particle size may e.g., be in the range of 10-425 pm. In embodiments, the mean particle size of the grinded and / or pulverized flour-like product is in the range of 5-500 pm, such as 5-450 pm, such as 5-400 pm, such as 5- 350 pm, such as 5-300 pm, such as 5-250 pm, such as 5-200 pm, such as 5-150 pm, such as 5- 100 pm, such as 5-50 pm, such as 5-40 pm, such as 5-30 pm, such as about 5-25 pm, more preferably around 5-20 pm, such as 5-15 pm, such as 10-20 pm, such as 10-15 pm, preferably 7- 15 pm. In particular embodiments, at least 10% of the particles are smaller than 10 pm, at least 50 % of the particles are smaller than 25 pm, and at least 90% of the particles are smaller than 125 pm.

[0094] The process may further comprise the step of d) fermenting the product obtained from step c). In embodiments, the fermentation is a solid-state fermentation, a dry fermentation, a liquid fermentation, or a submerged fermentation.

[0095] Additionally, step b) may in some instances be substituted with a fermentation step, wherein the microorganism used in such a fermentation expresses one or more hydrolytic enzymes as described herein.

[0096] By fermenting the product, the appearance, texture and flavour of the product significantly develops. During fermentation, the microorganisms convert carbohydrates to alcohol or organic acids, typically under anaerobic or aerobic conditions, depending on the microorganism used. Further, during fermentation the simple sugars, proteins and fatty acids may be converted into aromatic volatile compounds, which provides for a different gastronomic profile and may enhance the flavour of the product.

[0097] The fermentation may be solid-state fermentation, dry fermentation, liquid fermentation or submerged fermentation. Preferably, the fermentation is solid-state fermentation.

[0098] The microorganisms used for the fermentation may be selected from the list consisting of Aspergillus ssp., Rhizopus spp., Neurospora spp., Lactobacillus spp., Streptococcus spp., Acetobacter spp., Aaccharomyces spp., Pichia spp., Gluconobacter spp., Leuconostoc spp., Saccharomyces spp., Pleurotus ostreatus, Lentinula edodes, Kluyveromyces spp., Scheffersomyces spp. and Spathaspora spp. Preferably, the microorganism is selected from the list consisting of Aspergillus oryzae, Aspergillus luechuensis, Saccharomyces cerevisiae, Brettanomyces bruxellensis, Torulaspora delbrueckii, Bacillus coaguians, Gluconacetobacter liquefaciens, Lactobacillus nagelii, Lactobacillus mail, Komagataeibacter, acetic acid bacteria (AAB), Zygosaccharomyces, Lachancea and Starmereiia and Rhizopus oiigosporus.

[0099] In one embodiment, the microorganisms used for fermentation are selected from the list of Aspergillus ssp., Rhizopus spp., Neurospora spp., Lactobacillus spp., Streptococcus spp., Acetobacter spp., Aaccharomyces spp., Pichia spp., Gluconobacter spp., Leuconostoc spp., Pleurotus ostreatus, Lentinula edodes, Kluyveromyces spp., Scheffersomyces spp., and Spathaspora spp, preferably, wherein microorganism is selected from Aspergillus oryzae, Aspergillus luechuensis, Saccharomyces cerevisiae, Brettanomyces bruxellensis, Torulaspora delbrueckii, Bacillus coaguians, Gluconacetobacter liquefaciens, Lactobacillus nagelii, Lactobacillus mali, Komagataeibacter, acetic acid bacteria (AAB), Zygosaccharomyces, Lachancea and Starmereiia and Rhizopus oligosporus.

[0100] In embodiments, the microorganism(s) used for the fermentation is / are Pichia kiuyveri and / or Kluyveromyces marxianus. Numerous strains of these microorganisms are commercially available. For instance, the Global Catalogue of Microorganisms currently lists 436 Pichia kiuyveri strains and 797 strains of Kluyveromyces marxianus. Each of these strains can be utilized in embodiments of the present invention.

[0101] In embodiments, the microorganism(s) used for the fermentation is Saccharomyces cerevisiae.

[0102] Further, the fermentation may be conducted as a symbiotic fermentation culture, such as e.g., a scoby culture, where the fermentation is carried out as a symbiotic fermentation culture. A scoby culture is generally considered to be a symbiotic fermentation culture comprising co-culturing of bacteria and yeast. Preferably, such a symbiotic fermentation culture comprises a combination of lactic acid bacteria, acetic acid bacteria and yeast. In embodiments, the bacteria is lactic acid bacteria and / or acetic acid bacteria. In embodiments the lactic acid bacteria is selected from strains of the Lactobacillus genus. In embodiments the acetic acid bacteria is selected from strains of the Acetobacter genus, Gluconobacter genus or Komagataeibacter genus. In embodiments, the yeast is selected from species of the Saccharomyces genus or Zygosaccharomyces genus.

[0103] Additionally, the fermentation may be a lacto fermentation, using lactic acid bacteria such as e.g., one or more lactic acid bacteria selected from one or more strains of the Lactobacillus genus. The fermented product obtained from d) may be further dehydrated and optionally grinded and / or pulverized prior to further processing or consumption. Preferably, the fermented product obtained from d) is further dehydrated and optionally grinded and / or pulverized.

[0104] To develop the flavours of the product produced in step c) or d), the product may be roasted. Such roasting is usually performed at temperatures ranging from 50-300°C, and generally creates new flavors and / or aromas in the product.

[0105] Depending on the intended use, the fermented product may be directly used without any further processing steps, or it may be further dehydrated and optionally grinded and / or pulverized prior to consumption or prior to adding the obtained material to any other food products as an ingredient.

[0106] The product obtained from step b), c) and / or d) finds several uses in the preparation of foods and beverages, such as those disclosed herein. While several other uses will be evident to the skilled person, the present disclosure only provides non-limiting examples of the use of the product produced by the process described herein. The product obtained from step b), c) and / or d) of the process disclosed herein may for instance be used in a brewing process. For instance, the enzyme hydrolysed product may either be subjected to further treatment, such as in step c), but it may also be used directly after the enzymatic treatment. Preferably, the enzymes present in the enzymatically treated mixture are inactivated before use in a brewing process. Such inactivation may e.g., be heat inactivation. The product from step c) or the inactivated product from step b) may e.g., be added in the mashing step and / or fermentation step of the brewing process, or it may be added directly to the wort before fermentation. Usually, when preparing alcoholic beverages the degrees Plato (°P) is about 9-16 °P in the wort before initiating fermentation. Further, when preparing non-alcoholic beverages the degrees Plato (°P) is usually about 1-9 °P before initiating fermentation.

[0107] Typically, when used in a brewing process for preparing alcoholic beverages, the product from step b), c) or d) is added to the mashing step of the brewing process at a content of 1-90%. Normally the ingredients for a brewing process would be 1-90% of the product from step b), c) or d), 0- 75% malt, 5-10% sucrose, 10-80% water, 1-5% hops and 0.05-1% yeast / bacteria for the fermentation. Additionally, adjuncts such as e.g., oats or wheat may be added to enhance the mouthfeel and taste of the beer.

[0108] In embodiments, the microorganism(s) used for the brewing process for preparing alcoholic beverages are selected from the group consisting of Saccharomyces cerevisiae, Kluyveromyces spp., Brettanomyces bruxellensis, Torulaspora delbrueckii, Scheffersomyces spp., Pichia spp. and Spathaspora spp. In particular embodiments, the preparation of an alcoholic beverage comprise a first fermentation step of the flour-like product from step b) with A. oryzae in a solid state fermentation. The produced fermented with A. oryzae may then be roasted and / or grinded before further use in the preparation of an alcoholic beverage. As such the use of the roasted / grinded product in the further preparation of an alcoholic beverage is two-fold. In one embodiment, the roasted / grinded product is used in a maceration step following an alcoholic fermentation, to extract aromas and flavors from the product. In another embodiment, the product is used directly in the fermentation. Preferably for the preparation of an alcoholic beverage the fermentation is carried out by culturing of Saccharomyces cerevisiae in a suitable medium.

[0109] Additionally, low or non-alcoholic beverages may also be produced using the product obtained in step b), c) or d) of the disclosed process. In a brewing process for preparing non-alcoholic beverages, the product obtained in step b), c) or d) may be the primary source. In embodiments, the process for preparing non-alcoholic beverages does not comprise addition of malt. In other embodiments, the process for preparing non-alcoholic beverages comprise addition of minor abouts of malt, such as below 10% malt (w / w%). Preferably, the process for preparing nonalcoholic beverages comprise addition of less than 25% malt (w / w%), more preferably less than 10%, most preferably less than 1%. When preparing an alcoholic beverage, it is preferable that the yeast is a monosaccharide fermenting yeast, such as a glucose fermenting yeast. Additionally, it is preferable that the fermentation for preparing a non-alcoholic beverage is an aerobic fermentation. As such, many yeast strains suitable for carrying out the non-alcoholic fermentation will be known to the skilled person, but it is preferable that fermentation with oxygenation gives rise to a non-ethanol producing metabolic state in the yeast. The metabolic state obtained by oxygenation usually results in a preference of the yeast for monosaccharide fermentation over disaccharide fermentation, leading to low or no ethanol production. Yeasts that are especially suited for preparing low or non-alcoholic beverages are e.g., yeasts of the genus Pichia spp., preferably Pichia kluyveri, cf. the discussion above of this microorganism.

