Recovery process for 3-hydroxypropionate crystals and 3-hydroxypropionic acid

Abstract

To provide a process of recovering 3-hydroxypropionic acid and a crystal of the 3-hydroxypropionate, where the process includes the step of forming crystals of 3-hydroxypropionate.SOLUTION: The invention relates to a process of recovering 3-hydroxypropionic acid and a crystal of the 3-hydroxypropionate, where the process includes the steps of: forming crystals of 3-hydroxypropionate in a concentrate containing 3-hydroxypropionic acid of 300 g / L or more in the presence of an alkali metal salt; and separating the crystals of 3-hydroxypropionate from the concentrate and converting them into 3-hydroxypropionic acid.SELECTED DRAWING: None

Description

[Technical Field]

[0001] [Cross-reference of related applications]

[0002] This application claims priority rights under Korean Patent Application No. 10-2021-0006246 dated January 15, 2021, and Korean Patent Application No. 10-2021-0119997 dated September 8, 2021, and all content disclosed in the documents of said Korean Patent Applications is incorporated herein by reference.

[0003] The present invention relates to a process for recovering 3-hydroxypropionate crystals and 3-hydroxypropionic acid (3HP), and more specifically, to a process for recovering 3-hydroxypropionic acid using the crystallization of 3-hydroxypropionate and to the 3-hydroxypropionate crystals formed in the said process. [Background technology]

[0004] 3-hydroxypropionic acid (3HP) is a platform compound that can be converted into a variety of chemicals, including acrylic acid, methyl acrylate, and acrylamide. Since being selected as one of the Top 12 value-added biochemicals by the U.S. Department of Energy (DOE) in 2004, it has been actively researched in academia and industry.

[0005] The production of 3-hydroxypropionic acid is carried out by two main methods: chemical and biological. However, chemical methods are criticized for being unenvironmentally unfriendly due to the high cost of initial materials and the generation of toxic substances during the production process, leading to increased attention on environmentally friendly bio-processes.

[0006] During the production of organic acids by microbial fermentation, other by-products besides organic acids, such as 3-hydroxypropionic acid, are also produced during the fermentation process. Therefore, a process of extracting and separating the organic acids from the fermentation liquid is necessary. Methods for extracting and separating organic acids from microbial fermentation liquid include electrodialysis, reverse osmosis, and organic acid-organic solvent reaction extraction. In particular, the back extraction method using sodium hydroxide (NaOH) is widely used due to its high yield. However, these methods have the disadvantage of low purity because the products are in the form of organic acid salts, requiring further steps to convert them into organic acids.

[0007] Unlike other organic acids produced through fermentation, 3-hydroxypropionic acid exhibits high hydrophilicity, high solubility and reactivity in water. Therefore, conventional separation and purification processes for organic acids, such as precipitation or extraction, are difficult to apply.

[0008] Reaction extraction is a method of extracting organic acids using an active diluent such as an amine or alcohol that reacts well with the organic acid. This method allows for the selective extraction of organic acids and has relatively high extraction efficiency. Therefore, attempts have been made to apply this reaction extraction method to the separation and purification of 3HP. As an example, a method using trioctylamine (TOA) as the amine has been proposed, but it has the problem of low extraction efficiency for 3HP and the need for a large amount of organic solvent. Also, when tridecylamine is used as the amine, the extraction efficiency for 3HP is higher than that of TOA, but there is a problem of emulsion occurring during extraction, where the organic phase and aqueous phase do not separate.

[0009] Therefore, there is a need to develop a process for recovering 3-hydroxypropionic acid in high purity and high yield from raw material liquids containing 3-hydroxypropionic acid, such as microbial fermentation liquids. [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] The object of the present invention is to provide a 3-hydroxypropionic acid recovery process that includes a step of generating 3-hydroxypropionate crystals, and the 3-hydroxypropionate crystals. [Means for solving the problem]

[0011] The present invention provides a 3-hydroxypropionic acid recovery step comprising the steps of forming 3-hydroxypropionate crystals in a concentrated solution containing 300 g / L or more of 3-hydroxypropionic acid in the presence of an alkali metal salt, and separating the 3-hydroxypropionate crystals from the concentrated solution and converting them to 3-hydroxypropionic acid.

[0012] Furthermore, the present invention provides crystals of 3-hydroxypropionate represented by the following structural formula 1 or structural formula 2.

[0013] [Structural formula 1] Cation (3HP) n [Structural formula 2] Cation (3HP) n ·mH2O In the above structural formula, Cation is a positive ion (Cation), 3HP is 3-hydroxypropionic acid that binds to a cation. n is an integer greater than or equal to 1, where n is the number of 3HPs that bond with the cation. m is an integer greater than or equal to 1, representing the number of water molecules in the hydrate.

[0014] The following describes in more detail the 3-hydroxypropionic acid recovery process and the 3-hydroxypropionate crystals according to specific embodiments of the present invention.

[0015] Unless explicitly stated or otherwise specifically mentioned, the steps constituting a manufacturing method described herein are sequential or continuous, one step and another constituting a single manufacturing method shall not be construed as being limited to the order described herein. Therefore, the order of the steps constituting a manufacturing method can be changed to the extent that is easily understood by those skilled in the art, and in this case, any changes that are obvious to those skilled in the art are included within the scope of the present invention.

[0016] According to one embodiment of the present invention, a 3-hydroxypropionic acid recovery step is provided, which includes the steps of forming 3-hydroxypropionate crystals in a concentrated solution containing 300 g / L or more of 3-hydroxypropionic acid in the presence of an alkali metal salt, and separating the 3-hydroxypropionate crystals from the concentrated solution and converting them to 3-hydroxypropionic acid.

[0017] The inventors of the present invention have experimentally confirmed that when 3-hydroxypropionic acid is concentrated to a high concentration in the presence of an alkali metal salt, 3-hydroxypropionate crystals can be formed. They have also experimentally confirmed that the 3-hydroxypropionate crystals thus formed can be easily separated (and purified) from the liquid phase, and that high-purity 3-hydroxypropionate can be recovered with high efficiency, thus completing the invention.

[0018] More specifically, a concentrate containing 3-hydroxypropionic acid may contain 3-hydroxypropionic acid at concentrations of 300 g / L or more, 350 g / L or more, 400 g / L or more, 450 g / L or more, and 500 g / L or more, and may contain 3-hydroxypropionic acid at concentrations of 900 g / L or less, 850 g / L or less, and 800 g / L or less.

[0019] The presence or absence of 3-hydroxypropionic acid crystal formation appears to be influenced by factors such as the presence of alkali metal salts and the concentration of 3-hydroxypropionic acid in the concentrate.

[0020] Furthermore, if the concentration of 3-hydroxypropionic acid crystals in the concentrated solution is higher than the water solubility of 3-hydroxypropionic acid crystals, 3-hydroxypropionic acid crystals are formed more easily.

[0021] For example, since the water solubility of Ca(3HP)2, a crystalline form of 3-hydroxypropionic acid, is 450 g / L at room temperature, if the concentration of 3-hydroxypropionic acid in the concentrate exceeds 450 g / L, the formation of Ca(3HP)2 crystals is promoted. Also, since the water solubility of Mg(3HP)2, a crystalline form of 3-hydroxypropionic acid, is 250 g / L at room temperature, if the concentration of 3-hydroxypropionic acid in the concentrate exceeds 250 g / L, the formation of Mg(3HP)2 crystals is promoted.

