DK202300025Y3 - Mask-derived product and system and computer program product for its manufacture - Google Patents

Abstract

The present invention relates to a method of providing a food additive from moist mash, comprising adjusting the pH of the mash to 4.5 or below by adding a pH regulator.

Description

DK 202300025 Y3 1

Mask-derived product and system and computer program product for its manufacture

The scope of the invention

The present invention relates to grain-based foods, in particular food additives, including methods and systems for their production.

Specifically, the present invention relates to the provision of a food additive for human or animal consumption from mask.

The background of the invention

There is a general need for recycling and/or new use of waste products, including upgrading of waste products. This includes the supply of quality food for human consumption from e.g. waste products, especially used organic materials, including lignocellulosic material.

Brewing generates a waste product in the form of "mash", which consists of the solid remains of barley after mashing. Mask is the most widespread brewing by-product. It is believed to be ~85% of the by-products generated, ~31% of the original malt weight and ~20 kg per 1001 beer produced. Mask is initially wet with a short shelf life, but can be dried and treated to preserve it. In 2014, around 8 million t of mask were produced in the EU and USA alone. Mash is a residual product from one of the first steps in the brewing process 1 solubilization of malt and/or kernels of grain to ensure sufficient extraction afurten ("mashing"). It is a significant by-product in the overall brewing process, accounting for 30% - 60% of the biochemical oxygen demand and suspended solids from a typical brewery. Mask is commonly used as raw material. But mash contains some challenges, such as the existence of a complex outer layer (barley husks) that makes it difficult to separate and convert, and especially a high moisture content (-80-85%), which makes it susceptible to microbial growth and thus spoilage ( Buffington, 2014).

Industrial worm samples may include significant amounts of microorganisms such as molds, yeasts, aerobic and non-aerobic bacteria, e.g. lactic acid bacteria (LAB), which may also include pathogens. This seems to be facilitated by an only slightly acidic pH around pH 6, and a high water activity close to 1 and the presence of fermentable carbohydrates. As a consequence, mash does not appear to be suitable for direct human consumption, particularly due to the microbial risks posed by the uncontrolled microbial flora, as well as the risk of spoilage during storage. Therefore, in order to use mash for human food, such as a human food additive, which is desirable, especially because of its high protein and/or fiber content, mash is usually dried, ground and then

DK 202300025 Y3 2 aimed at a powder that can increase the fiber and protein content, while reducing the calorie content, capable of e.g. replace flour in baked goods and other foods.

WO 2018/202799 relates to a process for obtaining a food from mash containing the following steps: (a) comminution of mash, (b) heating of mash, (c) optionally: fermentation of mash, d) optionally: reduction of moisture content in mash .

DE 9420637U1 D1 describes a method for providing a mash-derived silage, where the water content in mash is reduced by approx. 50% before lactic acid fermentation, preferably after adding lactic, citric or acetic acid.

Preservation of mash using organic acids was investigated by A1 Hadithi et al. (1985) who found that organic acids prevented spoilage and preserved nutritional value for long periods.

US 4828846 relates to human foods made from dried distillers wet grain and solubles.

WO 2019/023647 Al deals with methods and systems for extracting protein-rich flour and fiber-rich flour from the brewery's mash.

Based on the risk and durability assessment, however, it can be suspected that Salmonella, in contrast to e.g. other pathogenic organisms, would still be a potentially problematic pathogen in pH-reduced products. Salmonella isolates are known to be able to grow and/or survive in conditions with pH values down to a pH of 3.7 to 4.2 depending on matrix type and other physical properties.

Surprisingly and unexpectedly, the inventors have developed a method to provide a food additive from moist mash which is microbially safe, possesses desirable organoleptic properties and is stable for an extended period of time despite its high water activity and/or high water content. Furthermore, the process is not dependent on e.g. drying and/or grinding, as the known processes, provision of e.g. cost, energy and/or CO savings. Such a food additive is suitable for human consumption. Furthermore, a food additive can be provided which does not include Salmonella.

DK 202300025 Y3 3

Summary of the Invention

The present invention was made in light of the prior art described above, and the object of the present invention relates to the provision of a consumer product derived from mash, such as feed or food additive, and/or a product comprising such a feed or food additive. The food or feed additive suitable for incorporation into a feed or food product. Furthermore, uses of the food/feed additive are disclosed, as well as systems for its provision, including modification of brewing systems, and/or computer program products for carrying out methods for providing such products and or control systems for their delivery.

In a first aspect, the present invention thus relates to a method for providing a food or feed additive from mash according to claim 1.

Such a method may comprise the following steps: (1) providing moist mask; (ii) adjusting the pH of the mask to 4.5 or below, such as a pH in the range of 3.5-4.5, by adding a pH regulator to provide a pH-adjusted product and/or food additive; and optionally (iii) packaging the pH-adjusted product to provide a packaged pH-adjusted product and/or food additive; and optionally (iv) subjecting the moist mask, pH adjusted product, packaged product or food additive to a viable cell count controlling treatment.

In another aspect, the present invention relates to a product obtained by a method according to the first aspect, such as a feed or food additive.

In a third aspect, the present invention relates to uses of a feed or food additive according to the first aspect or provided according to the second aspect.

In a fourth aspect, the present invention relates to a food product or feed product comprising e.g. 0.1-99.9, 1-90, 5-95 or 10-90% w/w (w/w) of a feed or food additive according to the second aspect.

In a fifth aspect, the present invention relates to a system for providing a feed or food additive according to the first or second aspect. Such a system may in particular include: (x)

DK 202300025 Y3 4 solid/liquid separation device (20), such as a lauter tun or mash filtration system for providing a moist mash from a mash; (y) preservative agents (50) for contacting the wet mask with at least one preservative (51, 52), said preservative comprising pH adjusting agents to provide a pH-adjusted moist mask, comprising, for example: (i) a pH monitoring device, such as pH meter, (11) a dosing device for adding one or more preservatives (51, 52), (iii) and optionally stirring means (58), such as a mixer for mixing the moist mash; and optionally (x) packaging means (70), such as a filling system for packaging the pH-adjusted and/or preserved mask in a sealable container and/or tank.

In a sixth aspect, the present invention relates to a computer program product comprising software code portions configured to, when executed in a computer's memory, perform the method of the first aspect and/or control the system of the fifth aspect. 1 a seventh aspect the present invention relates to a modification of a brewing system by incorporating a system according to the fifth aspect.

Brief description of the drawing

Figure 1. Schematic representation of the provision of a food additive and a food product according to some embodiments of the invention.

Figure 2. Schematic representation of the provision of a food additive and a food product according to some embodiments of the invention, where the food additive is transported.

Figure 3. Schematic representation of different possibilities for providing a food additive and a food product according to some embodiments of the invention, which may include different transport and/or storage steps.

Figure 4. Schematic representation of the provision of a food additive and a food product according to some embodiments of the invention, including storage before and/or after packaging.

Figure 5. Schematic representation of the provision of a food additive and a food product according to some embodiments of the invention, including storage prior to packaging.

Figure 6. Schematic representation of the provision of a food additive and a food product according to some embodiments of the invention, where the food additive is packaged directly and transported to the food production site.

DK 202300025 Y3

Figure 7. Schematic representation of the provision of a food additive and a food product according to some embodiments of the invention, where the moist mash is either pumped to a storage tank or subjected to one or more preservation steps to provide the food additive, which is then packaged.

Figure 8. Schematic representation of the provision of a food additive as shown in Figure 7, 5 comprising, inter alia, means for enabling the transfer of the moist mash from the storage tank to the preservation step.

Table I: Reference numbers

Mashing device

Solid/liquid separation

Brewing Fermentation device 40 Transport (e.g. Pump device pumping) 41-48 1.-8. flow control device (e.g. valve

Preservation device* sis2 | [1.2 preservative

Removal of O2 O» scavengers

Mixture Stirring device (60 [Storage ~~ |Storage device ~~

Storage Storage device

Packaging Packaging device 180 | Transportation | Transport device

Provision of a Device for providing a food product food product * preservation device (50) may comprise a container, stirring device such as a stirrer, pH measuring means such as a pH meter, and pH adjusting means such as a dosing device for dosing defined amounts of a preservative.

Detailed description of the invention

Definitions:

In the context of the present invention, the singular form of a word may include the plural and vice versa, unless the context clearly dictates otherwise.

Thus, references to "a" and "the" are generally inclusive of the plural of the respective terms.

For example, reference to “an ingredient” or “a method” may include a plurality of such “ingredients” or “methods”.

Similarly, "comprising", "comprising" and "comprehensive" shall be construed inclusively rather than exclusively. Embodiments provided by the present disclosure may lack any element not specifically described herein. Thus, a description of an embodiment defined using the term "comprising" is also a description of embodiments "consisting essentially of" and "consisting of the described components". Where used herein, terms such as “for

DK 202300025 Y3 6 example", "eg" or "such as", especially when followed by a list of terms, is merely exemplary and illustrative and should not be considered exclusive or exhaustive. Any embodiment described herein may be combined with any other embodiment described herein.

Unless otherwise indicated, all percentages herein are by weight of the total weight of the composition.

As used herein, "around" shall be understood to refer to numbers in a range of numbers, for example the range +/- 10,+-5,+/- 2, +/- 1, +/- 0.5, + /- 0 .1% of a referenced figure or value. Furthermore, all numeric ranges herein shall be understood to include all integers, whole or fractional, within the range. "Around" can also indicate the variations and/or uncertainties customary in the field. In some embodiments, "about" may also include +/- 20% of a referenced number or value.

The phrase "in some embodiments" can be used interchangeably with "in one or more embodiments."

An "individual" can e.g. be a human or an animal such as a mammal, bird, reptile, household or pet. The present disclosure should not be construed as being limited to a specific animal, human or demographic. "Food" shall be interpreted as a product for consumption and/or ingestion by an individual, such as a human or an animal, whereas "feed" refers to a product for consumption and/or consumption by an animal. In connection with the present invention, the term "foodstuffs" thus also includes "feed". "Malt" is germinated grain that has been dried in a process known as "malting", where the grain is made to germinate by soaking in water and then stopped from further germination by drying, usually with hot air.

Malting grains develop enzymes (a-amylase, b-amylase) necessary to modify grain starch into various types of sugars, including the monosaccharide glucose, the disaccharide maltose, the trisaccharide maltotriose, and higher sugars called maltodextrins. Malting grains also develop other enzymes, such as proteases, that break down the proteins in the grain into forms usable by yeast.

Malt is also commonly used to make beer or whiskey, but also malted milk, malted vinegar, sweets such as Maltesers and Whoppers, flavored drinks such as Horlicks, Ovaltine and Milo, and some baked goods.

DK 202300025 Y3 7

Various grains can be malted, although barley is the most common. There are many different types of malt, such as: Pilsner malt, Pale malt, Mild malt, Amber malt, Stout malt, Brown malt, Chocolate malt, Black malt, Crystal malt, Distiller's malt, Peated malt, Vienna malt, Miinchner malt, Rauchmalz and acid malt. Different types of malt can give e.g. different flavors and/or different colors for the beer or distilled product.

The terms "Brewer's used grain" (BSG), "mash", "BSG" or "draff" can be used interchangeably and refer to a by-product of brewing with grain malt, such as barley malt or wheat malt. Generally mash is the solid fraction after solid/liquid separation after wort production in a beer brewing process. Husks include grain hulls and parts of the pericarp and the seed coat layers of grains used for brewing, usually barley or wheat. Mask is usually rich in cellulose, hemicelluloses, lignin and protein. In addition, mash is also naturally rich in fibre, which makes it of great interest as a food additive, e.g. by replacing or supplementing low-fibre ingredients.

The type(s) or variety(s) of barley or other grains used may affect the composition and/or taste of the mask. In particular, the malting process can further influence the composition and/or taste of the mash, as well as the brewing process. Except for barley and/or, wheat, oats, rye, sorghum, millet, rice and maize used in the production of beer, whether malted or not, or any combination of malted and/or unmalted grains.

In brewing, the term "raw grain(s)" is commonly used to refer to unmalted grains/grains (such as corn, rice, rye, oats, barley, and wheat) or grain products used in brewing beer that supplement the main mash ingredient, which is usually malted barley. Raw fruit is often used for the purpose of reducing costs, but they can sometimes also be used to create one or more additional functions, such as better foam retention, flavoring or nutritional value, or additives. Generally, raw fruit is a source of carbohydrates.