[0110] Usually, by the end of the fermentation the alcoholic beverage contains about 1.5-12% ABV, while a non-alcoholic beverage usually contains about 0-0.5% ABV. As is disclosed in the examples, the process disclosed herein may result in an end non-alcoholic beverage, prepared from e.g., brewers spent grain, which is unique in the sense that it contains a very low amount of maltose, such as e.g., below 0.5 g / 100 g maltose, or lower. This effect is obtained specifically from the specific processing of the BSG disclosed herein. This is particularly beneficial, since a common issue with non-alcoholic beverages is the sweet taste due to disaccharides such as maltose. Accordingly, in some aspects the present disclosure also relates to a non-alcoholic beverage prepared using a flour-like product as defined herein may comprises less than 0.2 g / 100 g maltose. Accordingly, the disclosure also provides a fermented beverage wherein the fermentation broth at the end of the fermentation comprises less than 0.2 g / 10 0g maltose and less than 1% ABV of ethanol, and wherein the microorganism used in the fermentation is of the genus Pichia spp. Preferably the non-alcoholic fermentation is performed with the microorganism such as Pichia kluyveri, cf. the discussion above of this microorganism.

[0111] Another benefit of preparing non-alcoholic beverages from the product obtained in step b), c) or d) is that the presence of disaccharides is very low or even absent in the produced product. A common issue with non-alcoholic beverages is the sweet taste obtained from the presence of disaccharides, such as maltose. When starting the production process from malt and fermenting the produced wort using the non-disaccharide utilizing yeast strains, the disaccharides will often have to be removed via. additional processing and / or enzymatic steps. The product obtained in step b), c) or d) avoids this additional processing due to the initial absence of e.g., maltose in the enzymatically treated product, as can be seen in the table presented in Example 13. Thereby the enzymatically treated product disclosed herein provides the means for obtaining a less sweet nonalcoholic beverage, compare to that obtained from mashing of malt.

[0112] Additionally, the absence of disaccharides also minimize the risk of pathogen infection of the nonalcoholic beverage. As is evident, the use of the enzymatically treated product as described herein in a brewing process enables the upcycling of e.g., BSG, by reintroducing the spend grain into the brewing process, thereby reducing the need for grain in the brewing process. Such upcycling leads to a reduction in the overall cost of the brewing process.

[0113] The invention also relates to a process for producing chocolate-like product, the process comprising the steps of: 1) obtaining a product from the process as disclosed herein, followed by 2) optionally roasting the product of 1) at a temperature between 130-160 °C, 3) Refining the roasted product from step 2) with sugar and lecithin, 4) Adding 13-35% by weight fats to the refined product of step 3) to obtain a mixture, 5) Conching the mixture of step 4) for at least 6 hours and up to 30 hours, 6) Pouring the conched mixture of step 5) into moulds and 7) Cooling the mixture of step 6) to a temperature of 0-15°C.

[0114] The final product may be conveniently stored at a temperature of 0-25°C.

[0115] Conching the mixture enables a smaller particle size and thereby provides better smoothness of the flour-like product. This increases consumer acceptability and increases the inclusion rate of the flour-like product in the final formulation.

[0116] The butter may be selected from the list comprising mango butter, shea butter, coconut butter, dairy butter, such as salted or unsalted butter, ghee, brown butter, clarified butterm cultured butter, palm oil, illipe butter, sal butter, kokum butter, cacao butter, nut butters, such as almond nut butter, cashew butter, peanut butter, walnut butter, sunflower seed butter, brazil nut butter, hazelnut butter, macadamia butter, pecan butter, pistachio butter and pine nut butter.

[0117] A chocolate-like product may comprise 20-45% by weight flour-like product obtained at the end of step c) and / or fermented product obtained at the end of step d), 10-25% by weight sugar, 25- 50% by weight butter and 0.1-1.2% by weight lecithin, however the skilled person understands that variations may apply without departing from the scope of the invention.

[0118] Further, it is preferable that a chocolate like product may comprise 10-25% by weight sugar, 25- 50% by weight fats, 0.1-1.2% by weight lecithin and 0-0.01% by weight theobromine. Such a product preferably comprises 5-50% by weight of the flour-like product defined herein. To maintain its appeal it is preferable that the taste of the chocolate-like product is improved when a flour-like product obtained at the end of step c) and / or fermented product obtained at the end of step d) is used compared to addition of an unprocessed raw material.

[0119] Generally, chocolate is a well-defined product. In most parts of the world there is legally defined parameters for when and how a product can be defined as Chocolate. The legal definitions depend, among other factors, on levels and ratios of dried cocoa solids, cocoa fat and milk powder. Accordingly, chocolate based on cocoa beans has several characteristics, both legal, sensory and physical. The physical characteristics are generally a melting point of 30-50°C and a snap of 1-300 MPa along with its nutritional values i.e., the content of fat, sugar and protein. The sensory characteristics of chocolate are e.g., the flavor-palette, the mouthfeel (smooth, sandy, dusty) and the visual appearance of the coating. The more legal characteristics are e.g., the EU defined minimum content of cocoa powder of 16% dried cocoa powder and minimum content of fat of 25% (cocoa butter and milk fat).

[0120] As such, a chocolate-like product as disclosed herein may have both similar and different physical characteristics and still reach a similar sensory impression.

[0121] One characteristic of chocolate is the is the snap. The snap of real chocolate varies and depends on the ratio of cocoa butter, fats and level of dry products. It can be measured and defined by a Three- point bend, which is a standard test used to measure the flexural strength of a material. The process is as described by ISO 178, also mentioned above.

[0122] In brief, the three-point bending fixture is utilized to assess the rigidity properties of hard products, metrically positioned on the three-point bending fixture, including chocolate.

[0123] Generally, the sample tested such as a chocolate-like product, e.g., a bar, is symmetrical to ensure force is applied at the center of the sample. A compression of 2 cm is then applied at a rate of 0.5 mm / s to break the sample. This device TX-700 with 3 points bending fixture (Lamy Rheology, Champagne au Mont d'Or, France) enables the analysis of various hard products. The adjustable bench allows for the characterization of different types of chocolate bars (dark or white, thick or thin) using a single cell. For chocolate like products, it is preferable that the snap is in the range of 1-50N, when measured as described above.

[0124] In embodiments, a chocolate-like product described herein exhibits a snap at an applied pressure of 1-500 MPa when shaped as a product which is 70-100 mm long, 30-60 mm wide and 5-15 mm thick, such as a bar, and the snap is measured according to the three-point test of standard ISO 178. In particular embodiments, the snap is measured on a product, which is 95 mm long, 40 mm wide and 10 mm thick.

[0125] One limitation of real chocolate is, that it needs to be stored at a fixed temperature range I order to develop fat bloom or melt. The melting point is generally governed by a combination of particle size along with the fat content and fat type, and is generally for most chocolates in the range of 20- 40°C.

[0126] One of the benefits of a chocolate-like product as disclosed herein is that the melting point can be managed by addition of specific fats to the product, which increases the melting point, and enables the storage of the chocolate-like product at higher temperatures then what is possible for normal chocolate.

[0127] As is described in e.g., Example 14, the particle size of the flour-like product used to prepare the chocolate-like product greatly influences the mouthfeel of the produced product. Accordingly, it is preferable that when used for chocolate-like products the flour-like product has a mean particle size of less than 30um, where at least 90% of the particles are smaller than 125 pm, and at least 10% of the particles being smaller than 5 pm. In embodiments, at least 90% of the particles are smaller than 125 pm. In embodiments, at least 50% of the particles are smaller than 25 pm. In embodiments, at least 10% of the particles are smaller than 5 pm. The particle size may be evaluated using various method know to the skilled person, but is preferably evaluated using a Mastersizer apparatus. As is shown in the examples, the smaller particle size obtained following the enzymatic treatment correlated with an improved mouth feel for the chocolate-like product, compared to products using non-enzymatically treated flour-like products as ingredient.

[0128] Another benefit of the chocolate-like products described herein is that the nutritional values can be tailored, and the chocolate-like products may e.g., have a higher fiber content, a higher protein content etc., while maintaining the sensory characteristics of real chocolate. In particular embodiments, the chocolate-like product is a bar. Chocolate like bars is described in e.g., example 3, which describes the production of a chocolate-like bar, therein termed BSG-bar. A chocolate-like bar may be prepared using the product obtained from step b), c) or d) of the process described herein. Preferably, the chocolate-like product comprises 30% by weight flour-like product obtained at the end of step c) and / or fermented product obtained at the end of step d), 20% sugar, 32.6% mango butter, 16.7% shea butter and 0.7% lecithin.

[0129] The chocolate-like product resembles traditional chocolate in terms of taste, texture and consistency and provides the same or very similar sensory experience as traditionally prepared chocolate.

[0130] Besides resembling traditional chocolate, a chocolate-like product produced as described herein also exhibits several key differences compared with traditional chocolate. For instance, as is shown in Example 13, a chocolate-like product produced from the treated BSG contains no, or very little (less than 3mg / 100g) theobromine. Theobromine, also known as xantheose, is the principal alkaloid of the cacao plant, Theobroma cacao, and is the reason for traditional chocolate being poisonous to animals such as e.g., dogs, due to slow metabolism of theobromine. Especially dogs are often poisoned by ingestion of chocolate, since dogs express sweet taste receptors, and are therefore more likely to eat chocolate due to the sweet taste, as compared to cats which does not express such receptors. Accordingly, the absence of theobromine in the end product is highly favourable, since it allows for the use of the product in animal feed or snacks. Thus, the produced product may be used in animal feed and / or snacks. In some embodiments, the chocolate-like product as defined herein may be in the form of a pet treat, such as a dog treat.

[0131] An edible product may comprise 0.1-100% dry matter by weight flour-like product obtained at the end of step c) and / or the fermented product obtained at the end of step d).