[0022] Crystals of 3-hydroxypropionate can be formed from a concentrated solution containing an alkali metal salt and satisfying the above-mentioned concentration, and the alkali metal salt can be selected without limitation within the scope intended for forming 3-hydroxypropionate crystals, for example, the alkali metal salt is Na + Mg 2+ and Ca 2+ It may contain one or more cations selected from the group consisting of Mg 2+ or Ca 2+ When this is present, 3-hydroxypropionate crystals can be formed more effectively. For example, the alkali metal salt may be Ca(OH)2, Mg(OH)2, or a mixture thereof.

[0023] The alkali metal salt can be added and remain in the process of producing the 3-hydroxypropionic acid fermentation broth, or can be added during the process of forming the crystals of 3-hydroxypropionate in a concentrated solution containing 3-hydroxypropionic acid at 300 g / L or more. Also, the concentration of the alkali metal salt is 10% to 100%, or 30% to 90% of the concentration of 3-hydroxypropionic acid, for example, at a concentration of 10 to 900 g / L, 50 to 800 g / L, 100 to 700 g / L or 200 to 600 g / L, and may be present in the concentrated solution.

[0024] The crystals of 3-hydroxypropionate can be in the form of the following Structural Formula 1 or Structural Formula 2. That is, the crystals of 3-hydroxypropionate can contain 3-hydroxypropionate in the form of the following Structural Formula 1 or Structural Formula 2.

[0025] In the following Structural Formula 1 and Structural Formula 2, Cation is a cation, 3HP is 3-hydroxypropionic acid that binds to the cation, n is the number of 3HPs that bind to the cation, meaning an integer of 1 or more, and in the following Structural Formula 2, m is the number of water molecules that bind to Cation(3HP)n as a hydrate and is an integer of 1 or more. The cation can be, for example, Na + , Mg 2+ or Ca 2+ , but in the case of Mg 2+ or Ca 2+ , the crystals of 3-hydroxypropionate are formed more effectively.

[0026] [Structural Formula 1] Cation(3HP) n [Structural Formula 2] Cation(3HP) n ·mH2O

[0027] Also, the stage of forming the crystals of 3-hydroxypropionate may further include the stage of stirring the concentrated solution.

[0028] The stirring stages are as follows: 0-70°C, 0-60°C, 0-50°C, 0-40°C, 0-35°C, 0-30°C, 10-70°C, 10-60°C, 10-50°C, 10-40°C, 10-35°C, 10-30°C, 15-70°C, 15-60°C, 15-50°C, 15-40°C, 15-35°C, 15-30°C, 20°C This can be done at temperatures of ~70 degrees Celsius, 20-60 degrees Celsius, 20-50 degrees Celsius, 20-40 degrees Celsius, 20-35 degrees Celsius, or 20-30 degrees Celsius (e.g., room temperature) and / or under conditions of 100-2000 rpm, 100-1500 rpm, 100-1000 rpm, 100-500 rpm, 100-400 rpm, or 200-400 rpm (e.g., about 300 rpm).

[0029] Particle size distribution D of 3-hydroxypropionate crystals 50 These can be between 20 μm and 90 μm, between 25 μm and 85 μm, between 30 μm and 80 μm, or between 35 μm and 75 μm.

[0030] Furthermore, the particle size distribution D of the 3-hydroxypropionate crystals 10 The particle size distribution D of the 3-hydroxypropionate crystals is 5 μm to 40 μm, 8 μm to 35 μm, and 10 μm to 30 μm. 90 These can be between 50 μm and 200 μm, between 60 μm and 190 μm, between 65 μm and 180 μm, or between 70 μm and 175 μm.

[0031] Particle size distribution D 50 , D 10 , D 90 In the particle size distribution curve, these represent the particle sizes corresponding to 50%, 10%, and 90% of the cumulative volume, respectively. 50 , D 10 , D 90This can be measured, for example, using the laser diffraction method. The laser diffraction method can typically measure particle sizes ranging from the submicron region to several millimeters in size, and can yield highly reproducible and high-resolution results.

[0032] Particle size distribution D of 3-hydroxypropionate crystals 50 , D 10 , D 90 If the size is too large, impurities that must be removed during crystallization will be included in the crystal, reducing the purification efficiency. If it is too small, the permeability of the liquid will decrease during filtration of the crystal.

[0033] On the other hand, the crystals of 3-hydroxypropionate (D 90 -D 10 ) / D 50 It could be between 1.00 and 3.00, between 1.20 and 2.80, between 1.40 and 2.60, or between 1.60 and 2.40.

[0034] Furthermore, the crystals of 3-hydroxypropionate may have a volume-average diameter of 30 μm to 100 μm, 35 μm to 95 μm, or 40 μm to 90 μm; a number-average diameter of 1 μm to 30 μm, 3 μm to 25 μm, or 5 μm to 20 μm; and an area-average diameter of 10 μm to 70 μm, 15 μm to 60 μm, or 20 μm to 55 μm.

[0035] If the volume-average diameter, number-average diameter, and area-average diameter of 3-hydroxypropionate crystals are too large, impurities that must be removed during crystallization will be included in the crystals, reducing the purification efficiency. If they are too small, the permeability of the crystals during filtration will decrease.

[0036] Furthermore, the particle size distribution of 3-hydroxypropionate crystals (D 10 , D 50 , D 90In this case, the aspect ratio (LW ratio; length to width ratio) and the average aspect ratio may be between 0.50 and 3.00, between 0.70 and 2.80, and between 1.00 and 2.50, respectively. If the aspect ratio of the 3-hydroxypropionate crystals is too large, flow and clogging problems may occur during crystal transport, and if it is too small, the permeability will decrease during crystal filtration.

[0037] The water content of 3-hydroxypropionate crystals can be measured using the Karl Fischer method, and the water content of the 3-hydroxypropionate crystals may be between 200 ppm and 5000 ppm, between 250 ppm and 4800 ppm, between 300 ppm and 4600 ppm, or between 350 ppm and 4400 ppm.

[0038] In this case, the water contained in the 3-hydroxypropionate crystal refers to adhering water contained within the crystal, not crystal water (e.g., Ca(3HP)2·2H2O). Furthermore, if the water content in the 3-hydroxypropionate crystal is too high, it may be recovered in slurry form rather than as a crystalline solid, or impurities may be present in the water, leading to a decrease in purity.

[0039] In the recovery process of 3-hydroxypropionic acid according to one embodiment, 3-hydroxypropionic acid is produced by a process such as fermenting a bacterial strain having the ability to produce 3-hydroxypropionic acid, as described later, thereby the crystals of 3-hydroxypropionate are radioactive carbon isotopes ( 14 This may include C).

[0040] Radioactive carbon isotopes ( 14 C) is 10 carbon atoms in the Earth's atmosphere 12 Each atom contains almost one molecule, and its half-life is approximately 5700 years. Carbon stock is formed by cosmic rays and normal nitrogen. 14It can be abundant in the upper atmosphere due to nuclear reactions in which N) participates. On the other hand, in fossil fuels, radioactive carbon isotopes have already decayed. 14 The carbon ratio may be virtually zero. When bio-derived raw materials are used as the raw material for 3-hydroxypropionic acid, or when fossil fuels are used in combination with them, the radioactive carbon isotope content (pMC; percent modern carbon) and biocarbon content in 3-hydroxypropionic acid can be measured according to the ASTM D6866-21 standard.