In some embodiments, the mask is provided uniquely from malt, preferably barley malt. In some embodiments, mash may also be derived from grains other than barley, such as wheat, e.g. wheat and/or wheat malt from the production of "Weizenbier", also called "Hefeweizen" or

Weif3bier.

DK 202300025 Y3 8

In connection with the present invention, mash can also include (or be derived from) other cereals than barley or residues thereof, such as other malted or unmalted cereals, e.g. wheat, corn, rice, oats, rye, sorghum, millet, including any combination thereof. Thus, in some embodiments, the mask comprises raw fruit and/or residues thereof. In some other embodiments, the mask includes no raw fruit and/or residues thereof. "Brewer's spent grain" usually refers to a by-product of brewing using barley malt, while "distiller's grains" is a grain by-product of the distillation process. Distiller's grains commonly refers to a by-product that includes leftover corn/maize, rice, wheat and/or other grains. "Wet distillers grains" or "WDG" primarily contain unfermented grain residues (protein, fibre, fat and up to 70% moisture). WDG, like BSG, has a short shelf life, and is often dried to increase shelf life.

Dried distiller's grains with solubles (DDGS) is WDG that has been dried with the concentrated thin distillate to 10-12% moisture. DDGS has an almost indefinite shelf life and can be shipped to any market regardless of its proximity to an ethanol plant. Drying is expensive as it requires additional energy input. In the US, it is packaged and traded as a commodity product. In the context of the present invention, the term "mash" may include, at least in some embodiments, any of: distiller's grains, WDG and/or DDGS.

The terms "pH regulator", "acidity regulator" or "pH control agent" may be used interchangeably and are intended to include one or more compounds capable of changing or maintaining pH. They can be organic, mineral acids, bases, neutralizing agents or buffering agents, including salts of the organic or mineral acids, including mixtures of acid(s) and salts. A pH regulator can be a natural compound or a synthetic compound. In the context of the present invention, a pH regulator is usually "food grade", ie. an additive that is safe for human consumption.

Acidity regulators are often listed by their E number, such as E260 (acetic acid), or simply listed as "food grade acid" (see the comprehensive list of E numbers herein).

Acids

Mineral (inorganic) acids:

A mineral acid or inorganic acid is any acid derived from an inorganic compound that dissociates to produce hydrogen ions (H+) in water. Normally, mineral acids are very soluble in water.

Examples of mineral acids include, for example:

DK 202300025 Y3 9

Hydrochloric acid (HCl), Nitric acid (HNO3), Sulfuric acid (H2S04) and Sulfuric acid (H2S03). Phosphoric acid (H3P04) is also a commonly used acidulant in the food and beverage industry, e.g. used to produce cola drinks, which are massively sold around the world. This acid is known for its biting, harsh taste, which perfectly complements the taste of e.g. cola.

Organic acids

An organic acid can be described as an organic compound with acidic properties. The most common organic acids are the carboxylic acids, whose degree of acidity is linked to their carboxyl group -COOH.

Sulfonic acids containing the group -SO20H are relatively stronger acids. Alcohols with -OH can act as acids, but they are usually weak. The relative stability of the acid's conjugate base determines its acidity. Other groups can also give acidity, usually weak: the thiol group -SH, the enol group and the phenol group. Organic compounds containing these groups can also be referred to as organic acids.

Acetic Acid - Food grade acetic acid is known for its pungent smell and is found in vinegar. It is very commonly used in the pickling industry, as vinegar that ferments naturally has a variable pH, and therefore food grade acetic acid is used to create pickling liquid that has a specified acidity. It is also used in flavorings and confectionery.

Acetic acid can also be provided as sodium diacetate (NaH(C2H302)2), a mixture of acetic acid and sodium acetate, salt provided for example by half-neutralization of acetic acid followed by evaporation of the solution. In some embodiments, the term acetic acid may include acetic acid and sodium diacetate. In some embodiments, the term sodium acetate include sodium acetate and sodium diacetate.

Ascorbic Acid (Vitamin C) - Naturally occurring in lemon fruits, ascorbic acids have a number of uses as a natural food additive in commercial applications such as bakeries, beverage manufacturers, fruit processing plants, butchers and meat merchants.

Carboxylic acid

A carboxylic acid is one that contains a carboxyl group (C(FO)OH). The general formula for a carboxylic acid is R-COOH, where R refers to the alkyl group. Carboxylic acids occur widely.

Examples can e.g. includes the following acids:

Table II: List of carboxylic acid

DK 202300025 Y3 10

Atoms 6 [remy Pomanoic ad |CHACH2COOH 8 [Hepansre— Hepneiened [CHACH2-COOH 9 [chambers — Osmeleaed — | CHNCHDCOOH 8 [Penadecanore | Pendens ae — |CHNCHP)SCOOH — + [Hepadesm acid Hepidecanie aid —|CHNCHP)SCOOH 9 Ovtdscamyre — [Ocadesmoiraed —CHNCHDCOOH

A carboxylic acid may also include one or more C=C double bonds.

A carboxylic acid may also comprise an aromatic ring, such as e.g. benzoic acid.

A carboxylic acid may also comprise two or more carboxyl groups, such as oxalic acid or citric acid.

A carboxylic acid can also include one or more hydroxy groups (also called hydroxyl or OH groups), such as e.g. lactic or tartaric acid.

Citric acid - Citric acid, which is naturally found in fruits such as lemons, limes and grapefruit, is e.g. responsible for the sour taste found in many fruits.

Citric acid is very useful in commercial food applications as a preservative, as the acidic pH of the acid prevents bacteria from growing.

Apart from this, it can also be used to impart a sour taste to dry foods such as seasoning salts and powders.

DK 202300025 Y3 11

Fumaric Acid - This food acid that has a very strong taste.

Since it is not very soluble, it has only some limited uses in the food and beverage industry.

It is usually used in cheesecake mixes, dessert powders containing gelatin, and powdered beverages.

Fumaric acid's strong flavor and reasonable price make it an excellent choice for making animal feed.

Formic acid - also called methanoic acid is a carboxylic acid.

Lactic acid - Lactic acid is found naturally in products that go through fermentation, including cheeses, yogurt, and some types of bread.

In commercial settings, lactic acid is commonly used as an acidulant or preservative; this can include meats, cheeses, poultry, fish, beverages, dressings and more.

Malic acid - Malic acid is formed during fruit metabolism and is present in all fruits and many vegetables.

Able acid is used as a flavoring agent, and its tart flavor lends itself to confectionery, beverage processing, desserts, bakery products, and some medical consumables such as throat lozenges.

Oxalic acid - Its acid strength is much greater than that of acetic acid.

Oxalic acid can also act as a reducing agent, and oxalate is a chelating agent, e.g. for metal ions.

It occurs naturally in many foods.

Propionic acid - This acid is known to inhibit the growth of mold and some bacteria and is used as a preservative in both pet food and human food, despite its rather unpleasant smell.

Sorbic Acid - Sorbic acid and its salts, such as sodium sorbate, potassium sorbate, and calcium sorbate, are antimicrobial agents often used as preservatives in food and beverages to prevent the growth of molds, yeasts, and fungi.

In general, the salts are preferred over the acid form because they are more soluble in water, but the active form is the acid.

The optimal pH for antimicrobial activity is below pH 6.5.

Sorbates are generally used in concentrations of 0.025% to 0.10%.

However, adding sorbate salts to food will raise the pH of the food slightly, so the pH may need to be adjusted to ensure safety.

Tartaric Acid - Naturally occurring in some fruits, tartaric acid can be used as an additive to provide a sharp, sour flavor in fruit juices, confections and wine.

Trisodium Citrate - Often referred to as 'sodium citrate', trisodium citrate is most commonly used as a preservative or flavoring agent.

Commercial uses include use in soft drinks, ice cream, jams and candies.

Trisodium citrate can also be used as an emulsifier to make cheese sauces. pH regulators for use in food are regulated and common pH regulators have been assigned different "E numbers" (see

Table III),

Table III: pH regulators, their E-numbers and approved uses according to the Webgate food database (https:/Avebgate.ec.europa.eu/foods system/main/) (EM — [förbinese— [Authorized use [decodes (2626 ) | Nøvimeeter |] en Nee] https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=8633 https://webgate.ec.europa.eu/foods system /main/index.cfm?event=substance.view&identifier=116 https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=117 https://webgate.ec. europa.eu/foods system/main/index.cfm?event=substance.view&identifier=118

Carbon dioxide and carbonates https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=94 https://webgate.ec.europa.eu/foods system/main/index. cfm?event=substance.view&identifier=39 https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=222 https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=224 https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=222 https://webgate.ec .europa.eu/foods system/main/index.cfm ?event=substance.view&identifier=223 https://webgate.ec.europa.eu/foods system/main/index.cfm ?event=substance.view&identifier=223

Magnesium hydroxide carbonate https://webgate.ec.europa.eu/foods system/333main/index.cfm ?event=substance.view&identifier=224 https://webgate.ec.europa.eu/foods system/main/index.cfm? event=substance.view&identifier=224

Potassium bicarbonate https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=222

Phosphoric acid and its salts

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Aøpinseogdenske 6 |Ävreogdensete — p [Cronyeogdemsate — p [Seogdensae

D |ewompeogdenee 6 |sorbinsre benoroic acid andpropioneyre And its sweet sodium salt sodium salt potassium salt calcium salt sodium salt po Anger https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=96

Aer https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=224

Potassium hydroxide https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=224

Calcium hydroxide https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=243 https://webgate.ec.europa.eu/foods_system/main/index.cfm?event= substance.view&identifier=244 https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=245 https://webgate.ec.europa.eu/foods_system/main/index. cfm?event=substance.view&identifier=246 https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=247 https://webgate.ec.europa.eu/foods system /main/index.cfm?event=substance.view&identifier=97 https://webgate.ec.europa.eu/foods system/main/index.cfm?event=substance.view&identifier=98 https://webgate.ec. europa.eu/foods system/main/index.cfm?event=substance.view&identifier=99 https://webgate.ec.europa.eu/foods_system/main/index.cfm?event=substance.view&identifier=268

The first aspect of the invention relates to a method of providing a mash-derived feed or food additive, which method comprises the steps of: (1) providing moist mash; (ii) adjusting the pH of the mask to 4.5 or below, such as a pH in the range of 3.5-4.5, by adding a pH regulator to provide a pH-adjusted product and/or food additive; and optionally (111) packaging the pH-adjusted product to provide a packaged pH-adjusted product and/or food additive; and optionally (iv) subjecting the moist mask, pH adjusted product, packaged product or food additive to an additional viable cell count controlling treatment.

For the avoidance of doubt, "additional viable cell number controlling treatment" shall be understood as treatment, step or act other than acidification with a pH regulator according to step (11). This may be or include a preservation step, as described herein, particularly below. It may also be an additional addition of a pH regulator at another time.

In some embodiments, the moist mask, pH-adjusted product, packaged product, or food additive is subjected to an additional treatment other than step (ii), such as a preservation step or an additional addition of a pH regulator at a different time.

Steps (iii) and (iv) may be optional, and in some embodiments the method thus comprises steps (1), (ii) and (iii). In some embodiments, the method comprises steps (i), (ii) and (iv). In some embodiments, the method comprises steps (i), (ii), (iii) and (iv).

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The order of step (ii) and optional steps (111) and (iv) may differ: In some embodiments, the order of steps may be such that step (iv) is performed last. In some embodiments, the order of the steps may be such that step (111) is performed last. In some embodiments, the order of the steps may be such that step (ii) is performed last.

In some embodiments, two or more steps may be performed simultaneously or substantially at the same time.

In some embodiments, step (11) and step (111) are performed simultaneously or substantially at the same time. In some embodiments, step (ii) and step (iv) are performed simultaneously or substantially at the same time. In some embodiments, step (iii) and step (iv) are performed simultaneously or substantially at the same time. In some embodiments, steps (ii), (iii) and (iv) are performed simultaneously or substantially at the same time. "Carried out simultaneously" or "substantially at the same time" can e.g. include filling a container, e.g. while stirring and adding a pH regulator for pH adjustment during the filling process, thus "pH adjustment" (step ii) and "packing" (step iv) can be performed simultaneously or at the same time. If the pH regulator is further or comprises an organic acid or salt thereof in an active amount, such as lactic acid/lactate, malic acid/malate, acetic acid/acetate, sorbic acid/sorbate, steps (11), (iii) and (iv) can be carried out simultaneously or substantially at the same time.

