WO2024155184A2

WO2024155184A2 - Solid extraction process

Info

Publication number: WO2024155184A2
Application number: PCT/NL2024/050019
Authority: WO
Inventors: Carlos Iván CABRERA-RODRÍGUEZ; Francisco CAPARRÓS SALVADOR; Luuk ROOK; Joana Patricia CORREIA LEIRÓS
Original assignee: Greencovery BV
Current assignee: Greencovery BV
Priority date: 2023-01-17
Filing date: 2024-01-16
Publication date: 2024-07-25
Anticipated expiration: 2025-07-17
Also published as: EP4651961A2; WO2024155184A3; NL2033973B1

Abstract

The invention provides a method for converting a solid mixture (200) into a non- dissolved part (310), a precipitated part (320) and a supernatant part (330), the method comprising: an extraction stage (10) comprising dissolving at least part of the solid mixture (200) in an extraction liquid (400), thereby providing a liquid mixture (500) and the non- dissolved part (310), wherein the extraction liquid (400) comprises (a) an organic extraction material (410) selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of OH^-, (c) at least 0.01 M of a cation (430) selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH4⁺, and (d) water (700); a first separation stage (20) comprising separating the liquid mixture (500) from the non-dissolved part (310); and a precipitation stage (30) comprising providing CO₂(600) to the liquid mixture (500), thereby providing the precipitated part (320) and the supernatant part (330).

Description

Solid extraction process

FIELD OF THE INVENTION

The invention relates to a method for converting a solid mixture into parts. The invention further relates to a system for converting a solid mixture into parts. The invention additionally relates to a computer program product and to a data carrier. Further, the invention relates to a cheese extract.

BACKGROUND OF THE INVENTION

Methods for splitting a solid mixture into parts are known in the art. For instance, WO2009137934A1 relates to protein concentrates and protein isolates, in addition to processes for the production of protein concentrates and protein isolates. In particular, it describes a process for removing fiber from an oilseed meal, comprising: i) mixing an oilseed meal with a blending solvent, optionally water, saline solution, polysaccharide solution or protein containing solution, to form a mixture; ii) optionally adjusting the pH of the protein slurry to a pH of about 2 to about 10; and iii) separating the mixture to form a protein slurry comprising soluble and insoluble proteins and an insoluble fiber fraction.

US10113214B2 describes an alkali metal and/or alkali earth metal extraction method that allows repeated use of an aqueous solution that extracts an alkali metal and/or alkali earth metal from a solid. The alkali metal and/or alkali earth metal extraction method is a method for extracting an alkali metal and/or alkali earth metal from a solid containing the alkali metal and/or alkali earth metal, the method including an elution step in which the solid is added to a neutral amino acid-containing aqueous solution or an amino acid-containing mixed aqueous solution produced by mixing a pH adjusting agent with an aqueous solution containing at least one of a neutral amino acid, an acidic amino acid and a basic amino acid so as to elute the alkali metal and/or alkali earth metal in the neutral amino acid-containing aqueous solution or the amino acid-containing mixed aqueous solution.

JPS5828246A describes a method wherein a raw leaf of a stevia is extracted with water, hot water or a mixture of water with an alcohol, and if necessary the extract is then concentrated. A mixture of calcium hydroxide with calcium chloride in an amount of 0.5-2.0 times that of the solid content in the extract is then added to the extract or concentrated extract preferably while blowing gaseous carbon dioxide thereinto. Thus, impurities are precipitated in the form of a colloidal material in the extract, and separated by the filtration. W02018204061A1 describes methods of treating brown stock from a pulp mill to improve its delignification and/or bleaching response, including treating brown stock with an alkali alcohol/water co-solvent to yield a post-treatment solution, neutralizing the posttreatment solution, and recovering the treated brown stock.

SUMMARY OF THE INVENTION

Compounds of interest, such as amino acids (especially polypeptides) or natural fibers (especially soluble fibers), may often be present in solid mixtures. For instance, a compound may be present in a solid mixture obtained from a biomass stream, such as from a side stream, a waste stream, or a stream resulting from a fermentation process. However, the solid mixture may comprise a variety of other compounds, complicating the further use of the compounds of interest. It may hence be desirable to split the solid mixture into parts to access different compounds of interest.

Prior art processes for converting a solid mixture into parts may be complicated, may lack specificity for a particular compound, may be difficult to adapt to different compounds, may compromise the final product properties (such as by heating), may require extensive cleaning of materials, and/or may recover compounds at low concentrations (complicating downstream applications). Further, prior art processes may be costly in terms of specific reagents required, quantities of reagents required, the recovery of reagents after the process, and/or separation of the compound after the process. Additionally, prior art processes may not be sustainable, as such processes may be energy intensive, may generate large amounts of (undesirable) by-products, and/or may have large water utilization. Such prior art processes may moreover be inappropriate for food applications as the removal of flavor compounds and aroma compounds via water extraction may be complicated and/or limited.

Hence, it is an aspect of the invention to provide an alternative method for converting a solid mixture into parts, which preferably further at least partly obviates one or more of above-described drawbacks. The present invention may have as object to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

In a first aspect, the invention provides a method for converting (or “splitting”) a solid mixture, especially into a non-dissolved part, a precipitated part and a supernatant part. In general, the method comprises at least an extraction stage, a first separation stage, and a precipitation stage. The method may further comprise a second separation stage and/or a compound recovery stage. In embodiments, the extraction stage may comprise dissolving at least part of the solid mixture in an extraction liquid. Thereby a liquid mixture and the non- dissolved part may be provided. The extraction liquid may comprise an organic extraction material (or “first liquid” or “solvent” or “water-miscible organic material”). The organic extraction material may especially be selected from the group consisting of acetone and alcohols. The extraction liquid may further comprise at least 0.01 M of an anion, especially hydroxide (OH ). The extraction liquid may moreover comprise at least 0.01 M of a cation. The cation may especially be selected from the group comprising sodium (Na⁺), potassium (K⁺), calcium (Ca²⁺), magnesium (Mg²⁺), and ammonium (NH4⁺). Further yet, the extraction liquid may comprise water. The first separation stage may in embodiments comprise separating the liquid mixture from the non-dissolved part. In embodiments, the precipitation stage may comprise providing an acid, especially carbon dioxide (CO2), to the liquid mixture. Thereby, the precipitated part and the supernatant part may be provided. The second separation stage may in further embodiments comprise separating the precipitated part and the supernatant part. Moreover, in embodiments, the compound recovery stage may comprise separating a compound from any one of the non-dissolved part, the precipitated part and the supernatant part.

Therefore, in embodiments the invention provides a method for converting (or “splitting”) a solid mixture into a non-dissolved part, a precipitated part and a supernatant part, the method comprising: an extraction stage comprising dissolving at least part of the solid mixture in an extraction liquid, thereby providing a liquid mixture and the non-dissolved part, wherein the extraction liquid comprises (a) an organic extraction material selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of OH', (c) at least 0.01 M of a cation selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH4⁺, and (d) water; a first separation stage comprising separating the liquid mixture from the non-dissolved part, and a precipitation stage comprising providing CO2 to the liquid mixture, thereby providing the precipitated part and the supernatant part.

The invention may provide the benefit that the solid mixture may be split into parts comprising distinct compounds for recovery and further use while forming little waste product. In particular, the method of the invention may facilitate obtaining three parts comprising specific compounds of interest, especially via (i) separating insoluble compounds into a non-dissolved part, and (ii) the controlled precipitation of specific compounds from the liquid mixture into a precipitated part while other specific compounds remain in solution as a supernatant part. Hence, different varieties of compounds may be separated (depending on their pKa and solubility in acetone/alcohol) efficiently with the method of the invention. Especially, the extraction liquid may facilitate solubilizing compounds with a low pKa or poor solubility in acetone/alcohol. Further, the acid exposure, especially the CO2 exposure, may enable efficiently removing the cation and the OH' from the solution. Moreover, the execution of the method may be convenient and not require, for example, an ion exchange step.

In particular, during the extraction stage, the extraction liquid may comprise a salt, such as NaOH, which provides the Na⁺ cation as well as an OH' anion. The presence of the anion (such as OH') may substantially improve the solubility of certain compounds in the extraction liquid. For instance, an organic acid compound (or “organic acid”) may dissolve poorly in acetone/alcohol s/water or mixtures thereof, but the solubility of the organic acid compound may increase drastically in the presence of the anion. Hence, the salt, especially the cation and the anion, may facilitate solubilizing the organic acid compound from the solid mixture in the extraction liquid to provide a liquid mixture comprising the organic acid compound. As such, compounds such as an organic acid may (better) dissolve in the liquid mixture due to the anion, and the liquid mixture may thereby comprise compounds that would remain (partially) non-dissolved without the presence of the anion. A remainder of the solid mixture may largely consist of specific compounds that do not dissolve into the liquid mixture despite the presence of the anion. Hence, the invention initially facilitates converting (or “splitting”) a solid mixture into a dissolved part (comprised by the liquid mixture) and a nondissolved part.

Subsequently, during the precipitation stage, the addition of an acid (such as CO2) to the liquid mixture (as obtained in the first separation stage) may result in the cation and (at least part of) the acid forming a salt, which may precipitate, effectively removing the cation from the liquid mixture. Similarly, the addition of the acid to the liquid mixture may acidify the liquid mixture, resulting in a reduction of the anion (such as OH'), which may reduce the solubility of certain compounds in the liquid mixture (as described above). Additionally, the acidification of the liquid mixture may result in protonation of such compounds, which may (further) reduce the solubility of such compounds in the liquid mixture. Thereby, in embodiments, such compounds may precipitate upon addition of (sufficient) acid. Hence, following the acid addition, the liquid mixture may be separated into a precipitate and a supernatant. The precipitate may comprise carbonate (see below) and specific compounds that precipitated, such as e.g. a polypeptide. The supernatant may largely consist of the organic extraction material, water, and other specific compounds that remained in solution, such as e.g. a free amino acid. Hence, the invention further facilitates converting (or “splitting”) the dissolved part of the solid mixture into a precipitated part and a supernatant part. Therefore, as indicated above, the invention provides a method for converting, especially splitting, a solid mixture into a non-dissolved part, a precipitated part, and a supernatant part. Each of these three parts may comprise a (different) compound of interest from the solid mixture. The compound of interest may subsequently be recovered from any one of the different parts in a compound recovery stage. Hence, the method for converting a solid mixture into parts may provide access to a compound of interest for potential further use.

The term “mixture” may herein especially refer to a combination of two or more compounds, especially three or more compounds, such as two or more (or three or more) of e.g. an amino acid, a polypeptide, a fat, a soluble fiber, an insoluble fiber, and a (poly)saccharide.

In particular, a solid mixture may comprise a plurality of (different) compounds. The term “compound of interest” may herein refer to at least one compound targeted for recovery from such solid mixture via the method as described. The compound of interest may thus refer to a (single) compound targeted for recovery from the plurality of (different) compounds in the solid mixture. More especially, the compound of interest may refer to a subset of compounds targeted for recovery from the plurality of (different) compounds in the solid mixture. The subset of compounds targeted for recovery may comprise the same type of compound, i.e., in an embodiment the compound of interest may comprise a plurality of (different) free amino acids. However, the subset of compounds targeted for recovery may also comprise different types of compound, e.g., in an embodiment the compound of interest may comprise a free amino acid and an insoluble fiber. Hence, in embodiments, the compound of interest may comprise a mixture.

The (different) compounds of interest may be selected from the same type of compound, i.e., in embodiments two (different) compounds of interest may each comprise a (different) free amino acid. Yet further, the different compounds of interest may be selected from different types of compound, e.g., in embodiments two different compounds of interest may comprise a free amino acid and an insoluble fiber, respectively.

Specifically, with the current method, a solid mixture may be split into a plurality of parts, especially at least three different parts. The plurality of parts may in embodiments comprise at least a non-dissolved part, a precipitated part, and a supernatant part. The nondissolved part may especially comprise a subset of compounds from the solid mixture that were not solubilized by the extraction liquid into the liquid mixture during the extraction stage. The precipitated part may especially comprise a subset of compounds from the solid mixture that precipitated upon addition of an acid (such as CO2) to the liquid mixture during the precipitation stage. The supernatant part may especially comprise a subset of compounds from the solid mixture that remained in solution upon addition of an acid to the liquid mixture during the precipitation stage.

Hence, the solid mixture may comprise at least three different subsets of compounds that may be separated into the non-dissolved part, the precipitated part, and the supernatant part. For example, in an embodiment the solid mixture may comprise (i) an insoluble fiber as a first compound, (ii) a free amino acid as a second compound, and (iii) a monosaccharide as a third compound, which three compounds may end up in (i) the nondissolved part, (ii) the precipitated part, and (iii) the supernatant part, respectively. Hence, the at least three different parts may in embodiments provide access to at least three different compounds of interest.

The method may in embodiments comprise a plurality of different stages to split the solid mixture into at least three parts. The term “stage” and similar terms used herein may refer to a (time) period (also “phase”) of a method and/or an operational mode. In embodiments, the method may comprise at least an extraction stage, a first separation stage, and a precipitation stage. The method may further comprise one or more additional stages, such as a second separation stage and a compound recovery stage. The compound recovery stage may especially comprise one or more (sub)stages, such as a first compound recovery stage, a second compound recovery stage, and a third compound recovery stage. Specific embodiments may comprise additional stages, such as a preparation stage and an organic extraction material recovery stage. The different stages of the method may (partially) overlap (in time). For example, the extraction stage may, in general, be initiated prior to the compound recovery stage, but may partially overlap in time therewith. However, for example, the extraction stage may typically be completed prior to the precipitation stage.

It will be clear to the person skilled in the art how the stages may be beneficially arranged in time.

In embodiments, the method may comprise an extraction stage. The extraction stage may especially comprise dissolving at least part of the solid mixture in an extraction liquid. In particular, the extraction stage may comprise bringing the solid mixture and the extraction liquid into contact with each other. In practice, generally, the solid mixture and the extraction liquid may be arranged together in a container for a duration during the extraction stage, such that at least part of the solid mixture dissolves in the extraction liquid.

Hence, in embodiments, the extraction stage may comprise combining the solid mixture and the extraction liquid, to provide a liquid mixture and the non-dissolved part, especially wherein a dissolved part of the solid mixture is comprised by the liquid mixture. In embodiments, the extraction liquid may be selected to be suitable for dissolving a compound from the solid mixture, thereby providing a liquid mixture comprising the compound. Especially, the extraction liquid may be selected to be suitable for dissolving a second compound and a third compound. In particular, such (second or third) compound may have a solubility of at least 0.1 g/L in the extraction liquid, such as at least 0.5 g/L, especially at least 1 g/L, such as at least 5 g/L, especially at least 10 g/L.

However, in embodiments the extraction liquid may be selected to have a relatively low solubility for a (different) compound, as this may facilitate separating (different) compounds from the solid mixture into different parts. Especially, the extraction liquid may be selected to have a relatively low solubility for a first compound. Hence, such (first) compound may have a solubility of at most 5 g/L in the extraction liquid, such as at most 1 g/L, especially at most 0.5 g/L, such as at most 0.1 g/L, especially (essentially) no solubility.

For this purpose, the extraction liquid may comprise at least an organic extraction material, an anion, a cation, and water. The choice of the organic extraction material, the anion, the cation, and the respective amounts comprised by the extraction liquid, may influence the specificity for different compounds. In particular, the extraction liquid may be selected such that a second compound and/or a third compound are dissolved in the extraction liquid, and such that a first compound is not dissolved in the extraction liquid. Hence, during the extraction stage, a second compound and a third compound may dissolve in the extraction liquid, and a first compound may not dissolve in the extraction liquid.

The organic extraction material may in general be a water-miscible organic material, such as especially an organic solvent. The organic extraction material may be selected from the group consisting of acetone and alcohols, i.e., from the group consisting of acetone and of organic compounds comprising at least one hydroxyl functional group bound to a saturated carbon bond. In particular, the organic extraction material may be selected from the group consisting of acetone and compounds having the formula C_nH2n+iOH. In embodiments, the organic extraction material may be selected from the group consisting of alcohols, especially from the group consisting of water soluble alcohols, and/or especially from the group consisting of C1-C6 alcohols, such as from the group consisting of C1-C5 alcohols, especially from the group consisting of C1-C4 alcohols. Moreover, the alcohols may be primary alcohols, secondary alcohols or tertiary alcohols, especially primary alcohols, or especially secondary alcohols, or especially tertiary alcohols. In further embodiments, the group consisting of alcohols may especially be a group consisting of (water soluble) diols, such as the group consisting of C1-C6 diols, especially the group consisting of C1-C5 diols, such as the group consisting of C1-C4 diols. In certain embodiments, the organic extraction material may especially be selected from the group consisting of acetone, methanol, ethanol, and ethylene glycol. The term “organic extraction material” may herein also refer to a plurality of organic extraction materials, such as two or more (organic) organic extraction materials selected from the group consisting of acetone and alcohols. Thereby, the organic extraction material may be selected to facilitate solubilization of a soluble compound into the extraction liquid.

In embodiments, the alcohols may especially be liquid at room temperature, such as liquid at (about) 20 °C.