[0132] It is possible to combine the two products, i.e., the flour-like product obtained at the end of step c) and the fermented product obtained at the end of step d) in a single final edible product. In embodiments, the fermented product after step d) is a beverage, such as an alcoholic or nonalcoholic beverage. As is evident the present disclosure also relates to a process for preparing a beverage comprising fermentation by microorganisms of a carbohydrate-containing substrate, which comprises or consists of a flour-like product obtainable by the process as defined herein. In embodiments, the beverage is alcoholic, and the microorganisms convert carbohydrates in the substrate to produce ethanol as a metabolite. In other embodiments, the beverage is non-alcoholic and the microorganisms metabolize carbohydrates in the substrate to produce non-alcoholic metabolites. In particular embodiments, the beverage is a beer. In further embodiments, the beverage is a non-alcoholic beverage. In preferred embodiments, the non-alcoholic beverage is such as a non-alcoholic beer.

[0133] The edible product may the edible product is selected from the group comprising a chocolate or confectionary product; a spreadable spent grain or a chocolate-like coating or coverage; a cacao powder replacement product; a baked or cooked good such as a pasta, a dough or a bakery product; a beverage or drink powder; an alcoholic drink or beverage; a non-alcoholic drink or beverage; a frozen-foodstuff such as ice-cream; an infant nutritional formula; a canned soup, broth or product; a condiment; nutraceutical product such as an energy bar, a protein shake, a dehydrated complete meal package, a smoothie or smoothie bowl; an extruded product, such as a breakfast cereal product or pasta products; and a thickening agent, such as starch.

[0134] The skilled person understands that the flour-like product obtained at the end of step c) and / or the fermented product obtained at the end of step d) may be added to any food product to replace conventionally used materials, such as flour partly, or entirely, or to be used as additives in addition to conventionally prepared food products. Therefore, convenient uses for the flour-like product obtained at the end of step c) and / or the fermented product obtained at the end of step d) may be as additives to conventional food products or as replacement for traditional food products, such as flour in e.g., pasta or bakery products. Further, convenient uses for the product may be e.g., as a feed product or additive or replacement in a feed product, such as in infant formulas. The product obtained in step b), c) or d) further finds uses as flavor and / or aroma in different products, such as e.g., beer, bread, cookies, wine, spirits, soups, broths, emulsions, ice creams and sorbets, confectioneries, marinades, sausages and / or hydrolysates. Usually when used to confer flavor and / or aroma, the flour-like product is used in the range of 0.5-75% w / w of the intended product.

[0135] Therefore, present disclosure also relates to the use of the flour-like product obtained at the end of step c) and / or the fermented product obtained at the end of step d) as an additive and / or replacement for other conventional food ingredients or compounds. Further, the flour-like product may be for use in cosmetics, dietary supplements and / or nutraceuticals

[0136] EXAMPLES

[0137] EXAMPLE 1 - Flour-like product produced from brewer's spent grain

[0138] Pre-treatment of brewer's spent grain

[0139] Wet Brewer's spent grain (BSG) of "Grist bill" malting (consisting of 13 malts: Bonsak N special, Pilsner, Whey Cara red, Caramel, Chocolate, Black, Carafa special I, Carafa special II, Biscuit, Crystal, CM special, Ruby, and Cafe) was provided by the Carlsberg Research Laboratory (Copenhagen, Denmark).

[0140] Immediately after receiving the unroasted, fresh, and moist BSG, it was weighed to determine its moisture content. Then, it was vacuum packed at 99% (Henkelman® vacuum packaging machines, Neo model) and kept at -20°C for future use. The untreated sample was called Bo.

[0141] Dehydration

[0142] The wet BSG at room temperature was dehydrated in an Excalibur dehydrator machine at 60 ±5°C for 46 hours. To ensure homogeneous dehydration and preventing future microorganism growth, the BSG was placed in layers of 1.5 cm on the dehydrator trays (wax paper was placed on the dehydrating trays to avoid sticking) before dehydration.

[0143] Samples were weighed in triplicates before and after dehydration to determine the moisture lost; the following formula was applied:

[0144] (SW-FW) *100 / SW,

[0145] - where SW and FW are the starting and final BSG weights, respectively. According to the invention, the dehydration step is generally optional, but was carried out in this particular example.

[0146] Milling and sifting

[0147] After completing dehydration, 79.64% of humidity (SD 1.59) was lost in relation to the initial weight. Then, the dehydrated BSG was milled in a Komo Fidibus 21 mill and sifted with a fine metallic pastry sieve (40 mesh aperture or 0.425 mm). The non-sifted fibre was discarded from the samples to ensure the texture softness of the spent grain flour-like product. The BSG powder was vacuumed, packed at 99% with anti-humidity tablets (Silicasec; Ingenios, Albert y Ferran Adria, Spain) and stored in a dry place at room temperature. The particle sizes were between 10- 1,000 pm.

[0148] The flour that passed through the sieve had a darker colour and toasted aromas, compared to the non-sifted fibre, which had a lighter colour and a straw-like aroma. The non-sifted fibre may be remilled and re-sifted and subseguently added to the sifted batch in order to reduce waste.

[0149] Autohydrolysis

[0150] To perform the autohydrolysis step, the spent grain flour was mixed with H2O at a ratio of 1:3 respectively, then the resulting mixture was autoclaved at 2.7 bar and 140°C for 40 min (CertoClav, EL model). Common filtered water was used during the entire experiment. When the time was up, the sample was allowed to cool freely. The resulted sample, labelled as "Bl" was cooled and kept at -20°C for future use.

[0151] Enzymatic treatment

[0152] To perform the enzymatic hydrolysis, 30% (w / w) of water was added to the previously pre-treated material. This mixture was then placed into a Thermomix® to maintain it in constant agitation during the reaction (speed level 3). With the help of a micropipette, 2 ml (0.1-2% v / w per kg of mixture) of food grade enzyme concentrate Viscozyme® L (Beta-glucanase (endo-l,3(4)-, pectinases, hemicellulases and xylanases) and 2 ml of concentrate Ultraflo® Max (Xylanase (endo- 1,4-) Beta-glucanase (endo-l,3(4)-) (from Novozymes®, Bagsvaerd, Denmark) were added to each kg of mixture. Afterwards, the mixture was incubated for 5 h at 50°C with pH at 4.9 - 5. After this, the enzymes were inactivated by heating the mixture at 95°C for 15 min (according to the manufacturer's recommendation). The BSG after the enzymatic process was labelled as "B2".

[0153] Dehydration and grinding The dehydration step - and / or the step of subsequent grinding - can as explained above in some embodiments be omitted depending on the desired properties and intended / practical use of the end product; if the enzyme-treated product e.g. can be fed into a process or product without either or both of these steps need to be carried out, there is not a practical reason to dehydrate or grind. But in the present example, it is demonstrated that a flour-like product useful in a variety of settings can be prepared when dehydrating and grinding.

[0154] Dehydration

[0155] After the enzymatic hydrolysis, the BSG was dehydrated at range from 45 to 70 ±5°C (optimum was found to be around 55°C) for 4-8 h (stirring every 30 min to homogenize it) until it reached a consistency that simulated cooked grains. This dehydrated, flour-like product is typically used as a substrate for the solid fermentation stage, however the non-dehydrated product may also be used.

[0156] Grinding

[0157] The spent grain flour-like product is grinded for 30 minutes to 6 h in order to obtain a homogenous powder. If dehydration is employed prior to the dehydration step, the layers of spent grain flour-like product tend to coalesce into a block. Grinding helps re-obtain the powder form and ensures that the particle size remains in about 10-1000 pm.

[0158] Depending on the further use, the grain size may be for example 15-50 pm, or 5-20 pm for a chocolate-like product or may be in the range of 15-425 pm for other products, which allow for coarser particles.

[0159] EXAMPLE 2 - Solid state fermentation of the spent grain flour-like product

[0160] Solid state fermentation

[0161] For the solid-state fermentation, the flour-like product of Example 1 was used after dehydration.

[0162] Before fermentation, the stored dehydrated and grinded B2 sample of Example 1 was pasteurized at 110°C for 15 min and allowed to cool down to 30 °C before inoculating. This bagasse paste was divided into smaller batches of samples.

[0163] The sample labelled as "F2" resulted from the inoculation with 0.01% (w / w) koji spores (Aspergillus oryzae; white barley koji from Higuchi Matsunosuke Shoten Co., Osaka, Japan) and incubated during 36 h at 33°C, and relative humidity of 70% to 75%. The sample (labelled as "F3") resulted from the inoculation with 0.5% (w / w) tempeh spores Rhizopus oligosporus; tempeh starter from TopCultures Zoersel, Belgium) for 36 h at 33°C and relative humidity 70% - 75%.

[0164] Results The reducing sugar content of the samples (BO: raw or untreated, Bl: autoclave, B2: enzymes, and F2 or F3: fermentation with Aspergillus oryzae or Rhizopus oligosporus, respectively) was determined by using DNS method and HPLC analysis was performed to determine and quantify the concentration (mg / g) of glucose, fructose, and maltose present in each sample from the raw sample (BO) to the F2 and F3 samples. Other saccharides present in the samples (e. g., arabinose, xylose, mannose, etc.) were not detected due to limitations of standards of the calibration curve. The big difference between concentration on sugar reduction and the total concentration of glucose, fructose and maltose provides information about a significant level of cellulose degradation.