[0041] The measurement method involves, for example, converting the carbon atoms contained in the target compound into graphite or carbon dioxide gas form and measuring them using a mass spectrometer or liquid flash spectroscopy. At this time, along with the mass spectrometer, 14 C ions 12 By using an accelerator to separate the two isotopes from the C ion, the content and content ratio can be measured using a mass spectrometer.

[0042] Crystals of 3-hydroxypropionate may have a radiocarbon isotope content of 20 pMC (percent modern carbon) or more, 50 pMC or more, 90 pMC or more, or 100 pMC or more, as measured according to ASTM D6866-21 standards, and a biocarbon content of 20% by weight or more, 50% by weight or more, 80% by weight or more, 90% by weight or more, or 95% by weight or more.

[0043] The aforementioned radiocarbon isotope ratio (pMC) is the radiocarbon isotope contained in the crystal of 3-hydroxypropionate ( 14 C) and radioactive carbon isotopes of modern reference materials ( 14 This refers to the ratio of C), which indicates that the effects of the nuclear test program from the 1950s have not disappeared and are still present, and may be greater than 100%.

[0044] Furthermore, the biocarbon content refers to the amount of biocarbon relative to the total carbon content in the 3-hydroxypropionate crystal, and a higher value indicates a more environmentally friendly compound.

[0045] On the other hand, if the radiocarbon isotope content (pMC) and biocarbon content of 3-hydroxypropionate crystals are too low, their environmental friendliness decreases, and they are not considered to be bio-derived materials.

[0046] The crystalline state of 3-hydroxypropionate can be confirmed by peaks and other features in an X-ray diffraction (XRD) graph.

[0047] For example, when the 3-hydroxypropionate crystal is analyzed by X-ray diffraction (XRD), peaks appear between the crystal lattice when the 2θ value is in the range of 8 to 22 degrees.

[0048] For example, since the concentrate contains magnesium hydroxide (Mg(OH)2), if the formed 3-hydroxypropionate crystal is Mg(3HP)2, then during X-ray diffraction (XRD) analysis of Mg(3HP)2, peaks appear between the crystal lattice due to the 3-hydroxypropionic acid and magnesium bond in the 2θ range of 8 to 15 degrees. Such peaks show different results from X-ray diffraction (XRD) analysis results for magnesium hydroxide (Mg(OH)2) or magnesium sulfate (Mg(SO4)). If a specific peak appears in the X-ray diffraction (XRD) analysis results in the 2θ range of 8 to 15 degrees, it can be confirmed that Mg(3HP)2 crystals have been formed.

[0049] Specifically, during X-ray diffraction (XRD) analysis of Mg(3HP)2, three or more, four or more, or five or more peaks appear in the 2θ range of 8 to 15 degrees. For example, peaks appear in the ranges of 8.2 to 9.3 degrees, 9.5 to 11.0 degrees, 11.2 to 12.7 degrees, 12.9 to 13.3 degrees, and 13.5 to 14.8 degrees.

[0050] Furthermore, since the concentrate contains calcium hydroxide (Ca(OH)2), if the formed 3-hydroxypropionate crystals are Ca(3HP)2, X-ray diffraction (XRD) analysis of Ca(3HP)2 will show peaks between the crystal lattice due to the 3-hydroxypropionic acid and calcium bond in the 2θ range of 10 to 22 degrees. Such peaks show different results from X-ray diffraction (XRD) analysis results for calcium hydroxide (Ca(OH)2) or calcium sulfate (Ca(SO4)), and if a specific peak appears in the X-ray diffraction (XRD) analysis results in the 2θ range of 10 to 22 degrees, it can be confirmed that Ca(3HP)2 crystals have been formed.

[0051] Specifically, during X-ray diffraction (XRD) analysis of Ca(3HP)2, three or more, five or more, seven or more, and nine or more peaks appear in the 2θ range of 10 to 22 degrees. For example, peaks appear in the ranges of 10.0 to 11.0 degrees, 11.1 to 11.6 degrees, 11.6 to 12.5 degrees, 12.7 to 13.6 degrees, 13.8 to 16.0 degrees, 17.0 to 18.0 degrees, 19.0 to 19.8 degrees, 20.2 to 21.2 degrees, and 21.5 to 22.0 degrees.

[0052] On the other hand, the angle of incidence (θ) refers to the angle that X-rays make with a crystal plane when they are irradiated onto a specific crystal plane. The peak refers to a point on a graph in the xy-plane where the horizontal axis (x-axis) is twice the angle of incidence of the incident X-rays (2θ) and the vertical axis (y-axis) is the diffraction intensity, where the first derivative (slope of the tangent line, dy / dx) of twice the angle of incidence of the X-rays (2θ), which is the horizontal axis (x-axis), changes from a positive value to a negative value as the value of twice the angle of incidence of the X-rays (2θ), which is the horizontal axis (x-axis), increases in the positive direction, and the point where the first derivative (slope of the tangent line, dy / dx) becomes 0.

[0053] Furthermore, the interatomic spacing (d-value) in the crystals of 3-hydroxypropionate, as derived by X-ray diffraction (XRD) analysis, may be between 1.00 Å and 15.00 Å, between 1.50 Å and 13.00 Å, between 2.00 Å and 11.00 Å, or between 2.50 Å and 10.00 Å.

[0054] For example, if the crystal of 3-hydroxypropionate is Mg(3HP)2, the interatomic spacing (d value) in the crystal where the peak appears in the range of 8 to 15 degrees 2θ may be between 1.00 Å and 15.00 Å, between 2.00 Å and 13.00 Å, between 4.00 Å and 11.00 Å, or between 5.50 Å and 10.00 Å.

[0055] Furthermore, when the 3-hydroxypropionate crystal is Ca(3HP)2, the interatomic spacing (d value) in the crystal for peaks appearing in the 2θ range of 10 to 22 degrees may be between 1.00 Å and 15.00 Å, between 2.00 Å and 13.00 Å, between 3.00 Å and 10.00 Å, between 3.40 Å and 9.00 Å, and between 4.00 Å and 8.50 Å.

[0056] Furthermore, the 3-hydroxypropionate crystals may have a glass transition temperature of -55°C or higher and -30°C or lower, a melting point of 30°C or higher and 170°C or lower, and a crystallization temperature of 25°C or higher and 170°C or lower.

[0057] The glass transition temperature, melting point, and crystallization temperature of 3-hydroxypropionate crystals can be measured using a differential scanning calorimeter (DSC), with a heating rate of 1 to 20°C / min during measurement. The glass transition temperature of 3-hydroxypropionate crystals can be between -55°C and -30°C, between -50°C and -35°C, or between -45°C and -40°C. The melting point of 3-hydroxypropionate crystals can be between 30°C and 170°C, between 31°C and 160°C, or between 32°C and 150°C. The crystallization temperature of 3-hydroxypropionate crystals can be between 25°C and 170°C, between 27°C and 160°C, or between 30°C and 150°C. The crystallization stable range of 3-hydroxypropionate crystals can be between -40°C and 150°C.

