Methods for recovering mevalonic acid or salts or lactones thereof from aqueous solutions using water solvent crystallization and compositions thereof

Abstract

To provide methods for producing crystalline forms of mevalonolactone from an aqueous solution comprising a salt of mevalonic acid, and compositions comprising the mevalonolactone produced.SOLUTION: Methods for producing mevalonolactone from an aqueous solution comprising a salt of mevalonic acid (X-MVA) comprise subjecting the aqueous solution comprising the X-MVA to cation exchange thereby converting the aqueous solution comprising the X-MVA to an aqueous solution comprising mevalonolactone (MVL). Methods are also provided for producing mevalonolactone monohydrate crystals.SELECTED DRAWING: Figure 1B

Description

[Technical field]

[0001] This application claims the benefit of U.S. Provisional Application No. 62 / 892,760, filed August 28, 2019, and U.S. Provisional Application No. 63 / 012,492, filed April 20, 2020, each of which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to the field of organic acid separation techniques, and more particularly to a method for recovering organic acids and their salts or lactones from aqueous solutions using water solvent crystallization. [Background technology]

[0003] Known processes for isolating and purifying organic acids from solutions such as fermentation broths are highly complex and generally involve solvent extraction steps or solid phase absorption.

[0004] An important aspect for the industrial use of organic acids, such as carboxylic acids, produced by fermenting carbohydrate-containing substrates using various microorganisms is the cost-effectiveness and efficiency of removing and purifying the organic acids or their salts or lactones from such aqueous fermentation broths, which contain not only the organic acids or organic acid salts or lactones, but also residues of the substrate, such as sugars, proteins, lipids, and other trace organic and inorganic compounds, in addition to other organic acids, other fermentation by-products, microorganisms, and their components.

[0005] The classical method for purifying organic acids produced by microorganisms from fermentation broth involves protonating the target acid by lowering the pH of the broth and performing liquid / liquid extraction with a partially miscible organic solvent (Organic Laboratory Techniques, 3rd Edition pg. 49-67. Ralph Fessenden, Joan Fessenden, Patty Feist 2001, Brooks / Cole).

[0006] US2014 / 0371486 describes a method for purifying carboxylic acids from fermentation broth using solid-phase adsorption, which includes removing biomass and any solids present in the fermentation broth, fine-clarifying the biomass- and solid-free fermentation broth by nanofiltration, and removing carboxylic acids from the fine-clarified biomass- and solid-free fermentation broth by adsorption onto one or more solid phases bearing tertiary amino groups.KR20180070117 describes a method for producing mevalonolactone from biosynthesized mevalonic acid using phosphoric acid.

[0007] US Patent No. 5,034,105 describes a method for crystallizing succinic acid from an unsaturated solution of a succinic acid salt by subjecting the salt solution to electrodialysis to produce a supersaturated solution, and then crystallizing the supersaturated solution of succinic acid by adding to the solution an amount of acetic acid effective to promote crystallization of the succinic acid.

[0008] The disadvantage of these methods is that they provide additional substances to the process which should no longer be present in the desired product and which, if present in trace amounts in the desired product, may limit the quality and applicability of the product. The practical implementation of the methods may also involve considerable technical complexity and considerable energy consumption. Summary of the Invention [Problem to be solved by the invention]

[0009] Thus, it remains desirable to develop more efficient, reliable, environmentally friendly, and / or economically feasible processes for purifying and recovering organic acids and their salts and lactones, and for producing organic acids and their salts or lactones in crystalline form. [Means for solving the problem]

[0010] Compositions and methods are provided for producing crystalline forms of an organic acid or its salt or lactone from an aqueous solution. * Monohydrate (MVL * HO) and mevalonolactone * Monohydrate (MVL * A method for crystallizing HO is disclosed.

[0011] More specifically, in one aspect, the present specification provides a method for producing a crystalline form of a salt of mevalonic acid (also referred to as X-MVA) from an aqueous solution, the method comprising subjecting an aqueous solution containing the X-MVA to nanofiltration to produce a permeate, and crystallizing the X-MVA from the permeate by water solvent crystallization (FIG. 1A).

[0012] In another aspect, the present disclosure provides a method for producing water solubilized mevalonolactone from an aqueous solution, comprising: a) producing a crystalline form of a salt of mevalonic acid from an aqueous solution by subjecting an aqueous solution containing a salt of mevalonic acid (X-MVA) to nanofiltration to produce a permeate, and crystallizing the salt of mevalonic acid from the permeate by crystallization using water as a solvent to produce crystals of the salt of mevalonic acid; and b) producing a water solubilized salt of mevalonic acid by dissolving the crystals of (a) in water, and converting the water solubilized salt of mevalonic acid to water solubilized mevalonolactone by subjecting the solution to cation exchange (FIG. 1B).

[0013] In yet another aspect, the present disclosure provides a method for producing mevalonolactone from an aqueous solution containing a salt of mevalonic acid (X-MVA), comprising converting the aqueous solution containing a salt of mevalonate to an aqueous solution containing mevalonolactone (MVL) by subjecting the aqueous solution containing a salt of mevalonate to cation exchange; optionally producing highly pure (>90% pure) MVL from the aqueous solution by concentrating the solution; and producing mevalonolactone from the concentrated solution.* Monohydrate (MVL * HO) and mevalonolactone * Monohydrate (MVL * and obtaining a high purity MVL solution by dissolving crystals of MVL (H2O) in water (Figure 2).

[0014] Mevalonolactone * Monohydrate (MVL * Also provided are compositions and methods for producing crystalline forms of HO.

[0015] Further embodiments of the disclosed methods and compositions are provided herein.

[0016] The present disclosure will become more fully understood from the following detailed description and the accompanying drawings, which form a part of this application. [Brief description of the drawings]

[0017] [Figure 1A] 1 shows a schematic diagram of a method for producing a crystalline form of a salt of mevalonic acid (X-MVA) (such as, but not limited to, Na-MVA) from an aqueous solution, comprising subjecting an aqueous solution containing the salt of mevalonic acid (Na-MVA) to nanofiltration to produce a permeate, and crystallizing the salt of mevalonic acid from the permeate by aqueous crystallization, which comprises evaporating the permeate to obtain a syrup containing concentrated X-MVA, and then cooling the syrup to produce crystals of the salt of mevalonic acid (such as Na-MVA crystals). [Figure 1B]FIG. 1 shows a schematic diagram of a method for producing water-solubilized mevalonolactone (MVL) from an aqueous solution, the method comprising dissolving crystals of a salt of mevalonic acid (such as Na-MVA crystals) obtained by the method described in FIG. 1A in water to produce a water-solubilized salt of mevalonic acid (X-MVA), converting the water-solubilized salt of mevalonic acid (X-MVA) to water-solubilized mevalonolactone (MVL) by subjecting the liquid to cation exchange, and optionally concentrating the water-solubilized mevalonolactone by evaporation. [Diagram 2] 1 shows a schematic diagram of a method for producing mevalonolactone from an aqueous solution containing a salt of mevalonic acid (X-MVA), comprising converting the aqueous solution containing a salt of mevalonate to an aqueous solution containing mevalonolactone (MVL) by subjecting the aqueous solution containing a salt of mevalonate to cation exchange, and optionally concentrating the solution to a syrup that can produce mevalonolactone* monohydrate crystals upon storage at low temperatures below about 23° C. The mevalonolactone* monohydrate crystals can optionally be further dissolved in water and concentrated to produce a highly concentrated and pure MVL liquid (>90% purity). [Diagram 3] This photograph shows the typical shape of mevalonolactone* monohydrate (MVL*H2O) crystals. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] The present invention provides compositions and methods for recovering organic acids and their salts and lactones from aqueous solutions using crystallization with water as the solvent. * Monohydrate (MVL * HO) and mevalonolactone * Monohydrate (MVL * A method for crystallizing HO is disclosed.

[0019] More particularly, in one aspect, it is an object of the present invention to provide a method for recovering a salt of mevalonate (X-MVA) from a solution containing it. In another aspect, it is an object of the present invention to recover a salt of mevalonate (X-MVA), mevalonolactone (MVL ... * Monohydrate (MVL * The present invention provides a method for recovering mevalonolactone (MVL) from a solution containing a salt of mevalonate, a carboxylic acid salt, HO, or a combination thereof. The entire process for recovering mevalonate salts and / or recovering mevalonolactone can preferably be carried out in an aqueous solution without the use of organic solvents.

[0020] The methods described herein can result in a highly pure (>90% pure) MVL product that has a water-like appearance with little or no color formation. Thus, the MVL produced by the methods described herein is of particular interest to the personal care industry, where the highly purified MVL product can be scaled up for large-scale production and is highly valuable as it is clear and colorless with little or no colored contaminants present.

[0021] This detailed description is intended to acquaint others skilled in the art to which the applicant's invention pertains with the principles and practical application thereof, thereby enabling them to adapt and apply the invention in its numerous forms, as may best suit the requirements of a particular use. This detailed description and its specific examples, while indicating specific embodiments, are given for illustrative purposes only. Therefore, the present specification is not limited to the described embodiments, and various modifications may be made thereto.

[0022] This document is organized into sections for ease of reading, and the reader will understand that statements in one section may be applicable to other sections, and thus, the headings used for the different sections of this disclosure should not be construed as limiting.

[0023] The headings provided herein are not intended to limit the various aspects or embodiments of the present compositions and methods, which can be understood by reference to the specification as a whole, and thus, the terms defined immediately below are more fully defined by reference to the specification as a whole.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the compositions and methods belong. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present compositions and methods, representative illustrative methods and materials are now described.

[0025] All publications and patents cited in this specification are herein incorporated by reference to the same extent as if each individual publication and patent was specifically and individually indicated to be incorporated by reference, and are also herein incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0026] definition The following definitions are used throughout the specification, examples and claims.

[0027] The terms "mevalonic acid (MVA)" or "(R)-mevalonic acid" are used interchangeably herein and have the chemical formula CH 12 It refers to (3R)-3,5-dihydroxy-3-methylpentanoic acid having the molar mass of 148.16 g / mol and having the structure O4. The carboxylate anion of mevalonic acid is known as mevalonate. The mevalonic acid described herein is the biologically active R enantiomer.

[0028] The terms "salt of mevalonate" or "mevalonate salt" or "X-MVA" are used interchangeably herein and refer to a salt of mevalonic acid where X is a cation and MVA is the carboxylate anion of mevalonic acid (hence, X-MVA). The salt of mevalonate can be selected from the group consisting of Na-mevalonate (Na-MVA), K-mevalonate (K-MVA), ammonium mevalonate (NH4-MVA), lithium mevalonate (Li-MVA), any other monovalent salt of mevalonic acid, or any combination thereof.

[0029] The terms "mevalonolactone" or "(R)-mevalonolactone" or "MVL" are used interchangeably herein and refer to (R)-3-hydroxy-3-methyl-δ-valerolactone.

[0030] Aqueous solutions as described herein include solutions containing both an organic acid and its lactone, which convert into each other toward equilibrium concentrations. For example, mevalonic acid and mevalonolactone tend to equilibrate with each other. In this description, a solution containing an organic acid certainly includes its lactone form, unless otherwise specified.

[0031] Mevalonolactone monohydrate, MVL * It is also called H2O.

[0032] SAC refers to strong acid cation exchange resin.

[0033] WAC refers to weak acid cation exchange resin.

[0034] DS refers to the dry matter content expressed in % by weight. Dry matter can be determined by Karl Fischer water titration.

[0035] RDS refers to the refractometric dry substance content and is expressed as a percentage by weight according to the correlation between the refractive index of an aqueous solution of a sugar and the DS.

[0036] IX or IEX refers to the ion exchange process.

[0037] BV / h refers to the volumetric flow rate through the ion exchange material contained in a column or operating unit. BV refers to the bed volume, which is the volume of ion exchange material in a specified ionic form contained in a column or operating unit.

[0038] Purity is determined by the composition (Na-MVA, MVL, MVL * The area percentage calculation procedure reports the area of ​​each peak in a chromatogram (such as an HPLC chromatogram) as a percentage of the total area of ​​all peaks. For example, mevalonolactone having a purity of at least 90% refers to the percentage of the peak area corresponding to MVL (90 in this case) relative to the total area of ​​the peaks (100) using chromatographic analysis. Purity can be measured using HPLC (Rezex ROA-Organic Acid H+ (8%) column).