[0165] The results are shown in Table 1 below: Table 1. Glucose, fructose, and maltose concentration determined by HPLC and total reducing sugars concentration determined by DNS method of the samples taken from each step process of Stage I. Results of one-way ANOVA and post-hoc analysis (Tukey HSD) of each sample; means, standard deviation and p-value are showed. When comparing the raw sample (BO) with the autoclaved sample (autohydrolysis) (Bl), there is not a significant difference regarding glucose content, but there is a significant difference in the reduction of fructose and maltose after the autoclaving step. This is expected, as it was previously reported that sugar solutions subjected to high temperatures and pressures, the e.g., maltose content decreases while some organic acid contents increase (Woo et al., 2015) The total reducing sugar content was slightly increased (around 1.4 times) when the raw BSG was thermally treated in the autoclave step (Bl sample).

[0166] Significant differences were obtained between the Raw sample and the Enzymes sample. Eight times more glucose was obtained from raw spent grain after autohydrolysis and enzymatic hydrolysis; there is not significant difference in fructose concentration during enzymatic reaction because fructose is a monosaccharide. In contrast, a reduction of maltose after enzymatic treatment is significantly different compared to the Raw sample (0.35 mg / g). The reduction of maltose is in accordance with glucose concentration increment in Enzymes sample due to maltose is a glucose disaccharide. Another significative difference is regarding reducing sugars, after autoclave and enzymatic hydrolysis the reducing sugars' concentration improved three times (4.242 mg / g).

[0167] To conclude, due to the combined steps of autohydrolysis, enzymatic processes, and fermentation, it was possible to increase the nutritional value of BSG.

[0168] EXAMPLE 3 - Chocolate-like product and confectionary comprising the chocolate-like product

[0169] To create the BSG-Bar described below, the spent grain was considered a defatted Cocoa Powder. The prototypes were made with the Bl, F2 and F3 samples following the formulation showed in Table 2. B2 (enzymatically treated spent grain flour-like product from example 1) was dehydrated, milled and sifted to obtain a flour-like powder, F2 and F3 (fermentation with Aspergillus oryzae and Rhizopus oligosporus, respectively) were dehydrated after fermentation.

[0170] Before the fermentation, the stored sample may be pasteurized at 110°C for 15 min and allowed to cool down to 30°C before inoculating. The fermentation is typically carried out from about 24 to 36 hours (depending on the microorganisms employed) and typically at around 31°C and around 80% humidity. Table 2. Formulation of BSG BSG-Bar.

[0171] Amount % Ingredient

[0172] 30% BSG (Bi , F; or F;

[0173] 20% Sugar

[0174] 32.6% Mango buter

[0175] 16.7% Shea buter

[0176] 0.7% Lecithin

[0177] To develop the aromas, samples were roasted separately at 140°C for 3 min in an oven (stirring every 40 seconds). Then, each sample was refined with 30% of regular sugar into the Wonder Wet Grinder PG 503 by Premiere (200 - 240 V AC, 50 Hz, 1 phase, 180 W-1.5 L -110 V, 60 Hz). After about 50 min, soy lecithin (Lecite, by Ingenios Albert and Ferran Adria, Catalonia, Spain) was added and then mixed for 10 mins more (total time of refining 1 h). During this step, each sample is also refined with around 30% w / w sugar in a wet grinder.

[0178] Mango butter (by Hedenhus, Spottrup, Denmark) and shea butter (kindly provided by Fuji Oil holding Inc.) were melted separately over an electric cooktop up to 80°C, keeping the temperature for 30 min to erase fat crystallization memory. Subsequently, the mango butter was added slowly to the mixture in the conching machine and, immediately after, the shea butter was added in the same way. The whole mixture was left conching for 24 h. The resulted mixture was already the chocolate-like product.

[0179] The particle size of the chocolate-like product was analysed with a grindometer (ISO Standard One Channel QXD-100 by Baoshishan). The chocolate-like product was stirred until it reached a temperature of 32°C; subsequently, the mixture was poured into moulds and kept in a dry place at 4°C before demoulding. The resulted chocolate-like product was named "BSG-Bars".

[0180] Sensory analysis

[0181] Chocolate-like product

[0182] The sensory analysis of the BSG-bars was carried out with four samples of the BSG-bars, three of them with different concentrations of the mango and illipe butter, Recipe 1 - 38.6% Illipe butter + 10.7% mango butter - Recipe 2 - 32.6% illipe butter + 16.7% mango butter - Recipe 3: 38% illipe butter + 11.3% mango butter - and the fourth one with a mixture of shea, mango and illipe butter, Recipe 4 - 25% illipe butter, 14.3% mango butter + 10% shea butter. All the chocolates had been made following the recipe described earlier in the Brewer's spent grain chocolate section using the B2 BSG product. The study was conducted with 60 people from the restaurant Alchemist (Copenhagen, Denmark).

[0183] The samples were presented in semi-sphere shapes using a silicone mold, with cavities of 15 mm from (Silikomart, Venice, Italy). These samples had a determined weight of 4 grams each. Each chocolate had a coding based on 3-digit random numbers: 386 (Recipe 1), 971 (Recipe 2), 154 (Recipe 3) and 623 (Recipe 4). The order in which the samples were presented to the participants was also randomized. The results are presented in Table 3 below.

[0184] Table 3. Summary of the ANOVA carried out for the chocolate samples.

[0185] Note. Summary of the ANOVA test carried out for the four samples based on the sensory analysis responses, own elaboration. The scale was 1-9 points with 1 being 'extremely dislike' and 9 being 'extremely like'.

[0186] As an alternative to the sensory perception of the snap, the snap of the chocolate-like product may also be evaluated by a 3-point measurement, as described herein, where the actual snappoint is measured by applying force to the chocolate-like product, and comparing the obtained snap-point to traditional chocolate bars of the same or similar type.

[0187] EXAMPLE 4 - spreadable spent grain, such as hazelnut cocoa spread or coating / covering chocolate-like product

[0188] First, a chocolate-like product was prepared following the protocols of Example 3:

[0189] Solid state fermentation of the spent grain flour-like product

[0190] The spent grain flour-like product hydrated with 90% of its weight in distilled water and the hydrated mixture was pasteurized at 110°C for 15 min and allowed to cool down to 30°C before inoculating. The fermentation can carried out with different fungi and bacteria including but not limited to Aspergillus ssp., Rhizopus spp., Neurospora spp., Lactobacillus spp., Streptococcus spp., Acetobacter spp., Saccharomyces spp., Pichia spp., Gluconobacter spp., Leuconostoc spp., Pleurotus ostreatus, and Lentinula edodes.

[0191] The fermentation in this example was carried out with Pichia kiuyveri for 36 h at 25°C degrees, with rigorous shaking securing good aeration and Dissolved oxygen at >8 ppm and at about 80 % of humidity in incubation chamber.

[0192] Roasting

[0193] To develop the aromas, samples were roasted separately at 150°C for 20 min in an oven (occasionally mixing the powder securing the roasting is uniform)).

[0194] Stone-grinder / melanger / Conching

[0195] Then, each sample was refined with 0.1-60% (optimum about 30% w / w) of regular sugar in a Wet Grinder. After about 24 mins, soy lecithin was added and then mixed for about 25 mins more (total time of refining 1 h).

[0196] For this recipe, mango butter and shea butter were used (about 40% w / w comprising 66% mango butter and 33% shea butter).

[0197] The solid fats were melted separately over an electric cooktop up to 80°C, keeping the temperature for 30 min to erase fat crystallization memory. Subsequently, the mango butter was added slowly to the mixture in the conching machine and, immediately after, the shea butter was added in the same way. The whole mixture was left conching for 24 h. The resulting mixture was already the chocolate substitute.

[0198] In another recipe, in a concher, sucrose, trehalose, and skim milk powder were added for 1 h and the chocolate substitute and hazelnut pralines were added for another 1 h. To this mixture, oil and lecithin was further added and the mixture was conched for a further 5 h. The final product is kept at 4°C.

[0199] The amounts of ingredients for a spreadable spent grain, such as hazelnut cocoa spread are summarised below (w / w%):

[0200] - 25% Hazelnut praline

[0201] - 20% Chocolate substitute according to the invention

[0202] - 10% Sucrose / sugar

[0203] - 8% Trehalose

[0204] - 14% skim milk powder - 23% Sunflower oil

[0205] - 0.3% Lecithin.

[0206] The above amounts of ingredients are merely exemplary; for example, the amounts of fats or oils, sugars, milk powder, etc. are to be adapted to match the spent grain flour-like product concentration. Other suitable ingredients used in traditional or alternative chocolate-making or hazelnut cocoa spreads can also be incorporated.

[0207] Sensory analysis

[0208] The sensory analysis of the chocolate-like product-based hazelnut cocoa spread was carried out with two samples of the hazelnut cocoa spread, the two of them with different concentrations of the chocolate-like product base: Recipe 1: 45% and Recipe 2: 70%. All the chocolates-like product and hazelnut cocoa spread products have been made following the recipe described herein. The study was conducted with 40 people from the restaurant Alchemist (Copenhagen, Denmark).

[0209] The samples were presented in black plastic spoons. These samples had a pre-determined weight of five grams each. Each hazelnut cocoa spread product presented had a coding based on 3-digit random numbers: 271 (Recipe 1), and 103 (Recipe 2). The order in which the samples were presented to the participants was also randomized. The analysis consisted in two parts, first a 9- point hedonic scale based on the liking of the product and a Check-all-that-apply questionnaire (CATA). The results are tabulated in Table 4.

[0210] Table 4. Hazelnut cocoa spread liking results

[0211] Variable 103 271 p-values

[0212] TEXTURE 6.30±1.62 5.50±1.92 0.048

[0213] APPEARANCE 6.22±1.70 6.25±1.48 0.944

[0214] FLAVOR 6.90±1.43 7.34±0.93 0.081

[0215] Note. Summary of the ANOVA test carried out for the four samples based on the sensory analysis responses, own elaboration. The scale was 1-9 points with 1 being 'extremely dislike' and 9 being 'extremely like'.