In particular, in embodiments wherein a (second) compound is separated from the organic extraction material via precipitation, it may be beneficial for a (second) compound to have a poor solubility in the organic extraction material as such relative to its solubility in the extraction liquid. Hence, in embodiments, the solubility of a (second) compound in the extraction liquid may be at least three times the solubility of the (same) (second) compound in the organic extraction material, especially at least five times, such as at least ten times.

In embodiments, the extraction liquid may comprise at least 10 wt% of the organic extraction material (relative to the total weight of the extraction liquid), such as at least 15 wt%, especially at least 20 wt%. In further embodiments, the extraction liquid may comprise at least 25 wt% of the organic extraction material, especially at least 35 wt%, such as at least 40 wt%, especially at least 50 wt%. In further embodiments, the extraction liquid may comprise up to 99 wt% of the organic extraction material, such as up to 95 wt%, especially up to 90 wt%, such as up to 80 wt%, especially up to 70 wt%. Hence, the extraction liquid may comprise 10 - 99 wt% of the organic extraction material, such as 15 - 95 wt%, especially 20 - 90 wt%, such as 35 - 80 wt%, especially 50 - 70 wt%.

In further embodiments, the extraction liquid may further comprise water (H2O). In particular, the extraction liquid may comprise at least 1 wt% of water (relative to the total weight of the extraction liquid), such as at least 5 wt%, especially at least 10 wt%, such as at least 20 wt%, especially at least 30 wt%. In further embodiments, the extraction liquid may comprise up to 90 wt% of water, such as up to 85 wt%, especially up to 80 wt%, such as up to 75 wt%. In further embodiments, the extraction liquid may comprise up to 70 wt% of water, such as up to 65 wt%, especially up to 60 wt%, such as up to 50 wt%. Hence, the extraction liquid may comprise 1 - 90 wt% of water, such as 5 - 85 wt%, especially 10 - 80 wt%, such as 20 - 65 wt%, especially 30 - 50 wt%. Thereby, water-soluble compounds may be solubilized in the extraction liquid. The phrase “the extraction liquid may comprise at least 30 wt% of X”, and similar phrases, herein refers to at least 30 wt% of the extraction liquid consisting of X, i.e., X makes up at least 30 wt% of the extraction liquid.

In particular, in embodiments, the extraction liquid may comprise both water and organic extraction material. The weight ratio of organic extraction material to water in the extraction liquid may be selected from the range of 100: 1 - 1 : 10, such as 20: 1 - 1 :5, especially 10: 1 - 1 :3, such as 5:1 - 1 :2, especially 3:1 - 1: 1. Hence, both organic extraction materialsoluble and water-soluble compounds may be solubilized in the extraction liquid. The weight ratio of organic extraction material to water may especially be selected to influence the solubility of targeted compounds, such as a second compound or a third compound. In particular, the weight ratio of organic extraction material to water (at their respective weight percentages) may be selected to provide a relatively low solubility for a (second) compound, as this may facilitate separating the second compound from the liquid mixture at a later stage. Further, the weight ratio of organic extraction material to water may be selected to provide a relatively high solubility for a (third) compound, as this may facilitate retaining the third compound in the supernatant part at a later stage. Hence, the (second) compound may have a solubility of at most 10 g/L in the mixture of organic extraction material and water (or “solvent/water mixture”), especially at most 5 g/L, such as at most 1 g/L, especially at most 0.5 g/, such as at most 0.1 g/L, especially (essentially) no solubility. However, the (third) compound may have a solubility of at most 500 g/L in the mixture of organic extraction material and water, such as at most 100 g/L, especially at most 50 g/L, such as at most 10 g/L, especially at most 5 g/L. The (third) compound may in certain embodiments be (essentially) totally miscible in the mixture of organic extraction material and water. Further, the (third) compound may have a solubility of at least 0.1 g/L in the mixture of organic extraction material and water, such as at least 0.5 g/L, especially at least 1 g/L, such as at least 5 g/L, especially at least 10 g/L. It will be clear to the person skilled in the art that this solubility refers to the mixture of organic extraction material and water as such, i.e., not to the extraction liquid as applied in the extraction stage.

In particular, in embodiments wherein the (second) compound is separated from the mixture of organic extraction material and water via precipitation, it may be beneficial for the (second) compound to have a poor solubility in the mixture of organic extraction material and water as such relative to the solubility of the (second) compound in the extraction liquid. Hence, in embodiments, the solubility of the (second) compound in the extraction liquid may be at least three times the solubility of the (second) compound in the mixture of organic extraction material and water, especially at least five times, such as at least ten times.

In particular, in embodiments, the extraction liquid may further comprise an anion, most especially OH'. The extraction liquid may comprise at least 0.001 M of an anion, such as at least 0.005 M, especially at least 0.01 M, such as at least 0.05 M, especially at least 0.1 M, such as at least 0.2 M. In further embodiments, the extraction liquid may comprise at most 3 M of an anion, such as at most 1.5 M, especially at most 1 M, such as at most 0.5 M, especially at most 0.2 M. In further embodiments, the extraction liquid may comprise 0.01 - 1.5 M of an anion. Especially, the extraction liquid may comprise at least 0.001 M of OH' (relative to a total volume of the extraction liquid, including of the organic extraction material), such as at least 0.005 M, especially at least 0.01 M, such as at least 0.05 M, especially at least 0.1 M, such as at least 0.2 M. In further embodiments, the extraction liquid may comprise at most 3 M of OH', such as at most 1.5 M, especially at most 1 M, such as at most 0.5 M, especially at most 0.2 M. In further embodiments, the extraction liquid may comprise 0.01 - 1.5 M of OH'.

In embodiments, the extraction liquid may have a pH > 5.5, especially > 6, such as > 6.5, especially > 7. Such pH values may be suitable for a large range of compounds, such as acids or cations, especially organic acids. Such compounds may generally have relatively low pKa values. However, other compounds, such as bases and anions, may generally have relatively high pKa values. Hence, for (second or third) compounds with relatively high pKa values, an extraction liquid with relatively high pH may be selected. In particular, in further embodiments, the extraction liquid may have a pH> 8.5, such as > 9, especially > 9.5. In further embodiments, the extraction liquid may have a pH < 14, especially < 13, such as < 12.

In further embodiments, the compound, especially the second compound, or especially the third compound, may have a pKa Pi, and the extraction liquid may have a pH > Pi + 0.5, especially > Pi + 1, such as > Pi+ 1.5.

The term pH with respect to the extraction liquid may herein especially refer to 14+loglO([OH']), wherein [OH'] refers to the concentration of OH' in mol/L in the extraction liquid.

The pH of the extraction liquid may especially be determined using a pH meter, especially a pH strip.

The anion, especially OH', may especially be provided to the extraction liquid as a salt. Hence, in embodiments, the extraction liquid may further comprise a cation. The extraction liquid may comprise at least 0.001 M of a cation, such as at least 0.005 M, especially at least 0.01 M, such as at least 0.05 M, especially at least 0.1 M, such as at least 0.2 M. In further embodiments, the extraction liquid may comprise at most 3 M of a cation, such as at most 1.5 M, especially at most 1 M, such as at most 0.5 M, especially at most 0.2 M. The cation may be selected from the group comprising Li, Be, Al, Cr, Mn, Co, Ni, Cu, Zn, Rb, Sr, Pd, Ag, Cd, Cs, Ba, Na, K, Ca, Mg and NH4, especially from the group comprising Na, K, Ca, Mg and NH4. In embodiments, the cation may especially be selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH4⁺. In certain embodiments, the cation may comprise Na⁺.

In particular, the mentioned molarities for the cation in the extraction liquid may herein be defined relative to the total volume of the extraction liquid, i.e., the liquid volume defined by water and the organic extraction material. The same applies to the molarities for the anion in the extraction liquid mentioned above.

The cation may especially be selected to be suitable for forming a salt when exposed to an acid, such as for example forming a carbonate when exposed to CO2. Hence, the cation may together with an acid form a salt (in the precipitation stage; see below). Specifically, in the example of CO2, the CO2 may (together with water in the extraction liquid) form H2CO3, (i.e., carbonic acid), and the cation may react with HCOf or with COv' to form a carbonate. Further, the CO2 may (together with methanol as organic extraction material in the extraction liquid) form C2H4O3 (i.e., methyl carbonic acid). Moreover, the CO2 may (together with ethanol as organic extraction material in the extraction liquid) form C3H5O3 (i.e., ethyl carbonic acid). The cation may react with a carbonic acid to form a carbonate. Such embodiments may be particularly beneficial for separating the cation and the acid from the supernatant part, facilitating both purifying the third compound and recovering the organic extraction material (see below).

The cation and the anion may be provided together to the extraction liquid in the form of a salt. The salt may comprise the cation and the anion (especially OH'). Hence, in embodiments, the extraction liquid may comprise at least 0.001 M of a salt. The extraction liquid may comprise at least 0.001 M of a salt, such as at least 0.005 M, especially at least 0.01 M, such as at least 0.05 M, especially at least 0.1 M, such as at least 0.2 M. In further embodiments, the extraction liquid may comprise at most 3 M of the salt, such as at most 1.5 M, especially at most 1 M, such as at most 0.5 M, especially at most 0.2 M. The salt may especially be selected from the group comprising inorganic hydroxides, more especially from the group comprising sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, and ammonium hydroxide.

In particular, the method may thus comprise an extraction liquid preparation step, wherein the extraction liquid preparation step comprises adding a salt to a mixture of organic extraction material and water, wherein the salt comprises the cation and the anion (especially OH'), such as a salt selected from the group comprising NaOH, KOH, Ca(OH)2, Mg(OH)2 and NH4OH. Thereby, an extraction liquid comprising a mixture of organic extraction material and water with a defined weight ratio of organic extraction material to water and a defined amount of an anion and a cation may be provided with suitable characteristics for solubilizing a (second and third) compound in the extraction stage.

In certain embodiments, the extraction stage may comprise an extraction duration during which the solid mixture and the extraction liquid are in contact. The extraction duration may be selected from the range of 1 minute - 48 hours, such as from the range of 5 minutes - 24 hours, especially from the range of 10 minutes - 12 hours, such as from the range of 30 minutes - 4 hours. In certain embodiments, the extraction duration may be at least 1 minute, such as at least 5 minutes, especially at least 10 minutes, such as at least 30 minutes. In further embodiments, the extraction duration may be at most 48 hours, such as at most 24 hours, especially at most 12 hours, such as at most 4 hours.

In certain embodiments, the extraction stage may comprise controlling an extraction temperature of the extraction liquid. The extraction temperature may be at least 1 °C, such as at least 5 °C, especially at least 15 °C, such as at least 25 °C, especially at least 35 °C. The extraction temperature may especially be at most 100 °C, such as at most 75 °C, such as at most 50 °C. Thereby, the extraction stage may comprise controlling an extraction temperature of the extraction liquid at a temperature selected from the range of 1 - 100 °C, such as from the range of 5 - 100 °C, especially from the range of 15 - 100 °C, such as from the range of 25 - 75 °C, especially from the range of 35 - 50 °C.

The extraction duration and extraction temperature may be selected to be suitable for dissolving a compound from the solid mixture into the liquid mixture. Especially, the extraction duration and extraction temperature may be selected to be suitable for dissolving a second compound and a third compound. In particular, the extraction duration and extraction temperature may be selected such that during the extraction stage, a (second or third) compound may be dissolved to at least 0.1 g/L in the extraction liquid, such as at least 0.5 g/L, especially at least 1 g/L, such as at least 5 g/L, especially at least 10 g/L. That is, in embodiments, the extraction duration and extraction temperature may be selected such that during the extraction stage, at least 0.1 g, such as at least 0.5 g, especially at least 1 g, such as at least 5 g, especially at least 10 g, of the (second or third) compound may be dissolved per liter of extraction liquid.

However, the extraction duration and extraction temperature may be selected so that a (first) compound may dissolve relatively poorly into the extraction liquid, as this may facilitate separating (first) compounds from the solid mixture into different parts. Hence, the extraction duration and extraction temperature may be selected such that during the extraction stage, the (first) compound has a solubility of at most 5 g/L in the extraction liquid, such as at most 1 g/L, especially at most 0.5 g/L, such as at most 0.1 g/L, especially (essentially) 0.

Thereby, the extraction stage may provide the liquid mixture and the nondissolved part. The non-dissolved part may comprise the subset of compounds that are insoluble in the extraction liquid, especially at least the first compound. Vice versa, the liquid mixture may comprise the subset of compounds that are soluble in the extraction liquid, especially the second compound and the third compound. Further, the liquid mixture may comprise the organic extraction material, the anion, the cation, and the water.

In embodiments, the method may comprise a first separation stage following the extraction stage, i.e., the extraction stage may be followed by a first separation stage. The first separation stage may comprise separating the liquid mixture from the non-dissolved part. Hence, the first separation stage may comprise providing the liquid mixture and the nondissolved part (by separating these from each other). The first separation stage may, for instance, especially comprise filtration, sedimentation, hydrocyclone (separation), screw-press (separation), centrifugation, or a combination thereof. In embodiments, the first separation stage may comprise removing the non-dissolved part from the liquid mixture, especially by filtration, or especially by centrifugation.

In embodiments, the method may comprise a precipitation stage. The precipitation stage may generally follow the extraction stage or, if applicable, the first separation stage. The precipitation stage may comprise providing an acid, especially CO2, to the liquid mixture, especially the liquid mixture as obtained in the first separation stage. In particular, the precipitation stage may comprise bringing the acid and the liquid mixture into contact with each other. In practice, generally, the liquid mixture may be arranged in a container for a duration during the precipitation stage, to which the acid may be provided.

In embodiments, the acid may be selected from the group comprising carbon dioxide (CO2), hydrogen chloride (HC1), acetic acid (CH3COOH), other organic acids, and other inorganic acids, especially from the group comprising CO2, HC1, and acetic acid. Most especially, the acid may be CO2. An organic acid may be an organic compound (i.e., a compound comprising a carbon bonded to another element) comprising at least one acid group. An inorganic acid may be an inorganic compound (i.e., a compound comprising no carbon bonds) comprising at least one acid group. The acid may especially be a food-grade acid, such as a food-grade acid selected from the group comprising CO2, HC1, and acetic acid, especially CO2, or especially HC1. These acids may be widely available, cost-efficient, and commonly used in the food industry.

In embodiments, the acid may be selected from the group comprising acetic acid and (aqueous) HC1. Compared to CO2, these acids may be in liquid form at the temperature range that the method herein is performed. Hence, these acids may be relatively easier to handle than gaseous CO2, thereby simplifying the process.

The addition of an acid to the liquid mixture may result in the formation of protonated acid compounds. The protonated acid compound may behave as a protic acid and form an equilibrium with the deprotonated acid compounds in the liquid mixture. For instance, the addition of CO2 to the liquid mixture may result in the formation of carbonic acid (H2CO3), which may behave as a diprotic acid, forming an equilibrium with HCCh' and CCh²'. Thereby the liquid mixture may be acidified, in particular via a reduction of the amount of the anion (especially OH').

Especially, the acid (such as CO2) may be provided to the liquid mixture such that a substantial amount of the anion (especially OH') from the extraction liquid is neutralized. The acid may be provided to neutralize at least 5% of the anion from the extraction liquid, such as at least 10%, especially at least 20%, such as at least 30%, especially at least 50%. The acid may be provided to neutralize at least 75% of the anion from the extraction liquid. Moreover, in embodiments, the acid may be provided to the liquid mixture such that the liquid mixture is further acidified beyond neutralization of (essentially) all the anion (especially OH') from the extraction liquid. The acid may be provided (at an amount) of at most 500% of the amount of acid required to neutralize (essentially) all the anion from the extraction liquid, such as at most 250%, especially at most 150%. Thereby, in embodiments, the acid may be provided to the liquid mixture at an amount of 5 - 500% of the amount of acid required to neutralize (essentially) all the anion from the extraction liquid, such as 10 - 250%, especially 20 - 150%, such as 30 - 100%, especially 50 - 75%.

Thus the addition of the acid (such as CO2) to the liquid mixture may in certain embodiments especially result in a corresponding reduction of the pH of the liquid mixture. In embodiments, the pH of the liquid mixture may be reduced (by) at least 0.5 by providing the acid, such as at least 1.0, especially at least 1.5, such as at least 2.0, especially at least 2.5. In further embodiments, the pH of the liquid mixture may be reduced by at most 10.0 by providing the acid, such as at most 8.0, especially at most 6.0, such as at most 5.0, especially at most 4.0. Thus, the reduction in pH of the liquid mixture may be selected from the range of 0.5 - 10.0, such as 1.0 - 8.0, especially 1.5 - 6.0, such as 2.0 - 5.0, especially 2.5 - 4.0.