[0039] HPLC refers to high performance liquid chromatography.

[0040] The purity of sodium mevalonate refers to the amount of sodium mevalonate, assuming that mevalonic acid, mevalonate, and mevalonolactone are all in the form of sodium mevalonate, divided by the total amount of dry matter.

[0041] The mevalonolactone purity refers to the total amount of mevalonolactone, assuming that mevalonic acid, mevalonate, and mevalonolactone are all in the form of mevalonolactone, divided by the total amount of dry matter.

[0042] Mevalonate yield refers to the amount of mevalonate in a fraction of interest (such as the permeate or centrifuge cake from nanofiltration) divided by the amount of mevalonate in the feed fraction (such as the nanofiltration feed or centrifuge feed), assuming that all mevalonic acid, mevalonate, and mevalonolactone are in the mevalonate form.

[0043] Mevalonolactone yield refers to the amount of mevalonolactone in the fraction of interest (cation exchange product or centrifuge cake) divided by the amount of mevalonolactone in the feed fraction (cation exchange feed or centrifuge feed, etc.), assuming that all mevalonic acid, mevalonate, and mevalonolactone are in the mevalonolactone form.

[0044] Color refers to color based on the International Commission for Uniform Process of Sugar Analysis ("ICUMSA") sugar color grading system.

[0045] DSC thermograms were measured using a Mettler Toledo DSC822e differential scanning calorimeter. The measurements were performed in 40 μL aluminum crucibles under a nitrogen flow rate of 80 mL / min. The temperature range was 0 to 50°C, and the heating rate was 2°C / min.

[0046] Optical rotations were measured in aqueous solutions with mevalonolactone concentrations of 2 g / 100 mL using an Anton Paar MCP 300 Sucromet at a temperature of 20° C. using a 100 mm cuvette and a 589 nm Na light source.

[0047] The abbreviations are as follows: "sec" means seconds, "min" means minutes, "h" or "hr" means hours, "d" means days, "μL" means microliters, "mL" means milliliters, "L" means liters, "μM" means micromolar, μm means micrometer, "mM" means millimolar, "M" means molar, "mmol" means millimole, "μmole" means micromole, "kg" means kilogram, "g" means gram, "μg" means microgram, "ng" means nanogram, "U" means unit, "bp" means base pair, and "kb" means kilobase.

[0048] Starting materials The solutions used in the described methods include aqueous solutions containing an organic acid or its salt or lactone. More specifically, in one embodiment, the starting material is an aqueous solution containing at least MVA and / or one salt of mevalonate (X-MVA). In one embodiment, the starting material is mevalonolactone (MVL) or mevalonolactone monohydrate (MVL). * The starting material may be selected from, for example, a fermentation broth and / or an aqueous solution from a fermentation that contains MVA, a salt of mevalonate, mevalonolactone, mevalonolactone monohydrate, or any combination thereof.

[0049] In some embodiments, the aqueous solution comprises (or is derived in whole or in part from) a fermentation product. In some such embodiments, the aqueous solution is (or is derived in whole or in part from) a fermentation product used to produce the MVA, X-MVA, and / or MVL to be purified. In some embodiments, the fermentation comprises culturing a recombinant microorganism comprising at least one recombinant polynucleotide sequence encoding an enzyme (or enzymes) capable of producing MVA, X-MVA, and / or MVL in an aqueous medium comprising carbohydrates. The product of the fermentation process can be referred to as the fermentation "product" or "broth". The product typically comprises many components, such as, for example, mono- and divalent salts, sugars, oligosaccharides, monosaccharides, amino acids, polypeptides, proteins, organic acids, nucleic acids, etc., in addition to the X-MVA and / or MVL to be purified. The aqueous solution may contain some alcohol or other solvent derived from the fermentation.

[0050] Examples of enzymes that are often useful in the production of MVA include MvaE (acetyl-CoA acetyltransferase / HMG-CoA reductase) and MvaS (hydroxymethylglutaryl-CoA synthase). The mvaE gene encodes a polypeptide (MvaE) that has both thiolase and HMG-CoA reductase activity. The mvaS gene encodes a polypeptide (MvaS) that has HMG-CoA synthase activity. The enzyme capable of producing MVA (and the corresponding nucleotide sequence) can be derived from, but is not limited to, Listeria grayi, Enterococcus faecalis, (Streptococcus faecalis), Enterococcus faecium, Enterococcus gallinarum, and Enterococcus casseliflavus.

[0051] The fermentation broth can be a broth obtained by fermenting any organism capable of producing MVA or MVL. In some embodiments, the fermentation broth is a broth obtained by fermentation of Escherichia coli. In some aspects, the starting aqueous solution can be a fermentation broth with a neutral pH, such as, but not limited to, a fermentation broth obtained by fermentation of Escherichia coli, containing mevalonic acid as a salt. The starting aqueous solution can also be a fermentation broth with a low pH (pH 3-5), containing mevalonic acid partly in acid form and partly as a salt of mevalonate. The fermentation broth can be a clarified fermentation broth, the clarification being obtained by ultrafiltration using the fermentation broth as a feed.

[0052] In some embodiments, the organic acid to be purified is MVA and the MVA starting solution comprises (or is derived in whole or in part from) the product of a fermentation process comprising culturing in an aqueous broth a recombinant microorganism comprising recombinant polynucleotide sequences encoding MvaE and MvaS.

[0053] The fermentation broth can be clarified by removing the biomass and any insoluble solids from the fermentation broth by at least one of precoat filtration, microfiltration, centrifugation, or ultrafiltration. Ultrafiltration can be advantageous for removing large biomolecules such as endotoxins, proteins, nucleic acids, lipopolysaccharides, etc.

[0054] The cellular biomass can be separated from the fermentation product using, for example, filtration, centrifugation, sedimentation, and / or other processes suitable for removing the cellular biomass.

[0055] Nanofiltration As described herein, in one aspect, the object of the present invention is a purification method for producing a crystalline form of a salt of mevalonic acid (also referred to as X-MVA or X-mevalonate) from an aqueous solution, comprising subjecting an aqueous solution containing said salt of mevalonic acid (X-MVA) to nanofiltration to produce a permeate, and crystallizing said salt of mevalonic acid from the permeate by crystallization in water (see also FIG. 1A and Examples 1 to 4).

[0056] Nanofiltration (NF) is a pressure-driven membrane filtration-based process that results in two fractions: retentate and permeate.

[0057] In one aspect of the invention, the NF process aims to purify salts of mevalonate (such as, but not limited to, Na-mevalonate, K-mevalonate, Li-mevalonate, ammonium mevalonate, or other monovalent salts of mevalonate) to obtain a permeate (also called filtrate). The retentate (also called concentrate) from this process is a waste product containing antifoam, endotoxins, colorants, divalent salts, and larger molecules. In one embodiment of the invention, the aqueous solution is used as a feed for nanofiltration to obtain a permeate with a high content of mevalonate salts (at least 60% sodium mevalonate in the permeate, Examples 1-4) and only a small amount of waste materials.

[0058] The starting aqueous solution can be a fermentation broth with a neutral pH, such as, but not limited to, a fermentation broth obtained by fermentation of Escherichia coli, containing mevalonic acid as a salt. The starting aqueous solution can also be a fermentation broth with a low pH (pH 3-5), containing mevalonic acid partly in acid form and partly as a salt of mevalonate.

[0059] Nanofiltration according to the present invention can be carried out as a batch or continuous process.

[0060] Nanofiltration is usually carried out at temperatures in the range of 5-80°C, preferably in the range of 30-75°C, most preferably in the range of 50-70°C. The pressure of nanofiltration is usually in the range of 5-60 bar, preferably in the range of 10-50 bar, most preferably in the range of 20-45 bar. The pH can be in the range of 1-10, preferably in the range of 3-9, most preferably in the range of 6-9. The pH depends on the composition of the starting solution and the stability of the membrane used for nanofiltration and the components to be recovered. If necessary, the pH of the starting solution can be adjusted to the desired value before carrying out the nanofiltration.

[0061] Nanofiltration typically operates at flux rates of 1-100 l / m, depending on the concentration and viscosity of the nanofiltration feed. 2 h, preferably with a flux of 2-50 l / m2 h, most preferably with a flux of 3-12 l / m 2 It will be carried out in h.

[0062] The nanofiltration membrane used in the present invention can be selected from polymeric and inorganic membranes having a MgSO4 rejection rate of 50-99% (25°C, concentration 2g / l, 8 bar, pH 6), preferably 70-99% (25°C, concentration 2g / l, 8 bar, pH 6), more preferably 80-98% (25°C, concentration 2g / l, 8 bar, pH 6), and most preferably 90-98% (25°C, concentration 2g / l, 8 bar, pH 6). In one embodiment, the nanofiltration membrane is a nanofiltration membrane XN45 having a MgSO4 rejection rate of about 92-98%.

[0063] Nanofiltration membranes with 99% or greater MgSO4 rejection have excessively high rejection of monovalent salts and low MWCOs, resulting in high rejection of X-MVA salts. Membranes with <90% MgSO4 rejection are not as highly purified because they also allow small divalent salts to pass through.

[0064] In some embodiments, the membrane has a molecular weight cut-off (MWCO) in the range of about 100 to about 700 Daltons MWCO membranes, including but not limited to TriSep XN45 membranes. TriSep XN45 membranes are characterized by a MWCO of 300 to 500 Daltons. In some embodiments, the membrane has a molecular weight cut-off (MWCO) in the range of about 150 to about 400 Daltons. In some embodiments, the membrane has a molecular weight cut-off (MWCO) in the range of about 150 to about 300 Daltons, including but not limited to Suez duratherm EXL DL and DK membranes. Suez duratherm EXL DL and DK membranes are characterized by a MWCO of 150 to 300 Daltons. Both MWCO and MgSo4 rejection are important parameters in the selection of nanofiltration membranes. In one embodiment, the membranes for the processes described herein have a MWCO greater than 150 Da, but also have high rejection of divalent salts (>90% MgSO4), and are nanofiltration membranes such as, but not limited to, TriSep XN45 type membranes.

[0065] In some embodiments, the membrane has a molecular weight cut-off (MWCO) in the range of about 100 to about 900 Daltons and a MgSO4 rejection of about 50 to 99% at 25° C. In some embodiments, the membrane has a molecular weight cut-off (MWCO) in the range of about 150 to about 500 Daltons and a MgSO4 rejection of about 80 to 99% at 25° C. In some embodiments, the membrane has a molecular weight cut-off (MWCO) in the range of about 150 to about 300 Daltons and a MgSO4 rejection of about 98 to 99% at 25° C.

[0066] Nanofiltration membranes useful in the present invention can have a negative or positive charge. The membranes can be ionic membranes, i.e., they can contain cationic or anionic groups, but even neutral membranes are useful. Nanofiltration membranes can be selected from hydrophobic and hydrophilic membranes.

[0067] Typical forms of nanofiltration membranes include spiral wound membranes. Membrane configurations can also be selected from, for example, flat plates, tubes, and hollow fibers. "High shear" membranes such as vibrating and rotating membranes can also be used. Membranes can be tubular, spiral wound, or flat.

[0068] A nanofiltration installation useful in the present invention comprises at least one nanofiltration membrane element that divides the starting material (feed) into a retentate portion and a permeate portion. The nanofiltration installation also typically comprises means for controlling pressure and flow, such as pumps and valves, as well as pressure gauges and controls. The installation can also comprise several nanofiltration membrane elements arranged in parallel or series in different combinations within one pressure vessel.

[0069] The yield of X-MVA (a salt of mevalonate, e.g., Na-mevalonate) in nanofiltration is typically greater than 70%, preferably greater than 80%, and most preferably greater than 90% of the Na-mevalonate present in the starting material.

[0070] The Na-mevalonate content in the permeate is greater than 50% by DS, preferably greater than 60% by DS, more preferably greater than 70% by DS, and most preferably greater than 80% by DS.