[0216] EXAMPLE 5 - powders, such as cacao-powder substitutes (0.1-100% dry matter by weight spent grain flour-like product and / or fermented product)

[0217] First, a chocolate-like product was prepared following the protocols of Example 3, i.e. :

[0218] Solid state fermentation of the spent grain flour- like product (optional)

[0219] The spent grain flour-like product was pasteurized at 110°C for 15 min and allowed to cool down to 30°C before inoculating. The fermentation is carried out with different fungi and bacteria including but not limited to Aspergillus ssp., Rhizopus spp., Neurospora spp., Lactobacillus spp. Streptococcus spp., Acetobacter spp., Aaccharomyces spp., Pichia spp., Gluconobacter spp., Leuconostoc spp., Pleurotus ostreatus, and Lentinula edodes.

[0220] The fermentation was carried out from 24 to 36 h (depending on the microorganisms) at 31°C degrees, and at about 80 % of humidity.

[0221] Roasting (optional)

[0222] To develop the aromas, samples were roasted separately at 140°C for 3 min in an oven (stirring every 40 s).

[0223] Stone-grinder / melanger / Conching

[0224] Then, each sample was refined with 0.1-60% (optimum about 30 % w / w) of regular sugar into a Wet Grinder. After about 1-2 hours, milk powder, vanilla essence and lecithin was added and then mixed for about 10 mins more. The mixed product can then be dissolved (approx. 2-10 g I 200 mL) hot or cold milk or water for consumption.

[0225] The amounts of ingredients for an exemplary cacao-powder substitute are summarised below (w / w%):

[0226] - 50% Chocolate-like product according to the invention

[0227] - 33% Sucrose / sugar

[0228] - 16.5% milk powder

[0229] - 0.33% Vanilla essence

[0230] - 0.50% Lecithin.

[0231] EXAMPLE 6 - Alcoholic beverages

[0232] The present inventors have realised that the spent grain flour-like product surprisingly can also be used for the production of alcoholic beverages, such as spirits, beer and the like using alcoholic fermentation.

[0233] The spent grain flour-like product may directly be added to any alcoholic beverage for alcoholic fermentation.

[0234] The spent grain flour-like product obtained in Example 1 is in this example further processed by: Solid fermentation

[0235] Before fermentation, the stored sample was pasteurized at 110 °C for 15 min and allowed to cool down to 30°C before inoculating. The fermentation was carried out with Aspergillus oryzae. The fermentation is around 36 h (depending on the microorganisms) at is typically at around 31°C degrees, and 80 % of humidity.

[0236] Roasting

[0237] The spent grain flour-like product after the fermentation is roasted at 140°C for 2 to 10 mins to develop different flavours and aromas through the Maillard reaction.

[0238] Stone-grinder / Conching

[0239] The spent grain flour-like product is then grinded for 1 h in order to to obtain a homogenous powder. If dehydration is employed prior to the dehydration step, the layers of spent grain flourlike product tend to coalesce into a block. Grinding helps re-obtain the powder form and ensures that the particle size remains in about 10-1000 pm.

[0240] Once the fluor-like product is prepared as above, it may be used in the preparation of an alcoholic beverage, either in a maceration step (option 1, described below) to extract flavours to an alcoholic beverage or directly in the fermentation step (option 2, as described below).

[0241] Maceration (option 1)

[0242] The flour-like product may be mixed into alcohol at a ratio of 1:3 (w / w%) and kept at 25°C for 1 week in order to extract the aromas; the beverage can then be distilled.

[0243] Alcoholic fermentation (option 2)

[0244] The flour-like product and water were mixed at a ratio of 1:3 (w / w%) until the flour-like product was dissolved in the water. The fermentation may be carried out with Saccharomyces cerevisiae, The fermentation may take 10-14 days, depending on the microorganism used.

[0245] Filtration & Carbonation

[0246] The final beverage may be, as is typically done, filtered and may be carbonated if needed, using conventional methods known in the art.

[0247] The typical of ingredients for this example may be for example (w / w%): - 2-25% spent-grain flour

[0248] - 5-20% Sucrose / sugar

[0249] - 60-80% water

[0250] - 0.05-0.2% yeast / bacteria for the alcoholic fermentation

[0251] EXAMPLE 7 - Non-alcoholic fermentation

[0252] The spent grain flour-like product of Example 1 was further processed:

[0253] Roasting

[0254] The spent grain flour-like product may be roasted at 140°C for 2 till 10 min to develop different flavours and aromas through Maillard reaction.

[0255] Fermentation

[0256] A mature bacterial cellulose (BC) biofilm from a previously brewed kombucha culture (often called a "mother" or SCOBY, for Symbiotic Community of Bacteria and Yeast) was placed on top of the solution and was allowed to ferment for 10-14 days.

[0257] Filtration & Carbonation

[0258] The final beverage may be filtered and may be carbonated if needed, using conventional methods known in the art.

[0259] The typical of ingredients for this example may be for example (w / w%):

[0260] - 2-25% spent-grain flour

[0261] - 5-20% Sucrose / sugar

[0262] - 60-80% water

[0263] - 0.5-5% aromatics / spices / tea / herbs

[0264] - 0.05-19% yeast culture / bacteria / SCOBY / kefir culture

[0265] EXAMPLE 8 - Pasta or other baked goods (0.1-100° / o dry matter by weight spent grain flour-like product and / or fermented product)

[0266] "Baked goods" refer in the present example to perishable bakery products, such as bread, rolls, buns, pastries, cookies, muffins, tortillas, bagels, etc. The present example provides exemplary methods for preparing such bakery products. Solid-state fermentation

[0267] Before fermentation, the stored sample was pasteurized at 110 °C for 15 min and allowed to cool down to 30°C before inoculating. The fermentation was carried out with Aspergillus oryzae. The fermentation time is around 36 h and is typically done at a temperature of around 31°C degrees, and about 80 % humidity. For some uses the solid state fermentation step may be omitted.

[0268] Roasting

[0269] The spent grain flour-like product may be roasted at 140°C for 2 till 10 min to develop different flavours and aromas through Maillard reaction. For some uses the roasting step may be omitted.

[0270] Stone-grinder / Conching

[0271] The spent grain flour-like product, either provided directly from the grinding step, after fermentation or after roasting, may be grinded for 1 h in a coaching machine and afterwards mixed with the rest of the ingredients (e.g., semolina, flour, and pasteurized egg yolk as described below) in a Kitchen Aid mixer for 10 minutes. The product may be packed at 100% vacuum and stored for 12 h at 4°C.

[0272] The amounts of ingredients for an exemplary pasta product are summarised below (w / w%):

[0273] - 17.18% spent grain flour-like product

[0274] - 64.43% White flour

[0275] - 0.83% Olive oil

[0276] - 17.18% egg yolk

[0277] - 0.34%% Salt.

[0278] The amounts of ingredients for an exemplary cookie recipe are summarized below (w / w%):

[0279] - 12.42% spent grain flour-like product

[0280] - 20.70% flour

[0281] - 23.18% brown sugar

[0282] - 23.18% butter

[0283] - 0.50% salt

[0284] - 19.87% white sugar

[0285] - 0.17% vanilla essence. The above amounts of ingredients are merely exemplary; for example, the amount of flour, sugar or egg yolk etc. are to be adapted to match the spent grain flour-like product concentration.

[0286] Other dough and bakery products may also be prepared by utilizing the following steps: solid-state fermentation (optional), roasting (optional) and stone-grinding / conching the flour for 1 hour before mixing with the rest of the ingredients.

[0287] The inventors have shown that the spent grain flour-like product itself is an excellent substitute for conventional flour (or other flour products, such as maize flour, coconut flour, chickpea flour etc.) in e.g., bread, cookies, brownies, sponge cakes etc. In any recipe the skilled person may substitute part or all of the flour with the spent grain flour-like product; the taste, consistency and texture of the product resembles those made with conventional flour; however the ratio of flour (if used) and the spent gran flour-like product may be varied to create recipes that are not only visually appealing, but also are tasty.

[0288] EXAMPLE 9 - Frozen foodstuff - ice-cream (0.1-100% dry matter by weight spent grain flour-like product or fermented product)

[0289] Solid-state fermentation

[0290] Before fermentation, the stored sample was pasteurized at 110°C for 15 min and was allowed to cool down to 30°C before inoculating. The fermentation was carried out with different filamentous fungi and bacteria including but not limited Aspergillus oryzae, Aspergillus luechuensis. The fermentation is around 36 h (depending on the microorganisms) at typically at around 31°C degrees, and 80 % of humidity.

[0291] Roasting

[0292] The spent grain flour-like product may be roasted at 140°C for 2 to 10 min to develop different flavours and aromas through the Maillard reaction.

[0293] Stone-grinder / Conching

[0294] The spent grain flour-like product was grinded for 1 h in the coaching machine and afterwards was mixed with the rest of the ingredients in a mixer for 10 minutes.

[0295] The amounts of ingredients for this example are summarised below (w / w%):

[0296] 7% Spent grain flour-like product 60% Milk (3.5 fat%)

[0297] 12% Cream (35 fat%)

[0298] 2.35% Milk powder

[0299] 8.2% Dextrose

[0300] 10.05% Saccharose

[0301] 0.4% Neutro (ice-cream stabilizer)

[0302] Another exemplary ice-cream recipe may be as follows (w / w%):

[0303] - 11.85% flour-like product

[0304] - 14.22% sugar

[0305] - 26.07% milk

[0306] - 35-55% cream

[0307] - 0.47% salt

[0308] - 11.85% egg yolks.