A "preservation step" may thus include steps (ii) and/or (iv). "Preservative" may include means for pH regulation, such as a device and/or system for measuring pH and a device and/or system for the controlled addition or "dosing" of a pH regulator, such as an acid or its salt as described herein. Such devices or systems are believed to be known in the art. "Preservatives" may also include means to provide a viable cell number controlling treatment, such as one or more of heat treatments, removal of O,, packaging under N> atmosphere, reduction of air pressure, addition of one or more microorganisms that are generally recognized as safe (GRAS) and/or freezing, including any combination(s) thereof. Systems or devices for this are also believed to be known in the field. In some embodiments, the "preservation step" is an additional viable cell number controlling treatment as described herein, particularly above.

In some embodiments, the moist mask, pH-adjusted product, and/or packaged product is subjected to one or more additional viable cell count-controlling treatments). Such additional viable cell count controlling treatment(s) or measures may include one or more of: heat treatment, pasteurization, removal of O»; packaging under N> atmosphere, reduction of air pressure, addition of one or more micro-organisms generally recognized as

DK 202300025 Y3 16 safe (GRAS) and/or freezing, including any combination(s) thereof. Such treatment is believed to give the product an increased durability and/or increased microbial safety. In some embodiments, removal of O, is provided by allowing microorganisms in the mask to metabolize the available O). This process is usually facilitated by an airtight seal of the container in which the food additive is packed.

In some embodiments, the mask is fresh, ie. the mask is provided within 1 hour of filtration or less, such as within 45 minutes or less, or within 30 minutes or less. In some embodiments, the mask is provided within 20 minutes or less, such as within 15 minutes or less, within 10 minutes or less. In some embodiments, the mask is provided within 5 minutes or less, 4 minutes or less, 3 minutes or less, 2 minutes or less, or even 1 minute or less.

In some embodiments, where the mask is provided by one or more continuous solid-liquid separation steps or by one or more batch solid-liquid separation steps. Suitable solid/liquid separation systems may include a lauter tun and/or filtration systems such as a mash filtration system known in the art. In some embodiments, at least 2 solid/liquid separation steps are performed, such as 2 continuous solid-liquid separation steps, 2 batch separation steps, or a combination of a continuous and a batch separation step in any order.

Normally the mash has a higher temperature due to the temperature profile used for mashing.

In some embodiments, the mask has a temperature of at least 40, 45 or 50°C. In some embodiments, the mask has a temperature of at least 50, 55°C, 60, 65, or 70°C. A higher temperature is believed to improve the microbial safety and/or stability of the food additive. In addition to providing a mask with an elevated temperature, maintaining elevated temperatures throughout the various steps and/or unit operations seems advisable for product quality considerations, including microbial safety, stability and shelf life.

In some embodiments, the mask is fed continuously, fresh and at an elevated temperature for one or more of: pH adjustment, packaging and/or exposure to a viable cell count controlling treatment.

DK 202300025 Y3 17

Without wishing to be bound by any theory, it is believed that the use of fresh mash improves the quality of the food additive, e.g. in terms of microbial safety, especially when the mask remains at high temperatures throughout processing. In an industrial brewing process, the mash is usually filtered while still hot, resulting in a warm filter cake. It may be desirable to remove the mask while it is still warm, preferably in a continuous manner, such as by using devices and/or methods known in the art. Furthermore, maintaining the mask at elevated temperatures (eg, around or at least 40°C; 45°C; or 50°C, 55°C, 60, 65, or 70°C) may render an additional heating step through processing obsolete. This is believed to contribute to e.g. one or more of: microbial safety, cost reduction, time saving, energy saving; improved

CO2 footprint and/or product quality.

Generally, mash resulting from a solid/liquid separation of a mash contains a significant amount of moisture and/or has a high water activity. This, combined with the presence of fermentable carbohydrates, can be considered an ideal growth medium for a rich variety of microorganisms, including destructive microorganisms, which may even include pathogenic microorganisms (see, e.g., Examples 18-20).

In some embodiments, the mask in step (1) has a moisture content of 60-95%, 70-90%, 72-88%, or 75-85% (w/w); and/or a water activity of 0.98-1.0, 0.985-0.998 or 0.990-0.995 or about 0.992. In some embodiments, the mask in step (i) has a moisture content of 20-95%, such as 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80 -90%, 90-95% or more than 95% (w/w). In some embodiments, the mask has a moisture content of about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94 or 95% (w/w). In some embodiments, the mask has a moisture content of at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 , 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94 or 95% (w/w). In some embodiments, the mask has a water activity of 0.95, 0.96, 0.97, 0.98, 0.99, 0.995, or 0.998 or more. In some embodiments, the mask has a water activity in the range of 0.95-0.96, 0.96-0.97, 0.97, 0.98, 0.98-0.99, or 0.99-1.0.

In some embodiments, the wet mask has a water content of at least 20, 30, 40, 50, 60, 70, 72, 75, 78, 80, or 85% w/w or more. In some embodiments, the wet mask has a water content of at least 70, 72, 75, 78, 80, or 85% w/w or more.

DK 202300025 Y3 18

In some embodiments, the mask is fresh, i.e. the mask is provided within 1, 2, 3, 4, 5, 7, 10 or 15 minutes of herb separation or less; or within 20, 30, 45 or 60 minutes of herb separation or less. For the avoidance of doubt, "after herb separation" as used herein shall be understood to mean "after the mask has been separated from the herb". Herb separation is usually performed as a solid/liquid separation step involving the use of systems and/or devices customary in the industry.

In general, a high temperature and/or rapid feeding of the mash after herb separation is preferred, especially with regard to microbial safety, such as to minimize the presence of unwanted microorganisms such as spoilage and/or pathogenic microorganisms in the mash.

According to the first aspect, the pH of the mask is adjusted to pH 4.5 or below by adding a pH regulator. In some embodiments, the pH is adjusted to 4.4 or below; 4.3 or below; 4.2 or below; 4.1 or below; 4.0 or below; 3.9 or below; 3.8 or below; 3.7 or below; 3.6 or below; or 3.5 or below. In some embodiments, the pH is adjusted to pH 3.5-4.5 or pH 3.75-4.2. In some embodiments, the pH is adjusted to about pH 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, or 4, 5. Normally, the pH is determined by including temperature compensation and represents pH values at a reference temperature, such as 25°C.

The method of the first aspect includes the addition of one or more pH regulators. It may also be advisable to provide some agitation, such as stirring and the like, to provide adequate distribution of the pH regulator(s). In some embodiments, the pH regulator is or comprises one or more of: (i) one or more organic acids, (ii) one or more salts of an organic acid, (iii) one or more inorganic acids, and/or (iv) one or more salts of an inorganic acid, including any combination thereof. In some embodiments, the pH regulator is or comprises an acid and/or salt of an organic acid as described above, such as in the definitions section regarding acids. In some embodiments, the pH regulator is or comprises a compound of Table III.

In some embodiments, the pH regulator is one or more organic acid(s) selected from: acetic acid, lactic acid, malic acid, tartaric acid, and sorbic acid, including any combination thereof. In some embodiments, the pH regulator is one or more salts of one or more organic acids, such as one or more salts selected from: sodium acetate, sodium diacetate, potassium acetate, calcium acetate, sodium lactate, potassium lactate calcium lactate, sodium malate, sodium hydrogen malate, calcium malate, sodium tartrate, potassium tartrate, calcium tartrate , sodium sorbate, potassium sorbate and calcium sorbate, including any

DK 202300025 Y3 19 combination thereof. In some embodiments, the pH regulator is or comprises an acid and a salt of an acid, such as an organic acid and a salt of an organic acid. In some embodiments, the organic acid and/or the salt of the organic acid is a compound with E number 1701), 200, 201, 202, 203, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 260, 261(1), 262, 2621), 262(ii), 263, 270, 280, 281, 282, 283, 290, 296, 297, 300, 301, 302, 325, 326, 327, 330 , 331(1), 331(iii), 3321), 332(ii), 333 (ii), 334, 338, 339, 340, 341, 343, 3501), 350(i1) ), 352(ii), 355, 356, 363, 380, 450, 500), 500(iii), 501), 501(ii), 503(1), 503(ii), 5046), 504(h), 513, 5146), 514 (ii), 515(), 516, 524, 525, 526, 527, 528, 529, 530, 574, 575, 577, or 578.

Acidification of the mask helps to control the microbial flora. But with regard to e.g.

Salmonella, a pH of 4.2 or even as low as 3.7 may not be sufficient to prevent the survival or growth of this potentially pathogenic microorganism. In this context, selection of appropriate measures such as freshness and temperature of the mask as elucidated herein should be combined with the use of one or more appropriate pH regulator(s) to ensure microbial safety of the food additive and/or food additive comprising food or feed product .

In some embodiments, a food additive may be provided which does not include Salmonella, e.g. determined as zero colony forming units (CFU) per 10 or 25 g sample, a common food safety requirement in many countries (see, for example, the Salmonella criteria set out in Regulation (EC) 2073/2005, amended by Regulation (EC) ) 1441/2007, which lays down rules for sampling and testing and sets limits for the presence of Salmonella in specific food categories).

Such as. described in the examples, especially example 20, pH regulator(s) comprising acetic acid and or one of its salts (eg sodium acetate and/or sodium diacetate) can provide a Salmonella-free food additive/product. However, it may be desirable to use other organic acids and/or their salts, especially with regard to the organoleptic properties of the food additive, such as taste and/or smell.

Here it may be desirable to replace acetic acid at least partially with one or more less stringent organic acids, such as malic acid and/or lactic acid. Accordingly, in some embodiments, the pH regulator is or comprises a combination of acetic acid and/or acetate + lactic acid and/or lactate. In some embodiments, the pH regulator is or comprises a combination of acetic acid and/or acetate + malic acid and/or malate. In some embodiments, the pH regulator is or comprises a combination of acetic acid and/or acetate + tartaric acid and/or tartrate. In some embodiments, the pH regulator is or comprises a combination of acetic acid and/or acetate + sorbic acid and/or

DK 202300025 Y3 20 sorbate. In some embodiments, the pH regulator is or comprises a combination of lactic acid and/or lactate + malic acid and/or malate. In some embodiments, the pH regulator is or comprises a combination of lactic acid and/or lactate + tartaric acid and/or tartrate. In some embodiments, the pH regulator is or comprises a combination of lactic acid and/or lactate + sorbic acid and/or sorbate. In some embodiments, the pH regulator is or comprises a combination of malic acid and/or malate + tartaric acid and/or tartrate. In some embodiments, the pH regulator is or comprises a combination of malic acid and/or malate + sorbic acid and/or sorbate. In some embodiments, the pH regulator is or comprises a combination of tartaric acid and/or tartrate + sorbic acid and/or sorbate.

In some embodiments, the pH adjuster is or comprises acetic acid, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 0, 1.2, 1.4, 1.6, 1.8 or 2% by weight. /weight of acetic acid. In some embodiments, the pH regulator comprises, other than acetic acid at the above concentrations, one or more of: lactic acid, malic acid, and sodium diacetate; including any combination thereof. In some embodiments, the pH adjuster is or comprises lactic acid, such as at a concentration of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0, 9, 1.0, 1.2, 1.4, 1.6, 1.2 wt% or /v or w/w lactic acid. In some embodiments, the pH adjuster is or comprises malic acid, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 0, 1.2, 1.4, 1.6, 1.8 or 2.0% w/v. /w malic acid. In some embodiments, the pH adjuster is or comprises sodium diacetate, such as 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1, 2, 1.4, 1.6, 1.8 or 2.0% w/v sodium diacetate.

In some embodiments, the pH adjuster is or comprises: + —0.5% acetic acid + ~0.5% lactic acid + ~0.2% malic acid; + —0.5% acetic acid + —-0.2% lactic acid + ~0.5% malic acid; + —0.7% acetic acid + ~0.3% lactic acid + ~0.2% malic acid; + —0.7% acetic acid + ~0.3% lactic acid + ~0.2% malic acid + ~0.2% sodium diacetate; + —0.8% acetic acid + ~0.4% lactic and/or malic acid + ~0.2% sodium diacetate; + —0.4% acetic acid + ~0.8% lactic and/or malic acid + ~0.2% sodium diacetate; + —0.8% acetic acid + ~0.4% lactic and/or malic acid; + —0.4% acetic acid + ~0.8% lactic and/or malic acid; or e —1.2% acetic acid + —0.2% diacetate; where all % are weight/volume or weight/weight.