[0058] The 3-hydroxypropionic acid recovery process according to one embodiment may include the steps of: fermenting a strain having the ability to produce 3-hydroxypropionic acid to produce a 3-hydroxypropionic acid ferment liquor, before the step of forming crystals of 3-hydroxypropionate; and concentrating the ferment liquor to form a concentrated liquid containing 300 g / L or more of the 3-hydroxypropionic acid.

[0059] A bacterial strain capable of producing 3-hydroxypropionic acid may contain a gene encoding one or more proteins selected from the group consisting of glycerol dehydratase and aldehyde dehydrogenase, or both of the aforementioned proteins.

[0060] For example, a 3-hydroxypropionic acid-producing strain may further contain a gene (gdrAB) encoding glycerol dehydratase reactivating enzyme (GdrAB). For example, a 3-hydroxypropionic acid-producing strain may also be a strain capable of biosynthesizing vitamin B12.

[0061] Glycerol dehydratase may, but is not limited to, be encoded by the dhaB (GenBank accession no. U30903.1) gene. The dhaB gene may, but is not limited to, be an enzyme derived from Klebsiella pneumoniae. The gene encoding glycerol dehydratase may include genes encoding dhaB1, dhaB2, and / or dhaB3. The glycerol dehydratase protein and the gene encoding it may contain gene and / or amino acid sequence mutations within the range that maintains enzymatic activity in breaking down glycerol into 3-hydroxypropanal (3-HPA) and water (H2O).

[0062] The gene encoding aldehyde dehydrogenase (ALDH) (aldH) may, for example, be the aldH (GenBank Accession no. U00096.3; EaldH) gene derived from Escherichia coli or E. coli K12 MG1655 cell line, the puuC gene derived from Klebsiella pneumoniae, and / or the KGSADH gene derived from Azospirillum brasilense. The aldehyde dehydrogenase protein and the gene encoding it may contain gene and / or amino acid sequence mutations within a range that maintains activity for producing 3-hydroxypropionic acid from 3-hydroxypropanal.

[0063] The culture medium for fermentation broth production is selected without limitation within the desired range for 3-hydroxypropionic acid production. For example, the medium may contain glycerol as a carbon source. Further examples include, but are not limited to, crude glycerol and / or pre-treated crude glycerol. For example, the production medium may further contain vitamin B12.

[0064] In the step of producing a 3-hydroxypropionic acid ferment broth by fermenting a bacterial strain capable of producing 3-hydroxypropionic acid, the concentration of 3-hydroxypropionic acid contained in the 3-hydroxypropionic acid ferment broth may be 1-200 g / L, 10-150 g / L, 30-130 g / L, or 40-100 g / L.

[0065] Furthermore, the fermentation is neutral, and the pH during fermentation can be maintained in the range of 6-8, 6.5-8, 6-7.5, or 6.5-7.5, but is not limited to these ranges. The above pH range can be appropriately adjusted as needed. Alkali metal salts can be added for the neutral fermentation. The alkali metal salt is Mg 2+ Ca2 or a mixture thereof may be included. Furthermore, the alkali metal salt may be, but is not limited to, Ca(OH)2 or Mg(OH)2.

[0066] The 3-hydroxypropionic acid recovery process according to one embodiment may further include the steps of: producing a 3-hydroxypropionic acid fermentate; removing (separating) cells from the fermentate; purifying and / or decolorizing the fermentate and / or the fermentate from which cells have been removed; and / or filtering the fermentate and / or the fermentate from which cells have been removed.

[0067] Cell removal (isolation) can be performed using any method known in the industry, without limitation, within the scope of the purpose of cell (bacterial strain) removal. For example, cell isolation can be performed by centrifugation.

[0068] The step of purifying and / or decolorizing the fermentation liquid and / or the fermentation liquid from which the cells have been removed can be carried out by selecting, without limitation, any method known in the art to the extent necessary for the purpose of purifying the fermentation liquid, for example, by mixing activated carbon with the fermentation liquid and then removing the activated carbon.

[0069] The step of filtering the fermentation liquid and / or the fermentation liquid from which the cells have been removed may be carried out by any method known in the art to the extent of removing solid impurities, removing proteins and / or substances having hydrophobic functional groups, and / or decolorization, for example, by filter filtration and / or activated carbon filtration.

[0070] The 3-hydroxypropionic acid recovery process according to one embodiment may include the step of fermenting a strain having the ability to produce 3-hydroxypropionic acid to produce a 3-hydroxypropionic acid fermentate, followed by the step of concentrating the fermentate to form a concentrated solution containing 300 g / L or more of 3-hydroxypropionic acid.

[0071] The fermentation liquid can be concentrated by evaporating the fermentation liquid (for example, the liquid components of the fermentation liquid).

[0072] Concentration can be carried out by any means normally available for evaporating the liquid components of the fermentation broth, such as rotary evaporation, evaporation, vacuum concentration, and reduced pressure concentration, but is not limited to these.

[0073] For example, the concentration of 3-hydroxypropionic acid in the concentrated fermentation liquid may be 2 to 50 times, 2 to 40 times, 2 to 30 times, 2 to 20 times, 2 to 10 times, 5 to 50 times, 5 to 40 times, 5 to 30 times, 5 to 20 times, or 5 to 10 times higher than before concentration.

[0074] As described above, after the step of concentrating the fermentation liquid to form a concentrate containing 300 g / L or more of 3-hydroxypropionic acid, crystals of 3-hydroxypropionate can be formed in the concentrate containing 300 g / L or more of 3-hydroxypropionic acid in the presence of an alkali metal salt.

[0075] Furthermore, after the step of forming 3-hydroxypropionate crystals, the 3-hydroxypropionate crystals can be separated using a concentrated solution and converted to 3-hydroxypropionic acid.

[0076] The method for separating 3-hydroxypropionate crystals from the concentrate can be any method known in the art to which the present invention belongs, without limitation, within the scope of the purpose for separating the crystals. For example, the recovery of 3-hydroxypropionate crystals can be carried out by drying (e.g., heat drying) and / or filtration, but is not limited to these methods.

[0077] Furthermore, the method for converting (purifying) the separated 3-hydroxypropionate crystals to 3-hydroxypropionic acid can be any method known in the art within the scope of the purpose of purifying 3-hydroxypropionic acid, without limitation. For example, one or more of the methods listed below can be used, but are not limited to these.

[0078] For example, i) a method of protonating 3-hydroxypropionic acid by removing cations using a cation exchange resin, ii) Using liquid cation exchange, cations (e.g., Mg 2+ Ca 2+ A method for extracting (etc.) with an organic solvent and protonating 3-hydroxypropionic acid, and iii) A method of protonating 3-hydroxypropionic acid is to titrate it with an acid (e.g., sulfuric acid) to produce a salt (e.g., CaSO4(s) or MgSO4(s)).

[0079] In a 3-hydroxypropionic acid (3HP) recovery process according to one embodiment, the recovery rate of 3-hydroxypropionic acid can be calculated using the method shown in Formula 1 below.

[0080] [Formula 1] 3HP recovery rate (%) = {(3HP content in crystals) / (3HP content in fermentation liquid before crystallization)} × 100

[0081] As an example, the recovery rate of 3-hydroxypropionic acid in the 3-hydroxypropionic acid recovery step according to the present invention may be 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more, for example, 40-99.9%, 50-99.9%, 60-99.9%, 70-99.9%, 80-99.9%, 90-99.9%, 40-99%, 50-99%, 60-99%, 70-99%, 80-99%, 90-99%, 40-97%, 50-97%, 60-97%, 70-97%, 80-97%, 90-97%, 40-95%, 50-95%, 60-95%, 70-95%, 80-95%, or 90-95%, but is not limited to these. The aforementioned recovery rate may be calculated based on weight.