[0071] The nanofiltration permeate can be further concentrated by evaporation or any means known in the art for further concentrating mevalonate salts, including but not limited to evaporation under reduced pressure (vacuum evaporation), to produce a concentrated aqueous syrup. The Na-mevalonate content of the concentrated aqueous syrup can be greater than 60% by DS, preferably between 65% and 95% by DS.

[0072] The nanofiltration permeate may be subjected to further purification steps selected from ion exchange, evaporation, electrodialysis and filtration, which may be carried out before or after said membrane filtration.

[0073] Additionally, the recovered mevalonate salt fraction can be subjected to one or more additional steps, such as evaporation, concentration, filtration, ion exchange, activated carbon treatment, sterile filtration, crystallization, intermediate crystallization, nanofiltration, and chromatographic fractionation, etc. The recovered mevalonate salt fraction can be processed in different ways depending on the purity of the fraction.

[0074] In some embodiments, the permeate and / or concentrated aqueous syrup recovered from the nanofiltration is subsequently subjected to a crystallization step.

[0075] In one aspect, as described herein, a method is provided for producing a crystalline form of a salt of mevalonic acid (X-MVA) from an aqueous solution, the method comprising: subjecting the aqueous solution comprising the salt of mevalonic acid to a purification step, which produces a purified solution comprising X-MVA having a purity of at least 60%; and crystallizing the salt of mevalonic acid from the purified solution by crystallization in water, the purification step comprising subjecting the aqueous solution comprising the salt of mevalonic acid to nanofiltration to produce a permeate, the permeate comprising X-MVA having a purity of at least 60%.

[0076] Alternatively, in one aspect, the method is for producing a crystalline form of a salt of mevalonic acid (X-MVA) from an aqueous solution, comprising subjecting the aqueous solution containing a salt of mevalonic acid to a purification step, which produces a purified solution containing X-MVA having a purity of at least 60%, and crystallizing the salt of mevalonic acid from the purified solution by crystallization with water as a solvent, which purification step comprises subjecting the aqueous solution containing a salt of mevalonic acid to microfiltration, electrodialysis, ion exchange, filtration, activated charcoal treatment, evaporation, concentration, sterile filtration, chromatographic fractionation, or any combination thereof.

[0077] Cation Exchange In one aspect, the present specification provides a method for producing mevalonolactone dissolved in water from an aqueous solution, the method comprising: a) producing a crystalline form of the salt of mevalonic acid from an aqueous solution by subjecting an aqueous solution containing a salt of mevalonic acid (X-MVA) to nanofiltration to produce a permeate, and crystallizing the salt of mevalonic acid from the permeate by crystallization using water as a solvent to produce crystals of the salt of mevalonic acid; and b) dissolving the crystals of (a) in water to produce a water-solubilized salt of mevalonic acid, and subjecting the liquid to cation exchange to convert the water-soluble salt of mevalonic acid to water-soluble mevalonolactone (FIG. 1B and Examples 9-12).

[0078] In another aspect, the present disclosure provides a method for producing mevalonolactone from an aqueous solution containing a salt of mevalonic acid (X-MVA), comprising subjecting the aqueous solution containing a salt of mevalonate to cation exchange, thereby converting the aqueous solution containing a salt of mevalonate to an aqueous solution containing mevalonolactone (MVL) (FIG. 2).

[0079] Strong acid cation exchange resin (SAC resin) The SAC resin can have a styrene or acrylic backbone. In one embodiment of the invention, the resin is a sulfonated polystyrene-co-divinylbenzene resin. Other alkenyl aromatic polymer resins, such as those based on monomers such as alkyl-substituted styrenes or mixtures thereof, can also be applied. The resin may be crosslinked with other suitable aromatic crosslinking monomers, such as divinyltoluene, divinylxylene, divinylnaphthalene, divinylbenzene, or with aliphatic crosslinking monomers, such as isoprene, ethylene glycol diacrylate, ethylene glycol dimethacrylate, N,N'-methylenebisacrylamide, or mixtures thereof. The degree of crosslinking of the resin is typically about 1 to about 20%, preferably about 3 to about 8%, of the crosslinking agent, such as divinylbenzene (DVB).

[0080] The SAC resins for use in the ion exchange of the present invention can be in polyvalent, divalent, or monovalent cationic form. The monovalent cationic form is, for example, H + , Na + , and K + An example of a divalent cation form is Ca 2+ , Mg 2+ , Zn 2+ , Sr 2+ , and Ba 2+ An example of a trivalent cationic form is Al 3+ In a preferred embodiment of the present invention, the SAC resin for use in the ion exchange of the present invention is + The resin typically has a mean average particle size of 10 to 2000 μm, preferably 300 to 1200 μm.

[0081] In one embodiment, a monovalent SAC resin is used for ion exchange of the salt of mevalonic acid (X-MVA) to produce mevalonolactone (MVL), and the monovalent cation is H + It is a type.

[0082] Weakly acidic cation exchange resin (SAC resin) WAC resins are acrylic cation exchange resins with carboxyl functionality. Acrylic WAC resins are usually derived from the group consisting of acrylic acid esters, acrylonitrile, acrylic acid, and mixtures thereof. The acrylic acid esters are selected from the group consisting of methyl methacrylate, methyl acrylate, ethyl acrylate, and butyl acrylate. The matrix of the WAC resin may be other than acrylic. The active functional groups of the WAC resin may be other than carboxyl groups. These may be selected, for example, from other weak acids. WAC resins are also suitable for use in the preparation of cation exchange resins with H + , Na + , K + , Ca 2+ , or Mg 2+ It can be of the form H + Or Na + Other ionic forms may also be used.

[0083] The WAC resin is crosslinked with an aromatic crosslinker, preferably divinylbenzene (DVB). It can also be crosslinked with an aliphatic crosslinker, such as isoprene, 1,7-octadiene, trivinylcyclohexane, diethylene glycol divinyl ether, etc. The degree of crosslinking is 1-20%, preferably 3-8% DVB.

[0084] The average particle size is from 10 to 2000 μm, preferably from 300 to 1200 μm.

[0085] The ion exchange is preferably carried out using a cation exchange resin, in particular a strongly acidic cation resin, especially H + It is carried out using the ionic form.

[0086] In one embodiment, a monovalent WAC resin is used for the ion exchange of a salt of mevalonic acid (X-MVA) to produce mevalonolactone (MVL), and the monovalent cation is H + It is a type.

[0087] Crystallization using water as a solvent As described herein, in one aspect, the object of the present invention is a method for producing a crystalline form of a salt of mevalonic acid (also referred to as X-MVA or X-mevalonate) from an aqueous solution, comprising subjecting the aqueous solution containing said salt of mevalonic acid (X-MVA) to a purification step, such as, but not limited to, nanofiltration, to produce a high purity solution (permeate) from which said X-MVA can be crystallized, and crystallizing said salt of mevalonic acid from the permeate by crystallization using water as a solvent (FIG. 1A and Examples 5-8).

[0088] In another aspect of the present invention, the present disclosure provides a method for producing mevalonolactone from an aqueous solution containing a salt of mevalonic acid (X-MVA), comprising subjecting the aqueous solution containing a salt of mevalonate to cation exchange, thereby converting the aqueous solution containing a salt of mevalonate into an aqueous solution containing mevalonolactone (MVL), and then, optionally, concentrating the MVL solution and crystallizing the solution. * and obtaining a monohydrate crystal (FIG. 2 and Examples 13-20).

[0089] The term "aqueous crystallization" refers to crystallization carried out using an aqueous solvent such as water, without the use of any organic solvent.

[0090] Crystallization of X-MVA using water as a solvent The crystallization of X-MVA can be carried out by conventional methods such as cooling crystallization or precipitation crystallization at temperatures ranging from 10 to 80° C. The crystallization of X-MVA can also be advantageously carried out by the boiling crystallization method or the boiling and cooling crystallization method.

[0091] In one embodiment of the disclosure, the crystallization of X-MVA is carried out from a solution (feed) having a purity of X-MVA based on DS of greater than 60%, preferably greater than 70%, more preferably greater than 80%, most preferably greater than 90%, and especially greater than 95%. This crystallization typically results in a crystalline X-MVA product having a purity based on DS of greater than 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, preferably greater than 95%, and most preferably greater than 99%.

[0092] The solution containing X-MVA can first be evaporated to an appropriate dry matter content (e.g. DS of about 60-90%) depending on the X-MVA content of the solution. The DS concentration of the syrup needs to be high enough (supersaturated) to have the possibility of crystallization. The minimum DS content depends on the temperature and purity of the syrup. The higher the purity of the syrup, the lower the DS of the syrup needs to be. If the DS is too high, crystallization will be delayed or the suspension of crystals will be too concentrated to separate the crystals efficiently. In one embodiment of the present invention, the purity of the syrup containing Na-MVA is 65-99%.

[0093] The supersaturated solution may be added with seed crystals of X-MVA. When using seeds, the seeds are either in dry form or suspended in a solvent, preferably water, and the size of the crystals is preferably reduced, for example by pulverizing or milling. The evaporation temperature of the solution containing X-MVA may range from 30°C to 80°C. After the addition of the seeds, the crystallization mass is mixed simultaneously with cooling until the crystallization yield and viscosity are optimal for the separation of the crystals. The cooling time is preferably 10 to 60 hours. The temperature drop during cooling is preferably 5 to 40°C. The syrup containing the seeded Na-MVA can be cooled from about 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C, 65°C, 66°C, 67°C, 68°C, 69°C, 70°C, 71°C, 72°C, 73°C, 74°C to about 75°C or greater. Cooling can occur for about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or more hours. Cooling can be performed with continuous stirring or mixing. Mixing can be advantageous for controlling crystallization. Mixing prevents crystals from settling, maintains crystal growth, and reduces spontaneous crystal formation. Mixing also promotes advantageous heat and mass transfer. The crystallization feedstock can then be mixed at the final temperature for a period of time, preferably 0.5 to 24 hours, to reach maximum crystal yield. The crystals are separated from the mother liquor, for example, by filtration or centrifugation.

[0094] In one embodiment of the present invention, high purity X-MVA crystals having an X-MVA content of more than 97% by DS, preferably more than 98% by DS, more preferably more than 99% by DS are obtained in a single crystallization step (=single-step crystallization) from a solution having an X-MVA content of more than 65% by DS without dissolution and recrystallization steps. Single-step crystallization may include boiling and cooling steps, but does not include a recrystallization step.

[0095] In another embodiment of the invention, the crystallization of X-MVA includes a further washing step, which is usually carried out in conjunction with the separation of the crystals from the mother liquor. An additional wash can be carried out by mixing an aqueous wash solvent with the crystal cake and then separating the crystals. The washing solvent may be water. This embodiment of the invention typically results in X-MVA with a purity of greater than 98%.

[0096] Seed crystals can be generated by a variety of processes. In some embodiments, dried seeds are milled to a smaller particle size. The desired amount of seed crystals can depend, for example, on the size of the seed crystals. In some embodiments, crystallization is initiated without adding seed crystals to the supersaturated solution. In some such embodiments, for example, seeding is done using the natural generation of seeds.

[0097] In some embodiments, the initiation of crystallization (e.g., the addition of seed crystals) is performed when the dry solids content of the syrup is at least about 60% (by weight). In some embodiments, the initiation of crystallization (e.g., the addition of seed crystals) is performed when the dry solids content of the syrup is at least about 70% (by weight). In some embodiments, the initiation of crystallization (e.g., the addition of seed crystals) is performed when the dry solids content of the syrup is at least about 80% (by weight). In some embodiments, the initiation of crystallization (e.g., the addition of seed crystals) is performed when the dry solids content of the syrup is about 60 to about 90% (by weight). In some embodiments, the initiation of crystallization (e.g., the addition of seed crystals) is performed when the dry solids content of the syrup is about 70 to about 90% (by weight). In some embodiments, the initiation of crystallization (e.g., the addition of seed crystals) is performed when the dry solids content of the syrup is about 80 to about 90% (by weight). In some embodiments, the initiation of crystallization (eg, addition of seed crystals) is performed when the dry solids content of the syrup is about 80 to about 88% (by weight).