In some embodiments, the pH adjuster is or comprises: + 0.25-0.75% acetic acid + 0.25-0.75% lactic acid + 0.1-0.5% malic acid;

DK 202300025 Y3 21 + 0.25-0.75% acetic acid + 0.1-0.5% lactic acid + 0.25-0.75% malic acid; + 0.5-1.0% acetic acid + 0.1-0.5% lactic acid + 0.1-0.5% malic acid; + 0.5-1.0% acetic acid + 0.1-0.5% lactic acid + 0.1-0.5% malic acid + 0.1-0.5% sodium diacetate; + 0.5-1.0% acetic acid + 0.25-0.75% lactic and/or malic acid + 0.1-0.5% sodium diacetate; + 0.25-0.75% acetic acid + 0.5-1.0% lactic and/or malic acid + 0.1-0.5% sodium diacetate; + 0.5-1.0% acetic acid + 0.25-0.75% lactic and/or malic acid; + 0.25-0.75% acetic acid + 0.5-1.0% lactic and/or malic acid; or + 1.0-1.5% acetic acid + 0.1-0.5% diacetate; where all % are weight/volume or weight/weight.

In some embodiments, the pH regulator is or comprises: « acetic acid: lactic acid: malic acid in a ratio of about 1:1:0.4; 1:0.4:1; 0.7:0.3:0.2; 0.25-0.75:0.25-0.75:0.1-0.5; 0.25-0.75:0.1-0.5:0.25-0.75; or 0.5-1.0:0.1-0.5:0.1-0.5; » acetic acid : lactic acid : malic acid : sodium diacetate in a ratio of approx. 0.7:0.3:0.2:0.2; or 0.5-1.0:0.1-0.5:0.1-0.5:0.1-0.5; + acetic acid: lactic acid and/or malic acid: sodium diacetate in a ratio of about 0.8:0.4:0.2; or 0.4:0.8:0.2; or 0.5-1.0:0.25-0.75:0.1-0.5; or 0.25-0.75:0.5-1.0:0.1-0.5; « acetic acid: lactic and/or malic acid in a ratio of about 0.8:0.4; 0.4:0.8; 0.5-1.0:0.25- 0.75; or 0.25-0.75:0.5-1.0% « acetic acid: sodium diacetate in a ratio of about 1.2:0.2; or 1.0-1.5:0.1-0.5; where the ratios are w/w or w/v.

The term lactic acid and/or malic acid shall include lactic acid, malic acid or any combination of lactic acid and malic acid.

Regarding suitable concentrations of the pH regulator(s), they can e.g. be provided in a concentration of 0.1-5.0, 0.2-2.5, 0.4-2.0, 0.5-1.8, 0.6-1.6 or 0.8- 1.4% weight by volume (w/v) or weight/weight (w/w). In some embodiments, they may be provided at a concentration of about 1.9, 2.0, or more than 2.0, such as 2.5% or more (% w/v or w/w). In some embodiments, the above concentrations represent the total amount of all pH regulators used. In some embodiments, the above concentration(s) represent values for the individual pH regulators used. Generally, in most embodiments, the total concentration of pH regulators does not exceed 2.0 or 2.5% w/w or w/v. In some

DK 202300025 Y3 22 embodiments, the total concentration of pH regulators is greater than 2.5%, such as 2.5-5% w/w or w/v.

However, it is often desirable to minimize the amount of pH regulators for cost-effectiveness and/or organoleptic properties while ensuring microbial safety. This can e.g. is achieved with total pH regulator concentrations in the range of 0.4-2.0 or 0.5-1.5% w/w or w/v, especially when the pH regulator comprises a significant amount of acetic acid and or one or more of its salts , such as more than 0.2, 0.3 or 0.4% w/w or w/v.

According to the first aspect, the pH-adjusted product can be packaged. This may include any packaging known in the art and may include packaging the pH-adjusted mask or the pH-adjusted food additive in a suitable container such as a tank, tank container, tank truck, semi tank trailer, tank trailer, barrel, airtight big-bag, plastic bag, bottle, Tetra Pak, glass container, aluminum container or similar. Typically, packaging includes enclosing the pH-adjusted mask or the pH-adjusted food additive in a container or cover which is preferably waterproof, sealable and preferably airtight.

In some embodiments, the package size is in the range of 0.1-1 1; 1-100 1; 0.1-1 m3; 1-10 m3; or 10-50 m3. In some embodiments, the container is reusable. In some embodiments, the container is suitable for bulk shipping and/or transportation. In some embodiments, the pH-adjusted mask is packaged in a tank, such as a tank on a tanker truck, which enables easy transport of the food additive to, e.g. a production site for a food product.

As previously mentioned, one or more additional viable cell count control method(s) may be used. They can be applied or provided for before, during or after packaging.

This may include the provision of a GRAS microorganism, such as an acid-producing microorganism. A GRAS microorganism can also be any microorganism on the list of Qualified Biological Presumptions of Safety (QPS) biological agents intentionally added to food or feed maintained by the European Food Safety Authority (EFSA). Suitable GRAS microorganisms may include lactic acid bacteria (LAB), such as one or more of the genera: Lactobacillus, Pediococcus, Enterococcus, Lactococcus, Leuconostoc, Oenococcus,

Streptococcus, Tetragenococcus, Camobacterium, Vagococcus and Alkaliissbacterium. In some embodiments, the GRAS microorganism is selected from one or more of: Bacillus, Lactobacillus, or Lactococcus. In some embodiments, the one or more microorganisms are selected from a

DK 202300025 Y3 23 or more: (i) Lactobacillus, such as one or more of Lactobacillus acidophilus, Lactobacillus amylolyticus, Lactobacillus amylovorans, Lactobacillus animalis, Lactobacillus brevis, Lactobacillus acidophilus, Lactobacillus buchacillus, Lactobacillus buchneri, Lactilluobacillus, Brubacillus eckii,

Lactobacillus diolivorans, Lactobacillus farciminis, Lactobacillus farciminis, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus hilgardii, Lactobacillus lactis, Lacto bacillus mucosae, Lactobacillus paracillus, Lactobacillus paracillus, sus, Lactobacillus plantarum,

Lactobacillus reuteri, Lactobacillus paracasei, Lactobacillus sakei Lactobacillus salivarius, including any combination thereof; (ii) Lactococcus, such as Lactococcus lactis; or (iii) Pediococcus, such as one or more of Pediococcus acidilactici, Pediococcus parvulus, Pediococcus pentosaceus, including any combination thereof. In some embodiments, the GRAS microorganism is one or more microorganisms on the "QPS notification until 2019" list with positive QPS status for the species/family, e.g. available at: https://zenodo.org/record/3607184/files/Appendix%20D 2019 %200QPS notification until2019 xlsx?download=! .

Contrary to the prior art, where the mask is subjected to a drying step in order to increase durability and/or product stability, the method according to the first aspect cannot include a drying step.

Advantages of omitting the drying step include one or more of: energy savings, CO, savings, simplified process design and/or equipment, cost savings, improved organoleptic properties. Thus, according to some embodiments, the method of providing a food additive does not include a drying step.

In contrast to the prior art, where the mask is subjected to a comminution step, the method according to the first aspect cannot include a comminution step of the mask and/or pH-adjusted food additive.

Thus, according to some embodiments, the method of providing a food additive does not include a comminution step.

In contrast to the prior art, where the mask is subjected to an ensiling step, the method according to the first aspect cannot include such an ensiling step, i.e. an acidification achieved through the growth of microorganisms and fermentation, usually comprising lactic acid provided by lactic acid bacteria. Ensiling is also a time-consuming process, usually taking several days or weeks. According to some embodiments, the method of providing a food additive does not include ensiling. This is e.g. exemplified in Example 20, "microbial analysis", where it can be seen that the total cell count in cfu/g is below 100 at all times.

DK 202300025 Y3 24

In some embodiments, unlike the prior art where the mask is subjected to a heating step, the method of the first aspect may not include a heating step of the mask and/or pH adjusted food additive. Instead of a separate heating step, in some embodiments the temperature of the mask is not allowed to drop below a defined temperature, such as not below 40, 45, 50, 55, 60, 65 or 70°C throughout the treatment. In some embodiments, these temperatures differ from unit operation to unit operation, such as from solid/liquid separation to packaging.

In some embodiments, the food additive has a moisture content comparable to the moisture content of the mask. In some embodiments, the food additive has a water activity comparable to the moisture content of the mask. In some embodiments, the food additive has a moisture content of 20-60, 60-95%, 70-90%, 72-88%, or 75-85% (w/w); and/or a water activity of 0.98-1.0, 0.98-0.99, 0.985-0.998 or 0.990-0.995 or about 0.992. In some embodiments, the food additive has a moisture content of about 20-25, 25-30, 30-35, 35-40, 40-45, 45-50, 50-55, or 55-60% (w/w). In some embodiments, the food additive has a moisture content of about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, or 95% (w/w). In some embodiments, the food additive has a moisture content of at least 20, 25, 30, 35, 40, 45, 50, 55 or 60% (w/w). In some embodiments, the food additive has a moisture content of at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 , 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, or 95% (w/w).

In some embodiments, the food additive has a water activity of 0.95, 0.96, 0.97, 0.98, 0.99, 0.995, or 0.998 or more. In some embodiments, the food additive has a water activity in the range of 0.95-0.96, 0.96-0.97, 0.97-0.98, 0.98-0.99, or 0.99-1.0.

In another aspect of the invention, a feed or food additive according to the first aspect is provided.

This food additive can be packaged, e.g. as described herein. In some embodiments, the food additive has a shelf life of at least 5 days at room temperature (eg, 20 or 25°C) and/or at least 10 days under refrigeration (eg, at 4°C or 7°C). In some embodiments, the food additive may have a slightly shorter shelf life, such as at least 3 or 4 days at room temperature and/or at least 5, 6 or 8 days under refrigeration. In some embodiments, the food additive has a longer shelf life, such as at least 7, 10, 14, 20, 30 days or more at room temperature. In some embodiments, the food additive has a longer shelf life, such as at least 2, 4, 8, 10 or 12 weeks under refrigeration, such as at 7, 10, 14, 20, 30 days or more at room temperature. In some embodiments, the food additive has a shelf life of at least 3, 6 or 12 months under refrigeration.

DK 202300025 Y3 25

In some embodiments, the food additive has a shelf life of at least 3, 6, or 12 months without refrigeration, such as at room temperature.

In some embodiments, the food additive may be frozen, such as at -20°C or less.

In some embodiments, the food additive is used while frozen to provide a food product.

In some embodiments, the food additive may be classified as "pathogen free", "Sal monella free" and/or "Listeria free" at the end of shelf life. In some embodiments, the shelf life includes organoleptic stability, such as one or more of taste, texture, odor, and appearance.

In some embodiments, the food additive is for human consumption. In some embodiments, the food additive is for animal consumption. Different rules may apply to food and animal feed; such rules are known in the field. Without wishing to be bound by any theory, it is believed that a food additive as described herein may comply with such regulations.

In a third aspect, the present invention relates to uses of a feed or food additive according to the first aspect or provided according to the second aspect by providing a feed or food product.

In some embodiments, the food additive is used as a protein substitute or protein supplement in the provision of a food product. In some embodiments, the food additive is used as a fiber replacement or fiber supplement. In some embodiments, the food additive provides both protein and fiber to the food product.

Furthermore, in some embodiments, the food additive provides increased stability and/or increased shelf life.

Without wishing to be bound by any theory, this effect is believed to be caused by the addition of pH regulators and/or additional controlling properties of the number of viable cells, such as the presence of organic acid(s) and/or their salts. In some embodiments, this effect is believed to be related to the "E numbers" present in the food additive.

In some embodiments, the food product is for human consumption. In some embodiments, the food product is a feed for animal consumption. Without wishing to be bound by any theory, it is believed that

DK 202300025 Y3 26 that the use of food additives as described herein may be in accordance with the rules for animal feed and/or human food.