[0082] Furthermore, in the 3-hydroxypropionic acid (3HP) recovery process, the purity of the 3-hydroxypropionate contained in the 3-hydroxypropionate crystals can be calculated as the mass percentage (%) of the compound having structural formula 1 and / or structural formula 2 relative to the total mass of the recovered crystals. As an example, the purity of the 3-hydroxypropionate contained in the 3-hydroxypropionate crystals produced in the 3-hydroxypropionic acid recovery process according to the present invention may be, but is not limited to, 70% or more, 80% or more, 90% or more, 70-99.9%, 80-99.9%, 90-99.9%, 70-99%, 80-99%, or 90-99%.

[0083] According to another embodiment of the present invention, a crystal of 3-hydroxypropionate represented by structural formula 1 or structural formula 2 below is provided.

[0084] [Structural formula 1] Cation (3HP) n [Structural formula 2] Cation (3HP) n ·mH2O In structural formulas 1 and 2 above, Cation represents a cation, 3HP represents 3-hydroxypropionic acid that binds to the cation, and n represents the number of 3HP molecules that bind to the cation, meaning an integer of 1 or more. In structural formula 2 below, m represents the number of water molecules that bind to Cation(3HP)n in the hydrate, meaning an integer of 1 or more. The cation is, for example, Na. + Mg 2+ or Ca 2+ It is possible, but Mg 2+ or Ca 2+ In this case, 3-hydroxypropionate crystals are formed more effectively.

[0085] The 3-hydroxypropionate crystals are formed in a 3-hydroxypropionic acid recovery step according to one embodiment, and may be formed, for example, in a step where 3-hydroxypropionate crystals are formed in a concentrated solution containing 300 g / L or more of 3-hydroxypropionic acid in the presence of an alkali metal salt.

[0086] The 3-hydroxypropionate crystals have a particle size distribution D 50 The particle size distribution D is as follows: 20 μm to 90 μm, 25 μm to 85 μm, 30 μm to 80 μm, and 35 μm to 75 μm. 10 The particle size distribution D is 5 μm to 40 μm, 8 μm to 35 μm, and 10 μm to 30 μm. 90 The particle size can be between 50 μm and 200 μm, between 60 μm and 190 μm, between 65 μm and 180 μm, or between 70 μm and 175 μm.

[0087] Also, the crystals of 3-hydroxypropionate (D 90 -D 10 ) / D 50 It could be between 1.00 and 3.00, between 1.20 and 2.80, between 1.40 and 2.60, or between 1.60 and 2.40.

[0088] Furthermore, the crystals of 3-hydroxypropionate may have a volume-average diameter of 30 μm to 100 μm, 35 μm to 95 μm, or 40 μm to 90 μm; a number-average diameter of 1 μm to 30 μm, 3 μm to 25 μm, or 5 μm to 20 μm; and an area-average diameter of 10 μm to 70 μm, 15 μm to 60 μm, or 20 μm to 55 μm.

[0089] Furthermore, the particle size distribution of 3-hydroxypropionate crystals (D 10 , D 50 , D 90 The aspect ratio (LW Ratio; Length to width ratio) and the average aspect ratio may be between 0.50 and 3.00, between 0.70 and 2.80, and between 1.00 and 2.50, respectively.

[0090] The water content in 3-hydroxypropionate crystals may be between 200 ppm and 5000 ppm, between 250 ppm and 4800 ppm, between 300 ppm and 4600 ppm, or between 350 ppm and 4400 ppm.

[0091] Crystals of 3-hydroxypropionate may have a radiocarbon isotope content of 20 pMC (percent modern carbon) or more, 50 pMC or more, 90 pMC or more, or 100 pMC or more, as measured according to ASTM D6866-21 standards, and a biocarbon content of 20% by weight or more, 50% by weight or more, 80% by weight or more, 90% by weight or more, or 95% by weight or more.

[0092] Furthermore, when 3-hydroxypropionate crystals are analyzed by X-ray diffraction (XRD), peaks appear between the crystal lattice when the 2θ value is in the range of 8 to 22 degrees, and the interatomic spacing (d value) in the crystal derived from XRD analysis may be between 1.00 Å and 15.00 Å, between 1.50 Å and 13.00 Å, between 2.00 Å and 11.00 Å, or between 2.50 Å and 10.00 Å.

[0093] Furthermore, the crystals of 3-hydroxypropionate may have a glass transition temperature of -55°C to -30°C, a melting point of 30°C to 170°C, and a crystallization temperature of 25°C to 170°C.

[0094] Furthermore, the purity of 3-hydroxypropionate contained in the 3-hydroxypropionate crystals may be 70% or higher, 80% or higher, 90% or higher, 70-99.9%, 80-99.9%, 90-99.9%, 70-99%, 80-99%, or 90-99%.

[0095] Furthermore, the particle size distribution D of the 3-hydroxypropionate crystals mentioned above. 10 , D 50 , D 90 , (D 90 -D 10 ) / D 50 Volume average diameter, number average diameter, area average diameter, particle size distribution (D 10 , D 50 , D 90 The measurement conditions and specific explanations for the aspect ratio, average aspect ratio, water content, radiocarbon isotope content, biocarbon content, derivation results of XRD analysis (interatomic spacing in the crystal (d value), peaks appearing in a specific range of 2θ values), glass transition temperature, melting point, crystallization temperature, and purity are as described above in the 3-hydroxypropionic acid recovery process according to one embodiment.

[0096] According to yet another embodiment of the present invention, crystals of 3-hydroxypropionate are provided, comprising an alkali metal salt of 3-hydroxypropionic acid or its hydrate, with a biocarbon content of 20% by weight or more as measured according to ASTM D6866-21.

[0097] The alkali metal salt of 3-hydroxypropionic acid may be the calcium salt or magnesium salt of 3-hydroxypropionic acid.

[0098] Furthermore, the alkali metal salt of 3-hydroxypropionic acid is represented by structural formula 1 below, and the hydrate of the alkali metal salt of 3-hydroxypropionic acid is represented by structural formula 2 below. Cation is Mg 2+ or Ca 2+ It is possible.

[0099] [Structural formula 1] Cation (3HP) n [Structural formula 2] Cation (3HP) n ·mH2O

[0100] Furthermore, the 3-hydroxypropionate crystals may have a radiocarbon isotope content of 20 pMC (percent modern carbon) or more, 50 pMC or more, 90 pMC or more, or 100 pMC or more, as measured according to ASTM D6866-21 standards, and a biocarbon content of 20% by weight or more, 50% by weight or more, 80% by weight or more, 90% by weight or more, or 95% by weight or more.

[0101] The crystals of 3-hydroxypropionate may have a volume-average diameter of 30 μm to 100 μm, 35 μm to 95 μm, or 40 μm to 90 μm; a number-average diameter of 1 μm to 30 μm, 3 μm to 25 μm, or 5 μm to 20 μm; and an area-average diameter of 10 μm to 70 μm, 15 μm to 60 μm, or 20 μm to 55 μm.