[0309] The above amounts of ingredients are merely exemplary; the ingredients may be varied based on the desired taste / flavour / ingredients. For example, fruits (fresh or frozen), chocolate chips, colouring agents (natural or artificial) may further be added to the ice-cream. It is also possible to create vegan ice-cream using the spent grain flour-like product by replacing e.g., the dairy products with vegan alternatives (such as soy, almond, coconut, oat etc. milk or soy / oat cream or powder etc.) or the egg yolks with vegan alternatives.

[0310] EXAMPLE 10 - drink powder or other drink products, such as plant-based drinks or smoothie bowls (0.1-100° / o dry matter by weight spent grain flour-like product and / or fermented product)

[0311] Solid-state fermentation

[0312] Before fermentation, the stored sample was pasteurized at 110°C for 15 min and was allowed to cool down to 30°C before inoculating. The fermentation was carried out with different filamentous fungi and bacteria including but not limited to Aspergillus oryzae, Aspergillus luechuensis. The fermentation is around 36 h (depending on the microorganisms) at is typically at around 31°C degrees, and 80% of humidity.

[0313] Roasting

[0314] The spent grain flour-like product may be roasted at 140°C for 2 till 10 min to develop different flavours and aromas through Maillard reaction.

[0315] Stone-grinder / Conching The spent grain flour-like product was conched for at least l-2h, after which the rest of the ingredients were added and was mixed for 10 mins to obtain the drink powder.

[0316] Preparing the drink from the drink powder

[0317] 2-10 g of drink powder was dissolved in 200 ml hot milk, cold milk or water.

[0318] The amounts of ingredients for this example are summarised below (w / w%):

[0319] 25% spent grain flour-like product

[0320] 71.5% Sugar

[0321] 2% Lecithin

[0322] 1% Cinnamon

[0323] 0.5% Salt

[0324] To prepare other drink products, such as e.g., plant-based drinks, the powder may be mixed and subsequently filtered.

[0325] The amounts of ingredients for plant-based drinks may be e.g. : (w / w%):

[0326] - 12.98% flour-like product

[0327] - 86.54% water

[0328] - 0.35% lecithin

[0329] - 0.13% vanilla essence.

[0330] To prepare smoothie bowls, the following exemplary recipe may be used (w / w%):

[0331] - 5.45% flour-like product

[0332] - 36.35% sugar

[0333] - 0.15% milk or plant-based milk drink

[0334] - 0.05% vanilla essence or flavouring

[0335] - 42% banana

[0336] - 11.63% fresh mango

[0337] - 4.36% orange.

[0338] The above amounts of ingredients are merely exemplary; the ingredients may be varied based on the desired taste / flavour / ingredients. For example, for a chocolate flavoured drink, further ingredients may be added (e.g., the chocolate-like product of Example 3 or the spreadable hazelnut cocoa spread of Example 4 or even conventional chocolate). Various infusions (e.g., tea, coffee, coca-cola etc.) may also be prepared. EXAMPLE 11 - infant nutritional formula or breakfast cereals or nutraceutical products (0.1-100% dry matter by weight spent grain flour-like product and / or fermented product)

[0339] Nutraceutical products are food or components of food that provides health and medicinal benefits and may contribute to the prevention or treatment of certain conditions or diseases.

[0340] Roasting

[0341] The spent grain flour-like product may be roasted at 140°C for 2 till 10 min to develop different flavours and aromas through Maillard reaction.

[0342] Stone-grinder / Conching

[0343] The spent grain flour-like was grinded for 1 h in the coaching machine to reduce particle size and afterwards was mixed with the rest of the ingredients in a mixer for 10 minutes.

[0344] Enriching with proteins and vitamins

[0345] Typically, infant nutritional formulas and breakfast cereals are enriched (fortified) with proteins and vitamins. Any conventional fortifying techniques may be used to enrich the products with vitamins and proteins.

[0346] The amounts of ingredients for an exemplary energy var are summarised below (w / w%):

[0347] - 46.80% peanut butter

[0348] - 15.60% hazelnut

[0349] - 18.72% dates

[0350] - 15.60% almond milk

[0351] - 15.60% oats

[0352] - 24.96% flour-like product or chocolate-like product

[0353] - 9.36% flax seeds

[0354] - 0.16% salt.

[0355] The above amounts of ingredients are merely exemplary; the ingredients may be varied based on the desired taste / flavour / ingredients. EXAMPLE 12. - nutritional value of spent grain flour- like product

[0356] The nutritional value profile of the processed spent grain flour-like product of Example 1 was obtained prior to solid-state fermentation and any other processing steps and are tabulated below in table 5.

[0357] Table 5. Nutritional profile of the spent-grain flour-like product.

[0358] EXAMPLE 13 - Nutritional value profile of the processed spent grain flour- like product

[0359] A flour-like product prepared as described in Example 1 was analysed after dehydration and the nutritional value profile of the processed spent grain flour-like product and the unprocessed spent grain flour was obtained. In the present example the BSG used was from a traditional pilsner brew, in this example based on barley malt.

[0360] The data are presented in Table 6. Table 6

[0361] As can be seen from table 6, the flour-like product prepared according to the disclosure provides an improved content of several nutrients, such as e.g., protein and fats.

[0362] In particular, about 15-20% of the fibres in the BSG is digested into carbohydrates, leading to a release of about 30% more sugar in the processed BSG, with a carbohydrate / sugar ratio of 3.24 in the treated BSG compared to a carbohydrate / sugar ratio of 1.75 in the untreated BSG. An additional benefit of the treatment is the release of about 25% additional protein and about 33% more fat from the treated BSG, leading to an overall app. 15% increase in total energy content in the treated BSG.

[0363] Additionally, as can also be seen in table 6 theobromine along with caffeine are close to absent in the treated BSG, suggesting that the processed product as described herein is useful in chocolatelike products where it may be preferable that e.g., theobromine or caffeine are absent or close to absent in the final product, such as in particular certain snacks or in dog food or dog treats.

[0364] EXAMPLE 14 - Sensory analysis of particle size, malty taste and graininess of a Chocolate-like product

[0365] A particular challenge when preparing chocolate-like products, is to reach a similar mouth feel of the product, since this depends on factors such as particle size and melting temperature. While melting temperature is often governed by the specific fat composition, the particle size is governed by several of the components,

[0366] Samples of the chocolate-like product as described in Example 3 comprising either enzymatically treated BSG conched for 12 or 24 hrs, referred to as E-BSG12 and E-BSG24, respectively, and non-enzymatically treated BSG conched for 12 or 24 hrs, referred to as NE-BSG12 and NE-BSG24, respectively, were analysed for their particle size, malty taste and graininess. The two latter parameters are characterized by sensory analysis described below whereas the particle size is measured as described below using the Mastersizer apparatus.

[0367] Mastersizer analysis

[0368] Before analyzing the particle size distribution (Mastersizer apparatus) the samples were prepared by separation of the solid particles of the alternative chocolate from the fat phase. To do so, 60 grams of the samples were mixed with 240 grams of hot distilled water and stirred thoroughly until everything was dissolved. The samples were then transferred into centrifuge tubes and centrifuged at 4500 revolutions per minute (rpm) for 30 minutes. After the samples were run through the centrifuge, the liquid phase was discarded. Remaining fats were washed out by addition of a minor amount of distilled water to the pelleted solid particles, followed by a brief swirl of the tube, whereafter the water was discarded. The remaining pelleted material was then transferred into plastic tubes for particle size distribution analysis using a MasterSizer3000 Laser Diffraction Particle Size Analyzer (Malvern Instruments, Worcestershire, U.K.) with a Hydro 2000S sample. For the analysis, about 0.3g of solid samples were dissolved in 20ml milli-Q water to prepare a homogeneous solution. Sample dispersions were incrementally added to a Hydro-MV automatic wet dispersion unit until the obscuration reached between 10-15%. The particle refractive index of the protein particle was set at 1.45 and refractive index of the dispersant (water) was set to 1.33. The results from the Particle Size Distribution with the Mastersizer can be found in the graphs and table below. In the below results Dx (10) indicates that 10% of the particles in the sample are smaller than this size, thereby representing the finer fraction of the particle size distribution, Dx (50) also known as the median particle size, signifies that 50% of the particles are smaller than this size, and Dx (90) indicates that 90% of the particles are smaller than this size, representing the coarser fraction of the particle size distribution.

[0369] These values are used to analyze the spread and central tendency of the particle sizes within a sample. For example, if Dx (10) is 2 micrometers, Dx (50) is 10 micrometers, and Dx (90) is 50 micrometers, it means that 10% of the particles are smaller than 2 micrometers, 50% are smaller than 10 micrometers, and 90% are smaller than 50 micrometers.

[0370] The results are summarized in Table 7 and the particle distribution graphs are shown in figure 1.

[0371] From Table 7 and figure 1 it is clear that the process as described herein leads to formation of smaller particles, both in terms of mean particle size and width of the size distributions (see Fig. 1).

[0372] Sensory Analysis

[0373] A sensory analysis was conducted to evaluate the malty flavor and graininess of the barley spent grain (BSG) chocolate-like product using a panel of 11 evaluators with gastronomic expertise. The panelists were familiarized with the product characteristics through preparatory sessions. Evaluations took place in a controlled environment with both artificial and natural lighting, maintained at an average temperature of 24°C. Each sample was assigned a unique three-digit code, with no more than two identical digits, to ensure unbiased assessment. Panelists were provided with water to cleanse their palates between tastings.