Regarding the use of used from different brewing processes, such as including different types of malt such as described above, the food additive may taste differently and thus impart different flavors to the food product, such as a smoky flavor, which could be desirable in some meat or meat analog containing products, such as sausages. Likewise, different types of mask can provide other organoleptic properties.

In the fourth aspect, the present invention relates to a food or feed product comprising e.g. 0.1-99.9, 1.0-99, 5-95, or 10-90% w/w of a feed or food additive according to the first or second aspect. In some embodiments, the food product comprises 0.1-1%, 1-2%, 2-5%, 5-10%, 10-20%, 20-40%, 40-60%, 60-80%, 80-90 % or 90-99.9% w/w of the food additive.

In some embodiments, the food product is a meat product, such as sausage, bread, or a cooked or baked product. In some embodiments, the food product is selected from one or more of: snacks, especially cereal bars and/or snack bars, where the snack e.g. can be pressed, fried and/or roasted; baked product; baked goods, in particular bread, crispbread, pastry; and waffles, cookie; confectionery, e.g. cake; breakfast product; sauce; beverages, such as instant drink, smoothie; pasta; puree; spreads; filling; and product comprising minced meat or meat analogue, such as sausage, salted sausage, meat dough; soup; and instant food.

In some embodiments, the food product may be classified as one or more of: vegan, vegetarian, and/or organic, including any combination thereof.

In some embodiments, the food additive provides a stabilizing effect, such as a pH-regulating effect and/or a microbial flora-related effect (eg, controlling or providing an appropriate microbial flora) to the food or feed product. In some embodiments, the food product is stabilized through the addition of the food additive.

In some embodiments, the food additive is believed to provide a pro- and/or prebiotic effect.

DK 202300025 Y3 27

In some embodiments of the invention, the food additive can be used directly as human food or animal feed. In some embodiments, the food additive is suitable for immediate (ie, without incorporation into a food product) consumption by an individual (human or animal).

In a fifth aspect, the present invention relates to a system for providing a feed or food additive according to the first or second aspect. Such a system may in particular include: x. solid/liquid separation device (20), such as a lauter tun or mash filtration system for providing a moist mash from a mash; y. preservation device (50) for bringing the moist mask into contact with at least one preservative (51, 52), which preservation device comprises a pH adjusting device for providing pH-adjusted moist mask, comprising e.g. a pH monitoring device, such as a pH meter. a dosing device for adding one or more preservatives (51, 52), . and optional stirring device (58), such as a mixer, for mixing the moist mash; and possibly z. packaging device (70), such as a filling system for packaging the pH-adjusted and/or preserved mask in a sealable container and/or tank.

The mash can be provided using a mashing device (10) from grain, preferably a malted grain such as barley malt. The person skilled in the art will know how to select the appropriate mashing device (10).

In some embodiments, the mask is provided from an industrial scale brewing process, such as a large scale brewing process.

In some embodiments, said system includes one or more transport devices, such as a pipe and pump device (40), such as an auger or pneumatic pump, for transporting the wet mask, such as from one unit operation to the next, such as from a solid/liquid separation device to the preservation and/or — packaging device, as well as one or more flow control devices (41, 42, 43, 44, 45, 46, 47, 48), such as valves or other systems or devices customary in the industry.

In some embodiments, the pH adjusting means comprises stirring means (58) such as a mixer or the like to provide mixing of the mask to provide a better distribution of one or more agents such as pH regulator and/or preservative. A result of the mixing can also be a uniform pH in the food additive. Suitable pH measuring systems and/or devices

DK 202300025 Y3 28 are known in the field, such as pH meters, pH electrodes or spectroscopy-based devices.

Said pH measurement can be a direct measurement or an indirect pH measurement, such as pH predictions using spectroscopy, such as NIR, IR, FT-NIR or FT-IR spectroscopy, in combination with multivariate data analysis.

In some embodiments, the packaging device (70) is adapted to provide airtight packaging and/or protected atmosphere packaging, such as Nj. Suitable packaging devices can e.g. be a dispensing, filling or bottling station customary in the industry. The packaging device (70) may include a pump for removing oxygen.

The packaging device (70) may include a N 2 > source to displace the oxygen with Na. Thereby, the resulting packaging is isolated from the surrounding atmosphere and placed in an inert atmosphere.

The packaging device (70) may comprise a conveyor belt or the like that transports the material to a container.

In some embodiments, the system may include additional means for controlling the number of viable cells adapted to provide a treatment that controls the number of viable cells, such as one or more of a heating system or device for heat treatment, such as a pasteurization device known in the art for the area. In some embodiments, the system comprises an air and/or O> removal or replacement system, such as a vacuum pump, or a device or system for replacing air with a protective atmosphere, such as with an N> atmosphere, as described herein, which is believed to be customary within e.g. packaging. In some embodiments, a vacuum pump or vacuum system is used to reduce air pressure. In some embodiments, a dosing system is provided for providing one or more microorganisms. Microorganisms can be provided as freeze-dried powder or suspended in an aqueous medium, and devices and systems for this are customary in the art.

In some embodiments, the system is arranged to provide and/or maintain, when in operation, a defined temperature of the wet mask. This can e.g. be at least 40, 45, 50, 55, 60, 65 or 70°C from the solid/liquid separation device to the preservation device — and/or packaging device or any other unit operation. This can e.g. achieved by insulation or heating, such as by the use of hot water and/or steam. In some embodiments, heat sensors are used to determine or assess the heat of the spent material at various stages from filtration to packaging, transport and/or storage.

DK 202300025 Y3 29

Further embodiments of the system according to the first aspect are e.g. shown in Figures 1-8. In general, unless otherwise indicated or obviously incorrect, means and/or functions shown in one figure may also be present in any other figure.

Fig. 1 illustrates a system comprising mashing device (10) to provide a mash from malt, such as barley malt, optionally comprising other cereals, and filtering device (20) to separate the mash from the liquid fraction (the wort), which can be further processed by e.g. by fermentation agents (30). The fermentation device (30) is not necessarily an essential part of the system. The mask is subjected to a preservation step comprising preservation device (50) to provide the food additive as described herein. Preservation device (50) may include devices for dosing one or more preservatives (51, 52), such as one or more pH regulator(s) and/or one or more microbial strains. The preservation device (50) may include pH measuring means. The preservation device may also include means for removing O> (55). This can e.g. is achieved by replacing air such as in the headspace with N, as described herein. The system may include packaging device (70), which is not shown here. The food supply device (90), such as a food processing plant, is not necessarily an essential part of the system and is therefore not described in detail. The dosing device can be a valve. The valve can be computer controlled.

Fig. 2 illustrates a system similar to that described in fig. 1, except for a transport step comprising the transport device (80), such as truck, which transports the packaged food additive to a food production site (90). In one embodiment of the invention, the food additive is packaged by transferring the mask to a container, such as a tank on a tanker truck. Preservation step (50) can be at least partially carried out in the container, such as in the tank of a tanker, e.g. by adding one or more preservatives (50, 51) and/or removing O> by means or methods as described herein.

Fig. 3 and 4 illustrate further embodiments of a system as shown in FIG. I and 2. In particular, the pumping device (40) is shown to transport the mask from the filtering device (20) to the location (50) where one or more preservation steps are provided (50). After addition of the one or more pH regulators (51, 52) and/or removal of O, a first storage means (60) provides storage of the food additive prior to being subjected to packaging using the packaging device (70). After packaging — the — optional — additional storage device (65) provides storage after packaging prior to transport (80) of the packaged

DK 202300025 Y3 30 food additive for the food providing device (90). The pumping device (40), although not shown in FIG. 4, may be present. The first and second storage means (60, 65) may be a first and a second container or similar storage.

Fig. 5 and 6 illustrate further embodiments of a system as shown e.g. in fig. 3. In fig. 5, an embodiment is shown in which the pH-adjusted food additive is stored in the storage device (60) after the preservation step (50) and before packaging (70) and transport (80). In fig. 6, an embodiment is shown where the pH-adjusted food additive is packaged directly after preservation using the preservation device (50), such as in a tank, tank container, tanker or the like as described previously, and transported to the food provision device.

Fig. 7 and 8 illustrate further embodiments of a system according to the present invention.

Fig. 7 illustrates a system where the mash is provided by filtering (20) a mash provided in a mash tank (10) which is then pumped by means of the pump device (40) either to a storage member/tank (60) or to the preservation device (50). The mesh flow to either the storage device (60) or the preservation device (50) can be controlled by flow control devices (41, 42), such as valves. The preservation device includes the storage device (51, 52) for a 1st or 2nd preservative, such as a first and pH regulator as described herein, and the mixing device (58) to facilitate pH adjustment and/or preservation. The food additive is then packaged using the packaging device (70). This can be a container, tank or tanker or similar. Fig. 8 shows an additional connection between the storage device (60) and the preservation device (50) that can be controlled by an additional flow control device (46). In addition, additional devices (43, 44, 45, 47, 48) for flow regulation are shown.

The mixing device (58) may be a mixer.

In general, with respect to the embodiments illustrated and described herein, one or more additional pumping devices (40) may be present in a system according to the present invention. Pumping means may also include one or more pumps, use of gravity, air pressure and/or other known pumping and/or transport means suitable for transporting the mask, herb, food additive, pH regulator, etc. . Likewise, one or more flow control devices such as valves (41-48) may be present to control the flow of mash, herb, food additive, pH regulator, etc. Instead of or in combination with flow control devices, pump devices may also be used. Often, pumping devices and/or flow regulating devices are provided between different unit operations. With regard to the preservation device (50), they are also in some

DK 202300025 Y3 31 embodiments a container or container where the mask is brought into contact with a pH regulator.

Often pH measuring devices are also provided, often in combination with as dosing means for dosing the one or more pH regulators and/or to achieve a preset pH value. In some embodiments, means for removing or replacing oxygen, such as with nitrogen, may be provided. In some embodiments, an agitation device (58) is provided. This may be a simple agitator or pump or other suitable systems in the art for mixing a product of similar viscosity to wet mash.

As described herein, the system may include one or more pipes for transporting mash to a storage device (60) for storing mash, and one or more pipes for transporting mash to the preservation device (50), where the flow of mash can be directed to the storage device or preservation device by means of one or more flow control devices (41, 42). In some embodiments, the system includes a pipe for transporting mash from the storage device (60) to the preservation device (50), and optionally includes a flow control device (46) for regulating the flow of mash from

The chaser device (60) for the preservation device (50).

In some embodiments, the solid/liquid separation device (20) comprises two filtration units in series, where the first unit, when in operation, is adapted to supply wet mash with a first water content, such as a water content of about at least 60, 70, 80, 85% or more (w/w), and the second filtration provides a wet mask with a different water content, such as at least 20, 30, 40, 50 or 60%. Usually the first water content is higher than the second water content. It is believed that such a two-stage filtration system may be advantageous in providing mash with a lower water content.

The inventors have realized that an existing brewing system can be modified to allow the provision of a worm-derived food additive. By and large, this modification and/or adaptation can be accomplished using conventional components, materials, and/or equipment. Thus, in a further aspect of the invention, the invention relates to the modification of an existing brewing system by incorporating a system according to the fifth aspect as described herein.

In a sixth aspect, the present invention relates to a computer program product comprising software code portions configured to, when executed in a computer's memory, perform (A): the method according to the first aspect and/or (B) controlling the system according to the fifth aspect.

In some embodiments, the computer program product comprising software code portions is configured

DK 202300025 Y3 32 to, when run in a computer's memory, perform the method of providing a product according to the first aspect.

In a seventh aspect, the present invention relates to a modification of a brewing system by incorporating a system according to the fifth aspect. The invention relates to a modified brewing system in which a system according to the fifth aspect is incorporated.

Further details and/or embodiments related to the present invention can e.g. found in the following examples.

EXAMPLES

Example I - Provision of mask

Mash samples are harvested after mashing from brewing processes 1 industrial scale for different beer types and production methods. The mask is harvested without delay (e.g. within 15 minutes of solid-liquid separation while still warm (e.g. around 50-60 °C).

Example 2 - Analysis of water/moisture content and/or dry matter

Water content: ISO 712 4th edition 2009-11-15.

Principle: The sample is dried at 130°C until it is stable and the weight loss is determined by weighing.