[0102] Furthermore, the particle size distribution of 3-hydroxypropionate crystals (D 10 , D 50 , D 90 The aspect ratio (LW ratio; Length to width ratio) and average aspect ratio of the image may be between 0.50 and 3.00, between 0.70 and 2.80, and between 1.00 and 2.50, respectively.

[0103] The crystals of 3-hydroxypropionate have a particle size distribution D 50The particle size distribution D is as follows: 20 μm to 90 μm, 25 μm to 85 μm, 30 μm to 80 μm, and 35 μm to 75 μm. 10 The particle size distribution D is 5 μm to 40 μm, 8 μm to 35 μm, and 10 μm to 30 μm. 90 The particle size can be between 50 μm and 200 μm, between 60 μm and 190 μm, between 65 μm and 180 μm, or between 70 μm and 175 μm.

[0104] Also, the crystals of 3-hydroxypropionate (D 90 -D 10 ) / D 50 It could be between 1.00 and 3.00, between 1.20 and 2.80, between 1.40 and 2.60, or between 1.60 and 2.40.

[0105] The water content in 3-hydroxypropionate crystals may be between 200 ppm and 5000 ppm, between 250 ppm and 4800 ppm, between 300 ppm and 4600 ppm, or between 350 ppm and 4400 ppm.

[0106] Furthermore, when 3-hydroxypropionate crystals are analyzed by X-ray diffraction (XRD), peaks appear between the crystal lattice when the 2θ value is in the range of 8 to 22 degrees, and the interatomic spacing (d value) in the crystal derived from XRD analysis may be between 1.00 Å and 15.00 Å, between 1.50 Å and 13.00 Å, between 2.00 Å and 11.00 Å, or between 2.50 Å and 10.00 Å.

[0107] Crystals of 3-hydroxypropionate may have a glass transition temperature of -55°C to -30°C, a melting point of 30°C to 170°C, and a crystallization temperature of 25°C to 170°C.

[0108] Furthermore, the purity of 3-hydroxypropionate contained in the 3-hydroxypropionate crystals may be 70% or higher, 80% or higher, 90% or higher, 70-99.9%, 80-99.9%, 90-99.9%, 70-99%, 80-99%, or 90-99%.

[0109] The particle size distribution D of the crystals of the above-mentioned 3-hydroxypropionate 10 , D 50 , D 90 , (D 90 -D 10 ) / D 50 , volume average diameter, number average diameter, area average diameter, aspect ratio of particle size distribution (D 10 , D 50 , D 90 ), the measurement conditions for moisture content, radiocarbon isotope content, biocarbon content, XRD analysis results (interatomic spacing (d value) in the crystal, peaks appearing in a specific 2θ value range), glass transition temperature, melting point, crystallization temperature, purity, etc., and specific explanations are as described above in the recovery process of 3-hydroxypropionic acid according to one embodiment.

Advantages of the Invention

[0110] The crystals of 3-hydroxypropionate according to the present invention contain 3-hydroxypropionate with high purity, and due to having a specific particle size or shape, filtration from impurities such as fermentation by-products and / or additives is easy. Also, in the 3-hydroxypropionic acid recovery (separation, purification) process, fermentation by-products and / or additives can be easily separated by crystallization of 3-hydroxypropionate. Therefore, high-purity 3-hydroxypropionate can be recovered, the process can be carried out without an organic solvent, so the separation and purification process of 3-hydroxypropionic acid can be simplified, and the purification cost of 3-hydroxypropionic acid can be reduced.

Brief Description of the Drawings

[0111] [Figure 1] It is a graph showing the X-ray diffraction (XRD) analysis results of the crystals (Ca(3HP)2) of 3-hydroxypropionate in Example 1. [Figure 2] It is a graph showing the X-ray diffraction (XRD) analysis results of the crystals (Ca(3HP)2) of 3-hydroxypropionate in Example 1. [Figure 3]This graph shows the results of X-ray diffraction (XRD) analysis of the 3-hydroxypropionate crystal (Mg(3HP)2) from Example 2. [Figure 4] This graph shows the results of X-ray diffraction (XRD) analysis of the 3-hydroxypropionate crystal (Mg(3HP)2) from Example 2. [Figure 5] This graph shows the results of differential scanning calorimetry (DSC) analysis of the 3-hydroxypropionate crystal (Ca(3HP)2) from Example 1. [Figure 6] This graph shows the differential scanning calorimetry (DSC) analysis results for the 3-hydroxypropionate crystal (Mg(3HP)2) from Example 2. [Modes for carrying out the invention]

[0112] The present invention will be described in more detail below with reference to the following examples. However, the following examples are merely illustrative, and the content of the present invention is not limited to these examples.

[0113] Unless otherwise specified in this specification, all temperatures are expressed in Celsius. [Examples]

[0114] Preparation Example 1. Production of a strain for 3-hydroxypropionic acid production Recombinant vectors were prepared by introducing genes encoding glycerol dehydratase and aldehyde dehydrogenase, which are known to produce 3-hydroxypropionic acid (3HP) using glycerol as a substrate. The prepared recombinant vectors were introduced into E. coli W3110 strain to create a 3-hydroxypropionic acid-producing strain.

[0115] More specifically, a 3-hydroxypropionic acid-producing strain was produced by cloning the BtuR gene encoding adenosyltransferase into a plasmid pCDF containing the genes encoding glycerol dehydratase (dhaB), aldehyde dehydrogenase (aldH), and glycerol dehydratase reactivase (gdrAB), and then introducing the resulting pCDF_J23101_dhaB_gdrAB_J23100_aldH_btuR vector into strain W3110 (KCCM 40219) using electroporation with an electroporation device (Bio-Rad, Gene Pulser Xcell). The preparation process for the 3-hydroxypropionic acid-producing strain in Preparation Example 1, as well as the vector, primer, and enzyme used, were carried out with reference to Example 1 of Korean Published Patent No. 10-2020-0051375 (incorporated herein by reference).

[0116] Example 1. Preparation of Ca(3HP)2 crystals The 3-hydroxypropionic acid production strain prepared in Preparation Example 1 was fermented in a 5L fermenter at 35 degrees Celsius using unpurified glycerol as a carbon source to produce 3-hydroxypropionic acid. To prevent a decrease in pH due to 3-hydroxypropionic acid production, calcium hydroxide (Ca(OH)2), an alkali metal salt, was added to maintain a neutral pH during fermentation.

[0117] After fermentation culture, cells were removed by centrifugation (4000 rpm, 10 minutes, 4 degrees Celsius), and the primary fermentation broth was purified using activated carbon. Specifically, activated carbon was added to the fermentation broth from which the microbial cells had been removed by centrifugation and mixed thoroughly, and then the activated carbon was separated by centrifugation again. Subsequently, the fermentation broth from which the activated carbon had been separated was filtered using a vacuum pump with 0.7 μm filter paper to purify the 3-hydroxypropionic acid fermentation broth.

[0118] The concentration of 3-hydroxypropionic acid in the fermentation liquid after the primary purification was 50-100 g / L. The fermentation liquid was concentrated to a concentration of 600 g / L using a rotary evaporator (50 degrees Celsius, 50 mbar) to produce a concentrate, which was stirred at room temperature (300 rpm) to produce Ca(3HP)2 crystals. At this time, the concentration of the alkali metal salt in the concentrate was 493.3 g / L (based on Ca(OH)2).