In further embodiments, the precipitation stage may comprise providing the acid (such as CO2) to the liquid mixture to reduce the OH' concentration in the liquid mixture to at most 0.001 M, such as at most 10'⁴ M, especially to at most 10'⁵ M. In further embodiments, the precipitation stage may comprise providing the acid to the liquid mixture to reduce the OH' concentration in the liquid mixture to at most 10'⁶ M, such as at most 10'⁷ M, especially to at most 10'⁸ M. In further embodiments, the precipitation stage may comprise providing the acid to the liquid mixture to reduce the OH' concentration in the liquid mixture to at most 10'⁹ M, such as at most 10'¹⁰ M. In particular, in embodiments, the precipitation stage may comprise providing the acid to the liquid mixture to reduce the OH' concentration in the liquid mixture to at least 10'¹⁰ M, such as at least 10'⁹ M, especially to at least 10'⁸ M. For instance, in embodiments, the precipitation stage may comprise providing the acid to the liquid mixture to reduce the OH' concentration in the liquid mixture to a concentration selected from the range of 1 O'⁸ - 10'⁵ M.

Hence, in embodiments, the method may comprise providing the acid (such as CO2) to reduce the pH of the liquid mixture to a separation point, such as a precipitation point and/or a phase separation point of the (second) compound, especially to a precipitation point, or especially to a phase separation point.

The term “precipitation point” may herein especially refer to a pH at which a compound precipitates from a solution, i.e. the liquid mixture. The term “phase separation point” may herein especially refer to a pH at which a phase separation occurs, especially wherein the (second) compound is separated from a remainder of the liquid mixture during the phase separation, i.e., the phase separation may provide a compound fraction part which is enriched in a (second) compound relative to the liquid mixture. The compound fraction part may in embodiments be comprised by the precipitated part.

In particular, a (second) compound may precipitate from a solution particularly well at a pH at which the compound has a neutral charge. Hence, the precipitation point may lie close to an isoelectric point pl of the (second) compound. Hence, in embodiments, a (second) compound may have an isoelectric point pl (in the mixture of organic extraction material and water), wherein the precipitation point may be selected from the range of pl-3 - pI+3, especially from the range of pl-2 - pI+2, such as from the range of pl- 1 - pl+l, especially from the range of pl-0.5 - pI+0.5, such as from the range of pl-0.2 - pI+0.2.

In embodiments, the (second) compound may have an acid dissociation constant pKa, wherein the separation point, especially the precipitation point, or especially the phase separation, point, may be selected from the range of < pKa.

In particular, a (second) compound may have a pKa P2, and the pH of the liquid mixture may be reduced to a pH < P2+1.5, especially < P2+I, such as < P2+O.5. Further, a (third) compound may have a pKa P3, and the pH of the liquid may be reduced to a pH > P3+I.5, especially > P3+I, such as > P3+O.5. Thereby, the amount of acid (such as CO2) provided to the liquid mixture during the precipitation stage may be selected such that a (second) compound may precipitate while a (third) compound may remain dissolved.

In embodiments, the (second) compound may have a pKa selected from the range of 1.5 - 11, especially from the range of 2 - 10, such as from the range of 3-8, especially from the range of 4-6.

In further embodiments, the (second) compound may have a plurality of acid dissociation constants, such as a lower acid dissociation constant pKai and an upper acid dissociation constant pKa_u, wherein the precipitation point may be selected from the range of < pKa_u, and especially from the range of > pKai, or especially from the range of < pkai.

In general, recovering compounds with a low pKa may be challenging with prior art methods, especially for prior art methods relying on protonation of the (second) compound. The method of the invention may be particularly suitable for separating a (second) compound with a low pKa value from other compounds. Hence, in embodiments, the (second) compound may have a pKa < 5.0, such as < 4.5. In further embodiments, the (second) compound may have a pKa < 4.0, especially < 3.5, such as < 3.0. In further embodiments, the (second) compound may have a pKa < 2. In further embodiments, the (second) compound may have a pKa > 1, especially > 2, such as > 3.

In certain embodiments, the liquid mixture may comprise an (electrically charged) second compound, for example an amino acid, a polypeptide, or a long-chain fatty acid. In particular, the second compound may be soluble in the extraction liquid when the second compound is electrically charged. The second compound may especially be electrically charged in the liquid mixture as provided by the extraction stage prior to the precipitation stage. Thereby, the second compound may be electrically charged at a second anion (especially OH') concentration CE provided by the extraction liquid to the liquid mixture. However, the second compound may be electrically neutral in the liquid mixture at a second anion concentration CN, especially a second OH' concentration CN. The second anion (especially OH') concentration CN may be at least IO'¹⁰ M, such as at least 10'⁹ M, especially at least 10'⁸ M. The second anion (especially OH') concentration CN may be at most 10'⁴ M, such as at most 10'⁵ M, especially at most 10'⁶ M. Thereby the second anion (especially OH') concentration CN may be in the range of IO'¹⁰ - 10'⁴M, such as 10'⁹ - 10'⁵ M, especially 10'⁸ - 10'⁶ M. The second anion (especially OH') concentration CN may especially be (around) 10'⁷M.

In further such embodiments, the precipitation stage may comprise providing the acid (such as CO2) to reduce a charged anion (especially OH') concentration CE in the liquid mixture to (about) a second anion (especially OH') concentration CN. In particular, the precipitation stage may comprise reducing the charged anion concentration to at least 0.05 *CN, especially at least 0.1 *CN, such as at least 0.2*CN, especially at least 0.5*CN. In particular, the precipitation stage may comprise reducing the charged anion concentration to at most 2.0*CN, especially at most 1.5*CN, such as at most 1.2*CN, especially at most 1.0*CN. Hence, in embodiments, the precipitation stage may comprise reducing the charged anion concentration to a value selected from the range of 0.05 *CN - 2.0*CN, especially 0.1 *CN - 1.5*CN, such as 0.2*CN - 1.2*CN, especially 0.5*CN - 1.0*CN. In further embodiments, the precipitation stage may comprise reducing the charged anion concentration such that the second compound becomes (essentially) electrically neutral and precipitates.

Alternatively or additionally, the liquid mixture may comprise an (electrically charged) third compound, such as a (medium or short chain) carboxylic acid (salt), a saccharide (e.g., a monosaccharide, a disaccharide, an oligosaccharide, or a polysaccharide), a salt (e.g., a chloride salt), or a glycerol. In particular, the third compound may be solubilized in the extraction liquid when the third compound is electrically charged. The third compound may especially be electrically charged in the liquid mixture as provided by the extraction stage prior to the precipitation stage. Thereby, the third compound may be electrically charged at the charged anion (especially OH') concentration CE provided by the extraction liquid to the liquid mixture. The third compound may remain electrically charged at the second anion (especially OH') concentration CN described above. The value of the second anion (especially OH') concentration CN may especially be selected such that the third compound remains electrically charged. Hence, the third compound may be electrically charged in the liquid mixture before and after the precipitation stage, facilitating keeping the third compound (substantially) dissolved.

In certain embodiments, the precipitation stage may comprise providing the acid, especially CO2, via sparging or via pressurization. The acid may be provided in a gas state (such as for CO2) or in a liquid state (such as for HC1). The precipitation stage may comprise providing the acid at a precipitation temperature selected from the range of 1 - 50° C, such as from the range of 2 - 45° C, especially from the range of 5 - 40° C, such as from the range of 10 - 35° C, especially from the range of 15 - 30° C. Especially, the acid may be provided at an precipitation temperature of room temperature around 20° C. Further, the precipitation stage may comprise providing the acid at a precipitation pressure selected from the range of 0.01 - 10 bar, such as from the range of 0.05 - 5 bar, or such as from the range of 0.1 - 10 bar, especially from the range of 0.1 - 2 bar, such as from the range of 0.5 - 1.5 bar, especially from the range of 0.90 - 1.1 bar. In embodiments, the acid may be provided to the liquid mixture for a precipitation duration selected from the range of 1 - 240 minutes, such as from the range of 2 - 120 minutes, especially from the range of 5 - 30 minutes, such as from the range of 10 - 20 minutes. In certain embodiments, the acid may be provided to the liquid mixture for a precipitation duration of at least 1 minute, such as at least 2 minutes, especially at least 5 minutes, such as at least 10 minutes. In further embodiments, the acid may be provided to the liquid mixture for a precipitation duration of at most 24 hours, such as at most 2 hours, especially at most 30 minutes. The precipitation temperature, the precipitation pressure, and the precipitation duration may be selected to improve the efficiency of the precipitation of a (second) compound while keeping a (third) compound (substantially) dissolved.

Thereby, the precipitation stage may provide the precipitated part and the supernatant part. The precipitated part may comprise the subset of compounds (initially from the solid mixture) comprised by the liquid mixture that precipitated during the precipitation stage, especially the second compound. Vice versa, the supernatant part may comprise the subset of compounds (initially from the solid mixture) comprised by the liquid mixture that remained dissolved during the precipitation stage, especially the third compound. Moreover, the precipitated part and the supernatant part may both comprise compounds from the solid mixture and the liquid mixture that have reacted with other components during the extraction stage and precipitation stage, e.g., a compound from the solid mixture may have dissolved as a salt during the extraction stage and precipitated as a salt during the precipitation stage. Hence, the extraction stage provides the precipitated part and the supernatant part each comprising a (different) subset of compounds.

The precipitation stage may be followed by an optional second separation stage. The second separation stage may comprise separating the precipitated part from the supernatant part. This may be achieved using different properties between the precipitated part and the supernatant part. In general, the precipitated part may be in a solid state, and the supernatant may be in a liquid state, especially a liquid mixture of organic extraction material and water comprising dissolved components, such as the third compound. Hence, the second separation stage may comprise a separation step (as described above). The separation step may comprise separating the liquid components and the solid components (in the container comprising the precipitated part and the supernatant part), such as by filtration, or such as by centrifugation.

Thereby, the second separation stage may provide the separated precipitated part and the separated supernatant part for further use, such as a second compound recovery stage (for the precipitated part) or a third compound recovery stage (for the supernatant part).

Thus the present method provides the (separated) non-dissolved part, the (separated) precipitated part, and the (separated) supernatant part as a plurality of parts. The plurality of parts may comprise one or more of a compound of interest, such as a first compound (in the non-dissolved part), a second compound (in the precipitated part), and a third compound (in the supernatant part). Each of the plurality of parts may further comprise (trace amounts of) additional compounds used during the method, such as e.g. the organic extraction material, (a salt comprising) the anion, (a salt comprising) the cation, the acid, and water. Especially, the non-dissolved part may comprise (traces of) an organic extraction material and water. Moreover, the precipitated part may (substantially) comprise a cation and an organic acid (such as in the form of a salt comprising the cation and/or the organic acid). Further, the supernatant part may comprise the organic extraction material and water.

In certain embodiments, the (separated) non-dissolved part, the (separated) precipitated part, and/or the (separated) supernatant part may be provided as such. However, in further embodiments, the method may comprise an optional compound recovery stage. The compound recovery stage may comprise separating or enriching a compound of interest from the plurality of parts. Especially, the recovery stage may comprise separating or enriching one or more compounds of interest from any one of the plurality of parts. The one or more compounds of interest may be selected from any one of the first compound, the second compound, and the third compound.

The compound of interest (as a first compound, second compound, and/or third compound) may especially comprise an organic compound. The compound of interest may comprise an organic compound selected from a free amino acid, a polypeptide, a fat, a soluble fiber, an insoluble fiber, a saccharide, an organic acid, and an (organic) salt.

In embodiments, the compound of interest may comprise a natural compound, i.e., a naturally occurring compound. Further, the compound of interest may comprise a section of a natural compound, such as obtained after enzymatic treatment or chemical treatment. Further yet, the compound of interest may comprise a modified natural compound, such as a compound conjugated with a (synthetic) molecule. Alternatively, the compound of interest may comprise a non-naturally occurring compound.

The term “amino acid” may herein refer to an organic compound comprising an amino and a carboxyl functional group, along with a side chain. An amino acid may comprise a “free amino acid” that is not covalently bound to another amino acid.

In embodiments, the compound of interest may comprise a free amino acid. Such free amino acid may especially comprise a proteinogenic amino acid, such as an amino acid selected from the group comprising arginine, serine, asparagine, glutamine, glutamic acid, cysteine, selenocysteine, proline, alanine, tyrosine, histidine, isoleucine, leucine, methionine, phenylalanine, threonine, tryptophane, valine, lysine, aspartic acid, and glycine. Especially, the free amino acid may be selected from the group comprising glutamic acid, tyrosine, and leucine. In embodiments, the free amino acid may further comprise a non-proteinogenic amino acid, such as an amino acid selected from the group comprising betaine, 4-amino benzoic acid and gamma-amino butyric acid.

Further, the compound of interest may comprise a polypeptide, i.e., a chain of covalently bound amino acids. The polypeptide may comprise a plurality of different amino acids, especially a plurality of different proteinogenic amino acids.

The term “polypeptide” may herein refer to a sequence of amino acids of any length, such as a sequence of at least 2 amino acids, especially a sequence of at least 10 amino acids, more especially a sequence of 30 or more amino acids. The term polypeptide may herein thus also refer to a peptide, an oligopeptide, a protein, and a multiprotein complex. In certain embodiments, the polypeptide may especially comprise a peptide, such as a single peptide or oligopeptide, and the number of amino acids may be selected from the range of 2 - 60, especially from the range of 2 - 40, such as from the range of 3 - 40. Such peptide may comprise at least 2 amino acids, such as at least 3 amino acids, especially at least 5 amino acids, more especially at least 10 amino acids. In further embodiments, the polypeptide may especially comprise a protein, such as a protein or a multiprotein complex, and the number of amino acids may be selected from the range of 2 - 4000, such as from the range of 3-2500, especially from the range of 5-1000. Such protein may comprise at most 1000 amino acids, such as at most 500 amino acids, especially at most 100 amino acids, such as at most 50 amino acids.

Moreover, the compound of interest may comprise a fat. The term “fat” may herein refer to any ester bound to one or more fatty acids, such as a phospholipid. Especially, the fat may comprise a glyceride fat, i.e., a glycerol ester bound to one or more fatty acids, such as a monoglyceride or a diglyceride. More especially, the fat may comprise a triglyceride, i.e., a glycerol ester comprising three fatty acids.

Fatty acids are carboxylic acids with an aliphatic carbon chain and may be categorized by (i) the length of their carbon chain, (ii) the degree of saturation of their carbon chain, and (iii) the degree of branching of their carbon chain. The length of the carbon chain of a fatty acid is defined by the number of carbons in the carbon chain. The length of the carbon chain of a fatty acid may in embodiments comprise at least 5 carbons, such as at least 7 carbons, especially at least 9 carbons, especially at least 11 carbons. The length of the carbon chain of a fatty acid may in embodiments be up to 29 carbons, such as up to 27 carbons, especially up to 25 carbons, especially up to 23 carbons. Thereby, the fat may in embodiments comprise a shortchain fatty acid, a medium-chain fatty acid, a long-chain fatty acid, or a very-long-chain fatty acid.

The degree of saturation of a fatty acid is defined by the number of C=C double bonds between adjacent carbons in the carbon chain. A fatty acid is considered saturated when it contains only C-C single bonds between adjacent carbons in the carbon chain. Degrees of unsaturation increase with the number of C=C double bonds between adjacent carbons in the carbon chain. A monounsaturated fatty acid may comprise a single C=C double bond in the carbon chain. A polyunsaturated fatty acid may comprise two or more C=C double bonds in the carbon chain, such as three or more, especially four or more, especially five or more. Hence, the fat may in embodiments comprise a saturated fatty acid, a monounsaturated fatty acid, or a polyunsaturated fatty acid. Further, unsaturated fatty acids may be defined by the cis or trans configuration of the C=C double bond. Thereby, in embodiments, the fat may comprise an unsaturated fatty acid, such as a cis configured fatty acid or a trans configured fatty acid.

The degree of branching of a fatty acid is defined by the number of methyl branches on the carbon chain of a fatty acid. Especially, in embodiments, the fatty acid may comprise a linear carbon chain, i.e., a carbon chain comprising (essentially) no methyl branches. The fatty acid may comprise a branched carbon chain, i.e., a carbon chain comprising one or more methyl branches, especially two or more, such as three or more. Thereby, the fat may comprise a linear carbon chain fatty acid or a branched carbon chain fatty acid.

In embodiments, the compound of interest may comprise a fiber, especially a natural fiber. In general, a fiber is a natural or artificial structure that is substantially longer than it is wide. In particular, fibers may be integrated (or “arranged”) into a fibrous material.

In further embodiments, the fiber may comprise a naturally occurring fiber. A naturally occurring fiber may herein also be referred to as a “natural fiber”. Especially, the natural fiber may comprise a plant-derived fiber, a fungal -derived fiber, a microbial-derived fiber, or an animal-derived fiber, such as may be comprised by biomass from such aforementioned sources. A plant-derived fiber may comprise e.g. cellulose, hemicellulose, lignin, pectin, beta-glucan, milk oligosaccharide, etc. A fungal -derived fiber may especially comprise mycelium -based fibers, e.g., cellulose, chitin, etc. A microbial-derived fiber may comprise fibers from other sources that have been subjected to a microbial metabolic process such as fermentation. An animal-derived fiber may especially comprise protein-based fibers, e.g. collagen, chitin, sericin, fibroin, keratin, actin, etc.

In embodiments, the fiber may especially comprise a dietary fiber.