[0374] The panelists were presented with 10 different samples, including two reference samples for calibration. The first reference was a 1: 10 w / w solution of malt extract and filtered water, while the second was an infusion prepared by steeping BSG powder in 80°C water and filtering it through a coffee filter. The BSG samples included both untreated and enzymatically treated BSG, each with the same formulation but processed at varying conching times (12, 24, 36, and 48 hours) in a conching machine.

[0375] Panelists rated the samples on a 5-point scale for both malty flavor and graininess. Malty flavor levels were defined as follows: 1 = Not noticeable, 2 = Barely noticeable, 3 = Moderately malty, 4 = Strong malty flavor, and 5 = Very malty. Graininess levels were categorized as: 1 = Not noticeable, 2 = Barely noticeable, 3 = Dusty, 4 = Sandy, and 5 = Grainy. Data collection was facilitated through a questionnaire following subsequent data analysis. Statistical analysis, including a one-way ANOVA to determine the significance of the observed differences in sensory attributes among the samples.

[0376] The data presented in Table 8 below indicates a significant difference in mouthfeel between samples conched for 12 hours and those conched for 24 hours. It also suggests that the enzymatic treatment has contributed to making the samples feel smoother.

[0377] Interpreting the data in table 8 (sensory analysis) and comparing to the data in the particle size in table 7 and Figure 1 it is clear that enzymatically treated samples E-BSG12 & E-BSGE24 have significantly smaller particle sizes and a smoother appearance than that of the non-enzymatically treated samples NE-BSG12 & NE-BSG24.

[0378] EXAMPLE 15 - Upcycled material in alcoholic beer brewing

[0379] In the beer brewing industry, it is relevant to decrease the amount of malt going into the beer brewing process in order to cost optimize the brewing processes. This can be realized by utilization of the flour-like product from example 1 instead of malt or in combination with malt to reduce the use of malt in the beer brewing process. The BSG as described in Example may also be supplemented with e.g., wasted bread, or other starch containing waste streams, as a source of glucose for its further use. This will enhance the presence of glucose and improve the conditions for the beer brewing.

[0380] Generally, about 1-90% (w / w%) of the flour-like product of example 1 is dissolved in water and malt is added, then the mashing process can continue as per usual brewing procedures which will be known to the person skilled in the art. The mashing will be followed by lautering and subsequently the wort will be reduced aiming for an initial degrees Plato (°P) around 9-16 in the wort, whereafter the fermentation is initiated with addition of yeast and optionally hops. The fermentation is carried out with Saccharomyces cerevisiae. The fermentation can take 10-21 days, depending on the microorganism used and the type of beer produced.

[0381] The typical of ingredients for this example may be for example (w / w%):

[0382] 1-90% spent-grain flour

[0383] 0-75% malt

[0384] 5-10% sucrose

[0385] 10-80% water

[0386] 1-5 % Hops

[0387] 0.05-1.0 % yeast / bacteria for the alcoholic fermentation

[0388] Additionally, adjuncts like oats or wheat may be added to enhance mouthfeel and taste.

[0389] Filtration, Conditioning & Carbonation

[0390] The final beverage is typically filtered and may be conditioned, as it is also part of some brewing processes known to the person skilled in the art, such is the case for ales and lagers.

[0391] The resulting beer can be carbonated if needed, using conventional methods known in the art.

[0392] The final alcohol content of the alcoholic beer will be between 1.5-12% ABV. EXAMPLE 16 - Upcyded material in non-alcoholic beer brewing

[0393] Non- or low-alcoholic beverages can be made in a similar way using a similar yeast strain as described in Example 15 in respect of alcoholic beverages. It is generally recognized that a nonalcoholic beer is between 0-0.5% ABV.

[0394] For non-alcoholic brewing the use of malt can be omitted or limited and recycled BSG as the flourlike product from Example 1 can be used as the main substrate for the low / non-alcoholic beer aiming for an initial degrees Plato (°P) around 1-9 in the wort.

[0395] The typical of ingredients for this example may be for example (w / w%):

[0396] 1-90% spent-grain flour

[0397] X% glucose or fructose or malt or malt extract to a final0P of 1-9 1-5 % Hops 0.05-1.0 % yeast / bacteria (Pichia kluyveri) 0.5-10% yeast extract / yeast hydrolysate Water up to 100% volume

[0398] A minimum of 8 ppm of DO (Dissolved Oxygen) is recommended.

[0399] A more specific selection of ingredients are:

[0400] 10% spent-grain flour from Example 1

[0401] Glucose, fructose, malt or malt extract until a final plato of 1-9°P.

[0402] 1-5 % Hops

[0403] 0.05-1.0 % yeast / bacteria for the non-alcoholic fermentation (e.g. Pichia kluyveri) 1-10% yeast extract / yeast hydrolysate Water up to 100% volume

[0404] Non / low-alcoholic beer #1

[0405] In this case, the flour-like product from Example 1 is dispersed into room temperature water and taken to 80°C and held for 15 minutes, alternatively, the temperature can range from 70-90°C and for 5-30 minutes. After this time, the solids are separated from the liquid by techniques known in the art, and the liquid is placed back into the kettle for the boiling stage. At this stage, the sugar source of choice is added to achieve the desired °P alongside yeast extract / hydrolysate. The boiling process can run from 5 minutes to 2 hours, depending on the desired outcome. Additionally, bittering hops, flavoring hops and / or aromatic hops can be added at the different stages of the boiling of the wort to give the adequate characteristics as is known in the art of brewing. After boiling, the hops are strained off and the liquid is cooled down rapidly to 25°C. Subsequently, the liquid is inoculated with Pichia Kluyveri and placed in a vat with constant agitation and supplemented aeration, to avoid the production of alcohol by the microorganisms. The fermentation is carried out at 18-25°C depending on the desired outcome and for a period of 2-10 days. Additionally, the resulting nonalcoholic beer can be primed prior to bottling with extra dextrose or fructose -on a range of 2-15g per liter- for bottling conditioning. Optionally, the beer can be forced carbonated using methods known to those skilled in the art.

[0406] Non / low-alcoholic beer #2

[0407] In this case, untreated BSG has been grinded to a flour (5-150 microns) and dispersed into room temperature water and taken to 60°C. pH is adjusted using citric acid to 4-6, depending on the enzymes utilized. A mixture of carbohydrases is added at this stage, namely: Ultraflo® Prime, Attenuzyme Pro®, Cellic® CTec3 HS, in the amount specified by the producer. The mixture is then held at a temperature between 45-65°C for enough time to convert lignocellulosic material into fermentable sugars, normally 30-180 minutes. The process follows by lautering, where the solid portion is strained from the liquid through methods known to those skilled in the art. The resulting liquid is then returned to the kettle to be boiled and hopped. At this stage, the sugar source of choice is added to achieve the desired °P alongside yeast extract / hydrolysate. The boiling process can run from 5 minutes to 2 hours, depending on the desired outcome. Additionally, bittering hops, flavoring hops and / or aromatic hops can be added at the different stages of the boiling of the wort to give the adequate characteristics as known in the art. After boiling, the hops are strained off, and the liquid is cooled down rapidly to 25°C. Subsequently, the liquid is inoculated with Pichia Kluyveri and placed in a vat with constant agitation and supplemented aeration, to avoid the production of alcohol. The fermentation will be carried out at 18-25°C depending on the desired outcome and for a period of 2-10 days. Additionally, the resulting nonalcoholic beer can be primed prior to bottling with extra dextrose or fructose -on a range of 2-15g per liter- for bottling conditioning. Optionally, the beer can be forced carbonated using methods known to those skilled in the art.

[0408] General considerations

[0409] When hops are to be added this is to be done before the boiling of the wort, preferably and at least 30 min before the boiling of the wort. When brewing low / non-alcoholic beer some of the yeast have special requirements e.g., Pichia kluyveri often grow well at 15-20°C, and needs at least 8 ppm dissolved oxygen (DO) supplied to the fermentation as the strain will then respire and be metabolically active without producing any ethanol.

[0410] In addition, pH adjustment with e.g., lactic acid or phosphoric acid is needed to lower the pH. This is done prior to the addition of the yeast. Furthermore, when cultivating P. kluyveri extra mixing is needed e.g., by circulating the fermentation broth in the fermenter e.g., using a pump or internal impellers. Using Pichia kluyveri is especially practical for non-alcoholic brewing processes because it prefers to utilize monosaccharides like glucose as carbon and energy source, which, as can be seen from example 1 and 13 is the most abundant monosaccharide of the flour-like product from example 1.

[0411] The benefit of using the flour like substrate obtained according to the first aspect of the invention with P. kluyveri is that almost all of the glucose will be utilized, close to zero alcohol is produced (when oxygen is present) and since almost no other mono- or disaccharides (like maltose) are present in the substrate the produced beer will not have the sweet palate that most low / non- alcoholic beer otherwise can have if no additional disaccharide reduction steps are used in the process.

[0412] Thus, the low / non-alcoholic beer brewed in this way is expected to be more appealing to those who favor a non / low-sweet tasting beer. Additionally, the absence or low amount of mono- and disaccharides in the non / low alcoholic beer will also minimize the risk of pathogen infection of the beer and thus serve as a safer solution for non / low alcoholic beer in bottles, cans, kegs and containers and could make it possible to use a milder pasteurization step.

Claims

CLAIMS1. A process for preparing an upcycled material from a dry raw material with a moisture content of 0-30% w / w comprising lignocellulose and derived from spent cereal and / or pseudocereal grain and / or food waste or lignocellulosic biomass or a mixture thereof, the process comprising: a) a pre-treatment comprising the steps of: i) grinding and / or pulverizing the raw material to obtain a powder capable of passing through a sieve having a mesh size of 10-1000 pm and ii) performing autohydrolysis, followed by b) enzymatic hydrolysis of the of powder obtained from step ii), wherein the enzymatic hydrolysis comprises adding an amount of an enzyme mixture comprising at least a glucanase and a xylanase, preferably so as to add a controlled amount of enzyme(s), and c) a post-treatment comprising the step of grinding and / or pulverizing the enzymatically hydrolysed mixture obtained from step b) to obtain the upcycled material in the form of a flourlike product.