[0098] Evaporation can be continued after seeding if crystal growth capacity and viscosity permit. After evaporation, the crystallization feed is cooled and mixed at the same time until the crystal content and viscosity are optimal for crystal separation. The crystallization feed is usually cooled to a temperature of 10 to 75°C. The crystallization feed can then be mixed at the final temperature for a period of time, preferably 0.5 to 24 hours, until the maximum crystallization yield is reached, after which the crystals are separated, for example, by filtration or centrifugation. The process of the present invention usually includes washing of the crystals as a further step. Washing is usually performed in conjunction with the separation of the crystals from the mother liquor. An additional wash can be performed by mixing a washing solvent with the crystal cake and then separating the crystals. The washing solvent can be water.

[0099] In some embodiments, recrystallization is performed one or more times to increase X-MVA purity. Recrystallization can be performed, for example, by dissolving X-MVA crystals in water (usually deionized water), supersaturating the resulting solution with respect to X-MVA (e.g., by evaporation), seeding, and crystallizing using the cooling crystallization method described above.

[0100] In some embodiments, the yield is increased by performing a crystallization of the mother liquor produced in the first crystallization. This type of crystallization can be performed, for example, by supersaturating the mother liquor with respect to X-MVA (e.g., by evaporation), seeding, and crystallizing using the cooling crystallization method described above.

[0101] The crystallization described herein does not require the presence of an organic solvent in the solution. Organic solvent-free crystalline X-MVA is obtained. Crystalline X-MVA produced without the addition of an organic solvent in the crystallization step is essentially organic solvent-free.

[0102] Although organic solvents can be added to aqueous solutions to improve crystallization and crystal separation performance, such addition of organic solvents has disadvantages, including but not limited to, that the resulting crystals or purified products contain small amounts of such organic solvents, which are undesirable for commercial compositions, such as personal care compositions.

[0103] MVL * Crystallization of the monohydrate from water As described herein, it has been surprisingly and unexpectedly been found that a high purity syrup of MVL can be prepared by the methods described herein and, upon cooling at a temperature below 23° C. for several weeks, produces mevalonolactone monohydrate (MVL). * It was found that mevalonolactone monohydrate (MVL) crystals occur spontaneously. * HO) crystals were prepared using MVL as described herein. * It can be used as a seed material to further promote the crystallization of H2O. *Crystallization of HO is achieved at temperatures below the melting point, such as 23°C, below 22°C, below 21°C, below 20°C, below 19°C, below 18°C, below 17°C, below 16°C, below 15°C, below 14°C, below 13°C, below 12°C, below 11°C, below 10°C, below 9°C, below 8°C, below 7°C, below 6°C, below 5°C, below 4°C, below 3°C, below 2°C, below 1°C, and temperatures down to 0°C. MVL * Crystallization of HO can be carried out below 0° C., down to the freezing point of the solution. In one embodiment of the present invention, MVL * Monohydrate (MVL * The crystallization of MVL (H2O) is carried out from an aqueous solution having an MVL purity of more than 55% by DS, preferably more than 70%, more preferably more than 80%, most preferably more than 90%, in particular more than 95%. This crystallization usually results in crystalline MVL having a purity of more than 65%, more than 66%, more than 67%, more than 68%, more than 69%, more than 70%, more than 71%, more than 72%, more than 73%, more than 74%, more than 75%, more than 76%, more than 77%, more than 78%, more than 79%, more than 80%, more than 81%, more than 82%, more than 83%, more than 84%, more than 85%, more than 86%, more than 87%, more than 89%, more than 90%, more than 91%, more than 92%, more than 93%, more than 94%, preferably more than 95%, most preferably more than 99% by DS. * The H2O product is obtained.

[0104] In one embodiment, for practical reasons, MVL * Cooling crystallization of H2O is preferred over constant temperature crystallization, which requires very high supersaturation during seeding, making the process less controllable than with cooling. MVL * H2O releases a lot of heat during crystallization, so heat must be removed from the system to reach the desired crystallization yield. In some instances, such as when the seeding supersaturation is very high, crystallization may actually be faster and the temperature of the crystallization suspension may increase significantly. This can be overcome by using effective cooling means or by using lower seeding supersaturation and controlled cooling.

[0105] The solution containing MVL can first be evaporated to an appropriate dry matter content (e.g. DS about 65-90%) depending on the MVL content of the solution. The DS concentration of the syrup needs to be high enough (supersaturated) to have the potential for crystallization. The minimum DS content depends on the temperature and purity of the syrup. The purer the syrup, the lower the DS of the syrup needs to be. If the DS is too high, crystallization will be delayed or the suspension of crystals will be too concentrated for efficient separation. Supersaturated solutions contain MVL. * Seed crystals of H2O can be added. When using seeds, the seeds are either in dry form or suspended in a solvent, preferably water, and the size of the crystals is preferably reduced, for example by powdering or grinding. After seeding, the crystallization material is cooled and mixed at the same time until the crystallization yield and viscosity are optimal for the separation of the crystals.

[0106] MVL * The conditions for crystallizing HO are further described in Examples 13-20.

[0107] In one embodiment, mevalonolactone * Monohydrate (MVL * MVL (H2O) crystals can be warmed to a temperature (such as, but not limited to, room temperature, about 23°C to 30°C) without adding water, such that the crystals turn into a liquid. This type of liquid contains about 87% MVL and 13% water.

[0108] Mevalonolactone monohydrate (MVL * HO) crystals can be dissolved in water to produce highly purified (>90% purity) MVL that has a water-like appearance and viscosity and little or no color. This highly purified MVL solution can be further concentrated by evaporation while still retaining its water-like appearance.

[0109] MVA, X-MVA, MVL, and / or MVL * Compositions containing H2O Crystalline X-MVA or compositions comprising crystalline X-MVA can be used, for example, as ingredients in dietary supplements, infant and adult nutrition, pharmaceuticals, and cosmetics.

[0110] MVA, X-MVA, MVL, and / or MVL purified by the process herein * HO or MVA, X-MVA, MVL, and / or MVL as described herein * Compositions comprising HO can be used, for example, as ingredients in dietary supplements, personal care compositions (including, but not limited to, skin care, oral care, and hair care compositions), pharmaceuticals, and cosmetics.

[0111] For the purposes of using MVA or MVL in the personal care industry, a purified product that is clear and colorless and has little or no colored contaminants is of high value. As described herein, the highly purified MVL product obtained by the methods described herein is clear and colorless with little or no color remaining.

[0112] Non-limiting examples of the compositions and methods disclosed herein are given below: 1. A method for producing a crystalline form of an organic acid or a salt thereof from an aqueous solution, the method comprising: subjecting an aqueous solution containing the organic acid or a salt thereof to nanofiltration to produce a permeate; and crystallizing the organic acid or the salt thereof from the permeate by crystallization using water as a solvent.

[0113] 2. The method of embodiment 1, wherein the crystallization using water as a solvent comprises evaporating the permeate to obtain a syrup containing at least 60% DS, and then cooling the syrup to produce crystals of the organic acid or its salt.

[0114] 3. The method of embodiment 1, wherein the aqueous solution is a fermentation broth.

[0115] 4. The method of embodiment 3, wherein the fermentation broth is clarified by removing biomass and any solids from the fermentation broth by at least one of precoat filtration, microfiltration, or ultrafiltration.

[0116] 5. The method of embodiment 3, wherein the fermentation broth is a broth obtained by fermentation of Escherichia coli.

[0117] 6. The method of embodiment 2, further comprising removing said crystals of said organic acid or salt thereof from said syrup by filtration and centrifugation.

[0118] 7. The method of embodiment 1, wherein the organic acid is a carboxylic acid.

[0119] 8. The method of embodiment 7, wherein the carboxylic acid is selected from the group consisting of hydroxycarboxylic acids and dicarboxylic acids.

[0120] 9. The method of embodiment 8, wherein the hydroxycarboxylic acid is selected from the group consisting of malic acid, glycolic acid, isocitric acid, mandelic acid, lactic acid, tartronic acid, tartaric acid, citric acid, β-hydroxybutyric acid, mevalonic acid, and salicylic acid.

[0121] 10. A method for producing a crystalline form of a salt of mevalonic acid (X-MVA) from an aqueous solution, comprising subjecting the aqueous solution containing the salt of mevalonic acid to a purification step, which produces a purified solution containing X-MVA having a purity of at least 60%, preferably between 60 and 99%, and crystallizing the salt of mevalonic acid from the purified solution by crystallization using water as a solvent.

[0122] 11. The method of embodiment 10, wherein the purifying step comprises subjecting the aqueous solution comprising a salt of mevalonic acid to nanofiltration to produce a permeate, the permeate comprising X-MVA having a purity of at least 60%.

[0123] 11b. A method for producing a crystalline form of a salt of mevalonic acid from an aqueous solution, comprising: subjecting an aqueous solution containing the salt of mevalonic acid (X-MVA) to nanofiltration to produce a permeate; and crystallizing the salt of mevalonic acid from the permeate by crystallization using water as a solvent.

[0124] 12. The process of embodiments 10, 11 and 11b, wherein the crystallization using water as a solvent comprises evaporating the purified solution or permeate to obtain a syrup containing at least 60% DS, and then cooling the syrup to produce X-MVA crystals.

[0125] 12b. The method of embodiment 11b, wherein the X-MVA purity of the permeate is at least 60%, optionally between 65-95%.

[0126] 12c. The method of embodiment 12, wherein the X-MVA purity of the syrup is at least 60%, optionally between 65-95%.

[0127] 13. The method of embodiment 10, wherein the aqueous solution is a fermentation broth.

[0128] 14. The method of embodiment 13, wherein the fermentation broth is clarified by removing biomass and any solids from the fermentation broth by at least one of precoat filtration, microfiltration, or ultrafiltration.

[0129] 15. The method of embodiment 13, wherein the fermentation broth is a broth obtained by fermentation of Escherichia coli.

[0130] 16. The method of embodiment 12, further comprising removing the crystals from the syrup by filtration and centrifugation.

[0131] 16b. The method of embodiment 11, wherein the nanofiltration utilizes a nanofiltration membrane with a separation cut-off of about 150 Da to about 700 Da.

[0132] 17. The method of embodiment 10, wherein the nanofiltration utilizes a nanofiltration membrane capable of retaining at least 60-98% by weight of divalent salts while rejecting less than 30% by weight of monovalent salts.

[0133] 18. A method for producing water-solubilized mevalonolactone from an aqueous solution comprising: a) producing a crystalline form of a salt of mevalonic acid from an aqueous solution by subjecting an aqueous solution containing a salt of mevalonic acid (X-MVA) to nanofiltration to produce a permeate, and crystallizing the salt of mevalonic acid from the permeate by crystallization using water as a solvent to produce crystals of the salt of mevalonic acid; b) dissolving the crystals of (a) in water to produce a water-solubilized salt of mevalonic acid, and converting the water-solubilized salt of mevalonic acid to water-solubilized mevalonolactone by subjecting the liquid to cation exchange.

[0134] 18b. The method of embodiment 18, wherein the X-MVA purity of the permeate is at least 60%, optionally between 65-95%.

[0135] 18c. The method of embodiment 18, wherein the permeate is concentrated to produce a syrup prior to crystallizing the salt of mevalonic acid from the syrup by crystallization with water to produce crystals of the salt of mevalonic acid.

[0136] 18d. The method of embodiment 18c, wherein the X-MVA purity of the syrup is at least 60%, optionally between 65-95%.

[0137] 18e. The method of embodiment 18, wherein the purity of the water-solubilized mevalonolactone is at least 60%, optionally between 65-95%.

[0138] The method of embodiment 18, wherein the water-solubilized mevalonolactone of 18f.b) is further concentrated to produce a highly concentrated MVL liquid that is at least 90% DS.

[0139] 19. The method of embodiment 10 or embodiment 18, wherein the salt of mevalonic acid is selected from the group consisting of sodium mevalonate (Na-MVA), potassium mevalonate (K-MVA), ammonium mevalonate (NH4-MVA), lithium mevalonate (Li-MVA), any other monovalent salt of mevalonic acid, or any combination thereof.

[0140] 20. The method of embodiment 18, 18b, 18c, 18d, 18e, 18f, or 19, wherein the purity of the water-solubilized mevalonolactone is at least 90% (this percentage being the area of ​​the peak corresponding to MVL relative to the total area of ​​the peaks using HPLC analysis).