A fiber, especially a dietary fiber, may be categorized as a “soluble fiber” or as an “insoluble fiber”. A soluble fiber may in general be water-soluble (with solubility affected by factors such as pH, temperature, ionic strength, et cetera). An insoluble fiber may in general not be soluble in water. Dietary fibers may in particular consist of a chain of two or more polysaccharides. A dietary fiber may be a linear fiber. Further, a dietary fiber may be a branched fiber. A dietary fiber may be covalently bound to one or more of a non-polysaccharide molecule. The non-polysaccharide molecule may especially be selected from the group comprising a lignin, a free amino acid, and a polypeptide. A dietary fiber may at least partly resist digestion by human digestive enzymes. Further, a dietary fiber may (essentially) completely resist digestion by human digestive enzymes. Hence, upon human oral ingestion, a dietary fiber may pass through the stomach to the intestines. In the intestines, a dietary fiber may be partly digested by (digestive enzymes produced by) gut microbiota. Further, a dietary fiber may be (essentially) completely digested by (digestive enzymes produced by) gut microbiota. It should be noted herein that the categorization of a fiber as a soluble fiber or insoluble fiber is independent from the solubilization of the fiber into the liquid mixture during the extraction stage of the method of the present invention. Hence, in certain embodiments a soluble fiber may be solubilized during the extraction stage. In other embodiments, a soluble fiber may not be solubilized during the extraction stage. Further, in certain embodiments, an insoluble fiber may be solubilized during the extraction stage. In other embodiments, an insoluble fiber may not be solubilized during the extraction stage. Thereby, the compound of interest may comprise a soluble fiber or an insoluble fiber.

In embodiments, the compound of interest may comprise a carbohydrate. The carbohydrate may especially be a saccharide, such as e.g. a monosaccharide, a disaccharide, an oligosaccharide, and a polysaccharide. Most especially, the compound of interest may comprise a monosaccharide and/or a disaccharide. The compound of interest may in embodiments comprise a target acid compound, such as a target organic acid compound. Especially, the target organic acid compound may be selected from the group comprising carboxylic acid, lactic acid, caproic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, heptanoic acid, octanoic acid and galacturonic acid. Alternatively, the target compound may comprise an inorganic acid compound, such as sulfuric acid.

Further, the compound of interest may comprise a target (organic) salt. The target salt may comprise a cation selected from the group comprising sodium, potassium, calcium, magnesium, and ammonium. Thereby, the target salt may in embodiments be selected from the group comprising sodium salts, potassium salts, calcium salts, magnesium salts, and ammonium salts. Further, the target salt may comprise an anion. The anion may be an inorganic anion selected from the group comprising chloride and sulfuric acid. In particular, the anion may be selected from the group comprising chloride and sulfate. Thereby, the target salt may in embodiments be selected from the group comprising chloride salts and sulfate salts. The anion may especially be an organic anion selected from the group comprising carbonic acid, carboxylic acid, lactic acid, caproic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, heptanoic acid, octanoic acid and galacturonic acid. Further, the anion may be an organic anion selected from the group comprising carbonate, carboxylate, lactate, caproate, acetate, propionate, butyrate, isobutyrate, valerate, isovalerate, heptanoate, octanoate and galacturonate. Thereby, the salt may in embodiments be selected from the group comprising a carbonate, a carboxylate, a lactate, a caproate, an acetate, a propionate, a butyrate, an isobutyrate, a valerate, an isovalerate, a heptanoate, an octanoate, and a galacturonate. Especially, the target salt may be selected from the group comprising chloride salt, a sulfate, an acetate, a carbonate, a carboxylate, and a lactate.

In embodiments, the compound of interest (as described above) may be provided by the solid mixture. In certain embodiments, the solid mixture may comprise biomass. The term “biomass” may herein especially refer to a renewable organic material. The biomass may typically comprise one or more of a compound of interest. The (initial) biomass may be obtained from various sources, such as e.g. one or more of plant-derived material, fungal -derived material, microbial-derived material, animal-derived material, or food product-derived material. Hence, the solid mixture may comprise one or more of plant biomass, fungal biomass, microbial biomass, animal biomass, and a food product. Such biomass may, for instance, be obtained from the agricultural industry or the food industry. The production of agricultural products and food products may result in various streams of biomass suitable for the present invention. The biomass may comprise a primary product, a side stream, a waste stream, or a residual stream. Especially, the biomass may comprise a side stream. The present invention may provide a suitable alternative for the use of biomass material that would otherwise not be used rather than, for instance, relying on food-grade biomass. In certain embodiments, the present invention may provide a more economically attractive option than other alternative methods for the use of such biomass.

Primary products may be the dominant value products of a production process, e.g. plants may be grown and harvested with the express purpose of using said plants (or parts of said plants) as an agricultural product or food product. Such plants may especially include e.g. sunflowers, olive trees, oil palms, rapeseeds, flaxes, soybeans, oats, com, wheat, barley, coffee trees, cocoa trees, avocado trees, potatoes, peppers, tomatoes, etc. Said plants (or plant parts) may be used as primary products with limited processing. Said plants (or plant parts) may be further processed to produce primary products, such as e.g. sunflower oil, olive oil, palm oil, rapeseed oil, linseed oil, soy oil, soy milk, tofu, oat milk, corn syrup, corn starch, wheat flour, coffee beans, chocolate, processed flavors, processed tomatoes, etc. Further, domesticated animals may be bred, kept and slaughtered as livestock with the express purpose of using said animal (or parts from said animals) as an agricultural product or food product. Animal primary products may especially include meat (such as from cattle, pigs, poultry, farmed fish, etc.), milk (such as from cattle, goats, etc.), cheese (also see below), eggs (such as from poultry), honey (such as from bees), wool (such as from sheep), etc. Moreover, (primary) products obtained from plant sources and animal sources may be turned into fungal and microbial primary products through fermentation processes. Fungal and microbial primary products may especially include alcoholic beverages (such as beer, wine, liquors, spirits, etc.), fermented dairy products (such as cheeses, yoghurts, etc.), yeasted baked goods (such as breads, pastries, pizzas, etc.), flavoring compounds (such as yeast extract, etc.) and so forth.

Under certain circumstances, such as e.g. market forces, quality demands, decomposition (due to logistical issues, climate issues, or infestation), et cetera, the primary products may lose significant economic value. The primary product may thus become available as the residual stream of the production process and may, for instance, be suitable as biomass for the method of the invention. Hence, in embodiments, the biomass may comprise a residual stream.

Production processes from the agricultural industry and the food industry may further result in a side stream of secondary products. Secondary products may not comprise the dominant value products of a production process but may comprise by-products of the process that retain economic value. The production process may in certain processes include elements and steps to optimize or refine secondary products depending on their economic value. Such secondary products may especially include: brewer’s spent-grain, spent whiskey mash, distiller’s spent-grain, wine lees, et cetera from the production of alcoholic beverages; press cakes from the vegetable oil production process of sunflower oil, olive oil, palm oil, rapeseed oil, linseed oil, soy oil, et cetera; press cakes from the plant-based milk production process of oat milk, soy milk, mycelium milk, et cetera; cake from fermented sauce production; okara from soybean processing for soy milk and tofu; cheese rind from cheese products; and so on. Such secondary product may become available as the side stream of the production process and may be suitable as a source material for the biomass. Hence, in embodiments, the biomass may comprise a side stream.

Production processes from the food industry and agricultural industry may additionally result in a waste stream of waste products. Waste products may be by-products of the process that contain little to no economic value and are commonly discarded or disposed of. Such waste products may comprise e.g. peelings, cuttings, and scrapings from plants such as those described above, and non-edible parts of such plants that may be taken along during the harvesting process. Such waste products may especially include e.g. tomato skins, potato skins, cocoa husks, bean husks, coffee silver-skin, vegetable cell wall, etc. Such waste product may thus become available as the waste stream of the production process and may hence be suitable as a source material for biomass. Hence, in embodiments, the biomass may comprise a waste stream.

The use of biomass from a residual stream, a side stream, or a waste stream in the method of the invention may be particularly beneficial in view of a circular economy and sustainability.

In embodiments, the solid mixture (obtained from, e.g., biomass) may comprise one or more of a compound of interest, such as a first compound, a second compound, and/or a third compound. On a dry weight basis (i.e., without any of the liquid components of the solid mixture, such as water) the solid mixture may comprise a compound of interest as described above. In embodiments, the solid mixture may on dry weight basis comprise 0.1 - 55 wt% free amino acid, especially 0.5 - 35 wt%, such as 1 - 15 wt%. In further embodiments, the solid mixture may on dry weight basis comprise 0.1 - 35 wt% free amino acid, especially 0.5 - 25 wt%, such as 1 - 15 wt%.

In further embodiments, the solid mixture may on dry weight basis comprise 5 - 65 wt% polypeptide, especially 15 - 55 wt%, such as 25 - 45 wt%. In further embodiments, the solid mixture may on dry weight basis comprise 0 - 55 wt% fat, especially 0.1 - 45 wt%, such as 1 - 35 wt%. In further embodiments, the solid mixture may on dry weight basis comprise 0 - 40 wt% soluble fiber, especially 0.1 - 30 wt%, such as 1 - 20 wt%. In further embodiments, the solid mixture may on dry weight basis comprise 0 - 60 wt% insoluble fiber, especially 0.1 - 50 wt%, such as 1 - 75 wt%. In further embodiments, the solid mixture may on dry weight basis comprise 0 - 75 wt% insoluble fiber, especially 0.1 - 65 wt%, such as 1 - 50 wt%.

In further embodiments, the solid mixture may on dry weight basis comprise 0.1 - 15 wt% saccharide, especially 1 - 8 wt%, such as 2 - 5 wt%. In further embodiments, the saccharide may comprise a mono- or disaccharide, especially a monosaccharide, or especially a disaccharide. Hence, in embodiments, the solid mixture may on dry weight basis comprise 0.1 - 15 wt% mono/disaccharide, especially 1 - 8 wt%, such as 2 - 5 wt%.

Hence, in specific embodiments, the solid mixture may on dry weight basis comprise 0.1 - 55 wt% free amino acid; 15 - 55 wt% polypeptide; 0 - 55 wt% fat; 0 - 30 wt% soluble fiber; 0 - 50 wt% insoluble fiber; and 0 - 8 wt% mono-/disaccharide. In further embodiments, the solid mixture may on dry weight basis comprise 0.5 - 25 wt% free amino acid; 15 - 55 wt% polypeptide; 0 - 45 wt% fat; 0 - 30 wt% soluble fiber; 0 - 50 wt% insoluble fiber; and 1 - 8 wt% mono-/disaccharide.

Additionally, the solid mixture may comprise 0 - 20 wt% organic acid (compound), especially 0.1 - 15 wt%, such as 1 - 10 wt%. Further, the solid mixture may comprise 0 - 50 wt% (organic) salt, especially 0.1 - 35 wt%, such as 1 - 20 wt%. In further embodiments, the solid mixture may comprise at least 5 wt% of the compound of interest, such as at least 10 wt%, especially at least 20 wt%.

Examples of solid mixtures with corresponding typical compositions are hereby provided for reference. A hard cheese rind solid mixture may comprise 1 - 15 wt% free amino acid, 35 - 60 wt% polypeptide, 35 - 60 wt% fat, at most 5 wt% (soluble and/or insoluble) fiber, at most 5 wt% mono-/di saccharide, and at most 12 wt% organic acid compound. A vegetable cake solid mixture (e.g., from vegetable oil production) may comprise at most 5 wt% free amino acid, 15 - 60 wt% polypeptide, at most 15 wt% fat, at most 60 wt% (soluble and/or insoluble) fiber, at most 40 wt% mono-/disaccharide, and at most 5 wt% organic acid compound. A solid residue solid mixture (e.g., from the production of plant-based milk or beer) may comprise at most 5 wt% free amino acid, 8 - 28 wt% polypeptide, at most 15 wt% fat, 30 - 65 wt% (soluble and/or insoluble) fiber, 35 - 65 wt% mono-/di saccharide, and at most 5 wt% organic acid compound. A further solid residue solid mixture (e.g., from the production of vegetable flavoring) may comprise 8 - 60 wt% free amino acid, 30 - 60 wt% polypeptide, at most 5 wt% fat, at most 20 wt% (soluble and/or insoluble) fiber, at most 20 wt% mono-/di saccharide, and at most 10 wt% organic acid compound.

With the present method, the compound of interest in such solid mixture may be efficiently accessed for further use by converting (or “splitting”) the solid mixture (comprising the biomass) into different parts.

Examples of solid mixtures that may be split into parts in certain embodiments of the method of the invention are hereby provided for reference. It will be clear to the person skilled in the art that the compositions of the parts will depend on the composition of the solid mixture and on the embodiment of the method of the invention followed. For instance, from a hard cheese or hard cheese rind solid mixture the method of the invention may, for instance, provide a supernatant part comprising at least 1 wt% glutamic acid and less than 15 wt% fat (on dry weight basis). Further, from such hard cheese or hard cheese rind solid mixture the method may provide a precipitated part comprising at least 50 wt% polypeptide (on dry weight basis). From a vegetable cake solid mixture (e.g., from vegetable oil production) the method may provide a precipitated part comprising at least 50 wt% polypeptide (on dry weight basis), especially at least 60 wt%. Such vegetable cake solid mixture may further result (via the method of the invention) in a non-dissolved part comprising at least 35 wt% fiber (on dry weight basis), especially at least 50 wt%. A solid residue solid mixture (e.g., from the production of plantbased milk or beer) may result in a precipitated part comprising at least 35 wt.% polypeptide (on dry weight basis), especially at least 50 wt%. Such solid residue solid mixture may further result in a non-dissolved part comprising at least 50 wt.% fiber (on dry weight basis), especially at least 65 wt.%. A further solid residue solid mixture (e.g., from the production of vegetable flavoring) may result in a precipitated part comprising at least 50 wt.% free amino acid (on dry weight basis), especially at least 70 wt.%. Such further solid residue solid mixture may further result in a supernatant part comprising at least 35 wt.% free amino acid (on dry weight basis), especially at least 50 wt.%. A bacterial/fungal fermentation biomass solid mixture may result in a precipitated part comprising at least 30 wt.% polypeptide (on dry weight basis), especially at least 50 wt.%, and at least 50 wt.% fiber (on dry weight basis), especially at least 65 wt.%.

Hence, in embodiments, especially wherein the solid mixture comprises a hard cheese or hard cheese rind, the supernatant part may comprise at least 1 wt% glutamic acid and at most 15 wt% fat (on dry weight basis). Further, in such embodiments, the precipitated part may comprise at least 50 wt% polypeptide (on dry weight basis). In further embodiments, especially wherein the solid mixture comprises a vegetable cake solid mixture (e.g., from vegetable oil production), the precipitated part may comprise at least 50 wt% polypeptide (on dry weight basis), especially at least 60 wt%. Further, in such embodiments, the non-dissolved part may comprise at least 35 wt% fiber (on dry weight basis), especially at least 50 wt%.

In further embodiments, especially wherein the solid mixture comprises a solid residue solid mixture (e.g., from the production of plant-based milk or beer), the precipitated part may comprise at least 35 wt.% polypeptide (on dry weight basis), especially at least 50 wt%. Further, in such embodiments, the non-dissolved part may comprise at least 50 wt.% fiber (on dry weight basis), especially at least 65 wt.%.

In further embodiments, especially wherein the solid mixture comprises a (further) solid residue solid mixture (e.g., from the production of vegetable flavoring), the precipitated part may comprise at least 50 wt.% free amino acid (on dry weight basis), especially at least 70 wt.%. Further, in such embodiments, the supernatant part may comprise at least 35 wt.% free amino acid (on dry weight basis), especially at least 50 wt.%.

In further embodiments, especially wherein the solid mixture comprises a bacterial/fungal fermentation biomass solid mixture, the precipitated part may comprise at least 30 wt.% polypeptide (on dry weight basis), especially at least 50 wt.%, and at least 50 wt.% fiber (on dry weight basis), especially at least 65 wt.%.

In certain embodiments, the method may comprise a solid mixture preparation stage. The solid mixture preparation stage may comprise providing a solid mixture from an initial source (e.g., biomass) suitable for use with the present method. Thus the solid mixture preparation stage may in general precede the extraction stage. The solid mixture preparation stage may comprise exposing the initial source to one or more of drying, filtration, centrifugation, and hydrolysis, thereby providing the solid mixture.

In embodiments, the solid mixture preparation stage may comprise providing the solid mixture with a water content suitable for use with the method of the invention. The water content of the solid mixture may be at most 95 wt%, such as 90 wt%, especially 85 wt%, such as 75 wt%, especially 50 wt%. In particular, the water content of the solid mixture may be at most 95 wt%, such as at most 90 wt%, especially at most 85 wt%, such as at most 75 wt%, especially at most 50 wt%. The water content of the solid mixture may be at least 1 wt%, such as 2 wt%, especially 5 wt%, such as 10 wt%, especially 15 wt%. Alternatively or additionally, the water content of the solid mixture may be at least 1 wt%, such as at least 2 wt%, especially at least 5 wt%, such as at least 10 wt%, especially at least 15 wt%. Thereby, the water content of the solid mixture may be selected from the range of 1 - 95 wt%, such as from the range of 2 - 90 wt%, especially from the range of 5 - 85 wt%, such as from the range of 10 - 75 wt%, especially from the range of 15 - 50 wt%. Examples of typical water contents of solid mixtures are hereby provided for reference. A hard cheese rind solid mixture may comprise 10 - 35 wt% water. A vegetable cake solid mixture (e.g., from vegetable oil production) may comprise 2 - 15 wt% water. A solid residue solid mixture (e.g., from the production of plant-based milk or beer) may comprise 70 - 90 wt% water. A further solid residue solid mixture (e.g., from the production of vegetable flavoring) may comprise 35 - 60 wt% water.