2. The process according to claim 1, wherein the dry raw material is derived from a spent cereal or pseudocereal grain selected from the group consisting of millet, maize, sorghum, barley, oats, rice, rye, spelt, teff, triticale, wheat, wild rice, Brewer's spent grain, Distiller's spent grain, amaranth, buckwheat, chia, quinoa, wattle seeds, and acorn.

3. The process according to claim 1, wherein the dry raw material is food waste selected from the group consisting of Brewer's spent grain, Distiller's spent grain, coffee grounds, okara, wheat bran, rapeseed press cake, palm oil press cake, sunflower press cake, leftover liquorice root, tiger nut, cashew, hemp, coconut, carrot pomace, apple pomace grape pomace, olive pomace, tomato pomace, orange peels, banana peels pineapple core, rice hulls buckwheat hulls, barley husks and hulls, wheat bran oat bran malt rootlets, pumpkin seed, sunflower seed, hazelnut skins, almond skins, sorghum bagasse, coffee husks, cocoa husks, cocoa bean husks, cocoa pod husks, nut pulp leftover from plant-based drinks, such as almond pulp, oat pulps, nut husks, such as hazelnut husks, date seeds, pulses husks, such as chickpea husk, pulses hulls, such as pea hulls, corncob,sugar cane / sugar beet bagasse, fruit skins, vegetable skins, fruit pulp, vegetable pulp, cocoa chaff and coffee chaff and tea leaves.

4. The process according to any one of the preceding claims, wherein the raw dry material is a combined raw material from more than one of the raw materials defined in claim 2 or claim 3.

5. The process according to any of the preceding claims, wherein the raw dry material is a combination of Brewer's spent grain and cocoa husks.

6. The process according to any one of the preceding claims wherein the step a)i) of the pretreatment further comprises a dehydration step, wherein after the dehydration step, the moisture content of the raw material is reduced by 45-90% relative to initial moisture content of the raw material.

7. The process according to any one of the preceding claims, wherein step c) of the post-treatment further comprises a dehydration step before the grinding and / or pulverizing step, wherein after the dehydration step, the moisture content of the enzymatically hydrolysed mixture is reduced to 0- 30% w / w%.

8. The process according to any one of the preceding claims, wherein the mesh size in step a)i) is preferably 15-950 pm, such as 20-900 pm, such as 25-850 pm, such as 30-800 pm, such as 35- 750 pm, such as 40-650 pm, such as 50-600 pm, such as 60-550 pm, such as 70-500 pm, such as 80-450 pm, such as 90-400 pm, such as about 100-300 pm, more preferably around 15-200 pm, such as 15-180 pm, such as 15-170 pm, such as 15-160 pm, such as 15-150 pm, such as 15- 140 pm, such as 15-130 pm, such as 15-120 pm, such as 15-110 pm, such as 15-100 pm, more preferably around 15-90 pm, such as 15-80 pm, such as 15-70 pm, such as 15-60 pm, such as 15-50 pm, such as 15-40 pm, such as 15-30 pm, such as 15-20 pm, such as 15-15 pm, preferably between 15-30 pm , such as 16-29 pm , such as 17-28 pm, preferably around 70 pm.

9. The process according to any one of the preceding claims, wherein step a)i) of the pretreatment comprises the steps of I) milling the dry raw material and II) sifting the milled raw material through a sieve having a mesh size of 10-1000 pm and optionally, III) repeating the steps I) and II) steps at least once, preferably at least twice.

10. The process according to any one of the preceding claims, wherein step a)ii) comprises autoclaving or other heat or steam treatment or an acidic or alkaline treatment or ultrasound assisted hydrolysis.

11. The process according to any one of the preceding claims, wherein the glucanase is endo- l,3(4)-beta-glucanase and the xylanase is endo-l,4-beta-xylanase and wherein the enzymatic mixture further comprises a p-glucosidase.

12. The process according to any of the preceding claims, wherein the enzyme mixture further comprises one or more proteases, preferably one or more prolyl-specific proteases and / or one or more carboxy-specific protease.

13. The process according to any one of the preceding claims, wherein 0.1-2% v / w enzyme mixture is added in step b).

14. The process according to any one of the preceding claims, wherein the step c) of the posttreatment further comprises a dehydration step.

15. The process according to any one of the preceding claims, wherein the process further comprises the step of: d) fermentation of the product obtained from step c), wherein the fermentation is solid-state fermentation, dry fermentation, liquid fermentation or submerged fermentation.

16. The process according to claim 15, wherein microorganisms used for the fermentation are selected from the group consisting of Aspergillus ssp., Rhizopus spp., Neurospora spp., Lactobacillus spp., Streptococcus spp., Acetobacter spp., Aaccharomyces spp., Pichia spp., Gluconobacter spp., Leuconostoc spp., Saccharomyces spp., Pleurotus ostreatus, Lentinula edodes, Kluyveromyces spp., Scheffersomyces spp. and Spathaspora spp.

17. The process according to claim 15 or 16, wherein the fermented product obtained from d) is further dehydrated and optionally grinded and / or pulverized.

18. The process according to any one of the preceding claims, wherein the mean particle size of the grinded and / or pulverized flour-like product is in the range of 5-500 pm, such as 5-450 pm, such as 5-400 pm, such as 5-350 pm, such as 5-300pm, such as 5-250 pm, such as 5-200 pm, such as 5-150 pm, such as 5-100 pm, such as 5-50 pm, such as 5-40 pm, such as 5-30 pm, such as about 5-25 pm, more preferably around 5-20 pm, such as 5-15 pm, such as 10-20 pm, such as 10-15 pm, preferably 7-15 pm.

19. The process according to any one of the preceding claims, wherein at least 10% of the particles are smaller than 10 pm, at least 50 % of the particles are smaller than 25 pm, and at least 90% of the particles are smaller than 125 pm.

20. The process according to claim 15 or 16, wherein the fermented product after step d) is a beverage, such as an alcoholic or non-alcoholic beverage.

21. A process for preparing a beverage comprising fermentation by microorganisms of a carbohydrate-containing substrate, which comprises or consists of a flour-like product obtainable by the process according to any one of claims 1-14.

22. The process according to claim 21, wherein the beverage is alcoholic and the microorganisms convert carbohydrates in the substrate to produce ethanol as a metabolite.

23. The process according to claim 21, wherein the beverage is non-alcoholic and the microorganisms metabolize carbohydrates in the substrate to produce non-alcoholic metabolites.

24. The process according to claim 22 or 23, wherein the beverage is a beer.

25. A chocolate-like product comprising 20-45% matter by weight flour-like product obtained at the end of step c) and / or fermented product obtained at the end of step d), 10-25% by weight sugar, 25-50% by weight butter and 0.1-1.2% by weight lecithin.

26. A chocolate-like product comprising 10-25% by weight sugar, 25-50% by weight fats, 0.1- 1.2% by weight lecithin and 0-0.01% by weight theobromine.

27. The chocolate-like product according to claim 26, further comprising 5-50% by weight of the flour-like product as defined in any of claims 1-19.

28. The chocolate-like product according to claim 26 or 27 , wherein the taste of the chocolate-like product is improved when a flour-like product obtained at the end of step c) and / or fermented product obtained at the end of step d) is used compared to addition of an unprocessed raw material.

29. The chocolate-like product according to any one of claims 25-28, which exhibits a snap at an applied pressure of 1-500 MPa when shaped as a product, which is 70-100 mm long, 30-60 mm wide and 5-15 mm thick and the snap is measured according to the three-point test of standard ISO 178.

30. The chocolate-like product according to claim 29, where the snap is measured on a product, which is 95 mm long, 40 mm wide and 10 mm thick.

31. The chocolate-like product according to any one of claims 25-30, which is in the form of a pet treat, such as a dog treat.

32. An edible product comprising 0.1-100% dry matter by weight flour-like product obtained at the end of step c) and / or the fermented product obtained at the end of step d).

33. The edible product according to claim 32, wherein the edible product is selected from the group comprising a chocolate or confectionary product; a spreadable spent grain or a chocolatelike coating or coverage; a cacao powder replacement product; a baked or cooked good such as a pasta, a dough or a bakery product; a beverage or drink powder; an alcoholic drink or beverage; a non-alcoholic drink or beverage; a frozen-foodstuff such as ice-cream; an infant nutritional formula; a canned soup, broth or product; a condiment; a nutraceutical product such as an energy bar, an energy gel, a protein powder, a protein shake, a dehydrated complete meal package, a smoothie or smoothie bowl; an extruded product, such as a breakfast cereal product or pasta products; and a thickening agent, such as starch.

34. A flour-like product obtainable by the process defined in any of claims 1-19, for use in cosmetics, dietary supplements and / or nutraceuticals.

35. A non-alcoholic beverage prepared using a flour-like product as defined in any of claims 1-19, wherein the beverage comprises less than 0.2 g / 100 g maltose.

36. A fermented beverage, wherein the fermentation broth at the end of the fermentation comprises less than 0.2 g / 100 g maltose and less than 1% ABV of ethanol, and wherein the microorganism used in the fermentation is of the genus Pichia spp., such as Pichia kluyveri.

37. The fermented beverage according to claim 36, wherein the beverage is a non-alcoholic beverage, such as a non-alcoholic beer.

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