[0141] 21. A method for producing mevalonolactone from an aqueous solution containing a salt of mevalonic acid (X-MVA), comprising subjecting said aqueous solution containing a salt of mevalonate to cation exchange, thereby converting said aqueous solution containing a salt of mevalonate to an aqueous solution containing mevalonolactone (MVL).

[0142] 22. The method of embodiment 21, wherein the salt of mevalonic acid is selected from the group consisting of Na-mevalonate (Na-MVA), K-mevalonate (K-MVA), ammonium mevalonate (NH4-MVA), lithium mevalonate (Li-MVA), any other monovalent salt of mevalonic acid, or any combination thereof.

[0143] 23. The method of embodiment 21, further comprising concentrating the aqueous solution containing mevalonolactone to provide a liquid containing at least 65% to 90% DS.

[0144] 24. Mevalonolactone monohydrate (MVL) * 24. The method of embodiment 23, further comprising cooling the liquid to 0-25° C. to obtain HO) crystals.

[0145] 25. The MVL * 25. The method of embodiment 24, further comprising dissolving / solubilizing the HO crystals in water to obtain a dilute liquid of highly purified mevalonolactone, wherein the dilute liquid has a mevalonolactone purity of at least 90% (this percentage being the area of ​​the peak corresponding to MVL relative to the total area of ​​the peaks using HPLC analysis).

[0146] 25b. The method of embodiment 25, wherein said dilute liquid containing highly purified mevalonolactone is further concentrated to produce a highly concentrated MVL liquid that is at least 90% DS.

[0147] 26. The method of embodiment 21, wherein the aqueous solution is a fermentation broth.

[0148] 27. The method of embodiment 26, wherein the fermentation broth is clarified by removing biomass and any solids from the fermentation broth by at least one of precoat filtration, microfiltration, or ultrafiltration.

[0149] 28. The method of embodiment 26, wherein the fermentation broth is a broth obtained by fermentation of Escherichia coli.

[0150] 29. A pharmaceutical or home care or nutritional or personal care or cosmetic composition comprising MVL or a solubilized form thereof produced by any one of embodiments 20-26.

[0151] 30. Crystalline X-MVA obtainable by the method of any one of embodiments 1 to 17.

[0152] 31. Crystalline mevalonolactone obtained by the method of embodiment 24 * Monohydrate (MVL * H2O).

[0153] 32.Molecular formula C6H 10 O3 *Crystalline mevalonolactone with HO * Monohydrate (MVL * H2O).

[0154] 33. Crystalline mevalonolactone having a melting point of less than 25°C * Monohydrate (MVL*H2O).

[0155] 34. Mevalonolactone monohydrate (MVL) * A composition comprising crystalline HO.

[0156] 35. Solubilized mevalonolactone monohydrate (MVL * A composition comprising crystalline HO. EXAMPLES

[0157] In the following examples, parts and percentages are by weight and degrees are Celsius unless otherwise specified. It should be understood that these examples, which illustrate embodiments of the present disclosure, are provided for illustrative purposes only. From the above discussion and these examples, those skilled in the art can make various changes and modifications to adapt the present disclosure to their usage and circumstances. Such modifications are also intended to be encompassed by the appended claims.

[0158] Example 1 Nanofiltration (NF) of sodium mevalonate (Na-MVA) solution using tight nanofiltration membranes In this example, the nanofiltration of an aqueous solution containing an organic acid or its salt, for example, sodium mevalonate (Na-MVA) solution, using a highly dense nanofiltration membrane, is described.

[0159] The treatment equipment included a plate and frame filtration unit (Alfa Laval Labstak M20), feed and diafiltration pumps, heat exchanger, chiller, a 70 L feed tank, as well as inlet and outlet pressure gauges and pressure control valves. The total membrane area was 0.65 m. 2The membranes used were Desal-5 DL (Suez, molecular weight cutoff of about 150-300 daltons, MgSO4 rejection >98% at 25°C) and XN45 (TriSep (registered trademark) / Microdyn Nadir, molecular weight cutoff of about 300-500 daltons, MgSO4 rejection of 90-96% at 25°C). The MgSO4 rejection was determined at a concentration of 1-2 g / l, 7-8 bar, 25°C, pH 6-8, and a recovery rate of 10-25%.

[0160] The permeate of an ultrafiltration process derived from the fermentation of sodium mevalonate using Escherichia coli as the raw material was used as the feed, with the aim of passing the sodium mevalonate while retaining the impurities (other salts, colouring substances, antifoaming agents, proteins).

[0161] 63.4 kg of feed was fed to a 70 liter feed tank. The feed had a dry matter concentration of 6.8 g / 100 g based on sucrose RDS, a conductivity of 17.7 mS / cm, and a pH of 7.8. Permeate was collected from the DL membrane and permeate from the XN45 membrane was recycled to the feed tank. The feed had the composition shown in Table 1. HPLC and IC analyses are shown in % / sucrose RDS.

[0162] [Table 1]

[0163] The feed liquid was maintained at 50-68°C, and water was used for diafiltration. The filtration pressure was set to 20-33 Bar, and the DS (dry matter) concentration of the concentrate was controlled to a flux of 7 kg / m 2 / h (the minimum flux point observed).

[0164] After batch filtration, three permeate fractions and a final retentate fraction were collected. The results, including HPLC and IC analyses (% / sucrose RDS basis) for the permeate and final retentate fractions, are shown in Table 2.

[0165] [Table 2]

[0166] The overall yield of mevalonate was determined to be 49.9%. The average sodium mevalonate purity of the permeate was 75.6%.

[0167] The sulfate removal efficiency was determined to be 78.1% and the phosphate removal efficiency was 71.0% from feed and concentrate samples. The average sodium mevalonate rejection measured at five points for the DL membrane was 76.7%. The mevalonate rejection of DL and XN45 was measured in parallel at the fourth sampling point after collecting 41 kg of permeate, and the rejections were 71.0% and 41.8% for DL ​​and XN45, respectively.

[0168] Example 2 Nanofiltration of Sodium Mevalonate Solution Using an Open Nanofiltration (NF) Membrane In this example, the nanofiltration of an aqueous solution containing an organic acid or its salt, e.g., the nanofiltration of a sodium mevalonate solution using an open nanofiltration membrane, is described.

[0169] The treatment equipment included a plate and frame filtration unit (Alfa Laval Labstak M20), feed and diafiltration pumps, heat exchanger, chiller, a 100 litre feed tank, as well as inlet and outlet pressure gauges and pressure control valves. The total membrane area was 0.72 m. 2 The membrane used was XN45 (TriSep (registered trademark) / Microdyn Nadir, molecular weight cutoff of about 300-500 daltons, MgSO4 rejection rate of 90-96% at 25°C). The MgSO4 rejection rate was determined at a concentration of 1-2 g / l, 7-8 bar, 25°C, pH 6-8, and a recovery rate of 10-25%.

[0170] The permeate of an ultrafiltration process derived from the fermentation of sodium mevalonate using Escherichia coli as the raw material was used as the feed, with the aim of passing the sodium mevalonate while retaining the impurities (other salts, colouring substances, antifoaming agents, proteins).

[0171] 80.0 kg of feed solution was added to a 100 L feed tank. The concentration of the feed solution was 6.6 g / 100 g based on the RDS of sucrose. The pH was adjusted to 8.8 with NaOH. The concentration of antifoam agent, Foamblast 882, was measured to be 1337 mg / L. The feed solution had the composition shown in Table 3. HPLC and IC analyses are given on a % / sucrose RDS basis.

[0172] [Table 3]

[0173] The feed liquid was maintained at 46-51°C. The filtration pressure was 10-25 Bar and the flux was 7 kg / m 2 / h (minimum observed flux value).

[0174] After batch filtration, two permeate fractions and a final retentate fraction were collected. The results, including HPLC and IC analyses (% / sucrose RDS basis) for the permeate and final retentate fractions, are shown in Table 4.

[0175] [Table 4]

[0176] The overall yield of mevalonate was determined to be 82.1%. The antifoam concentration was determined by evaporation of a 29.5% concentration permeate sample to be 84.4 mg / L, which means that 99% of the antifoam was removed from the product permeate fraction. The antifoam concentration in the final concentrate was determined to be 13230.0 mg / L. The average sodium mevalonate rejection measured at three points was 23.6%.

[0177] Example 3 Nanofiltration of sodium mevalonate solutions using low density nanofiltration (NF) membranes This example describes the nanofiltration of an aqueous solution containing an organic acid or its salt, for example, a sodium mevalonate solution, using a low density nanofiltration membrane.

[0178] The treatment equipment included a plate and frame filtration unit (Alfa Laval Labstak M20), feed and diafiltration pumps, heat exchanger, chiller, a 100 litre feed tank, as well as inlet and outlet pressure gauges and pressure control valves. The total membrane area was 0.72 m. 2 The membrane used was XN45 (TriSep (registered trademark) / Microdyn Nadir, molecular weight cutoff of about 300-500 daltons, MgSO4 rejection rate of 90-96% at 25°C). The MgSO4 rejection rate was determined at a concentration of 1-2 g / l, 7-8 bar, 25°C, pH 6-8, and a recovery rate of 10-25%.

[0179] The permeate of an ultrafiltration process derived from the fermentation of sodium mevalonate using Escherichia coli as the raw material was used as the feed, with the aim of passing the sodium mevalonate while retaining the impurities (other salts, colouring substances, antifoaming agents, proteins).

[0180] 79.7 kg of feed solution was fed to a 100 liter feed tank. The feed concentration was 7.0 g / 100 g. The pH was adjusted to 8.25 with 50 grams of 30% NaOH. The feed solution had the composition shown in Table 5. HPLC analysis is shown on a % / sucrose RDS basis (Table 6).

[0181] [Table 5]

[0182] The feed liquid was maintained at 46-51°C. The filtration pressure was 10-25 Bar and the flux was 7 kg / m 2 / h (minimum observed flux).

[0183] After batch filtration, the permeate and final retentate fractions were collected. A portion of the total permeate was evaporated to a dry solids of 28.7%. The results, including HPLC analysis (% / sucrose RDS basis) for the permeate and final retentate fractions, are shown in Table 6.

[0184] [Table 6]

[0185] The total mevalonate yield was determined to be 86.8%.

[0186] Example 4 Nanofiltration of Sodium Mevalonate Solution Using a Low Density Spiral Wound Nanofiltration (NF) Membrane In this example, the nanofiltration of an aqueous solution containing an organic acid or its salt, for example, the nanofiltration of a sodium mevalonate solution using a low density spiral wound nanofiltration membrane, is described.

[0187] The treatment system included a plate and frame filtration unit (Alfa Laval Labstak M20), feed and diafiltration pumps, heat exchangers, chillers, a 1000 litre feed tank, as well as inlet and outlet pressure gauges and pressure control valves. The total membrane area was 14.8 m. 2 The membrane used was a 4040 XN45 (TriSep® / Microdyn Nadir, molecular weight cutoff approximately 300-500 Daltons, MgSO4 rejection 90-96% at 25°C) with a 31 mil spacer. The MgSO4 rejection was determined at a concentration of 1-2 g / l, 7-8 bar, 25°C, pH 6-8, and 10-25% recovery.

[0188] The permeate of an ultrafiltration process derived from the fermentation of sodium mevalonate using Escherichia coli as the raw material was used as the feed, with the aim of passing the sodium mevalonate while retaining the impurities (other salts, colouring substances, antifoaming agents, proteins).

[0189] 600 kg of feed solution was fed to a 1000 liter feed tank. The feed solution had a concentration of 6.7 g / 100 g. The pH was adjusted to 8.5 with 30% NaOH. The feed solution had the composition shown in Table 7. HPLC analysis values ​​are given on a % / sucrose RDS basis.

[0190] [Table 7]

[0191] The feed liquid was maintained at 45-52°C. The filtration pressure was 10-25 Bar and the flux was 5 kg / m 2 / h (minimum observed flux value).

[0192] After batch filtration, the permeate and final retentate fractions were collected. The total permeate was evaporated to a dry solid of 39.1%. The results, including HPLC analysis (% / sucrose RDS basis) for the permeate and final retentate fractions, are shown in Table 8.

[0193] [Table 8]

[0194] The overall yield of mevalonate was determined to be 87.5%.