In certain embodiments, upon the addition of the acid to the liquid mixture during the precipitation stage, the acid and the cation may form a salt. For example, in certain embodiments, the CO2 and the cation may form a carbonate. The term “carbonate” may herein especially refer to a compound comprising the carbonate anion (CO₃ ^2-), especially to a salt, ester, or mineral comprising the carbonate anion. For instance, the term “carbonate” may herein refer to any one of HCO₃‘ H2CO3, HNaCO3,Na₂CO₃, K₂CO₃, CaCO₃, (NH₄)₂CO₃, and MgCO₃. In further embodiments, the HC1 and the cation may form a chloride salt. As for the term “carbonate”, the term “chloride salt” may herein especially refer to a compound comprising the chloride anion (Cl“). For example, the term “chloride salt” may herein refer to any one of NaCl, CaCl₂, KC1, NH4CI, and MgCl₂. In other embodiments, the acetic acid and the cation may form an acetate. Like for “carbonate” and “chloride salt”, the term “acetate” may herein especially refer to a compound comprising the acetate anion (C₂H₃O₂ ). For example, the term “acetate” may herein refer to any one of CH₃COONa, Ca(C₂H₃O₂)₂, CH3COOK, NH4CH3COO, and Mg(C₂H₃O₂)₂. In further embodiments, the acid and the cation may form an acid salt, ester, or mineral comprising the corresponding anion of the acid.

Hence, the precipitation stage may comprise providing the acid to the liquid mixture such that the acid and the cation form a salt. Such formed salts, especially carbonates, may generally have a (relatively) poor solubility. Thereby, the formed salt (e.g., a carbonate) may precipitate during the precipitation stage. In embodiments, the carbonate precipitation stage may comprise providing the acid to the liquid mixture such that at least 30% of the cation precipitates (as an acid salt, especially a carbonate), especially at least 50%, such as at least 70%. In further embodiments, the precipitation stage may comprise providing the acid to the liquid mixture such that at least 80% of the cation precipitates (as an acid salt, especially a carbonate), especially at least 90%, such as at least 95%. Hence, the precipitated part may comprise the formed salt, such as a carbonate. In embodiments, the liquid mixture may comprise m M of a cation, such as Na⁺, and the precipitation stage may comprise providing M of an acid, such as CO2, wherein is at least 0.1 *m, such as at least 0.2*m, especially at least 0.25*ni. In particular, even adding a relatively small amount of CO2 relative to the cation may already provide a desired effect on the solubility of the salt. In further embodiments, may be at least 0.5*m, such as at least 1 *m. In further embodiments, may be at least 1.5*m, such as at least 2*m, especially at least 3*m. In further embodiments, may be at most 1000*m, such as at most 500*ni, especially at most 200*ni. In further embodiments, may be at most 100*ni, such as at most 50*m, especially at most 10*ni. In further embodiments, may be at most 5*m, such as at most m, especially at most 0.5*m. For instance, in specific embodiments, the cation may comprise sodium, and the carbonate may comprise sodium carbonate.

In certain embodiments, the solubility of a (second) compound in the liquid mixture may further decrease if the pH is lowered further. Hence, in embodiments, the precipitation stage may comprise providing the acid (especially CO2) to the liquid mixture to (gradually) reduce the pH of the liquid mixture to a precipitation point of a (second) compound.

Hence, in such embodiments, the precipitation stage may comprise a first salt precipitation stage and a second precipitation stage. The first precipitation stage may comprise adding the acid to remove the cation (and the anion, especially OH') via precipitation of a formed salt. The second precipitation stage may comprise adding additional acid (especially CO2) to further reduce the pH of the liquid mixture to precipitate a (second) compound.

The first precipitation stage and the second precipitation stage may, in such embodiments, be temporally separated, and/or may be executed at different locations. For instance, the first precipitation stage may be executed in a first vessel, after which the liquid mixture is provided from the first vessel to a second vessel, especially via a sieve, and the second precipitation stage may be executed in the second vessel.

In further embodiments, the first precipitation stage may smoothly transition to the second precipitation stage, especially wherein providing of the acid (especially CO2) to the liquid mixture is not interrupted between the two stages.

In embodiments, the optional compound recovery stage may comprise one or more substages selected from the first compound recovery stage, the second compound recovery stage, and the third compound recovery stage. These compound recovery stages may especially comprise separating the compound from a part or enriching the compound in the part. In separation, the compound may be separated from other compounds in the part. In enrichment, one or more other compounds may be removed from the part. In certain embodiments, the method may comprise a first compound recovery stage. The first compound recovery stage may comprise separating or enriching a first compound from the non-dissolved part. The first compound may comprise one or more selected from the group comprising a free amino acid, a polypeptide, a fat, a soluble fiber, and an insoluble fiber. The first compound may comprise a free amino acid, especially glutamate. The first compound may comprise a soluble fiber, especially pectin and/or beta-glucan. The first compound may comprise an insoluble fiber, especially cellulose or hemicellulose.

In further embodiments, the method may comprise a second compound recovery stage. The second compound recovery stage may comprise separating or enriching a second compound from the precipitated part. The second compound may comprise one or more selected from the group comprising a free amino acid, a polypeptide, and a salt. The second compound may comprise a free amino acid, especially tyrosine and/or leucine. The second compound may comprise a salt, especially a sulfate, a lactate and/or a formed salt (e.g., a carbonate, a chloride salt, an acetate salt) from the cation reacting with the acid during the precipitation stage.

Additionally or alternatively, the method may comprise a third compound recovery stage. The third compound recovery stage may comprise separating or enriching a third compound from the supernatant part. The third compound may comprise one or more selected from the group comprising a free amino acid, a polypeptide, a soluble fiber, a saccharide, an organic acid, and an (organic) salt. The third compound may comprise a free amino acid, especially glutamate. The third compound may comprise an organic acid, especially carboxylic acid and/or lactic acid. The third compound may comprise an (organic) salt, especially a carboxylate salt and/or a lactate salt.

In certain embodiments, the method may further comprise an organic extraction material recovery stage following the precipitation stage. The organic extraction material recovery stage may comprise subjecting the supernatant part to an organic extraction material recovery step, especially to one or more of distillation, drying, adsorption, and filtration. Thereby, at least part of the organic extraction material (provided by the extraction liquid) may be separated from the supernatant part. Further, a substantial part of the organic extraction material may be separated from the supernatant part, such as at least 30 wt%, especially at least 50 wt%, further at least 70 wt%. In some embodiments, (essentially) all the organic extraction material may be separated from the supernatant part, such as at least 90 wt%, especially at least 95 wt%, further at least 98 wt%. Hence, the organic extraction material recovery stage may provide a recovered organic extraction material part and a supernatant residue part for further use. The recovered organic extraction material part may, e.g., be utilized to provide at least part of the organic extraction material of the extraction liquid in another run of the process. The supernatant residue part may, e.g., provide a (third) compound without organic extraction material for further use.

The organic extraction material recovery step may be selected from the group comprising distillation, drying, adsorption, and filtration. One or more organic extraction material recovery steps may be combined (sequentially) to provide a recovered organic extraction material part suitable for further use. A further organic extraction material recovery step may comprise subjecting the recovered organic extraction material part to an organic extraction material recovery step to provide a further recovered organic extraction material part and a further supernatant residue part. The organic extraction material recovery step may especially comprise at least one organic extraction material recovery step comprising distillation.

In certain embodiments, the method may further comprise subjecting the liquid mixture to a liquid removal step, especially to one or more of drying, adsorption, and filtration. The liquid removal step may comprise separating at least part of the mixture of organic extraction material and water from the liquid mixture. Especially, the liquid removal step may comprise separating at least part of the mixture of organic extraction material and water from the liquid mixture. At least part of the mixture of organic extraction material and water may be separated from the liquid mixture, such as at least 1 wt%, especially at least 2 wt%, such as at least 5 wt%, especially at least 10 wt%, such as at least 15 wt%. Further, a substantial part of the mixture of organic extraction material and water may be separated from the liquid mixture, such as at least 20 wt%, especially at least 35 wt%, such as at least 50 wt%, especially at least 65 wt%, such as at least 80 wt%. In some embodiments, (essentially) all the mixture of organic extraction material and water may be separated from the liquid mixture, such as at least 90 wt%, especially at least 95 wt%, such as at least 98 wt%. Hence, the liquid removal step may provide a liquid mixture with decreased organic extraction material and water content and may thereby be more suitable for efficient use in a following stage in the current method.

The liquid removal step may be selected from the group comprising drying, adsorption, and filtration. In certain embodiments of the liquid removal step, (at least part of) both the organic extraction material and the water may be separated from the liquid mixture. The organic extraction material and the water may be separated in equal measure from the liquid mixture, but may also be differentially separated, e.g., a liquid removal step may remove twice as much organic extraction material compared to water from the liquid mixture, or vice versa. In other embodiments of the liquid removal step, only (part of) the organic extraction material may be separated from the liquid mixture. In further embodiments, only (part of) the water may be separated from the liquid mixture.

One or more liquid removal steps may be combined (sequentially) to provide a liquid mixture suitable for use in the current method. In particular, one liquid removal step may separate (at least part of) the organic extraction material from the liquid mixture, and another liquid removal step may separate (at least part of) the water from the liquid mixture. Especially, in certain embodiments, the liquid mixture may be subjected to at least two liquid removal steps. One of the at least two liquid removal steps may comprise drying.

In certain embodiments, the method may further comprise subjecting one or more of the plurality of parts (e.g. the non-dissolved part, the precipitated part, the supernatant part, the recovered organic extraction material part, etc.) to a purification step. The purification step may comprise separating at least part of the components used in the current method from one or more of the plurality of parts. Thereby, the purification step may comprise separating at least part of the organic extraction material, the anion, the cation, and/or water from one or more of the plurality of parts. At least part of the components used in the current method may be separated from one or more of the plurality of parts, such as at least 1 wt%, especially at least 2 wt%, such as at least 5 wt%, especially at least 10 wt%, such as at least 15 wt%. Further, a substantial part of the components used in the current method may be separated from the one or more of the plurality of parts, such as at least 20 wt%, especially at least 35 wt%, such as at least 50 wt%, especially at least 65 wt%, such as at least 80 wt%. In some embodiments, (essentially) all the components used in the current method may be separated from one or more of the plurality of parts, such as at least 90 wt%, especially at least 95 wt%, such as at least 98 wt%. Hence, the purification step may provide one or more of the plurality of parts without components used in the current method, thereby the one or more of the plurality of parts may be more suitable for further use.

The purification step may be selected from the group comprising distillation, drying, adsorption, crystallization, filtration, membrane purification, washing, and extrusion. In certain embodiments of the purification step, (at least part of) all the components used in the current method may be separated from the one or more of the plurality of parts. The components used in the current method may not be separated in equal measure from the one or more of the plurality of parts, e.g., a purification step may remove twice as much anion compared to cation from the plurality of parts, or vice versa. In other embodiments of the purification step, only (part of) one of the components used in the current method may be separated from the one or more of the plurality of parts. In further embodiments, (part of) two of the components used in the current method may be separated from the one or more of the plurality of parts. In specific embodiments, (part of) three of the components used in the current method may be separated from the one or more of the plurality of parts.

One or more purification steps may be combined (sequentially) to provide one or more of the plurality of parts suitable for further use. Especially, one purification step may separate (at least part of) one component (from the components used in the current method) from the one or more of the plurality of parts, and another purification step may separate another component (from the components used in the current method) from the one or more of the plurality of parts. Especially, in certain embodiments, the one or more of the plurality of parts may be subjected to two or more purification steps. One of the two or more purification steps may comprise washing, and another of the two or more purification steps may comprise drying.

The purification step may be combined or overlap with an organic extraction material recovery stage and/or a compound recovery stage. Especially, the one or more purification steps may be combined or overlap with a first compound recovery stage, a second compound recovery stage, and/or a third compound recovery stage. Thereby, the one or more purification steps may provide a first compound, a second compound, and/or a third compound suitable for further use.

In certain embodiments, the precipitation stage may further comprise providing additional organic extraction material to the liquid mixture. An increased weight ratio of organic extraction material to water may reduce the solubility of a (second) compound. The weight ratio of organic extraction material to water in the liquid mixture may thus be selected from the range of 250: 1 - 1 :3, such as from the range of 100: 1 - 2:3, especially from the range of 20:1 - 1 :2, such as from the range of 10 : 1 - 1 : 1 , especially from the range of 5 : 1 - 2 : 1. Hence, the addition of organic extraction material to the liquid mixture may facilitate reaching the precipitation point of a (second) compound.

In further embodiments, the (second) compound may comprise two or more (different) (second) compounds, wherein the precipitation stage comprises providing the acid (such as CO2) to the liquid mixture to successively reduce the liquid mixture to the precipitation points of the two or more (second) compounds to separate the two or more (second) compounds. Hence, the method, especially the precipitation stage, may comprise first providing the acid to the liquid mixture to reduce the liquid mixture to the precipitation point of one (second) compound of the two or more (second) compounds to separate the one (second) compound, and subsequently providing the acid to the liquid mixture to reduce the liquid mixture to the precipitation point of a further (second) compound of the two or more (second) compounds to separate the further (second) compound.

In particular, in embodiments, the one (second) compound may have a first isoelectric point pH, and the further (second) compound may have a second isoelectric point pI2, wherein pI2 < PI1, and the method, especially the precipitation stage, may comprise first providing the acid (such as CO2) to the liquid mixture to reduce the liquid mixture to a pH selected from the range of pIl-0.5 - pl 1+0.5, and subsequently providing the acid to the liquid mixture to reduce the liquid mixture to a pH selected from the range of pI2-0.5 - pI2+0.5.

In certain embodiments, the non-dissolved part may be utilized for at least one more run of the method of the invention. The non-dissolved part may be utilized for at most two more runs of the method of the invention. Each run may comprise the extraction stage and the precipitation stage of the method of the invention. Each run may provide another plurality of parts. Each run may be a different run with different conditions, e.g., different components of the extraction liquid. Hence, the plurality of parts obtained from a first run may be different from the plurality of parts obtained from a second run. Thereby different compounds of interest in the non-dissolved part of the solid mixture may be accessed.

In a further aspect, the invention may provide a system for converting (or “splitting”) a solid mixture into a non-dissolved part, a precipitated part, and a supernatant part. The system may comprise one or more of an inlet, a liquid supply, an extraction unit, a precipitation unit, and an acid supply. In certain embodiments, the system may comprise a control system (described further below).

The invention may especially provide a system for converting (or “splitting”) a solid mixture into a non-dissolved part, a precipitated part and a supernatant part, the system comprising an inlet, a liquid supply, an extraction unit, a precipitation unit, and an acid supply, and wherein the system has an operational mode, wherein: the inlet is configured to provide the solid mixture to the extraction unit; the liquid supply is configured to provide an extraction liquid to the extraction unit; wherein the extraction liquid comprises (a) an organic extraction material selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of OH' , (c) at least 0.01 M of a cation selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH₄ ⁺, and (d) water; the extraction unit is configured to dissolve at least part of the solid mixture and (i) to provide a liquid mixture to the precipitation unit and (ii) to provide the non-dissolved part; the acid supply is configured to supply an acid, especially CO2, to the liquid mixture hosted in the precipitation unit thereby forming a precipitated part and a supernatant part; the precipitation unit is configured to provide the precipitated part and the supernatant part. In further embodiments, the system, especially the control system, may have an operational mode. The term “operational mode” may also be indicated as “controlling mode”. The system, or apparatus, or device may execute an action in a “mode” or “operational mode” or “mode of operation”. Likewise, in a method an action, stage, or step may be executed in a “mode” or “operation mode” or “mode of operation”. This does not exclude that the system, or apparatus, or device may also be adapted for providing another operational mode, or a plurality of other operational modes. Likewise, this does not exclude that before executing the mode and/or after executing the mode one or more other modes may be executed. However, in embodiments a control system may be available, that is adapted to provide at least the operational mode. Would other modes be available, the choice of such modes may especially be executed via a user interface, though other options, like executing a mode in dependence of a sensor signal or a (time) scheme, may also be possible. The operation mode may in embodiments also refer to a system, or apparatus, or device, that can only operate in a single operation mode (i.e. “on”, without further tunability).

In embodiments, the operational mode may especially comprise one or more of an extraction stage, a first separation stage, and a precipitation stage. In further embodiments, the operational mode may comprise one or more of a first separation stage, a compound recovery stage, a first compound recovery stage, a second compound recovery stage, a third compound recovery stage, a first precipitation stage, a second precipitation stage, and an organic extraction material recovery stage.