[0119] The generated crystals were washed three times with ethanol (EtOH), dried in an oven at 50 degrees Celsius, and finally recovered. The weight of the crystals was measured, and the crystals were dissolved in water. The amount of 3-hydroxypropionic acid (3HP) was quantified using high-performance liquid chromatography (HPLC) to determine the amount of 3-hydroxypropionic acid in the crystals, and the recovery rate was measured by comparing it with the amount of 3-hydroxypropionic acid measured by HPLC in the fermentation liquid (Equation 1).

[0120] [Formula 1] 3HP recovery rate (%) = {(3HP content in crystals) / (3HP content in fermentation liquid before crystallization)} × 100 The recovery rate of 3-hydroxypropionic acid recovered in crystal form was approximately 92%.

[0121] Example 2. Preparation of Mg(3HP)2 crystals The fermentation process was carried out in substantially the same manner as in Example 1, but Mg(OH)2 was used instead of Ca(OH)2 as the alkali metal salt, and the mixture was concentrated to produce and recover Mg(3HP)2 crystals. Table 1 below shows the recovery rate of Mg(3HP)2 crystals at different concentrations of each concentrate. [Table 1]

[0122] As can be seen from Table 1 above, more than 90% of 3-hydroxypropionic acid was recovered as a result of crystallization after concentration, confirming that crystallization yields excellent recovery rates for 3-hydroxypropionic acid.

[0123] Comparative Example 1. Titration with sulfuric acid after fermentation. After carrying out the fermentation process in the same manner as in Example 1, the fermentation solution was titrated to pH 2 using sulfuric acid (H2SO4). Gypsum (Ca(SO)4) generated during the titration process was removed, and 3-hydroxypropionic acid was protonated.

[0124] The fermented liquid from which the gypsum had been removed was purified and concentrated in substantially the same manner as in Example 1, and the presence or absence of crystal formation was confirmed.

[0125] However, when the concentration was increased to 600 g / L (based on 3 HP), no crystals were formed, and even when the concentration was increased to 1000 g / L (based on 3 HP), no crystal formation was observed.

[0126] <Example Test> 1. Purity analysis of 3-hydroxypropionate in crystals Ion chromatography (IC) analysis (Dionex Aquion; Eluent: 20 mM Methanesulfonic Acid; Column: Dionex IonPac CS12A; Flow Rate: 1.0 ml / min; Suppressor: Dionex CERS 500 4 mm; Current: 59 mA) was performed to analyze the cations of the 3-hydroxypropionate crystals obtained in Example 1 and Example 2, and the purity of each 3-hydroxypropionate in the crystals was confirmed.

[0127] Table 2 below shows the cation analysis results for Ca(3HP)2 crystals in Example 1, and Table 3 below shows the purity measurement results for 3-hydroxypropionate at different concentrations of each concentrated solution of Mg(3HP)2 crystals in Example 2. [Table 2]

[0128] [Table 3]

[0129] Referring to Tables 2 and 3 above, it was confirmed that the purity of 3-hydroxypropionate in the crystals in Examples 1 and 2 was 92.9% or higher. In the case of the purity of Example 1, the Ca in Table 2 above is 2+ When the purity of Ca(3HP)2 was checked using this ratio, it was confirmed to have an excellent purity of over 98%.

[0130] 2. Analysis of 3-hydroxypropionic acid in crystals High-performance liquid chromatography (HPLC) was used to analyze the 3-hydroxypropionic acid content in the 3-hydroxypropionate crystals obtained in Examples 1 and 2. The specific HPLC conditions are shown in Table 4 below.

[0131] [Table 4]

[0132] Table 5 below shows the HPLC results for the crystals of Example 1 and Example 2. In Table 5 below, 3-hydroxypropionate in the crystals refers to Ca(3HP)2 in Example 1 and Mg(3HP)2 in Example 2.

[0133] [Table 5]

[0134] Referring to Table 5 above, the HPLC results for the crystals of Example 1 and Example 2 confirmed that the purity of the 3HP salt in the crystals was 93.20% or higher.

[0135] 3.Grain size analysis of 3-hydroxypropionate crystals Using a particle size and shape analyzer (Size and Shape Particle Analyzer; Microtrac TurboSync), the D of the crystals of 3-hydroxypropionate (Ca(3HP)2) in Example 1 and the crystals of 3-hydroxypropionate (Mg(3HP)2) in Example 2 10 , D 50 , D 90 , the volume average diameter, number average diameter, and area average diameter were measured respectively, and the results are shown in Table 6 below.

[0136] Also, using the particle size analyzer, the D of the crystals of 3-hydroxypropionate in Example 1 and Example 2 10 , D 50 and D 90 The aspect ratio (LW Ratio; Length to width ratio) and average aspect ratio of each were measured and shown in Table 7 below.

[0137]

Table 6

Table 7

[0138] [[ID=,32]] Aspect ratio (LW Ratio; Length to width ratio) Referring to Table 6 and Table 7 above, the particle size distribution D of Example 1 and Example 2 10 is 13.68 - 27.41 μm, the particle size distribution D 50 [[ID=,39]]is 36.50 - 71.40 μm, the particle size distribution D 90 is 74.02 - 170.40, (D 90 - D 10 ) / D 50 is 2.00 for Example 1 and 1.65 for Example 2, the volume average diameter is 41.30 - 88.71 μm, the number average diameter is 5.13 - 18.99 μm, the area average diameter is 24.81 - 53.45 μm, and the aspect ratio (LW Ratio; Length to width ratio) of D 10 is 13.68 - 27.41, D50 The aspect ratio is 36.50 to 71.40, D 90 We confirmed that the aspect ratios ranged from 74.02 to 170.40, with an average aspect ratio of 41.30 to 88.71.

[0139] 4. Analysis of the water content of 4,3-hydroxypropionate crystals The water content of the 3-hydroxypropionate crystals (Ca(3HP)2) from Example 1 and the 3-hydroxypropionate crystals (Mg(3HP)2) from Example 2 was analyzed using a Karl Fischer titrator manufactured by Metrohm, and the results are shown in Table 8 below. [Table 8]

[0140] Referring to Table 8 above, it was confirmed that the moisture content of Examples 1 and 2 was between 351 and 4370 ppm.

[0141] 5. Analysis of biocarbon content in 5,3-hydroxypropionate crystals The radiocarbon isotope ratio (pMC) and biocarbon content of the 3-hydroxypropionate crystals (Ca(3HP)2) from Example 1 and the 3-hydroxypropionate crystals (Mg(3HP)2) from Example 2 were analyzed using ASTM D6866-21 (Method B), and the results are shown in Table 9 below.

[0142] The biocarbon content refers to the biocarbon content contained in the 3-hydroxypropionate crystals of Example 1 and Example 2, respectively, and the radiocarbon isotope ratio (pMC) refers to the radiocarbon isotopes contained in the 3-hydroxypropionate crystals. 14 C) and radioactive carbon isotopes of modern reference materials ( 14 This refers to the ratio of C).

[0143] [Table 9]

[0144] -pMC: percent Modern Carbon Referring to Table 9 above, it was confirmed that all pMC of the 3HP salt crystals in Example 1 and Example 2 were 101 or higher, and that the biocarbon content was also 100%.