Example 3 - Analysis of water activity

NMKL 168.

Principle: Water activity (aw) is a measure of the proportion of water in a sample that is available for microbiological growth. It describes the relative humidity of the enclosed sample after equilibrium has been reached with respect to temperature and humidity. The water activity is influenced by the composition, temperature and water content of the sample and can be defined as: aw= p/po where p is the equilibrium water vapor pressure above the food and po is the saturated vapor pressure above deionized water at the same time. Temperature. The water activity can therefore also be defined as: aw= ERH/100 where ERH is the equilibrium for relative humidity.

Example 4 - pH measurement

NMKL 179.

DK 202300025 Y3 33

Principle: pH is defined as the negative log of the concentration of H+ ions (pH = - log [H+]).

The pH of the sample is measured by measuring the potential between a pH-selective electrode and a reference electrode after calibration using buffers of known pH. Temperature compensation, preferably automatic temperature compensation is used if necessary with 25°C as reference temperature.

Example 5 - Microbial analysis

Bacillus cereus ISO 7932

Principle: Presumptive Bacillus cereus is enumerated on the surface of a selective and differential medium.

Clostridium botulinum ISO 17919

Principle: A horizontal method for the molecular detection of Clostridia carrying botulinum neurotoxin

A, B, E and F genes - which are the types that are pathogenic to humans - by real-time PCR.

Salmonella NMKL 71

Principle: Salmonella are counted on the surface of a selective and differential medium after pre-enrichment and enrichment in non- and selective media.

Staphylococcus aureus ISO 6888-1

Principle: Staphylococcus aureus is counted on the surface of a selective and differential medium.

Pathogenic E coli ISO 16649-2

Principle: E. coli is counted using a colony counting technique at 44 °C on a solid medium containing a chromogenic ingredient to detect the enzyme — glucuronidase. -glucuronidase- positive Escherichia coli grow with typical blue colonies.

Total cell count 30°C: ISO 4833

Principle: The total number of bacteria grown at 30°C is enumerated using a non-selective medium.

Yeast and mold: NMKL 98 mod.

Principle: Yeast and mold are enumerated on the surface of a selective and differential medium.

Sulphur-reducing bacteria ISO 15213

Principle: Anaerobic sulphite-reducing bacteria are detected and enumerated on iron sulphite agar, where they form typical black colored colonies.

The black color of the colonies and the surrounding zone is due to the formation of iron (Il) sulphide as a result of the reaction between sulphide ions and trivalent iron [Fe(III)] present in the medium.

Aerobic and anaerobic spores NMKL 189

Principle: Aerobic and anaerobic spores are counted on a non-selective medium after heat treatment of the sample to kill all vegetative cells.

The samples are incubated under both aerobic and anaerobic conditions.

DK 202300025 Y3 34

Example 6 - Organoleptic analysis

NMKL procedure 6, ISO 8585, ISO 8586

Principle: Evaluation of the organoleptic quality of food by assessing the various properties based on the basic tastes sweet, sour, salty and bitter. The organoleptic properties are assessed in relation to normal characteristics of the samples.

Example 7

Stability analysis

Samples are supplied in an airtight container with minimal head space and stored refrigerated (4 or 7°C) and at room temperature (20 or 25°C) and analyzed at regular intervals until spoiled, e.g. according to example 5 - microbial analysis and/or example 6 - organoleptic analysis.

If necessary, the headspace can be replaced with Na.

Example 8

Provision of stock solutions

Stock solutions of inorganic and organic acids and salts are prepared according to standard laboratory methods. Depending on sample size such as desired pH and buffer capacity of the samples, e.g. 1 M stock solution may be used directly or diluted accordingly if necessary to facilitate accurate titration.

Example 9

Titration

Titration and/or addition of various acids/salts for pH adjustment is performed with stirring using conventional laboratory equipment. Care is taken to avoid microbial contamination.

In general, samples must not cool below 40°C.

Example 10

Provision of moist mask sample A

Mask is obtained according to example 1 from a lager brewing process exclusively from malt without sparging. Different parameters are analyzed according to examples 2-6.

Example 11

Provision of moist mask preve B = Example 10 + sparging (ie washing the filter cake with water to extract additional sugars)

Example 12

Provision of moist mash sample C = Example 10, but from a "high gravity" brewing process

Example 13

Provision of moist mash sample D = Example 10, but brewed according to "Reinheitgebot".

Example 14

Provision of moist mash sample E = Example 10, but brewed with malt and raw fruit.

Example 15

Provision of various food additives (FA 1-35) using a single pH regulator

FA Sample pH set to Inorganic | Acetic acid/ | Lactic acid/ | Malic acid/s | Tartaric acid/salt | Sorbic acid # a/b/c k acid salt salt all e/salt 6 | A| 454038 | none | = the egg | none | none | X| none 8 | B | none | none | none | none | none | none | none 9 | B | 454038 | x | none | none | none | none | none 10 | B | 454038 | none | x | none | none | none | none 11 | B | 454038 | none | = the egg | x | none | none | none 12 | B | 454038 | none | none | none | x | none | none 13 | B | 454038 | none | none | none | none | X| none 14 | B | 454038 | none | none | none | none | none | x 22 | D | none | none | none | none | none | none | none 23 | D | 454088 | x | none | none | none | none | none 24 | D | 454038 | none | x | none | none | none | none | D | 454038 | none | = the egg | x | none | none | none 26 | D | 454038 | none | none | none | x | none | none 27 | D | 454038 | none | none | none | none | X| none 28 | D | 454038 | none | none | none | none [| none | x none: compound not added x: compound added

Stability analysis is performed in accordance with Example 7.

Using an organic acid to reduce the pH has a stronger antimicrobial effect than reducing to the same pH using an inorganic acid such as HCl. This is believed to be due to the combined antimicrobial effect of the lower pH and the direct antimicrobial effect of the undissociated organic acid.

The proportion of undissociated organic acid is inversely proportional to pH: The lower the pH, the higher the proportion of the organic acid in undissociated form. In general, the lower the pH, the greater the additional benefit of using an organic acid compared to using an inorganic acid

Example 16

Provision of different food additives (FAIOI-138) using two pH regulators

FA £ Sample | pH set Inorganic | Acetic acid | Lactic acid/ | Cider | Tartaric acid/s Sorbinsy tila/b/c k acid e/salt salt e/salt alt re/salt 100 [A [4540 [x Ix J 00000 000 0000p 000] jus [E [4540 0 [x 0 Ix LP 000] none: compound not added x: compound added

Stability analysis is performed according to Example 7.

Using a combination of organic acids further strengthens the antimicrobial effect due to differences in the specific mode of action of the different acids. e different effects in the mode of action are linked to the differences in the molecular structure of the acids, which can lead to synergistic interactions.

The combination of two organic acids can provide microbial stability at a higher pH than a single organic acid.

Example 17

Preparation of various food additives (FA201-238) by addition of LAB (lactic acid bacteria)

FA Mask pH LAB Inorganic | Acetic acid/ | Lactic acid | Cider | Tartaric acid/ | Sorbinsy # sample set to | strain I/Il | acid salt e/salt e/salt salt re/salt a/ble 208 |B [4540 [x none [none [none [none [none [none 209 |B [4540 |x [x none [none [none [none | no 210 |B [4540 [x no |x [no [no [no [no (211 |B [4540 [x no [no |X [no [no [no 1212 [B [4540 |x [no [no [ none |x [none [none 1213 |B [4540 [x none [none [none [none [X [none 1214 |B [4540 |x [none [none [none [none [none |x 1222 |D [4540 [ x no [no [no [no [no [no 1223 |D [4540 [x |x no [no [no [no [no 1224 |[D [4540 |x [no |x [no [no [no [no 225 |D [4540 [x — none [none |X [none [none [none 1226 |D [4540 |x [none [none [none |x [none [none 1227 |D [45/40 [x — [none [none [none [none [X [none 228 |D [4540 |x [none [none [none [none [none |x

LAB I = Lactobacillus plantarum

LAB II = Lactococcus lactis

Stability analysis is performed according to Example 7.

DK 202300025 Y3 38

The use of live lactic acid bacteria in connection with reduced pH by the addition of one or ganic acid gives a triple antimicrobial effect: Lowered pH + undissociated organic acid + competitive effect of another microorganism.

The competitive effect is particularly effective when a food-approved microorganism is used that is well adapted to the lowered pH, such as the lactic acid bacterium.

Furthermore, using a live lactic acid bacteria culture can increase the nutritional value of the product and/or provide a pre- and/or probiotic effect.

Example 18

Microbial analysis of a worm sample (CFU/g)

Parameter Method Result Unit

Total number ~~ NMKL86 — 32x10° CFU/g

Yeast NMKL 98 mod. 3.7x 10" CFU/g

Mold fungus NMKL 98 mod. < 100 CFU/g

S-ed. bakers ISO 15213 < 400% CFU/g

Aerobic spores NMKL 189 < 400% CFU/g

Anaerobic spores NMKL 189 < 100 CFU/g aW NMKL 168 0.99 per unit pH NMKL 179 6.02 per unit * A number of organisms below the detection limit have been detected

Example 19

Microbial analysis of another worm sample (CFU/g) over time

Week Total number of LAB Mold — Yeast — 0 0 17x10% — 18x10% ~~ <10 <10 2 21 x 109 19 x 10° n.a. reach. 4 60 x 10° 55x 10° n.a. reach. 6% 3.0 x 10° 2.5 x 10° n.a. reach. 10 54 x 10° 39 x 10° <10 > 1500 14 22 x 10° 21 x 109 n.a. reach. 18 63 x 10° 86 x 10° n.a. reach.

Reference methods: Total number: ISO 4833-1; LAB: FVST 08:2007; yeasts and molds; NMKL 98:2005; reach. not analyzed *) week 6 appears to be an outlier

DK 202300025 Y3 39

Example 20

DURABILITY AND RISK ASSESSMENT OF VAD MASK

Product description, processing and handling

The product wet mash is initially a by-product from beer production. After the mashing process in ale production, the malt is filtered and possibly pressed. The mask will be transferred directly to a packing station, where addition of organic acids (and diacetate) will be mixed into the product before packaging (see e.g. Figures 1-8). It is then sent directly to food manufacturers as a protein-rich ingredient for us age e.g. in the bakery industry or as a partial meat substitute in various types of sausages. In beer production, the mash is heated to over 70 °C, and most often kept at 76 °C for at least 30 min.

In the mashing process, barley malt is granulated, dissolved in water and heated at different temperature levels. In the last step, the temperature is kept above 75 °C for approx. 30 min., which corresponds to pasteurisation. After this process, most of the soluble carbohydrates, mostly starch, originally present in the barley malt are dissolved and enzymatically converted to mainly maltose and maltotriose. The water phase, called wort, is filtered and used for beer production. The remaining solids, called wet mash, consist mainly of husks from barley malt and are very rich in protein and cellulose. It is expected that some residual sugars from the water phase are present on the surface of the shells.

It may be desirable for the pH-reduced product to be packaged in various sizes of airtight bags with a minimum of headspace. In some cases, the product is packaged in larger containers where an inert gas such as nitrogen is used as a cover for the headspace.

Purpose and background

Normally, for such a product, wet mash, which is not pH-reduced, a cooling process immediately after production will be necessary to ensure the safety and durability of the product within a certain shelf life. Such a process will require high energy consumption, which will negatively affect the value of the product. The results listed here must document that cooling can be replaced by a pH reduction without risks in terms of the product's safety and organoleptic properties.

Based on a previous risk and shelf-life assessment, it was clear that the non-pH lowered product was very unstable with respect to the growth of microorganisms. The product was also concluded to be harmless and can support significant growth of a number of pathogenic bacteria in a short time. DNA-based analyzes showed the presence of DNA from pathogenic organisms (date not shown). In particular, DNA from B. cereus was found in the product. The conclusion of the previous risk - and durability as

DK 202300025 Y3 40 assessment was that all disease-causing organisms could be prevented by reducing the pH to below 4.2 using organic acids, except for Salmonella.

Salmonella is known to be a potentially problematic pathogen in pH-reduced products.

Salmonella isolates can grow in conditions with pH values down to a pH of 3.7 to 4.2 depending on the type of matrix.