In the extraction stage, the inlet may (be configured to) provide the solid mixture to the extraction unit. Moreover, the liquid supply may be configured to provide an extraction liquid to the extraction unit during the extraction stage. In further embodiments, the precipitation unit may be configured to receive a liquid mixture from the extraction unit after the extraction stage.

The term “extraction liquid” may herein especially refer to a liquid provided to the extraction unit, whereas the term “liquid mixture” may herein especially refer to the liquid after having been in contact with a solid mixture in the extraction stage. Hence, the liquid supply may be configured to provide an extraction liquid to (a solid mixture in) the extraction unit to provide a liquid mixture.

The extraction liquid may especially comprise an organic extraction material selected from the group consisting of acetone and alcohols. In embodiments, the extraction liquid may comprise at least 0.1 M of an anion, especially OH'. The extraction liquid may further comprise at least 0.1 M of a cation selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH4⁺, especially a monovalent cation, such as a monovalent cation selected from the group comprising Na⁺, K⁺, and NH4⁺, or especially a bivalent cation, such as a bivalent cation selected from the group comprising Ca²⁺, Mg²⁺. The extraction liquid may additionally comprise water.

In the extraction stage, the liquid supply may (be configured to) provide an extraction liquid to the extraction unit, especially to the solid mixture. The extraction unit may (be configured to) dissolve at least part of the solid mixture. Thereby the extraction unit may (be configured to) to provide a liquid mixture comprising a (second and third) compound to the precipitation unit, and to provide a non-dissolved part.

In embodiments, during a first separation stage, the liquid mixture and the nondissolved part may be separated in the extraction unit, or alternatively in a first separation unit.

In embodiments, the extraction unit may be configured to separate the liquid mixture from the non-dissolved part, to provide the non-dissolved part and the liquid mixture (wherein the liquid mixture may be provided to the precipitation unit). Hence, in embodiments, the extraction unit may comprise one or more of a filter (e.g. a sieve), a sedimentation apparatus (e.g. a sedimentation tank), a hydrocyclone, a screw-press, and a centrifuge, configured to separate the liquid mixture from the non-dissolved part. Especially, the extraction unit may comprise one or more of a filter and a centrifuge. In embodiments, the liquid mixture may be transferred from the extraction unit to the precipitation unit, i.e., the extraction unit may be configured to provide the liquid mixture to the precipitation unit. Hence, in embodiments, the extraction unit may comprise a outlet, and the precipitation unit may comprise an inlet, wherein the outlet of the extraction unit may be (fluidically) connected to the inlet of the precipitation unit. Additionally or alternatively, the extraction unit may comprise a fluid transfer device, such as e.g. a pump, wherein the fluid transfer device may be configured to transfer the liquid mixture (from the extraction unit) to the precipitation unit.

In (other) embodiments, the extraction unit and the precipitation unit may be comprised by the same vessel, i.e., the same vessel may serve as the extraction unit (during the extraction stage) and the precipitation unit (during the precipitation stage). In such embodiments, the phrase “provide a liquid mixture to the precipitation unit”, and similar phrases, may indicate a removal of the non-dissolved part from the vessel (e.g. by filtration or centrifugation) to provide the liquid mixture suitable for the precipitation stage (executed in the precipitation unit). Yet, especially, the extraction unit may be (configured) separate from the precipitation unit. Further, in embodiments, the separation of the liquid mixture and the nondissolved part may be performed in a separate unit, such as especially in a first separation unit. Hence, in embodiments, the system may comprise a first separation unit, and the extraction unit may be configured to provide the (combined) liquid mixture and non-dissolved part to the first separation unit. In embodiments, the first separation unit may comprise one or more of a filter, a sedimentation apparatus, a hydrocyclone, a screw-press, and a centrifuge, such as especially one or more of a filter and a centrifuge, and may be configured to separate the liquid mixture from the non-dissolved part to provide the liquid mixture and the non-dissolved part (wherein the first separation unit may further be configured to provide the liquid mixture to the precipitation unit).

After the extraction stage, the extraction unit (or first separation unit) may (be configured to) recover a (first) compound from the non-dissolved part. The first compound recovery may be achieved especially via one or more of evaporation, distillation, crystallization, filtration, esterification, and acidification. Hence, in embodiments, the extraction unit (or first separation unit) may comprise one or more of an evaporation unit, a distillation unit, a crystallization unit, a filtration unit, an esterification unit and an acidification unit.

In the precipitation stage, the acid supply may (be configured to) provide the acid (especially CO2) to the precipitation unit, especially to the liquid mixture in the precipitation unit. Especially, (at least part of) the acid and the cation may form a formed salt, especially wherein the formed salt precipitates. In particular, the solubility of the formed salt may depend on the cation. Hence, the cation may be selected in view of the solubility of the corresponding formed salt, and in view of the to be recovered (second and third) compound. Thereby the precipitation unit may (be configured to) provide a precipitated part and a supernatant part.

In further embodiments, during a second separation stage, the precipitated part and the supernatant part may be separated in the precipitation unit, or alternatively in a second separation unit.

Hence, in embodiments, the precipitation unit may be configured to separate the precipitated part from the supernatant part. In (such) embodiments, the precipitation unit may thus comprise one or more of a filter (e.g. a sieve), a sedimentation apparatus (e.g. a sedimentation tank), a hydrocyclone, a screw-press, and a centrifuge, configured to separate the precipitated part from the supernatant part. Especially, the precipitation unit may comprise one or more of a filter and a centrifuge. Further, in embodiments, the precipitation unit may comprise a precipitate outlet and a supernatant outlet, wherein the precipitation unit may be configured to provide (i) the precipitated part via the precipitate outlet, and (ii) the supernatant part via the supernatant outlet.

In (other) embodiments, the separation of the precipitated part from the supernatant part may be performed in a separate unit, such as especially in a second separation unit. Hence, in embodiments, the system may comprise a second separation unit, and the precipitation unit may be configured to provide the (combined) precipitated part and supernatant part to the second separation unit. In embodiments, the second separation unit may comprise one or more of a filter, a sedimentation apparatus, a hydrocyclone, a screw-press, and a centrifuge, such as especially one or more of a filter and a centrifuge, and may be configured to separate the precipitated part from the supernatant part to provide the precipitated part and the supernatant part.

After the precipitation stage, the precipitation unit (or second separation unit) may (be configured to) recover a (second) compound from the precipitated part. The precipitation unit (or second separation unit) may further (be configured to) recover a (third) compound from the supernatant part. The second compound recovery and third compound recovery may be achieved especially via one or more of evaporation, distillation, crystallization, filtration, esterification, and acidification. Hence, in embodiments, the precipitation unit (or second separation unit) may comprise one or more of an evaporation unit, a distillation unit, a crystallization unit, a filtration unit, an esterification unit and an acidification unit.

In embodiments, the system may further comprise a control system, especially wherein the control system is configured to control one or more of the inlet, the liquid supply, the extraction unit, the precipitation unit, and the acid supply.

The term “controlling” and similar terms herein may especially refer at least to determining the behavior or supervising the running of an element. Hence, herein “controlling” and similar terms may e.g. refer to imposing behavior to the element (determining the behavior or supervising the running of an element), etc., such as e.g. measuring, displaying, actuating, opening, shifting, changing temperature, etc.. Beyond that, the term “controlling” and similar terms may additionally include monitoring. Hence, the term “controlling” and similar terms may include imposing behavior on an element and also imposing behavior on an element and monitoring the element. The controlling of the element can be done with a control system. The control system and the element may thus at least temporarily, or permanently, functionally be coupled. The element may comprise the control system. In embodiments, the control system and the element may not be physically coupled. Control can be done via wired and/or wireless control. The term “control system” may also refer to a plurality of different control systems, which especially are functionally coupled, and of which e.g. one control system may be a master control system and one or more others may be slave control systems.

In certain embodiments, the precipitation unit may comprise a first precipitation unit and a second precipitation unit, especially wherein the first precipitation unit is configured to receive a liquid mixture from the extraction unit. In such embodiments, in the operational mode, especially in the first precipitation stage, the acid supply may (be configured to) provide the acid (especially CO2) to the first precipitation unit, wherein the first precipitation unit (is configured to) separate the precipitated formed salt and (remaining) liquid mixture, and to provide the (remaining) liquid mixture to the second precipitation unit. In further embodiments, in the operational mode, especially in the second precipitation stage, the second precipitation unit may (be configured to) provide the precipitated part and the supernatant part. In particular, in further embodiments, in the second precipitation stage, the acid supply may (be configured to) provide the acid to the second precipitation unit, especially to (gradually) reduce the pH of the liquid mixture to a precipitation point of the (second) compound, and/or especially to protonate the (second) compound.

In further embodiments, the system may comprise an organic extraction material recovery unit. The organic extraction material recovery unit may (be configured to) receive the supernatant part from the (second) precipitation unit. The organic extraction material recovery unit may subject the supernatant part to an organic extraction material recovery step, especially distillation, to separate at least part of the organic extraction material from the supernatant part. Thereby, the organic extraction material recovery unit may provide a recovered organic extraction material part and a supernatant residue part.

In embodiments, the system may be configured to execute the method of the invention. The operational mode of the system may comprise executing the method of the invention, especially in continuous operation, batch operation, or countercurrent operation. Continuous operation may imply a system where specific units may be (configured to be) in an operational state at all times. Especially, the inlet and liquid supply may provide a continuous source of solid mixture and extraction liquid to the extraction unit, which together with the precipitation unit may be in continuous operation. Batch operation may imply a system (configured to be) executing the method on a single batch of solid mixture and extraction liquid with units being turned off in between batches. Countercurrent operation may imply a system (configured to be) executing the method such that the liquid mixture flows in opposite direction to the solid mixture, thereby enriching the liquid mixture in a (second and/or third) compound. This may provide for a more effective use of extraction liquid. Especially, in embodiments, the system may be configured for continuous operation of the precipitation stage, such that the acid (such as CO2) and the liquid mixture are both provided continuously to the precipitation unit. Such configuration may especially enhance mixing.

Especially, the control system may be configured to (have the system) execute the method of the invention.

As indicated above, in embodiments, cheese (rind) may be used as the solid mixture. Hence, in embodiments, the method of the invention may be a method for converting a cheese (rind) into a non-dissolved part, a precipitated part, and a supernatant part. In such embodiments, the supernatant part may especially be (used to provide) a cheese extract. That is, in embodiments, the supernatant part may (directly) provide the cheese extract, without further processing steps. Alternatively, the supernatant part may be processed to provide the cheese extract. For instance, the organic extraction material (and optionally the water) may be removed from the supernatant part (e.g. by distillation) to provide the cheese extract.

Hence, according to a further aspect, the invention may provide a cheese extract. The cheese extract may be obtainable, especially be obtained, using the method of the invention. Further, the cheese extract may comprise at least part of the supernatant part (of the method of the invention). In embodiments, the cheese extract may have a solubility in water selected from the range of > 70%, such as from the range of > 80%, especially from the range of > 90%. Further, on a dry weight basis, the cheese extract may comprise (one or more of) (i) > 0.5 wt% free glutamic acid, such as > 1 wt% free glutamic acid, especially > 1.5 wt% free glutamic acid (ii) 0.2-20 wt% fat, such as 0.5-15 wt% fat, especially 1-10 wt% fat, (iii) 30-95 wt%, such as 40-90 wt%, especially 45-85 wt%, polypeptide, (iv) < 7.5 wt%, such as < 5 wt%, especially < 4 wt%, carbohydrate, wherein the cheese extract may comprise < 3 wt%, such as < 2 wt%, especially < 1.5 wt%, of lactose, and (v) 2-30 wt%, such as 4-25 wt%, especially 5-20 wt%, organic acid, wherein the organic acid may comprise one or more of acetic acid, lactic acid, butyric acid, and propionic acid, such as especially one or more of butyric acid and propionic acid. Especially, in embodiments, the cheese extract may have a solubility in water selected from the range of > 90%. Further, on a dry weight basis, the cheese extract may comprise (one or more of) (i) > 1 wt% free glutamic acid, (ii) 1-10 wt% fat, (iii) 45-85 wt% polypeptide, (iv) < 5 wt% carbohydrate, wherein the cheese extract may comprise < 2 wt% of lactose, and (v) 5- 20 wt% organic acid, wherein the organic acid may comprise one or more of acetic acid, lactic acid, butyric acid, and propionic acid, such as especially one or more of butyric acid and propionic acid. Hence, in specific embodiments, the invention provides a cheese extract obtained using the method as defined herein, wherein the cheese extract comprises at least part of the supernatant part, wherein the cheese extract has a solubility in water selected from the range of > 90%, and wherein on a dry weight basis the cheese extract comprises: (i) > 1 wt% free glutamic acid; (ii) 1-10 wt% fat; (iii) 45-85 wt% polypeptide; (iv) < 5 wt% carbohydrate, wherein the cheese extract comprises < 2 wt% of lactose; and (v) 5-20 wt% organic acid, wherein the organic acid comprises one or more of acetic acid, lactic acid, butyric acid, and propionic acid. Such a cheese extract may have (per gram of dry product) a relatively stronger cheese flavor and/or taste compared to the (original) cheese (rind), yet may comprise a relatively lower fat concentration. Hence, such a cheese extract may be suitable for imparting a cheese flavor and/or taste to low-fat (or diet) products. Further, such a cheese extract may facilitate imparting a cheese flavor and/or taste to liquid products, as the cheese extract (unlike the parent cheese rind) may have a high solubility in water (at room temperature).

In embodiments, the solubility of a substance (such as a single (pure) compound, a mixture, etc.) in water may be determined as follows: (a) the substance is placed in a suitable container; (b) water having a temperature of 20 °C is added to the container, wherein 100 ml of water is added per 5 grams of the substance (resulting in a 5% (w/v) solution), wherein the water is brought into contact with the substance, and the combination of water and substance is optionally stirred; (c) after 60 minutes, the solution is checked for the presence of a sediment, and the transparency (or cloudiness) of the solution is recorded; (d) the solution is passed through a filter, and the filtrate is (completely) dried and the remaining solids weighed to determine the amount of substance dissolved. The procedure for determining solubility may typically be performed at atmospheric pressure.

In embodiments, the solubility of the cheese extract (of the invention) may be > 70%, such as > 80%, especially > 90%, like > 95%, including 100%. That is, assuming a solubility of > 90%, and using the test method above, over 4.5 g (per 100 ml of water) of dried cheese extract may be recovered from the filtrate after step (d). Yet, in embodiments, the solubility of the cheese extract may be < 99%, such as < 98%, especially < 97%.

In embodiments, the cheese extract may comprise a free amino acid (see above), such as especially (free) glutamic acid. Especially, on a dry weight basis, the cheese extract may comprise > 0.5 wt%, such as > 1 wt%, especially > 1.5 wt%, (free) glutamic acid. Further, in embodiments, on a dry weight basis, the cheese extract may comprise > 2 wt%, such as > 2.5 wt%, especially > 3 wt%, (free) glutamic acid. Additionally or alternatively, on a dry weight basis, the cheese extract may comprise < 10 wt%, such as < 8 wt%, especially < 6 wt%, (free) glutamic acid.

Further, on a dry weight basis, the cheese extract may comprise > 0.2 wt% fat, such as > 0.5 wt%, especially > 1 wt%, like > 1.5 wt%. Additionally or alternatively, in embodiments, on a dry weight basis, the cheese extract may comprise < 20 wt% fat, such as < 15 wt%, especially < 10 wt%, like < 8 wt%. Hence, in embodiments, on a dry weight basis, the cheese extract may comprise 0.2-20 wt%, such as 0.5-15 wt%, especially 1-10 wt%, like 1.5-8 wt%, fat. The fat content of (mixture of) compound(s) may herein especially be determined by the fraction of the compound soluble in petroleum benzine after acid hydrolysis. That is, a weighed amount of the (dry) compound (e.g. the cheese extract) may be subjected to acid hydrolysis, optionally neutralized and/or dried, dissolved in petroleum benzine, and filtered to remove any non-dissolved matter from the petroleum benzine extract. The petroleum benzine may subsequently be evaporated from the petroleum benzine extract, and the weight of the (solid or oily) residue may be determined and divided by the starting weight of the compound to provide the fat content (of the compound).