[0195] Example 5 Crystallization of sodium mevalonate (Na-MVA) in water In this example, the aqueous crystallization of an organic acid or its salt, for example, the aqueous crystallization of sodium mevalonate (Na-MVA) and the preparation of sodium mevalonate seed crystals are described.

[0196] The crystallization feedstock was an aqueous syrup containing sodium mevalonate. The sodium mevalonate purity of the syrup was 96% by DS. An aqueous syrup containing ≧95% sodium mevalonate by DS can be produced by dissolving a known commercially available sodium mevalonate salt in deionized water or by mixing a known commercially available sodium mevalonate salt with the nanofiltration permeate containing sodium mevalonate prepared according to Examples 1-4.

[0197] The raw syrup was evaporated (Rotavapor R-151) at a temperature of 70°C. Crystals were formed by spontaneous nucleation when the DS (dry matter) was about 54%. After nucleation, evaporation was continued at 70°C for 2 hours until the DS of the crystallization raw material was 81.8%.

[0198] The evaporated crystallization material was transferred to a 2 L cooling crystallizer. The crystallization material was maintained at 70° C. for 1 hour and then cooled to a temperature of 40° C. within 20 hours with continuous stirring.

[0199] The resulting crystallization material (930 g) was centrifuged with 20 mL of wash water (batch centrifugation, basket diameter 22.5 cm, 3500 rpm, 5 min). The mevalonate yield from centrifugation was 45%, the sodium mevalonate purity of the centrifuge cake was ≥98% by DS, and the sodium mevalonate purity of the centrifuge mother liquor was 94% by DS.

[0200] The crystal cake was dried for 19 hours in a heating chamber at 60° C. The moisture content of the dried cake was ≦0.2 wt.%.

[0201] Sodium mevalonate crystals prepared according to this process were used as seed crystals for Examples 6, 7, and 8. The crystals were ground in a porcelain grinder before use.

[0202] Example 6 Crystallization of sodium mevalonate (Na-MVA) in water In this example, the crystallization of an organic acid or its salt in water, for example, the crystallization of sodium mevalonate (Na-MVA) in water, is described.

[0203] The crystallization feedstock was the evaporated nanofiltration permeate prepared according to Example 4. Before crystallization, the syrup was treated with activated carbon to remove coloring substances. Carbon treatment was carried out by dosing 76 kg of evaporated syrup with 600 g of Norit DX 1 carbon powder (approximately 20 g of carbon powder per kg of DS). The syrup was heated to a temperature of 50° C. and left under continuous stirring at a constant temperature for 45 minutes. The carbon powder was then separated from the syrup by filtration using a filter aid (Seitz depth filter, 1.1 kg of Kenite 300 filter aid). The sodium mevalonate purity of the obtained syrup was 73% by DS, the color number was 6300 ICUMSA and the DS was 24.1%.

[0204] 41 kg of carbon-treated raw syrup was evaporated to a DS of 86.0% (Luwa thin film evaporator NL3-210 / 1600 / 10 and Rotavapor R-153 evaporator). 4 g of 10% Struktol J 650 antifoam was added during evaporation to prevent foaming. 11.6 kg of the resulting syrup was transferred to a 10 liter cooling crystallizer and seeded twice at a temperature of 70° C.: first with 0.3 g of sodium mevalonate dry seeds (prepared according to Example 5) and then 2.0 g of sodium mevalonate dry seeds 0.6 hours after the first seeding. The first seeding produced very fine crystals.

[0205] The seeded syrup was cooled with continuous stirring within 16 hours to a temperature of 48° C. After cooling to 48° C., the crystallization material was maintained at 48° C. for 2 hours.

[0206] The resulting crystallization material (10.7 kg) was centrifuged with 170 mL of wash water (batch centrifuge, basket diameter 40.5 cm, 2050 rpm, 10 min). The yield of mevalonate from centrifugation was 33%. The sodium mevalonate purity of the centrifuge cake was 94% by DS, the wet cake moisture content was 5.3 wt.%, and the color value was 5200 ICUMSA. The sodium mevalonate purity of the centrifuge mother liquor was 68% by DS and the color value was 82000 ICUMSA.

[0207] Example 7 Crystallization of sodium mevalonate (Na-MVA) in water In this example, the crystallization of an organic acid or its salt in water, for example, the crystallization of sodium mevalonate (Na-MVA) in water, is described.

[0208] The crystallization feed was the evaporated nanofiltration permeate prepared according to Example 4. It was treated with activated carbon according to the procedure described in Example 6 (20 g Norit DX 1 carbon powder per kg DS, contact time 45 min, contact temperature 50° C., carbon powder was separated together with Kenite 300 filter aid using a Seitz depth filter). The sodium mevalonate purity of this feed was 73% by DS and the color number was 6300 ICUMSA.

[0209] The raw syrup was evaporated (Luwa thin film evaporator NL3-210 / 1600 / 10 evaporator and Rotavapor R-153 evaporator) until DS was 86.2%. 11.7 kg of the resulting syrup was transferred to a 10 liter cooling crystallizer and 2.0 g of sodium mevalonate dry seeds (prepared according to Example 5) were added at a temperature of 69° C. The seeded syrup was cooled to a temperature of 44° C. within 16 hours under continuous stirring. The crystallization feed was then diluted with deionized water to a DS of 85.4%. The diluted crystallization feed was cooled to a temperature of 40° C. within 3 hours and maintained at 40° C. for 2 hours.

[0210] The resulting crystallization material (10.9 kg) was centrifuged with 170 mL of wash water (batch centrifuge, basket diameter 40.5 cm, 2050 rpm, 10 min). The yield of mevalonate from centrifugation was 37%. The sodium mevalonate purity of the centrifuge cake was 95% by DS, the wet cake moisture content was 7.0 wt.%, and the color value was 1100 ICUMSA. The sodium mevalonate purity of the centrifuge mother liquor was 67% by DS and the color value was 14000 ICUMSA.

[0211] Example 8 Crystallization of sodium mevalonate (Na-MVA) in water This example describes the aqueous crystallization of an organic acid or its salt, e.g., the aqueous crystallization of sodium mevalonate (Na-MVA) from a syrup prepared by dissolving the crystal cake in deionized water.

[0212] The sodium mevalonate crystals from Example 6 and Example 7 were combined and dissolved in deionized water. The resulting solution was treated with activated carbon according to the procedure described in Example 6 (20 g Norit DX 1 carbon powder per kg DS, contact time 45 min, contact temperature 50° C., carbon powder was separated by Buchner filtration with Kenite 300 filter aid), filtered through a 0.2 μm sterile filter, and evaporated (Rotavapor R-151 evaporator) until the DS was 83.5%. The sodium mevalonate purity of the syrup was 96% by DS and the color value was 1300 ICUMSA.

[0213] 5.6 kg of the resulting syrup was transferred to a 6 liter cooling crystallizer and 1.6 g of sodium mevalonate dry seeds (prepared according to Example 5) were added at a temperature of 68° C. The seeded syrup was cooled to a temperature of 40° C. within 18 hours under continuous stirring. After cooling to 40° C., the crystallization material was maintained at 40° C. for 4 hours.

[0214] The resulting crystallization material (5.1 kg) was centrifuged with 150 mL of wash water (batch centrifuge, basket diameter 40.5 cm, 2050 rpm, 10 min). The yield of mevalonate from centrifugation was 42%. The sodium mevalonate purity of the centrifuge cake was ≥98% by DS, the wet cake moisture content was 1.5 wt.%, and the color value was 100 ICUMSA. The sodium mevalonate purity of the centrifuge mother liquor was 94% by DS and the color value was 3400 ICUMSA.

[0215] Example 9 Cation exchange from crystallized Na-MVA to mevalonolactone (MVL) In this example, we describe the cation exchange of a solution containing dissolved Na-MVA crystals to produce a dilute liquid containing mevalonolactone (MVL), which can be optionally concentrated by evaporation (see also FIG. 1B).

[0216] The treatment equipment included a jacketed glass column with an internal diameter of 45 mm and a total length of 1000 mm, a feed vessel placed above the column, a peristaltic pump at the column outlet, and a laboratory rotary evaporator, Büchi rotavapor R200. The column was packed with 1 liter of Dowex 88 strong acid cation exchange resin, giving a bed length of approximately 700 mm. The resin was backwashed and then washed with 4 liters (4 bed volumes) of 5% H2SO4 at a flow rate of 2 BV / h and H2SO4 at a temperature of 25°C. + Regenerated into a mold.

[0217] Sodium mevalonate crystals with a DS purity of about 98% obtained by a crystallization process with water were dissolved in water to produce a 24.2% solution, with the aim of exchanging the sodium cation to produce pure mevalonic acid or mevalonolactone syrup.

[0218] 957 grams of feed was passed through the column at a flow rate of 1 BV / h at room temperature (approximately 25° C.) and elution was continued with water. The pH of the feed was 9.78 and the conductivity was 39.4 mS / cm. Product collection was started when the column effluent brix was 0.4% and stopped when the effluent brix was 1.2%. 1322 grams of slightly colored product were collected. The column was again eluted with H + The mould was regenerated and the feed was passed through at a rate of 1 BV / h at room temperature (approximately 25°C). Product collection was started when the column effluent brix was 3.0% and was stopped when the effluent brix was 1.4%. 1641 grams of colourless product were collected. The material was rotary evaporated to KF-DS 99.1%. The DS purity of the MVL was 95.5%.

[0219] The results, including HPLC analysis (% dry basis) of the feed and ion exchange (IEX) products, are shown in Table 9.

[0220] [Table 9]

[0221] The overall yield of mevalonolactone from the IEX process was determined to be 95.6%.

[0222] The specific rotation of the mevalonolactone-containing syrup produced according to this process was -34.7° (water, c=1, 20°C).

[0223] Example 10 Cation exchange from Na-MVA solution to mevalonolactone (MVL) In this example, we describe the cation exchange of an aqueous solution containing Na-MVA to produce an aqueous solution containing mevalonolactone (MVL), which can optionally be concentrated by evaporation (see also FIG. 2).

[0224] The treatment equipment included a 100 liter tank, a Seitz depth filter, three jacketed glass columns with an internal diameter of 130 mm and a total length of 1500 mm, a feed vessel positioned above the columns, a peristaltic pump at the column outlet, and a LUWA thin film evaporator. Each column was packed with 13 liters of Dowex 88 strong acid cation exchange resin (Dow) to give a bed length of approximately 1000 mm. The resin was backwashed and eluted with 130 liters (approximately 3.3 bed volumes) of 5% H2SO4 at a flow rate of 2 BV / h and a temperature of 25°C. + Regenerated into a mold.

[0225] The purpose was to exchange the sodium cation to produce a mevalonic acid or mevalonolactone syrup for further purification.Potential colorants that precipitated from the IEX feed were removed using powdered activated carbon.

[0226] The nanofiltration permeate with a Na-MVA purity of 75.5% / DS and a color value of 10986 ICUMSA units was evaporated to 39.1% brix. 76.2 kg of this solution was heated to a temperature of 50-60 °C and Norit DX 1 powdered activated carbon with 2% / DS was added and mixed for 45 minutes. The suspension was passed through a Seitz depth filter with a filtration area of ​​0.56 m 2 T2600 filter sheet (Pall) and Kenite 300 (IMERYS Filtration) diatomaceous earth filter aid 1 kg / m as a precoat 2 The mixture was filtered using a 24.1% brix filter. 110 litres of the filtrate was evaporated to give a solution with a brix of 53% and a colour value of 6496 ICUMSA units. 14.8 kg of this material was diluted to 39.8% brix for the IEX feed and heated to 50°C.

[0227] 20.05 kg of feed was passed through the column at 50° C. at a rate of 2 BV / h and elution continued with water. Product collection was started when the final column effluent brix was 0.5% and stopped when the effluent brix was 4.0% and pH was 2.5. The product was collected in two fractions. The fractions were combined and evaporated. The combined evaporated IEX product had a color value of 2362 ICUMSA units and an MVL purity of about 76% / DS.

[0228] The results for the feed and IEX products are listed in Table 10.