Nine different blends of organic acids and diacetate are used to make 9 different variations of the pH-reduced wet mask product. In a pre-test, these mixtures have been selected based on their organoleptic properties and their functionality as a preservative (pre-test results not shown). Five of these 9 mixtures are selected to produce 5 variants of the pH-reduced product and used for a Salmonella challenge test. The results of this test are used as documentation for the safety of the product within a shelf life of 7 days at 25°C.

Sample data

Item description: Wet mask from Biograin ApS

Sample/batch number customer: Biograin 08/07/2020

Sample no. ISI Food Protection: P-1892

Sample received: 08.07.2020

Temperature: Warm

Condition of the sample on arrival: Still warm, delivered directly after production

Storage in ISI Food Protection Freezer at -18°C

Analysis performed on arrival

Parameter Method Results (cfu/g)

Total cell count, 30 °C aerobic NMKL 86 <100

Total cell count, 30 °C anaerobic NMKL 86 <100

Gives NMKL 98 mod. <100

Mold fungus NMKL 98 mod. <100

Durability test

Nine different pH-reduced variants of the product + 1 reference product are supplied. All acids and diacetate are premixed in water. Water is added to the product (30% V/w in the mix) to improve the mixing and distribution of acids and diacetate in the wet mash.

DK 202300025 Y3 41

The following combinations are produced, where the percentage [w/v] accounts for the use of pure undissolved acids and diacetate: 1) 0.5% acetic acid + 0.5% lactic acid + 0.2% malic acid 2) 0.5% acetic acid + 0.2 % lactic acid + 0.5% malic acid 3) 0.7% acetic acid + 0.3% lactic acid + 0.2% malic acid 4) 0.7% acetic acid + 0.3% lactic acid + 0.2% malic acid + 0.2% diacetate 5) 0.8% acetic acid + 0.4% lactic acid + 0.2% diacetate 6) 0.4% acetic acid + 0.8% lactic acid + 0.2% diacetate 7) 0.8% acetic acid + 0.4% lactic acid 8) 0.4% acetic acid + 0.8% lactic acid 9) 1.2% acetic acid + 0.2% diacetate 10) Reference (only water is added)

After production, the pH and the water activity, aw, are analyzed.

The products are packed in sealed cups of 100 ml and stored at 25°C. An airtight partition is placed in the lid and used to measure the gas composition in the headspace. The gas composition is measured for 12 days. It is believed that production of CO2 indicates microbial growth and metabolism.

After 7 and 30 days of storage, the total bacterial cell count and yeast/mold cell count are analyzed for each sample.

After 12 days of storage, 2 individual laboratory technologists are 1 time assessing the organoleptic properties of the samples by describing the properties of the samples, especially smell.

Salmonella challenge test

Overview of the recipes for the produced product variants included in the challenge test. Sample No. refers to the same number used in the durability test:

Sample preparation

Sample Wet mask Acetic acid Lactic acid Able acid Diacetate Water Total [g] [ml] [ml] [ml] [e] [e] [ml] [g] 2 210.0 15.0 0.8 1.5 0.0 72.8 300.0 3 210.0 21.0 1.1 0.6 0.0 67.3 300.0 6 210.0 12.0 3.0 0.0 0.6 74.4 300.0 8 210.0 12.0 3.0 0.0 0.0 75.0 300.0 9 210.0 36.0 0.0 0.0 0.6 53.4 300.0

DK 202300025 Y3 42

Five of the pH-reduced variants of the product from the shelf-life test were selected for a Salmonella challenge test (see table above).

Indicator strains and their manufacture

A pool of approximately equal amounts of the following six Salmonella strains is provided from fresh overnight cultures in BHI, grown at 37°C for 18 hours: . Salmonella 4.12 1- from salami (ISI 2344) . Salmonella 4.5, 12:1- from salami (ISI 2346) . Salmonella Choleraesuis from salami (ISI 1158) - Salmonella Enteritidis from pork (ISI 2938). Salmonella Derby from barn, pig (ISI 173) . Salmonella Typhimurium DTI 2 from barn, pig (ISI 166)

The strains are stored in the ISI FOOD PROTECTION ApS (Aarhus, DK) strain collection as frozen cultures at -80 °C. The six test strains are prepared separately and mixed before use in approximately equal concentrations based on cell counts from each individual strain.

A contamination rate of 500 cfu/g is aimed for when adding 100 ml of inoculum to 200 g of each type of product. After addition, the samples are thoroughly mixed for 2 min.

The samples are stored in air-sealed cups at 25 °C.

Sampling plan

Analyzes performed during the challenge test on each pH-reduced product variant.

Day 0 Day 7

Total bacterial count, anaerobic, at TSA-SB, NMKL 189 1 1

Quantitative enumeration of Salmonella on XLD-selective media 1 3

Determination of pH value, NMKL 179 3 3

Determination of water activity, aw, NMKL 168 1 1

RESULTS

Durability test

Intrinsic and extrinsic parameters analyzed during durability tests at 25°C and 30 days of storage. pH values are averages of triplicates.

Sample Day 0 Day 7 Day 12 Day 21 Day 30 pb gg gee ee gd gb gg ee ed

Ce amr us 05 ser ead x6 oy AND I A343

The pH reduced samples 1-9 had a stable pH during storage of up to 30 days at 25°C. The pH value in sample 10 was decreasing during storage. This indicates microbial growth in which some species were acid-producing microorganisms.

Water activity (aw) was stable in all samples at 0.98-0.99 approx. for 7 days.

The production of CO2 was less in samples 1-9, while a large amount of CO2 was produced in sample 10.

This indicated that microbial growth was stabilized in samples 1-9 and not in sample 10 where microorganisms produced CO2 during respiration and growth. The small decrease in CO2 concentration from day 7 to day 12 was most likely a consequence of the septum not being completely gas-tight after the first penetration on day 7.

Organoleptic properties

On day 12 during storage, the organoleptic properties were assessed. Samples 1-9 all had a mild fresh sour smell. Sample 10 had an unpleasant, rotten smell. This indicates that samples 1-9 will not introduce off-odors when used as an ingredient in a food product.

DK 202300025 Y3 44

Microbial analyses

Microbial analyzes during 30 days of storage at 25°C

Total number | Mold | Total number of Mold Total number of Mold cells [cfu/g] and yeast [cfu/g] cells [cfu/g] and yeast [cfu/g] cells [cfu/g] and yeast [cfu/g] tv ft le LE TE EE See : 1m | SHo : wp : Samant : seams — amet : <a : microbial analyzes showed no microbial growth in the pH-reduced variants (sample 1-9). A significant outgrowth of bacteria and some growth of yeast was found in reference sample 10. This was consistent with the results for pH, gas production and organoleptic properties.

The results of the microbial analysis are presented above.

Summary of Durability Tests

Analyzes during shelf life tests show that the pH-reduced samples were stabilized, while the reference sample is spoiled. The background flora detected in the 9 pH-reduced variants of the product does not introduce any safety or spoilage risks were consumed (data on microbiome composition using NGS not shown).

Salmonella Challenge Test

Salmonella inoculum count

Inoculum target 1x 106 cfu/ml

Seed cell count on XLD 1.1 x 106 cfu/ml

Calculated inoculation rate in samples 550 cfu/g

Average inoculum recovery rate 72 cfu/g

Log 10 recovery rate 1.86 Logl0 cfu/g

The Salmonella recovery cell count was calculated to be 550 efu/g based on the inoculum cell count. After inoculating the pool, it took up to 1 hour for the sample to be diluted and analyzed. The low recovery rate analyzed, 72 cfu/g, could indicate that the low pH values had a "killing effect" on

DK 202300025 Y3 45

The Salmonella during the first 1 hour until analysis. The low pH and combinations of acids were expected to introduce a possible "kill" effect of Salmonella. Acid adaptation during the preparation of the inoculum could have minimized this "killing effect" after the addition of acids. However, Salmonella strains that may be present in the product's production environment will not be acid-adapted and therefore this was not performed with the inoculum.

After 7 days, no Salmonella cfu were detected in any of the pH-reduced samples (see below).

Thus, none of the 5 product variants produced supported the growth of Salmonella in this test.

Test result of Salmonella challenge test during 7 days of storage at 25°C

Salmonella on XLD Slog 10 Total number of [cfu/g] triplicates (from day 0) TSA-SB (single) 2 IQ 38 <30 % SUED 23a IO 5 C30 3.38 v30 8 SEH SHES <YO a 39 138 <30

Intrinsic parameters in the produced samples

SAMPLE | DAY a | DAY 7 fripdicates) {shade} | foripdicates) {singled

Nn cass | AN can en DEB EO nae

DISCUSSION AND CONCLUSION

The pH of the product was successfully lowered by preparing 9 different varieties of the products using 9 different combinations of organic acids and diacetate.

The impact on the organoleptic properties was less. 30% addition of water was used to ensure proper mixing of acids with the product.

The shelf life test indicates that the 9 different pH-reduced variants of the product were stable within a period of 30 days. Microbial analyzes document that the product was completely stable within 7 days at 25°C (data not shown). The background flora was completely suppressed by

DK 202300025 Y3 46 the reduction of pH and poses no risks when consumed. The non-pH-reduced reference product was completely destroyed after 7 days of storage at 25°C.

Salmonella was identified as the most important pathogen for the safety of the pH-lowered product if the pH is lowered to pH 4.2 or below.

A Salmonella test was performed on 5 of the 9 different varieties of the pH lowered product to test this issue. Here, it was found that Salmonella was unable to grow in all 5 of the tested pH-reduced variants of the product. The Salmonella cell count decreased in the product matrix after inoculation. Indication of a kill of Salmonella was seen during recovery within the first hour after inoculation. After 7 days of storage at 25°C, Salmonella was not detected (detection limit: <10 cfu/g) in any of the 5 different variants of the product included in the test.

List of references

Buffington, J. (2014) The Economic Potential of Brewer's Spent Grain (BSG) as a Biomass

Feed stock. Advances in Chemical Engineering and Science, 4, 308-318.

Claims (54)