In embodiments, the cheese extract may further comprise polypeptides. Especially, the cheese extract may comprise (on a dry weight basis) > 30 wt% polypeptide, such as > 40 wt%, especially > 45 wt%, like > 50 wt%. Additionally or alternatively, in embodiments, on a dry weight basis, the cheese extract may comprise < 95 wt%, such as < 90 wt%, especially < 85 wt%, like < 80 wt%, polypeptide. Hence, on a dry weight basis, the cheese extract may comprise 30-95 wt%, such as 40-90 wt%, especially 45-85 wt%, like 50-80 wt%, polypeptide. The polypeptide content of a compound may especially be determined using the Total Nitrogen (TN) content of the compound. The Total Nitrogen content may, for instance, be measured using kit LCK 238 as available from HACH LANGE GMBH, Germany, following the default protocol, and the value may be multiplied by 6.25 to obtain the polypeptide content. In embodiments, the cheese extract may (further) comprise (partially) hydrolyzed milk protein. Hydrolyzed milk protein herein refers to the products obtained after (enzymatic) hydrolysis of milk proteins (e.g. casein) by enzymes found (and/or added) in e.g. the cheesemaking process, and may comprise a combination of shorter-chain peptides (compared to the parent protein) and free amino acids. Hydrolyzed milk protein may be identified by identifying the peptides found in a substance (such as the cheese extract) and comparing the found peptides to reference peptides of milk proteins, such as e.g. casein. For instance, the peptides may be identified through (a) peptide sequencing, (b) using affinity probes (e.g., antibodies), or (c) using mass spectroscopy. Such methods are known to the person skilled in the art. The cheese extract may in embodiments have a relatively low carbohydrate content. Especially, the cheese extract may comprise (on a dry weight basis) < 10 wt% carbohydrate, such as < 7.5 wt%, especially < 5 wt%. Further, the cheese extract may comprise (on a dry weight basis) < 4 wt% carbohydrate, such as < 2.5 wt%, especially < 1.5 wt%. Additionally or alternatively, the cheese extract may comprise (on a dry weight basis) > 0.1 wt% carbohydrate, such as > 0.5 wt%, especially > 0.75 wt%. In embodiments, the carbohydrate may comprise lactose, such as in a concentration selected from the range of 10-90%, especially from the range of 25-75%. Hence, in embodiments, the cheese extract may comprise lactose. Especially, the cheese extract may comprise (on a dry weight basis) < 4 wt% lactose, such as < 3 wt%, especially < 2 wt%. Further, the cheese extract may comprise (on a dry weight basis) < 1.5 wt%, such as < 1 wt%, especially < 0.5 wt%, lactose. Additionally or alternatively, the cheese extract may comprise (on a dry weight basis) > 0.01 wt%, such as > 0.05 wt%, especially > 0.01 wt%, lactose.

In embodiments, during the cheesemaking process, such as especially during fermentation, lactose (and/or other carbohydrates) may be converted into organic acids by bacteria added to the milk. Hence, in embodiments, the cheese extract may comprise an organic acid. Especially, on a dry weight basis, the cheese extract may comprise > 1 wt%, such as > 2 wt%, especially > 4 wt%, like > 5 wt%, organic acid. Additionally or alternatively, in embodiments, the cheese extract may comprise on a dry weight basis < 35 wt%, such as < 30 wt%, especially < 25 wt%, like < 20 wt%, more especially < 15 wt%, organic acid. Hence, on a dry weight basis, the cheese extract may comprise 1-35 wt%, such as 2-30 wt%, especially 4- 25 wt%, like 5-20 wt%, more especially 5-15 wt%, organic acid. In embodiments, the organic acid may be present in the cheese extract as a free anion and/or as a salt. Further, in embodiments, the organic acid (in the cheese extract) may comprise one or more organic acids selected from the group comprising acetic acid, lactic acid, butyric acid, and propionic acid, especially one or more of butyric acid and propionic acid. Especially, in embodiments, the organic acid may comprise propionic acid in a concentration of < 15 wt% (compared to the total weight of the organic acid), such as in a concentration of < 10 wt%, especially in a concentration of < 8 wt%, though larger concentrations are herein not excluded. Further, the organic acid may comprise propionic acid in a concentration of > 1 wt%, such as in a concentration of > 2 wt%, especially in a concentration of > 5 wt%. Additionally or alternatively, the organic acid (of the cheese extract) may comprise butyric acid in a concentration of < 20 wt% (compared to the total weight of the organic acid), such as in a concentration of < 15 wt%, especially in a concentration of < 10 wt%, though larger concentrations are herein not excluded. Further, the organic acid (of the cheese extract) may comprise butyric acid in a concentration of > 2 wt%, such as in a concentration of > 5 wt%, especially in a concentration of > 7.5 wt%.

In a further aspect, the invention may provide a computer program product comprising instructions for execution on a control system functionally coupled to a system, wherein the instructions, when executed by the control system, cause the system to carry out the method of the invention. In a further aspect, the invention may provide a data carrier, carrying thereupon program instructions which, when executed by a control system functionally coupled to a system, cause the system to carry out the method of the invention.

Especially, the invention may provide a computer program product comprising instructions for execution on a control system functionally coupled to the system of the invention (as defined above), wherein the instructions, when executed by the control system, cause the system to carry out the method of the invention. In a further aspect, the invention may provide a data carrier, carrying thereupon program instructions which, when executed by a control system functionally coupled to the system of the invention (as defined above), cause the system to carry out the method of the invention.

In yet a further aspect, the invention (thus) provides a software product, which, when running on a computer is capable of bringing about (one or more embodiments of) the method as described herein.

The embodiments described herein are not limited to a single aspect of the invention. For example, an embodiment describing the method may, for example, further relate to the system, especially to an operational mode of the system, or especially to the control system. Similarly, an embodiment of the system describing an operation of the system may further relate to embodiments of the method. In particular, an embodiment of the method describing an operation (of the system) may indicate that the system may, in embodiments, be configured for and/or be suitable for the operation.

BRIEF DESCRIPTION OF THE DRAWINGS

Embodiments of the invention will now be described, by way of example only, with reference to the accompanying schematic drawings in which corresponding reference symbols indicate corresponding parts, and in which: Fig. 1 schematically depicts embodiments of the method of the invention. Fig. 2 schematically depicts embodiments of the system of the invention. The schematic drawings are not necessarily on scale.

DETAILED DESCRIPTION OF THE EMBODIMENTS Fig. 1 schematically depicts an embodiment of the method of the invention. Specifically, Fig. 1 depicts a method for converting a solid mixture 200 into a non-dissolved part 310, a precipitated part 320, and a supernatant part 330. The solid mixture 200 may comprise a first compound 110, a second compound 120, and a third compound 130. In the depicted embodiment, the method comprises an extraction stage 10, a first separation stage 20, a precipitation stage 30, a first compound recovery stage 41, a second compound recovery stage 42, a third compound recovery stage 43, and an organic extraction material recovery stage 50.

In certain embodiments, the solid mixture 200 may comprise one or more of plant biomass, fungal biomass, microbial biomass, animal biomass, and a food product. In embodiments, the solid mixture may comprise 5 - 85 wt% water 800. On dry weight basis the solid mixture may comprise: 0.1 - 55 wt% free amino acid, 05 - 65 wt% polypeptide, 0 - 55 wt% fat, 0 -40 wt% soluble fiber, 0 - 75 wt% insoluble fiber, 0.1 - 15 wt% mono-/disaccharide, 0 - 20 wt% organic acid, and 0 - 50 wt% salt.

The extraction stage 10 may comprise dissolving at least part of the solid mixture 200 in an extraction liquid 400, thereby providing a liquid mixture 500 and the non-dissolved part 310. The extraction liquid 400 may comprises (a) an organic extraction material 410 selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of an anion 420, especially OH', (c) at least 0.01 M of a cation 430 selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH⁴⁺, and (d) water 700.

The extraction stage 10 may comprises (a) an extraction duration selected from the range of 5 - 30 minutes, and (b) controlling an extraction temperature of the extraction liquid 400 at a temperature selected from the range of 15 - 100 °C.

Further, the organic extraction material 410 may be selected from the group consisting of acetone, methanol, ethanol, and ethylene glycol. The cation may especially comprise Na⁺. The extraction liquid 400 may comprise 20-90 vol.% of the organic extraction material 410 and 10-80 vol.% of water 700.

A first separation stage 20 may follow the extraction stage 10. The first separation stage 20 may comprise separating the liquid mixture 500 from the non-dissolved part 310.

A precipitation stage 30 may follow the first separation stage 30. The precipitation stage 30 may comprise providing an acid 600, especially CO2610, to the liquid mixture 500. Thereby, the precipitated part 320 and the supernatant part 330 may be provided.

In the depicted embodiment, the liquid mixture 500 comprises a second compound 120 that is electrically neutral in the liquid mixture 500 at a second OH' concentration CN, for example at an OH' concentration of 10'⁷ M. The precipitation stage 30 may comprise providing the CO2 610 to the liquid mixture 500 to reduce the OH' concentration from a first OH' concentration CE to a value close to CN, such as to a value selected from the range of 0.01 *10'⁷ M - 100* 10'⁷ M for above example, such that the second compound 120 precipitates. The precipitated part 320 thereby comprises the second compound 120.

In further embodiments such as depicted in Fig. 1, the precipitation stage 30 may comprise providing the CO2 610 to the liquid mixture 500 such that the CO2 610 and the cation 430 form a carbonate 440. The carbonate 440 may precipitate and the precipitated part 320 thereby comprises the carbonate 440.

A first compound recovery stage 41 may follow the first separation stage 20. The first compound recovery stage 41 may comprise separating or enriching a first compound 110 from the non-dissolved part 310, especially separating it from the components of the extraction liquid 400. The first compound 110 may especially be selected from the group comprising a free amino acid, a polypeptide, a fat, a soluble fiber, and an insoluble fiber.

A second compound recovery stage 42 may follow the precipitation stage 30. The second compound recovery stage 42 may comprise separating or enriching a second compound 120 from the precipitated part 320, especially separating it from the components of the extraction liquid 400. The second compound 120 may especially be selected from the group comprising a free amino acid, a polypeptide, and a salt.

A third compound recovery stage 43 may follow the precipitation stage 30. The third compound recovery stage 43 may comprise separating the third compound 130 from the supernatant part 330 or enriching the third compound 130 in the supernatant part 330, especially separating it from the components of the extraction liquid 400. The third compound 130 may especially be selected from the group comprising a free amino acid, polypeptide, soluble fiber, insoluble fiber, organic acid, and salt.

An organic extraction material recovery stage 50 may follow the precipitation stage 30. The organic extraction material recovery stage 50 may comprise subjecting the supernatant part 330 to an organic extraction material recovery step, especially distillation, to separate at least part of the organic extraction material 410 from the supernatant part 330.

In certain embodiments, the method may further comprise subjecting the liquid mixture 500 to a liquid removal step, the liquid removal step comprising separating at least part of the organic extraction material 410 or water 700 from the liquid mixture 500.

In further embodiments, one or more of a plurality of parts 300 comprising the non-dissolved part 310, the precipitated part 320 and the supernatant part 330 may be subjected to a purification step. The purification step may comprising separating at least part of the organic extraction material 410, OH' 420, the cation 430, or water 700 from one or more of the plurality of parts 300.

Fig. 2 schematically depicts an embodiment of the system 2000 for converting a solid mixture 200 into a non-dissolved part 310, a precipitated part 320 and a supernatant part 330. In the depicted embodiment, the system 2000 comprises an inlet 2100, a liquid supply 2200, an extraction unit 2300, a precipitation unit 2400, and an acid supply 2500. In further embodiments, the system 2000 may comprise a control system 3000 configured to control the system 2000, especially one or more of the inlet 2100, the liquid supply 2200, the extraction unit 2300, the precipitation unit 2400, and the acid supply 2500.

In embodiments, the system 2000, especially the control system 3000, may have an operational mode. The operational mode may comprise executing the method of the invention in continuous operation, batch operation, or countercurrent operation.

In embodiments, in the operational mode, the inlet 2100 may (be configured to) provide the solid mixture 200 to the extraction unit 2300. The liquid supply may (be configured to) provide the extraction liquid 400 to the extraction unit 2300. The extraction liquid 400 may comprise (a) an organic extraction material 410 selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of an anion 420 comprising OH', (c) at least 0.01 M of a cation 430 selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH⁴⁺, and (d) water 700.

In further embodiments, in the operational mode, the extraction unit 2300 may (be configured to) dissolve at least part of the solid mixture 200 into the extraction liquid 400. Thereby it may provide a liquid mixture 500 and a non-dissolved part 310. The liquid mixture 500 may be provided to the precipitation unit 2400. The non-dissolved part 310 may provide a first of the plurality of parts 300 comprising a first compound 110.

In further embodiments, in the operational mode, the acid supply 2500 may be configured to provide the acid 600 to the precipitation unit 2400. Especially, the acid 600 may be provided to the liquid mixture 500 hosted in the precipitation unit 2400. Thereby, a precipitated part 320 and a supernatant part 330 may be provided. The precipitated part 320 may provide a second of the plurality of parts 300 comprising a second compound 120. The supernatant part 330 may provide a third of the plurality of parts 300 comprising a third compound 130.

In certain embodiments, in the operational mode, the precipitation unit 2400 may subject the supernatant part 330 to an organic extraction material recovery step comprising distillation to provide an organic extraction material 410. Experiments

Embodiments of the method of the invention were experimentally evaluated with different solid mixtures 200, extraction liquids 400 and conditions during the stages. These experiments are briefly described herein.

Unless specified otherwise, the experiments were performed using the following conditions.

Chemicals - Several chemicals were used to execute the method of the invention in the experiments, such as ethanol absolute (Merck), ethanol 70% vol (VWR) and ethanol 96% vol (VWR), sodium hydroxide (98% SigmaAldrich), L-leucine (>98% SigmaAldrich), sodium L-glutamate monohydrate (>99% Scharlau). Solutions were prepared by diluting the chemicals with Milli-Q water and, afterwards, all solutions were filtrated using 0.2 pm filters.

Carbon dioxide - CO2 (>99.7 %) was provided by Linde in a pressurized bottle.

Analysis - Amino acids analysis was done using in a Agilent 1290 Infinity II UHPLC system, equipped with a UV/Visible Detector. Separation occurred on a ACQUITY BEH Cl 8 1.7pM column. The free amino acid content of the different samples was analyzed in UHPLC following the protocol described by Meussen et al., “A Fast and Accurate UPLC Method for Analysis of Proteinogenic Amino Acids”, Food Analytical Methods, 2014, which is hereby herein incorporated by reference. Total Nitrogen content was measured using the Hach kit LCK 238, the value was multiplied by 6.25 to obtain the polypeptide content using TN kit. The polypeptide extraction yield (compared to the total amount of polypeptide in the solid mixture 200) during the extraction stage 10, the polypeptide precipitation yield (compared to the total amount of polypeptide in the liquid mixture 500) during the precipitation stage 30, and the total amount of polypeptide recovered (compared to the total amount of polypeptide in the solid mixture 200) during the extraction and precipitation stages were calculated based on polypeptide content determination. Fat content was determined by a gravimetric method in which the solid samples were extracted with hexane.

Experiment 1 - Solubilization of pure amino acids and precipitation with CO2.

A solid mixture 200 comprising a defined amount of free amino acid was suspended in a vessel with 15 mL of extraction liquid 400. The extraction liquid 400 consisted of a mixture of ethanol (as organic extraction material 410), NaOH (to provide the anion 420 OH' and the cation 430 Na⁺), and water 700. After an extraction stage 10 of 30 minutes - 1 hour, the non-dissolved part 310 and the liquid mixture 500 were separated. The free amino acid composition of the liquid mixture 500 was determined using UHPLC. Table 1. Characteristics and components of the extraction liquid 400 and solid mixture 200

The liquid mixture 500 was then subjected to a precipitation stage 30 comprising sparging the liquid mixture 500 with CO2 610 as the acid 600 until (most of) the anion 420 OFF was neutralized to provide a precipitated part 320 and a supernatant part 330. The amino acid composition of the non-dissolved part 310, precipitated part 320 and the supernatant part 330 was determined using UHPLC.

Table 2. Free amino acid recovery (in wt%) in the non-dissolved part 310, the precipitated part 320 and the supernatant part 330.

Table 3. Specific free amino acid recovery (in wt.%) in the non-dissolved part 310, the precipitated part 320 and the supernatant part 330.

Different free amino acids seem to be enriched in the plurality of parts 300 in a specific amino acid or amino acids group by changing the organic extraction material 410 concentration and the anion 420 concentration of the extraction liquid 400. The enrichment of free amino acid in the plurality of different parts 300 may thereby be tuned relatively easily by the skilled person. The higher the anion 420 concentration, the higher the amount of free amino acid that seems to be dissolved. The higher the organic extraction material 410 concentration, the lower the amount of free amino acid that seems to be dissolved. For example, tyrosine seems to (at least partly) stay in the non-dissolved part 310 unless a high NaOH concentration is used, as tyrosine may be one of the most insoluble amino acids. If tyrosine is dissolved, it may be concentrated in the precipitated part 320 (due to the low solubility in the absence of the anion 420). Glutamate seems to dissolve for middle-low ethanol concentrations (<90%) and may just be partially precipitated afterwards, due to the high solubility of glutamate in water. Experiment 2 - Extraction of umami flavor from cheese.

A solid mixture 200 comprising biomass obtained from cheese rinds was cut and grinded using a kitchen blender. The solid mixture 200 was subjected to an extraction stage 10 with extraction liquid 400 comprising 70% ethanol as the organic extraction material 410 with 0.1 M sodium hydroxide (NaOH) to provide the anion 420 and the cation 430 with a solid mixture 200 load of 70 g/L. The extraction stage 10 was performed at room temperature with stirring for 1.5 hours.

After extraction, the non-dissolved part 310 and liquid mixture 500 were separated by filtration. Both parts were neutralized during a precipitation stage 30 by sparging with an acid 600 comprising CO2 610. After neutralization, part of the liquid mixture 500 precipitated, thereby providing the precipitated part 320 and the supernatant part 330. The three parts were analyzed for total nitrogen content (to estimate polypeptide content), free amino acid content, and fat content.