[0229] [Table 10]

[0230] Example 11 Cation exchange from Na-MVA solution to mevalonolactone (MVL) In this example, we describe the cation exchange of an aqueous solution containing Na-MVA to produce an aqueous solution containing mevalonolactone (MVL), which can optionally be concentrated by evaporation (see also FIG. 2).

[0231] The treatment system included a 100 liter tank, a Seitz depth filter, three jacketed glass columns with an internal diameter of 130 mm and a total length of 1500 mm, a feed vessel positioned above the columns, a peristaltic pump at the column outlet, and a LUWA thin film evaporator. Each column was packed with 13 liters of Dowex 88 strong acid cation exchange resin (Dow) to give a bed length of approximately 1000 mm. The resin was backwashed and eluted with 130 liters (approximately 3.3 bed volumes) of 5% H2SO4 at a flow rate of 2 BV / h and a temperature of 25°C. + Regenerated into a mold.

[0232] The purpose was to exchange the sodium cation to produce a mevalonic acid or mevalonolactone syrup for further purification. Potential colorants that precipitated from the IEX feed were removed using powdered activated carbon.

[0233] The nanofiltration permeate, with a Na-MVA purity of 78.8% / DS and a color value of 8887 ICUMSA units, was evaporated to 42.0% brix. 41.4 kg of this solution was heated to a temperature of 55-58 °C and Norit DX 1 powdered activated carbon, 2.2% / DS, was added and mixed for 70 minutes. The suspension was passed through a Seitz depth filter with a filtration area of ​​0.28 m 2 T2600 filter sheet (Pall) and Kenite 300 (IMERYS Filtration) diatomaceous earth filter aid 1 kg / m as a precoat 2 The mixture was filtered using a filtration filter. 56 liters of filtrate was obtained with brix 23.8% and color value of 2124 ICUMSA units. The material was evaporated to brix 31.1% for IEX feed.

[0234] 35.7 kg of feed was passed through the column at a rate of 2 BV / h at 25° C. and elution was continued with water. Product collection was started when the final column effluent had a brix of 0.5% and was stopped when the effluent had a brix of 2.0% and a pH of 2.5. Product and residual fractions were collected. The MVL purity of the evaporated IEX product was about 78-83% / DS. The results for the feed and IEX product are listed in Table 11.

[0235] [Table 11]

[0236] Example 12 Cation exchange from Na-MVA solution to mevalonolactone (MVL) In this example, we describe the cation exchange of an aqueous solution containing Na-MVA to produce an aqueous solution containing mevalonolactone (MVL), which can optionally be concentrated by evaporation (see also FIG. 2).

[0237] The processing equipment included three jacketed glass columns with an internal diameter of 130 mm and a total length of 1500 mm, a feed vessel placed above the columns, and a peristaltic pump at the column outlet. Each column was packed with 13 liters of Dowex 88 strong acid cation exchange resin (Dow) to give a bed length of approximately 1000 mm. The resin was backwashed with 130 liters (approximately 3.3 bed volumes) of 5% H2SO4 at a flow rate of 2 BV / h and a temperature of 25°C. + Regenerated into a mold.

[0238] The purpose was to exchange the sodium cation to produce a mevalonic acid or mevalonolactone syrup for further purification.

[0239] The mother liquor obtained after Na-MVA crystallization, with a Na-MVA purity of 68.2% / DS and a color value of 82952 ICUMSA units, was diluted to 39.4% brix.

[0240] 17.0 kg of feed was passed through the column at a rate of 2 BV / h at 25° C. and elution continued with water. Product collection was started when the final column effluent brix was 0.8% and stopped when the effluent brix was 2.0%. The product was collected in three fractions. The fractions were combined and evaporated. The MVL purity of the combined and evaporated IEX product was approximately 70-73% / DS.

[0241] The results for the feed and IEX products are listed in Table 12.

[0242] [Table 12]

[0243] Example 13 Aqueous crystallization and MVL of mevalonolactone monohydrate * Preparation of HO seed crystals This example describes the crystallization of mevalonolactone monohydrate from water and the preparation of mevalonolactone monohydrate seed crystals.

[0244] A syrup of high purity MVL was prepared by the methods described herein (FIGS. 1A and 1B, Examples 1-9). Unexpectedly and surprisingly, when this high purity MVL syrup was stored at low temperature (approximately 6° C.) for several weeks, it was found that mevalonolactone monohydrate (MVL) * Spontaneous formation of HO crystals was observed.

[0245] The MVL purity of this syrup is 98% by DS (HPLC, H + The water content is about 6% w / w, which is the MVL * It was high enough to crystallize the H2O.

[0246] The crystals were filtered and the mother liquor was removed from the crystal surface by centrifugation (570 g, approx. 15° C. for 2 min) in a filter tube. The crystals and mother liquor were then analyzed. The water content of the crystals was 10.8% (by Karl Fischer method), which was slightly lower than the theoretical value of 12.1% due to the crystallization conditions. The water content of the mother liquor was 3.2%, which means that the mother liquor was concentrated during the formation of the monohydrate crystals. The MVL purity of the crystals was 98.8% by DS and that of the mother liquor was 97.7% / DS. These MVL * H2O crystals were used as seed crystals in Example 14. The crystals were ground in a porcelain grinder before use.

[0247] Example 14 Crystallization of mevalonolactone monohydrate in water This example describes the aqueous crystallization of mevalonolactone monohydrate from mevalonolactone syrup produced by subjecting sodium mevalonate crystallization mother liquor to cation exchange.

[0248] The centrifuged mother liquor containing sodium mevalonate obtained in Example 6 was treated with cation exchange in accordance with Example 12 to obtain a liquid containing mevalonolactone. The mevalonolactone purity of the obtained syrup was 71% in terms of DS and the color value was 34,000 ICUMSA.

[0249] The raw syrup was evaporated to a DS of 93.7% (Rotavapor R-153 evaporator). 2.1 kg of the resulting syrup was transferred to a 2 liter cooling crystallizer and 0.4 g of mevalonolactone monohydrate dry seeds (prepared according to Example 13) were added at a temperature of 11° C. The seeded syrup was cooled to a temperature of 7° C. within 16 hours under continuous stirring. After cooling to 7° C., stirring was continued at 7° C. for 50 hours. While the temperature was constant, the crystallization feed was diluted with deionized water in three portions as follows: (1) 40 g added 23 hours after seeding, (2) 50 g added 43 hours after seeding, and (3) 10 g added 47 hours after seeding. The DS of the resulting crystallization feed was 89.1%.

[0250] 1.7 kg of crystallization raw material was centrifuged without using washing water by batch centrifugation (basket diameter 22.5 cm, 4000 rpm, 3 min). The yield of mevalonolactone by centrifugation was 52%. The mevalonolactone purity of the centrifugation cake was 94% by DS, the moisture content of the wet cake was 10.8% by weight, and the color value was 5200 ICUMSA. The mevalonolactone purity of the centrifugation mother liquor was 56% by DS, and the color value was 52000 ICUMSA.

[0251] Mevalonolactone monohydrate crystals prepared according to this process were used as seed crystals in Example 15. The crystals were ground in a porcelain grinder before use.

[0252] Example 15 Crystallization of mevalonolactone monohydrate in water In this example, we describe the aqueous crystallization of mevalonolactone monohydrate from a mevalonolactone-containing syrup produced by using nanofiltration followed by a cation exchange step (FIG. 2).

[0253] The nanofiltration permeate containing sodium mevalonate prepared according to Example 4 was subjected to a cation exchange treatment according to Example 10 to obtain a liquid containing mevalonolactone. The mevalonolactone purity of the obtained syrup was 76% by DS and the color value was 2400 ICUMSA.

[0254] The raw syrup was evaporated to a DS of 90.7% (Rotavapor R-153 evaporator). 6.3 kg of the resulting syrup was transferred to a 6 liter cooling crystallizer and 0.5 g of mevalonolactone monohydrate dry seeds (prepared according to Example 14) were added at a temperature of 12° C. The seeded syrup was cooled to a temperature of 6° C. within 17 hours under continuous stirring. After cooling to 6° C., stirring was continued at 6° C. for 5 hours. While the temperature was constant at 6° C., the crystallization raw material was diluted with deionized water in three portions as follows: (1) 50 g added 18 hours after seeding, (2) 40 g added 19 hours after seeding, and (3) 70 g added 20 hours after seeding. The resulting crystallization raw material had a DS of 88.1%.

[0255] 5.2 kg of the resulting crystallization material was centrifuged in four batches using wash water in an amount of 27-33 mL / kg DS mass (batch centrifugation, basket diameter 22.5 cm, 3500 rpm, 3 min). A crystal cake sample and a mother liquor sample were collected from the first centrifugation. The yield of mevalonolactone from the first centrifugation was 71%. The mevalonolactone purity of the first centrifugation cake was 93% by DS, the moisture content of the wet cake was 12.7 wt%, and the color value was 560 ICUMSA. The mevalonolactone purity of the first centrifugation mother liquor was 55% by DS and the color value was 5700 ICUMSA.

[0256] Mevalonolactone monohydrate crystals prepared according to this process were used as seed crystals in Examples 16 and 17. The crystals were ground in a porcelain grinder before use.

[0257] Example 16 Crystallization of mevalonolactone monohydrate in water In this example, we describe the aqueous crystallization of mevalonolactone monohydrate from a mevalonolactone-containing syrup produced by using nanofiltration followed by a cation exchange step (FIG. 2).

[0258] The nanofiltration permeate containing Na-MVA prepared according to Example 3 was subjected to a cation exchange treatment according to Example 11 to produce a liquid containing MVL. The mevalonolactone purity of the resulting syrup was 81% by DS and the color value was 1200 ICUMSA.

[0259] The raw syrup was evaporated to a DS of 84.1% (Rotavapor R-153 evaporator). 6.2 kg of the resulting syrup was transferred to a 6 liter cooling crystallizer and 1.0 g of mevalonolactone monohydrate dry seeds (prepared according to Example 15) were added at a temperature of 16° C. The seeded syrup was maintained at 16° C. for 3 hours and then cooled to a temperature of 6° C. within 11 hours with continuous stirring. After cooling to 6° C., stirring was continued at 6° C. for 7 hours.

[0260] 5.5 kg of the obtained crystallization raw material was centrifuged in five batches using wash water in an amount of 72-79 mL / kg DS mass (batch centrifugation, basket diameter 22.5 cm, 3500 rpm, 3 min). A crystal cake sample and a mother liquor sample were collected from the first centrifugation. The yield of mevalonolactone from the first centrifugation was 62%. The mevalonolactone purity of the first centrifugation cake was ≥97% by DS, the moisture content of the wet cake was 11.6 wt%, and the color value was 61 ICUMSA. The mevalonolactone purity of the first centrifugation mother liquor was 64% by DS and the color value was 2700 ICUMSA.

[0261] Example 17 Crystallization of mevalonolactone monohydrate in water This example describes the aqueous crystallization of mevalonolactone monohydrate from a mixture of centrifugation mother liquor and diluted crystallization feed.

[0262] The crystallization feed was an aqueous syrup containing mevalonolactone, obtained by combining the centrifugation mother liquor of Example 16 with the diluted crystallization feed recovered by washing the equipment with deionized water. The mevalonolactone purity of the obtained syrup was 70% by DS and the color value was 2400 ICUMSA.

[0263] The raw material was evaporated to a DS of 85.3% (Rotavapor R-153) and 2.8 kg of the resulting syrup was transferred to a 2 L cooling crystallizer. The syrup was seeded twice: once with 1.0 g of mevalonolactone monohydrate dry seeds (prepared according to Example 15) at a temperature of 13° C., and then with 1.0 g of mevalonolactone monohydrate dry seeds at a temperature of 10° C. The first seeding produced very fine crystals.

[0264] The seeded syrup was maintained at 10° C. for 35 minutes and then cooled to a temperature of 4° C. within 16 hours with continuous stirring. After cooling to 4° C., stirring was continued at 4° C. for 2 hours.

[0265] The resulting crystallization material (2.5 kg) was centrifuged in two batches (batch centrifuge, basket diameter 22.5 cm, 3500 rpm, 3 min). The amount of washing water in the first centrifugation was 90 mL / kg DS. The yield of mevalonolactone obtained by centrifugation was 42%. The mevalonolactone purity of the centrifuge cake was ≥ 97% by DS, the moisture content of the wet cake was 14.7 wt%, and the color value was 33 ICUMSA. The mevalonolactone purity of the centrifuge mother liquor was 58% by DS and the color value was 4200 ICUMSA.