DK 202300025 Y3 47 REQUIREMENTS 1. pH-adjusted product and/or food additive provided by a method comprising the steps of: (a) providing moist mash; (b) adjusting the pH of the moist mash to 4.5 or below, such as a pH in the range of 3.5-4.5, by adding a pH regulator to provide a pH adjusted product and/or food additive; and optionally (c) packaging the pH-adjusted product to provide a packaged pH-adjusted product and/or food additive. 2. Product according to claim 1, wherein the method further comprises the step of: (d) subjecting the wet mask, pH-adjusted product, packaged product or food additive to a further treatment other than step (b), such as a preservation step or a further addition of a pH regulator at another time. 3. Product according to claim 1 or 2, wherein the moist mask has a water content of at least 20, 30, 40, 50, 60, 70, 72, 75, 78, 80 or 85% w/w (w/w) or more . 4. Product according to any one of the preceding claims, wherein the wet mask has a water content of at least 70, 72, 75, 78, 80 or 85% by weight or more. 5. Product according to any one of the preceding claims, wherein the mask is fresh, ie. the mask is provided within 1, 2, 3, 4, 5, 7, 10 or 15 minutes of herb separation or less; or within 20, 30, 45 or 60 minutes of herb separation or less 6. Product according to any one of the preceding claims, wherein the mask is provided by one or more continuous solid-liquid separation steps or by one or more batch solid-liquid separation steps. 7. Product according to claim 6, wherein the method comprises 2 solid-liquid separation steps, such as 2 continuous solid-liquid separation steps, 2 batch separation steps or a combination of a continuous and a batch separation step in any order. DK 202300025 Y3 48 8. Product according to any one of the preceding claims, wherein the mask has a temperature of at least 40°C; 45°C; or 50°C. 9. Product according to any one of the preceding claims, wherein the mask has a temperature of at least 50°C; 55°C, 60°C, 65°C; or 70°C. 10. Product according to any one of the preceding claims, wherein the mask has a moisture content of 20-95%, 20-30%, 30-40%, 40-50%, 50-60%, 60-95%, 60 -80%, 70-90%, 72-88% or 75-85% or 80-95% w/w. A product according to any one of the preceding claims, wherein the mask has a water activity of 0.98-1.0, 0.985-0.998 or 0.990-0.995 or about 0.992. 12. The product of any one of the preceding claims, wherein the pH is adjusted to 4.4 or below; 4.3 or below; 4.2 or below; 4.1 or below; 4.0 or below; 3.9 or below; 3.8 or below; 3.7 or below; 3.6 or below; or 3.5 or below. 13. Product according to any one of the preceding claims, wherein the pH is adjusted to pH 3.5-4.5 or pH 3.75-4.2; and/or about pH 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, or 4.5. 14. Product according to any one of the preceding claims, wherein the pH regulator(s) is provided in a concentration of 0.4-2.0, 0.5-1.8, 0.6-1.6 or 0.8-1.4% weight per volume (weight/volume) or weight/weight ; and/or about 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1, 5, 1.6, 1.7, 1.8, 1.9, or 2.0% w/v or w/w. 15. Product according to any one of the preceding claims, wherein the pH regulator is or comprises one or more of: (i) one or more organic acids, (ii) one or more salts of an organic acid, (iii) a or more inorganic acids and/or (iv) one or more salts of an inorganic acid; including any combination(s) thereof. 16. Product according to claim 15, where the organic acid or acids are selected from: acetic acid, lactic acid, malic acid, tartaric acid and sorbic acid, including any combination thereof; and/or where the salts of the organic acid(s) are selected from: sodium acetate, sodium diacetate, potassium acetate, calcium acetate, sodium lactate, potassium lactate, calcium lactate, sodium malate, sodium hydrogen malate, DK 202300025 Y3 49 calcium malate, sodium tartrate, potassium tartrate, calcium tartrate, sodium sorbate, potassium sorbate and calcium sorbate, including any combination thereof. 17. Product according to any one of claims 15 or 16, wherein the organic acid and/or the salt of the organic acid is a compound with E number 170(1), 200, 201, 202, 203, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 260, 261(1), 262, 2621), 262(ii), 263, 270, 280, 281, 282, 283, 290, 296, 297 , 300, 301, 302, 325, 326, 327, 330, 33 161), 331 (iii), 332(1), 332(ii), 333(iii), 334, 338, 339, 340, 341, 343 , 350(), 350(ii), 352(i1), 355, 356, 363, 380, 450, 500(i), 500Gii), 5011), 501(ii), 503( 1), 503(ii) , 5046), 504(ii), 513, 5146), 514Gi), 51561), 516, 524, 525, 526, 527, 528, 529, 530, 574, 575, 577, or 578. 18. Product according to any preceding claim, wherein the pH regulator is or comprises acetic acid, such as 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 , 0.9, 1.0, 1.2, 1.4, 1.6, 1.8, or 2.0% w/v or w/w acetic acid. 19. Product according to claim 18, further comprising one or more of: lactic acid, malic acid and sodium diacetate; including any combination thereof. 20. Product according to any preceding claim, wherein the pH regulator is or comprises lactic acid, such as 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 , 0.9, 1.0, 1.2, 1.4, 1.6, 1.8 or 2.0% w/v or w/w lactic acid. 21. Product according to any preceding claim, wherein the pH regulator is or comprises malic acid, such as 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 , 0.9, 1.0, 1.2, 1.4, 1.6, 1.8, or 2.0% w/v or w/w malic acid. 22. Product according to any preceding claim, wherein the pH regulator is or comprises sodium diacetate, such as 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 , 0.9, 1.0, 1.2, 1.4, 1.6, 1.8 or 2.0% w/v or w/w sodium diacetate. 23. Product according to any one of the preceding claims, wherein the pH regulator is or comprises: + -0.5% acetic acid + ~0.5% lactic acid + -0.2% malic acid; + —0.5% acetic acid + ~0.2% lactic acid + —0.5% malic acid; + —0.7% acetic acid + ~0.3% lactic acid + —0.2% malic acid; DK 202300025 Y3 50 + ~0.7% acetic acid + ~0.3% lactic acid + ~0.2% malic acid + ~0.2% sodium diacetate; + —0.8% acetic acid + ~0.4% lactic and/or malic acid + ~0.2% sodium diacetate; + —0.4% acetic acid + ~0.8% lactic and/or malic acid + ~0.2% sodium diacetate; + —0.8% acetic acid + ~0.4% lactic and/or malic acid; «-0.4% acetic acid + ~0.8% lactic and/or malic acid; or + —1.2% acetic acid + —0.2% diacetate; where all % are weight/volume or weight/weight. 24. A product according to any one of the preceding claims, wherein the pH regulator is or comprises: + acetic acid: lactic acid: malic acid in a ratio of about 1:1:0.4; 1:0.4:1; 0.7:0.3:0.2; 0.25-0.75:0.25-0.75:0.1-0.5; 0.25-0.75:0.1-0.5:0.25-0.75; or 0.5-1.0:0.1-0.5:0.1-0.5; « acetic acid: lactic acid: malic acid: sodium diacetate in a ratio of about 0.7:0.3:0.2:0.2; or 0.5-1.0:04-0.5:0.1-0.5:04-0.5; « acetic acid: lactic acid and/or malic acid: sodium diacetate in a ratio of about 0.8:0.4:0.2; or 0.4:0.8:0.2; or 0.5-1.0:0.25-0.75:0.1-0.5; or 0.25-0.75:0.5-1.0:0.1-0.5; «acetic acid: lactic and/or malic acid in a ratio of about 0.8:0.4; 0.4:0.8; 0.5-1.0:0.25-0.75; or 0.25-0.75:0.5-1.0%; or « acetic acid: sodium diacetate in a ratio of about 1.2:0.2; or 1.0-1.5:0.1-0.5; + where said ratio is weight/weight or weight/volume. 25. Product according to any one of the preceding claims, wherein the pH-adjusted product is packaged in a sealable container, such as a tank container, tank truck, semi tank trailer, tank trailer, barrel, airtight big-bag, plastic bag, Tetra Pak, glass container, metal container, aluminum container, barrel or can. 26. Product according to any one of the preceding claims, wherein the package size is in the range 0.1-11; 1-1001; 0.1-1 m3; 1-10 m3; or 10-50 m3. 27. Product according to any one of the preceding claims, wherein the additional viable cell count controlling treatment comprises one or more of: heat treatment, pasteurization, removal of 0»; packaging under N» atmosphere, reduction of air pressure, addition of one or more micro-organisms that are generally recognized as safe (GRAS) and/or have a positive QPS status on the "QPS notification until 2019" list. 28. Product according to claim 27, where the GRAS microorganism is an acid-producing microorganism, such as lactic acid bacteria, e.g. from the genus: Lactobacillus, Pediococcus, Enterococcus, DK 202300025 Y3 51 Lactococcus, Leuconostoc, Oenococcus, Streptococcus, Tetragenococcus, Carnobacterium, Vagococcus, Weissella and Alkalibacterium. 29. Product according to claim 27 or 28, where the microorganism is selected from one or more: (1) Lactobacillus, such as one or more of Lactobacillus acidophilus, Lactobacillus amylolyticus, Lactobacillus amylovorans, Lactobacillus animalis, Lactobacillus brevis, Lactobacillus buchneri, Lactobacillus casei, Lactobacillus cellobiosus, Lactobacillus delbrueckii, Lactobacillus diolivorans, Lactoba cillus farciminis, Lactobacillus farciminis, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus hilgardii, Lactobacillus lactis, Lactobacillus mucosae, Lactobacillus paracasei, Lactobacillus parafarraginis, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus paracasei, Lactobacillus sakei, Lactobacillus salivarius, including any combination thereof; (ii) Lactococcus, such as Lactococcus lactis; or (iii) Pediococcus, such as one or more of Pediococcus acidilactici, Pediococcus parvulus, Pediococcus pentosaceus, including any combination thereof. 30. Product according to any one of the preceding claims, wherein the method does not comprise a drying step. 31. Product according to any one of the preceding claims, wherein the method does not comprise a comminution step. 32. Product according to any one of the preceding claims, wherein the method does not include ensiling. 233. The product of any one of the preceding claims, wherein the food additive has a moisture content of 70-90%, 72-88% or about 75-85% w/w; or a moisture content of 20-30%, 30-40%, 40-50%, 50-60%, 60-80% or 80-95% w/w. 34. A product according to any one of the preceding claims, wherein the food additive has a water activity of 0.98-1.0, 0.98-0.99, 0.985-0.998 or 0.990-0.995 or about 0.992. 35. Product according to any one of the preceding claims, which is a food additive. 36. Food additive according to claim 35, where the additive is for human or animal consumption. DK 202300025 Y3 52 37. Use of a moist mask-derived food additive according to claim 35 or 36 for providing a food product. 38. Use according to claim 37, where the food additive is used as a protein substitute or protein supplement; and/or as a fiber replacement or fiber supplement. 39. Use according to claim 37 or 38, where the food product is for human consumption. 40. Use according to claim 37 or 38, where the food product is a feed product for animal feed. 41. Food product comprising 0.1-99.9, 1.0-99, 5-95 or 10-90% by weight of a food additive according to claim 35 or 36. 42. Food product according to claim 41, where the food product is selected from one or more snacks, especially cereal bars and/or snack bars, where the snack e.g. can be pressed, fried and/or roasted; baked product; baked goods, in particular bread, crispbread, pastry; and waffles, cookie; confectionery, e.g. cake; breakfast product; sauce; beverages, such as instant drink, smoothie; pasta; puree; spreads; filling; and product comprising minced meat or meat analogue, such as sausage, cured sausage and meat dough; soup and instant food. 43. Feed product comprising 0.1-99.9, 1.0-99, 5-95 or 10-90% w/w of a food additive according to claim 36. 44. System for providing a feed or food additive according to claim 35 or 36 and/or a system for carrying out the method according to any one of claims 1-34, which system comprises: a. solid/liquid separation device (20), such as a lauter tun or mash filtration system to provide a wet mash from a mash; b. preservation device (50) for bringing the moist mask into contact with at least one preservative (51, 52), which preservation device comprises a pH adjustment device for providing pH-adjusted moist mask, comprising e.g. (1) a pH monitoring device, such as pH meter (11) a dosing device for adding one or more preservatives (51, 52), DK 202300025 Y3 53 (iii) and optional stirring device (58), such as a mixer for mixing the wet mask; and optionally c. packaging device (70), such as a filling system for packaging the pH-adjusted and/or preserved mask in a sealable container and/or tank. 45. System according to claim 44, further comprising one or more transport devices, such as a pipe and pump device (40), such as an auger or pneumatic pump for transporting the moist mash from the solid/liquid separation device to the preservation device and/or packaging device, as well as a or more flow control devices (41, 42, 43, 44, 45, 46, 47, 48), such as valves. 46. System according to claim 44 or 45, wherein the packaging device (70) is adapted to provide an airtight packaging, optionally under a protected atmosphere, such as No. 47. The system of any one of claims 44-46, further comprising viable cell count control means adapted to provide a viable cell count control treatment, such as one or more of heat treatment, pasteurization, removal of O,, packing under N, atmosphere, reduction of air pressure and/or provision of one or more microbial strains. 48. System according to any one of claims 44-47, adapted to provide and/or maintain, when in operation, a temperature of the wet mask of at least 40, 45, 50, 55, 60, 65 or 70 °C from the solid/liquid separation device to the preservation device and/or and packaging device. 49. A system according to any one of claims 44-48, further comprising one or more tubes for transporting mask to a storage device (60) for storing the mask, and one or more tubes for transporting the mask to the preservation device (50), wherein the flow of the mesh can be directed to the storage device or preservation device by means of one or more flow control devices (41, 42). 50. System according to claim 49, further comprising a pipe for transporting mesh from the storage device (60) to the preservation device (50), and optionally comprising a flow regulating device (46) for regulating the flow of the mesh from the storage device (60) to the preservation device (50) ). DK 202300025 Y3 54 A system according to any one of claims 44-50, wherein the solid/liquid separation device (20) comprises two filtration units in series, the first unit, when in operation, being adapted to provide moist mesh with a first water content, such as a water content of at least about 60, 70, 80, 85% or more (w/w), and the second filtration provides a wet mask with a different water content, such as at least 20, 30, 40, 50 or 60%, wherein the first water content is higher than the second water content. 52. A modified brewing system in which the system of any one of claims 44-51 is incorporated. 53. A computer program product comprising software code portions configured to, when executed in a computer's memory, perform the method of providing a product according to any one of claims 1-34. 54. A computer program product comprising software code portions configured to, when executed in a computer's memory, control the system of any one of claims 44-51.