[0145] 6. X-ray diffraction (XRD) analysis of 6,3-hydroxypropionate crystals The crystals produced in Example 1 and Example 2 were irradiated with Cu-Kα rays at a wavelength of 1.54 Å, and the reflection mode X-ray diffraction (XRD) patterns were measured.

[0146] The measurement device used was a Bruker AXS D4 Endeavor XRD. The operating voltage and current were 40kV and 40mA, respectively, and the optical elements (optics) and detector used are as follows.

[0147] -Primary (incident beam) optics: motorized divergence slit, soler slit 2.3 degrees -Secondary(diffracted beam)optics:soller slit 2.3 degrees -LynxEye detector(1D detector)

[0148] Figures 1 and 2 are graphs showing the results of X-ray diffraction analysis of the 3-hydroxypropionate crystals of Example 1, and Figures 3 and 4 are graphs showing the results of X-ray diffraction analysis of the 3-hydroxypropionate crystals of Example 2.

[0149] Table 10 shows the 2θ values ​​and grid plane spacing (d values) of the numbered peaks in the XRD graphs of Figures 1 and 2, and Table 11 shows the 2θ values ​​and grid plane spacing (d values) of the numbered peaks in the XRD graphs of Figures 3 and 4. [Table 10] [Table 11]

[0150] Tables 10 and 11 above confirm that the 3-hydroxypropionate crystals produced in Example 1 were Ca(3HP)2, and the 3-hydroxypropionate crystals produced in Example 2 were Mg(3HP)2.

[0151] 7. Differential scanning calorimeter (DSC) analysis of 7,3-hydroxypropionate crystals The melting points (Tm) of the crystals in Example 1 and Example 2 were determined using a differential scanning calorimeter (TA Instruments, "DSC(Q2000)"). In Example 1, the glass transition temperature and crystallization temperature were measured at a heating rate of 10°C / min in the range of -60°C to 150°C. In Example 2, the glass transition temperature and crystallization temperature were measured at a heating rate of 10°C / min in the range of -70°C to 30°C. The results are shown in Table 12 below.

[0152] Figure 5 is a graph showing the differential scanning calorimeter (DSC) analysis results for the 3-hydroxypropionate crystal (Ca(3HP)2) of Example 1, and Figure 6 is a graph showing the differential scanning calorimeter (DSC) analysis results for the 3-hydroxypropionate crystal (Mg(3HP)2) of Example 2.

[0153] [Table 12]

[0154] Referring to Table 12 above, it was confirmed that the glass transition temperature of Example 1 and Example 2 was -45 to -40°C, the melting point was 32 to 155°C, and the crystallization temperature was 30 to 150°C.

Claims

1. The biocarbon content measured according to the ASTM D6866-21 standard is 20% by weight or more. Crystals of 3-hydroxypropionate represented by structural formula 1 or structural formula 2 below, with a volume-average diameter of 30 μm or more and 100 μm or less, a number-average diameter of 1 μm or more and 30 μm or less, and an area-average diameter of 10 μm or more and 70 μm or less: [Structural formula 1] Cation(3HP) ｎ [Structural formula 2] Cation(3HP) ｎ ・mH ２ O In the above structural formula, A cation is a cation (cation) that is Mg²⁺. 3HP is 3-hydroxypropionic acid that binds to a cation. n is the number of 3HP cells that bond with the cation, and is an integer greater than or equal to 2. m is an integer greater than or equal to 1, representing the number of water molecules in the hydrate.

2. The 3-hydroxypropionate crystals have a particle size distribution D ５０ The size is 20 μm or more and 90 μm or less, (D ９０ -D １０ ) / D ５０ A crystal of 3-hydroxypropionate according to claim 1, wherein the ratio is 1.00 or more and 3.00 or less.

3. The 3-hydroxypropionate crystal according to claim 1 or 2, wherein the 3-hydroxypropionate crystal has an average aspect ratio (LW Ratio; Length to width ratio) of 0.50 or more and 3.00 or less.

4. The 3-hydroxypropionate crystals according to any one of claims 1 to 3, wherein the radioactive carbon isotope content measured according to ASTM D6866-21 is 20 pMC (percent modern carbon) or more.

5. The 3-hydroxypropionate crystal according to any one of claims 1 to 4, wherein the interatomic spacing (d value) in the crystal, as derived by X-ray diffraction (XRD) analysis, is 1.00 Å or more and 15.00 Å or less.

6. The 3-hydroxypropionate crystal according to any one of claims 1 to 5, wherein, when analyzed by X-ray diffraction (XRD), interlattice peaks appear in the range of 2θ values ​​from 8 to 22 degrees.

7. The 3-hydroxypropionate crystals described above have a glass transition temperature of -55°C or higher and -30°C or lower when analyzed by differential scanning calorimeter (DSC), as described in any one of claims 1 to 6.

8. The 3-hydroxypropionate crystals described in any one of claims 1 to 7 have a melting point of 30°C or higher and 170°C or lower when analyzed by differential scanning calorimeter (DSC).

9. The 3-hydroxypropionate crystal according to any one of claims 1 to 8, wherein the purity of the 3-hydroxypropionate contained in the 3-hydroxypropionate crystal is 70% or more.

10. It contains the magnesium salt of 3-hydroxypropionic acid or its hydrate, Crystals of 3-hydroxypropionate having a biocarbon content of 20% by weight or more as measured according to the ASTM D6866-21 standard, a volume-average diameter of 30 μm to 100 μm, a number-average diameter of 1 μm to 30 μm, and an area-average diameter of 10 μm to 70 μm.

11. The crystal of the 3-hydroxypropionate has a particle size distribution D ５０ of 20 μm or more and 90 μm or less, and (D ９０ - D １０ ) / D ５０ is 1.00 or more and 3.00 or less. The crystal of 3-hydroxypropionate according to claim 10.

12. The 3-hydroxypropionate crystal according to claim 10 or 11, wherein the 3-hydroxypropionate crystal has an average aspect ratio (LW Ratio; Length to width ratio) of 0.50 or more and 3.00 or less.

13. The 3-hydroxypropionate crystals according to any one of claims 10 to 12, wherein the radioactive carbon isotope content measured according to ASTM D6866-21 is 20 pMC (percent modern carbon) or more.

14. The 3-hydroxypropionate crystal according to any one of claims 10 to 13, wherein the interatomic spacing (d value) in the crystal, as derived by X-ray diffraction (XRD) analysis, is 1.00 Å or more and 15.00 Å or less.

15. The 3-hydroxypropionate crystal according to any one of claims 10 to 14, wherein, when analyzed by X-ray diffraction (XRD), interlattice peaks appear in the range of 2θ values ​​from 8 to 22 degrees.

16. The 3-hydroxypropionate crystal according to any one of claims 10 to 15, wherein the 3-hydroxypropionate crystal has a glass transition temperature of -55°C or higher and -30°C or lower when analyzed by differential scanning calorimeter (DSC).

17. The 3-hydroxypropionate crystals described above have a melting point of 30°C or higher and 170°C or lower when analyzed by differential scanning calorimeter (DSC), as described in any one of claims 11 to 16.

18. The 3-hydroxypropionate crystal according to any one of claims 10 to 17, wherein the purity of the 3-hydroxypropionate contained in the 3-hydroxypropionate crystal is 70% or more.

Images