[0266] The second centrifugation was performed without using wash water. The yield of mevalonolactone obtained by centrifugation was 54%. The mevalonolactone purity of the centrifuge cake was ≥97% by DS, the moisture content of the wet cake was 11.7% by weight, and the color value was 120 ICUMSA. The mevalonolactone purity of the centrifuge mother liquor was 53% by DS, and the color value was 4600 ICUMSA.

[0267] Mevalonolactone monohydrate crystals prepared according to this process were used as seed crystals in Examples 18 and 19. The crystals were ground in a porcelain grinder before use.

[0268] Example 18 Crystallization of mevalonolactone monohydrate in water In this example, recrystallization of mevalonolactone monohydrate crystals dissolved in deionized water using water as a solvent is described.

[0269] The crystallization feedstock was an aqueous syrup containing mevalonolactone, obtained by dissolving and diluting 2.2 kg of the centrifuge cake from Example 15 with the addition of deionized water. The mevalonolactone purity of the resulting syrup was 92% by DS and the color value was 560 ICUMSA.

[0270] The raw syrup was evaporated to a DS of 80.8% (Rotavapor R-153), and 2.3 kg of the resulting syrup was transferred to a 2 L cooling crystallizer. The syrup was cooled without seeding under continuous stirring in two stages: (1) from 20° C. to 15° C. within 2 hours, and (2) from 15° C. to 10° C. within 10 hours. After cooling to 10° C., stirring was continued at 10° C. for 6 hours, which ensured that no spontaneous crystal formation occurred.

[0271] To this syrup, 0.5 g of mevalonolactone monohydrate dry seeds (prepared according to Example 17) were added at a temperature of 10° C. The cooling water temperature of the crystallizer jacket was kept constant at 7° C. Crystal formation started immediately after the addition of the seeds. The temperature of the crystallization feedstock first increased from 10° C. to 16° C. within 50 minutes of the seed addition due to the heat of crystallization, and then decreased from 16° C. to 12° C. within 3 hours. After decreasing to 12° C., the cooling water temperature was increased from 7° C. to 12° C. and stirring was continued at 12° C. for 1 hour.

[0272] The resulting 2.2 kg of crystallization material was centrifuged in two batches (batch centrifugation, basket diameter 22.5 cm, 3500 rpm, 3 min) using wash water in an amount of 65-66 mL / kg DS mass. A crystal cake sample and a mother liquor sample were collected from the first centrifugation. The yield of mevalonolactone from the first centrifugation was 52%. The mevalonolactone purity of the first centrifugation cake was ≥98% by DS, the moisture content of the wet cake was 13.6 wt%, and the color value was 35 ICUMSA. The mevalonolactone purity of the first centrifugation mother liquor was 87% by DS and the color value was 1100 ICUMSA.

[0273] Example 19 Crystallization of mevalonolactone monohydrate in water This example describes the recrystallization of mevalonolactone monohydrate crystals dissolved in deionized water using water as a solvent.

[0274] The crystallization feed was an aqueous syrup containing mevalonolactone, obtained by combining 2.0 kg of centrifuge cake from Example 16 with 320 g of centrifuge cake from Example 17. Prior to mixing, the mevalonolactone product from Example 16 was diluted with deionized water to a DS of 80.9%, and the resulting syrup was treated with activated carbon according to the procedure described in Example 6 (20 g Norit DX 1 carbon powder per kg DS, contact time 1 hour, contact temperature 50° C., carbon powder was separated by Buchner filtration with Kenite 300 filter aid). The mevalonolactone purity of the crystallization feed syrup was ≧97% by DS and the color was 34 ICUMSA.

[0275] The raw syrup was evaporated to a DS of 78.3% (Rotavapor R-151 evaporator). 2.6 kg of the resulting syrup was transferred to a 5 liter cooling crystallizer and 0.4 g of mevalonolactone monohydrate dry seeds (prepared according to Example 17) were added at a temperature of 17° C. The seeded syrup was maintained at 17° C. for 1.5 hours and then cooled to a temperature of 13° C. within 15 hours with continuous stirring. After cooling to 13° C., stirring was continued at 13° C. for 5 hours.

[0276] The resulting 2.4 kg of crystallization material was centrifuged in two batches (batch centrifugation, basket diameter 22.5 cm, 3500 rpm, 3 min) using wash water in an amount of 65-71 mL / kg DS mass. A crystal cake sample and a mother liquor sample were collected from the first centrifugation. The yield of mevalonolactone from the first centrifugation was 50%. The mevalonolactone purity of the first centrifugation cake was ≥ 98% by DS, the moisture content of the wet cake was 12.1 wt%, and the color value was 3 ICUMSA. The mevalonolactone purity of the first centrifugation mother liquor was 96% by DS and the color value was 70 ICUMSA.

[0277] Mevalonolactone monohydrate crystals prepared according to this process were used as seed crystals in Example 20. The crystals were ground in a porcelain grinder before use.

[0278] Example 20 Crystallization of mevalonolactone monohydrate in water This example describes the crystallization of mevalonolactone monohydrate in water and the preparation of a high purity aqueous syrup of mevalonolactone.

[0279] The crystallization feed was an aqueous syrup containing mevalonolactone, obtained by combining the second centrifuge cake from Example 17 with the centrifuge mother liquors from Examples 18 and 19. The syrup was diluted to a DS of 51.2% by adding deionized water, and the resulting solution was treated with activated carbon according to the procedure described in Example 6 (20 g Norit DX 1 carbon powder per kg DS, contact time 1 h, contact temperature 50° C., carbon powder separated by Buchner filtration together with Kenite 300 filter aid). The mevalonolactone purity of the resulting syrup was 93% by DS and the color value was 170 ICUMSA.

[0280] The carbon treated raw syrup was evaporated to a DS of 80.3% (Rotavapor R-153 evaporator). 2.6 kg of the resulting syrup was transferred to a 2 liter cooling crystallizer and 0.6 g of mevalonolactone monohydrate dry seeds (prepared according to Example 19) were added at a temperature of 18° C. The seeded syrup was maintained at 18° C. for 1 hour and then cooled to a temperature of 13° C. within 15 hours with continuous stirring. After cooling to 13° C., stirring was continued at 13° C. for 4 hours.

[0281] 2.3 kg of the resulting crystallization material was centrifuged in two batches using wash water in an amount of 67-68 mL / kg DS mass (batch centrifugation, basket diameter 22.5 cm, 3500 rpm, 3 min). A crystal cake sample and a mother liquor sample were collected from the first centrifugation. The yield of mevalonolactone from the first centrifugation was 51%. The mevalonolactone purity of the first centrifugation cake was ≥98% by DS, the moisture content of the wet cake was 12.3 wt%, and the color value was 3 ICUMSA. The mevalonolactone purity of the first centrifugation mother liquor was 87% by DS and the color value was 370 ICUMSA.

[0282] DSC analysis of mevalonolactone monohydrate crystals prepared according to this process showed an endothermic peak with a maximum at 22.6°C.

[0283] The centrifuge cakes were combined and deionized water was added to a DS of 57.4% to give a liquid mevalonolactone product. The resulting syrup was evaporated to a DS of ≥ 97% (Rotavapor R-153 evaporator). After evaporation, the syrup had a mevalonolactone purity of ≥ 98% by DS and a color value of 2 ICUMSA.

[0284] MVL * Exemplary conditions for crystallizing HO are shown in Tables 13A-13E.

[0285] [Table 13A]

[0286] [Table 13B]

[0287] [Table 13C]

[0288] [Table 13D]

[0289] [Table 13E]

[0290] Furthermore, MVL * H2O can be crystallized at temperatures below 0°C up to the freezing point of the mother liquor, provided that cooling equipment is available and the viscosity of the crystallization feedstock is tolerable.

[0291] Example 21 Mevalonolactone monohydrate (MVL * Characterization of HO Formula C6H 10 O3 * Crystalline mevalonolactone monohydrate (MVL) with HO * HO) was further characterized as follows:

[0292] AX-ray crystal structure MVL * X-ray diffraction measurements of H2O were carried out.

[0293] B. Melting point MVL-monohydrate (MVL * The melting point (mp) of HO) is 20-25°C, preferably 21-24°C, depending on the purity of the crystals. Crystallization is only possible below the mp. The melting point was determined using the peak temperature of a differential scanning calorimeter (DSC). DSC thermograms were measured using a Mettler Toledo DSC822e differential scanning calorimeter. The measurements were carried out in 40 μL standard aluminum crucibles under a nitrogen gas flow with a flow rate of 80 mL / min. The temperature range was 0-50°C, and the heating rate was 2°C / min.

[0294] C. Moisture content of crystals The theoretical water content of MVL monohydrate calculated from the molecular weight is 12.15% (DS content 87.85%) (MVL* The molecular weight of H2O is 148.16 g / mol). This corresponds to crystals crystallized under conditions with excess water. If the water present in the crystallization syrup is less than the water of crystallization content, the mother liquor is concentrated during crystallization and the crystals formed have a lower water of crystallization content than the theoretical value. The water of crystallization content of the forming MVL monohydrate crystals was observed to be 10.8% (by Karl Fischer method) as described in Example 13. A water of crystallization content of 10-11% was also observed for the MVL monohydrate crystals formed in high DS (96 and 89) syrups.

[0295] D.Water soluble Solubility is one of the properties that characterize crystalline compounds. Crystallization is only possible when the concentration of a supersaturated syrup exceeds the solubility. Solubility determines the minimum concentration of DS that will crystallize at a given temperature. Solubility can be determined by standard methods such as analyzing the equilibrium concentration from a crystal suspension.

[0296] MVL * The water solubility of H2O was calculated by using crystals with a purity of 93% and converting it to a purity of 100. The DS content at equilibrium was measured and calculated in the range of 0 to 20°C (Table 14).

[0297] [Table 14]

[0298] E. Crystal shape Figure 3 shows the results of the study of mevalonolactone monohydrate (MVL) * The figure shows the typical shape of a HO crystal. The crystals grow easily, large and quickly, releasing a large amount of heat of crystallization.

Claims

1. 1. A method for producing mevalonolactone from an aqueous solution containing a salt of a mevalonate (X-MVA), the method comprising subjecting the aqueous solution containing the salt of mevalonate to cation exchange, thereby converting the aqueous solution containing the salt of mevalonate to an aqueous solution containing mevalonolactone (MVL).

2. The salts of mevalonic acid include sodium mevalonate (Na-MVA), potassium mevalonate (K-MVA), ammonium mevalonate (NH ４ 2. The method of claim 1, wherein the monovalent salt of mevalonic acid is selected from the group consisting of lithium mevalonate (Li-MVA), lithium mevalonate (Li-MVA), any other monovalent salt of mevalonic acid, or any combination thereof.

3. 10. The method of claim 1, further comprising concentrating the aqueous solution containing mevalonolactone to a liquid containing at least 65% to 90% DS.

4. Mevalonolactone monohydrate (MVL ＊ H ２ The method according to claim 1 or claim 3, further comprising: O) cooling said aqueous solution or liquid to 0-25° C. to obtain crystals.

5. In order to obtain a dilute liquid containing mevalonolactone, the mevalonolactone purity of which is at least 90%, ＊ H ２ 5. The method of claim 4, further comprising dissolving the O crystals in water.

6. The method of claim 1 , wherein the aqueous solution is a fermentation broth.

7. 7. The method of claim 6, wherein the fermentation broth is clarified by removing biomass and any solids from the fermentation broth by at least one of precoat filtration, microfiltration, or ultrafiltration.

8. 7. The method of claim 6, wherein the fermentation broth is a broth obtained by fermentation of Escherichia coli.

9. A pharmaceutical or homecare or nutritional or personal care or cosmetic composition comprising MVL produced according to any one of claims 1 to 6 or a solubilized form thereof.

10. Crystalline mevalonolactone obtained by the method according to claim 15. ＊ Monohydrate (MVL ＊ H ２ O).

11. Mevalonolactone monohydrate (MVL ＊ H ２ O) A composition comprising crystals.

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