The extraction efficiency of the extraction stage 10 was determined to be 64% on dry weight basis, i.e., 64% of the total mass of the solid mixture 200 on dry weight basis was extracted into the liquid mixture 500. Further, the liquid mixture 500 comprised 91% of the total polypeptide content on dry weight basis of the solid mixture 200. The precipitation efficiency of the precipitation stage 30 was determined to be 48% on dry weight basis, i.e., on dry weight basis 48% of the total mass originally from the solid mixture 200 was precipitated from the liquid mixture 500 into the precipitated part 320. Further, this comprised 64% of the total polypeptide content on dry weight basis comprised by the liquid mixture 500.

Table 3. Composition (in wt.%) of free amino acid, polypeptide, and fat of the solid mixture 200, precipitated part 320 and supernatant part 330 on dry weight basis.

The extracted parts 320,330 seem low in fat content relative to the solid mixture 200. The supernatant part 330 seems to comprise most of the free amino acid. The polypeptide content seems similar between the precipitated part 320 and the supernatant part 330.

The non-dissolved part 310 was mainly composed of fat (74 wt%). The supernatant part 330 contained 4-10 wt% organic acid. The composition of the organic acid is provided in Table 4, wherein the wt% indicates a weight percentage relative to the total weight of the organic acid. Table 4. composition (in wt%) of organic acid in the supernatant part 330, on dry weight basis.

Experiment 3 - Solubilization of pure soy protein and precipitation with CO2.

A solid mixture 200 comprising a known amount of commercial soy protein isolate was suspended in 15 mL of extraction liquid 400 with different components. The organic extraction material consisted of a mixture of ethanol EtOH (as the organic extraction material 410), NaOH (to provide the anion 420 and the cation 430), and water 700. Different ratios of solid/liquid, temperatures, and durations during the extraction stage 10 were evaluated.

Table 4. Extraction liquid 400 components and extraction stage 10 conditions.

After the extraction stage 10, the non-dissolved part 310 was separated, and the polypeptide content of the liquid mixture 500 was determined. The liquid mixture 500 was then sparged with CO2 610 as the acid 600 until (most of) the OH' as the anion 420 was neutralized to provide a precipitated part 320 and supernatant part 330. The polypeptide content of the supernatant part 330 was determined. Thereby the polypeptide extraction yield and the polypeptide precipitation yield were calculated.

Table 5. Polypeptide extraction yield and polypeptide precipitation yield

Different polypeptide extraction yields and polypeptide precipitation yields were obtained with varying concentrations of solid mixture 200, organic extraction material 410, anion 420 and cation 430 in the extraction liquid 400, and varying conditions of duration and temperature of the extraction stage 10. Higher organic extraction material 410 concentrations seem to decrease polypeptide extraction yield and increase polypeptide precipitation yield. Higher extraction temperature seems to increase polypeptide extraction yields, but decrease polypeptide precipitation yields. The effect of the tested differences in extraction duration (from 30 min to 1 h) seems limited.

Experiment 4 - Hydrolysis of pure soy protein, solubilization and precipitation with CO2 610.

A solid mixture 200 comprising a known amount of commercial soy protein isolate was subjected to a solid mixture preparation step comprising hydrolysis. To achieve this, the solid mixture 200 was exposed to 120 °C for a pre-determined amount of time (see Table 6). During the duration of this pre-treatment, the sample was agitated (via magnetic stirrer) for examples 4.1-4.6. Afterwards, the solid mixture 200 was subjected to an extraction stage 10 with varying extraction liquid 400 components (comprising at least ethanol as organic extraction material 410 and water 700) for one hour to provide a liquid mixture 500 and a non- dissolved part 310.

Table 6. Hydrolysis as solid mixture preparation step, and extraction stage conditions

After the extraction stage 10, the non-dissolved part 310 was separated, and the polypeptide content of the liquid mixture 500 was determined using TN kit. The liquid mixture 500 was then subjected to a precipitation stage 30 comprising sparging CO2 610 as the acid 600 to provide a precipitated part 320 and a supernatant part 330. The polypeptide content of the supernatant part 330 was determined. Thereby the polypeptide extraction yield and the polypeptide precipitation yield were calculated.

Table 7. Polypeptide extraction yield and polypeptide precipitation yield

Different polypeptide extraction yields and polypeptide precipitation yields were obtained with varying conditions of hydrolysis and water content. An increase in the hydrolysis duration seems to result in increased (polypeptide) solubilization during the extraction stage and increased (polypeptide) precipitation during the precipitation stage. Thereby the solid mixture preparation step may facilitate obtaining (different) compounds of interest in the (different) parts of the plurality of parts 300.

Experiment 5 - Solubilization of brewers spent grain protein and precipitation with CO2 610 and HC1

Solid mixture 200 obtained from brewer’s spent grain from beer production was suspended in extraction liquid 400 with varying compositions. The solid mixture 200 had a polypeptide content of 25 wt% dry weight basis and a water content of 77%. The extraction liquid 400 consisted of a mixture of ethanol EtOH as organic extraction material 410, NaOH to provide the anion 420 and the cation 430, and water 700. Differences in solid/liquid ratio were evaluated. The extraction duration was 24 hours at an extraction temperature of room temperature. After the extraction stage 10 the non-dissolved part 310 was separated.

During the precipitation stage 30, the OH- was partially or totally neutralized with HC1 or CO2 610 to provide a precipitated part 320 and a supernatant part 330.

Table 8. Extraction stage and precipitation stage components

The polypeptide content of the liquid mixture 500 after the extraction stage 10 and of the supernatant part 330 after the precipitation stage 30 was determined. Thereby the polypeptide extraction yield, the polypeptide precipitation yield and the total amount of polypeptide recovered were calculated. The method may facilitate enriching the non-dissolved part 310 in soluble fiber and non-soluble fiber, while further enriching the precipitated part 320 in polypeptide, with organic acids going to the supernatant part 330.

Table 9. Polypeptide extraction yield, polypeptide precipitation yield, and total polypeptide recovered.

Different polypeptide extraction yields, polypeptide precipitation yields, and total polypeptide recovery were obtained with varying concentrations of solid mixture 200, organic extraction material 410, anion 420 and cation 430 in the extraction liquid 400, and varying acid 600 used in the precipitation stage 30. Higher anion 420 concentrations seem to increase polypeptide extraction yield but decrease polypeptide precipitation yield. Due to the high water content of brewer’s spent grain, the organic extraction material 410 concentration had little effect on polypeptide extraction yield.

Experiment 6 - Solubilization of polypeptide from oil press cakes and precipitation with CO2 610.

Solid mixture 200 obtained from press cake from different vegetable oil production processes were subjected to an extraction stage 10 for one hour. The solid mixture 200 was suspended in extraction liquid 400 at a ratio of 8.7 g press cake per 100 g of extraction liquid 400. The extraction liquid 400 consisted of 30 wt% ethanol EtOH to provide the organic extraction material 410, 0.1 M of NaOH to provide the anion 420 and the cation 430, and water 700.

Table 10. Solid mixture 200 from vegetable oil press cake characteristics

Table 11. Polypeptide extraction yield, polypeptide precipitation yield, and total polypeptide recovered.

The extraction of polypeptide from rapeseed and flaxseed seems higher than from sunflower, despite similar initial polypeptide concentration. The polypeptide precipitation yield seems independent of the polypeptide extraction yield. The polypeptide precipitation yield seems dependent on the polypeptide concentration of the solid mixture 200.

The term “plurality” refers to two or more. Furthermore, the terms “a plurality of’ and “a number of’ may be used interchangeably. The terms “substantially” or “essentially” herein, and similar terms, will be understood by the person skilled in the art. The terms “substantially” or “essentially” may also include embodiments with “entirely”, “completely”, “all”, etc. Hence, in embodiments the adjective substantially or essentially may also be removed. Where applicable, the term “substantially” or the term “essentially” may also relate to 90% or higher, such as 95% or higher, especially 99% or higher, even more especially 99.5% or higher, including 100%. Moreover, the terms ’’about” and “approximately” may also relate to 90% or higher, such as 95% or higher, especially 99% or higher, even more especially 99.5% or higher, including 100%. For numerical values it is to be understood that the terms “substantially”, “essentially”, “about”, and “approximately” may also relate to the range of 90% - 110%, such as 95%-105%, especially 99%-l 01% of the values(s) it refers to.

The term “comprise” also includes embodiments wherein the term “comprises” means “consists of’. The term “and/or” especially relates to one or more of the items mentioned before and after “and/or”. For instance, a phrase “item 1 and/or item 2” and similar phrases may relate to one or more of item 1 and item 2. The term "comprising" may in an embodiment refer to "consisting of but may in another embodiment also refer to "containing at least the defined species and optionally one or more other species".

Furthermore, the terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.

The devices, apparatus, or systems may herein amongst others be described during operation. As will be clear to the person skilled in the art, the invention is not limited to methods of operation, or devices, apparatus, or systems in operation.

The term “further embodiment” and similar terms may refer to an embodiment comprising the features of the previously discussed embodiment, but may also refer to an alternative embodiment. It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and that those skilled in the art will be able to design many alternative embodiments without departing from the scope of the appended claims. In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim.

Use of the verb "to comprise" and its conjugations does not exclude the presence of elements or steps other than those stated in a claim. Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, “include”, “including”, “contain”, “containing” and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. The article "a" or "an" preceding an element does not exclude the presence of a plurality of such elements.

The invention may be implemented by means of hardware comprising several distinct elements, and by means of a suitably programmed computer. In a device claim, or an apparatus claim, or a system claim, enumerating several means, several of these means may be embodied by one and the same item of hardware. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage.

The invention also provides a control system that may control the device, apparatus, or system, or that may execute the herein described method or process. Yet further, the invention also provides a computer program product, when running on a computer which is functionally coupled to or comprised by the device, apparatus, or system, controls one or more controllable elements of such device, apparatus, or system.

The invention further applies to a device, apparatus, or system comprising one or more of the characterizing features described in the description and/or shown in the attached drawings. The invention further pertains to a method or process comprising one or more of the characterizing features described in the description and/or shown in the attached drawings. Moreover, if a method or an embodiment of the method is described being executed in a device, apparatus, or system, it will be understood that the device, apparatus, or system is suitable for or configured for (executing) the method or the embodiment of the method, respectively.

The various aspects discussed in this patent can be combined in order to provide additional advantages. Further, the person skilled in the art will understand that embodiments can be combined, and that also more than two embodiments can be combined. Furthermore, some of the features can form the basis for one or more divisional applications.

Claims

CLAIMS:

1. A method for converting a solid mixture (200) into a non-dissolved part (310), a precipitated part (320) and a supernatant part (330), the method comprising: an extraction stage (10) comprising dissolving at least part of the solid mixture (200) in an extraction liquid (400), thereby providing a liquid mixture (500) and the nondissolved part (310), wherein the extraction liquid (400) comprises (a) an organic extraction material (410) selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of OH', (c) at least 0.01 M of a cation (430) selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH⁴⁺, and (d) water (700); a first separation stage (20) comprising separating the liquid mixture (500) from the non-dissolved part (310); and a precipitation stage (30) comprising providing CO2 (610) to the liquid mixture (500), thereby providing the precipitated part (320) and the supernatant part (330).

2. The method according to claim 1, wherein the solid mixture (200) comprises one or more of plant biomass, fungal biomass, microbial biomass, animal biomass, and a food product; wherein the solid mixture (200) comprises 5 - 85 wt% water (700); and wherein on dry weight basis the solid mixture (200) comprises:

0.5 - 25 wt% free amino acid;

15 - 55 wt% polypeptide;

0 - 45 wt% fat;

0 - 30 wt% soluble fiber;

0 - 50 wt% insoluble fiber; and

1 - 8 wt% mono-/disaccharide.

3. The method according to any one of the preceding claims, wherein the extraction stage (10) comprises controlling an extraction temperature of the liquid mixture (500) at a temperature selected from the range of 15 - 100 °C.

4. The method according to any one of the preceding claims, wherein the precipitation stage (30) comprises providing the CO2 (610) to the liquid mixture (500) such that the CO2 (610) and the cation (430) form a carbonate (440), wherein the carbonate (440) precipitates, wherein the precipitation stage (30) comprises providing the CO2 (610) (a) at a precipitation temperature selected from the range of 5 - 40° C, (b) at a precipitation pressure selected from 0.1 - 10 bar, and (c) for a precipitation duration selected from the range of 5 - 30 min.

5. The method according to any one of the preceding claims, wherein the method further comprises a first compound recovery stage (41), the first compound recovery stage (41) comprising separating a first compound (110) from the non-dissolved part (310), wherein the first compound (110) is selected from the group comprising a free amino acid, a polypeptide, a fat, a soluble fiber, an oligosaccharide, a polysaccharide, and an insoluble fiber.

6. The method according to any one of the preceding claims, wherein: the liquid mixture (500) comprises a second compound (120), wherein the second compound (120) is electrically neutral in the liquid mixture (500) at a second OH' concentration CN, and wherein the precipitation stage (30) comprises providing the CO2 (610) to reduce the OH' concentration in the liquid mixture (500) to a value selected from the range of 0.1 *CN - 1.5*CN such that the second compound (120) precipitates.

7. The method according to any one of the preceding claims, wherein the method further comprises a second compound recovery stage (42), the second compound recovery stage

(42) comprising separating the second compound (120) from the precipitated part (320), wherein the second compound (120) is selected from the group comprising free amino acid, polypeptide, and a salt.

8. The method according to any one of the preceding claims, wherein the method further comprises a third compound recovery stage (43), the third compound recovery stage

(43) comprising separating a third compound (130) from the supernatant part (330), wherein the third compound (130) is selected from the group comprising free amino acid, polypeptide, soluble fiber, a monosaccharide, an organic acid, and a salt.

9. The method according to any one of the preceding claims, wherein the method further comprises an organic extraction material recovery stage (50) following the precipitation stage (30), the organic extraction material recovery stage (50) comprising subjecting the supernatant part (330) to distillation to separate at least part of the organic extraction material

(410) from the supernatant part (330).

10. The method according to any one of the preceding claims, wherein the organic extraction material (410) is selected from the group consisting of acetone, methanol, ethanol, and ethylene glycol, wherein the cation (430) comprises Na⁺, wherein the extraction liquid (400) comprises 20-99 vol.% of the organic extraction material (410) and 1-80 vol.% of water (700).

11. The method according to any one of the preceding claims, wherein the method further comprises subjecting the liquid mixture (500) to a liquid removal step, the liquid removal step comprising separating the organic extraction material (410) or water (700) from the liquid mixture (500).

12. The method according to any one of the preceding claims, wherein one or more of a plurality of parts (300) comprising the non-dissolved part (310), the precipitated part (320) and the supernatant part (330) is subjected to a purification step, the purification step comprising separating the organic extraction material (410), OH', the cation (430), or water (700) from one or more of the plurality of parts (300).

13. A system (2000) for converting a solid mixture (200) into a non-dissolved part (310), a precipitated part (320) and a supernatant part (330), the system (2000) comprising an inlet (2100), a liquid supply (2200), an extraction unit (2300), a precipitation unit (2400), and an acid supply (2500), and wherein the system (2000) has an operational mode, wherein: the inlet (2100) is configured to provide the solid mixture (200) to the extraction unit (2300); the liquid supply (2200) is configured to provide an extraction liquid (400) to the extraction unit (2300); wherein the extraction liquid (400) comprises (a) an organic extraction material (410) selected from the group consisting of acetone and alcohols, (b) at least 0.01 M of OH', (c) at least 0.01 M of a cation (430) selected from the group comprising Na⁺, K⁺, Ca²⁺, Mg²⁺ and NH⁴⁺, and (d) water (700); the extraction unit (2300) is configured to dissolve at least part of the solid mixture (200) and (i) to provide a liquid mixture (500) to the precipitation unit (2400) and (ii) to provide the non-dissolved part (310); the acid supply (2500) is configured to supply an acid (600) to the liquid mixture (500) hosted in the precipitation unit (2400) thereby forming a precipitated part (320) and a supernatant part (330); the precipitation unit (2400) is configured to provide the precipitated part (320) and the supernatant part (330).

14. A computer program product comprising instructions for execution on a control system (3000) functionally coupled to a system (2000), wherein the instructions, when executed by the control system (3000), cause the system (2000) to carry out the method according to any one of the preceding claims 1-12.

15. A data carrier, carrying thereupon program instructions which, when executed by a control system (3000) functionally coupled to the system (2000) according to claim 13, cause the system (2000) to carry out the method according to any one of the preceding claims 1-12.

16. A cheese extract obtainable using the method according to any one of claims 1- 12, wherein the cheese extract comprises at least part of the supernatant part (330), wherein the cheese extract has a solubility in water selected from the range of > 90%, and wherein on a dry weight basis the cheese extract comprises:

> 1 wt% glutamic acid;

1-10 wt% fat;

45-85 wt% polypeptide;

< 5 wt% carbohydrate, wherein the cheese extract comprises < 2 wt% of lactose; and

5-20 wt% organic acid, wherein the organic acid comprises one or more of acetic acid, lactic acid, butyric acid, and propionic acid.