WO2011043443A1 - Method for producing aliphatic dicarboxylic acid - Google Patents
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- WO2011043443A1 WO2011043443A1 PCT/JP2010/067694 JP2010067694W WO2011043443A1 WO 2011043443 A1 WO2011043443 A1 WO 2011043443A1 JP 2010067694 W JP2010067694 W JP 2010067694W WO 2011043443 A1 WO2011043443 A1 WO 2011043443A1
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- aliphatic dicarboxylic
- dicarboxylic acid
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/44—Polycarboxylic acids
- C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
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- the present invention relates to a method for producing an aliphatic dicarboxylic acid such as succinic acid using a microorganism.
- the present invention particularly relates to aliphatic dicarboxylic acids from aqueous solutions containing aliphatic dicarboxylic acids (salts) and saccharides, amino acids, proteins, inorganic salts, and the like obtained from microorganisms such as glucose, sucrose, and cellulose, which are biological materials. Relates to a method for producing an aliphatic dicarboxylic acid.
- Aliphatic dicarboxylic acids are widely used as raw materials for polymers such as polyesters and polyamides, particularly biodegradable polyesters, and as raw materials for foods, pharmaceuticals and cosmetics.
- a high-purity aliphatic dicarboxylic acid is required for maintaining the degree of polymerization of the polymer and preventing coloring.
- succinic acid is expected in recent years as a raw material for biodegradable polymers together with lactic acid.
- Aliphatic dicarboxylic acids have been produced industrially from petroleum-derived raw materials.
- succinic acid has been produced by a hydrogenation reaction of maleic acid, a raw material derived from petroleum.
- various aliphatic dicarboxylic acids can be produced from plant-derived materials with a high carbon yield from a wide range of biological materials.
- succinic acid, adipic acid and the like can be produced by fermentation.
- the fermentation broth is treated by centrifugation and filtration to remove solids such as bacterial cells, and then desalted with an ion exchange resin or the like, and crystallized from the solution.
- a method of purifying an aliphatic dicarboxylic acid by column chromatography is used.
- Patent Document 1 discloses a method of purifying an aliphatic dicarboxylic acid with an H + type strongly acidic cation exchange resin after centrifuging cells.
- Patent Document 2 as a method for removing microbial cells, the microbial cell is aggregated by adjusting the suspension of the microorganism to pH 4 or lower and dissolving sodium polyacrylate in the microbial suspension.
- a method for separating bacterial cell agglomerates is disclosed.
- the raw materials are generally sugars, such as glucose, sucrose, and cellulose.
- the raw material saccharide may not be completely assimilated by the microorganism, and the saccharide may be mixed into the aliphatic dicarboxylic acid.
- amino acids are by-produced and amino acids are mixed into aliphatic dicarboxylic acids.
- proteins derived from microorganisms and inorganic salts used in fermentation may be mixed in aliphatic dicarboxylic acids.
- a method using electrodialysis (for example, see Patent Document 6) is also generally known.
- electrodialysis increases in proportion to the production scale, the scale merit is small and the cost is high even in industrial scale production.
- an object of the present invention is to improve the purification efficiency of an aliphatic dicarboxylic acid from a suspension containing an aliphatic dicarboxylic acid obtained by reacting a biological material and a microorganism with microbial cells, thereby producing an aliphatic dicarboxylic acid.
- the object is to provide a method capable of efficiently producing an acid.
- the extraction method is performed by adding a phase-separable solvent to the aqueous solution containing the aliphatic dicarboxylic acid.
- a solid content that does not dissolve in either the aqueous phase or the solvent phase is generated.
- the subsequent processes are adversely affected and the purity of the aliphatic dicarboxylic acid is deteriorated.
- an aliphatic solution obtained by microbial conversion from glucose, sucrose, cellulose, or the like, which is a biological material is brought into contact with an aqueous solution composed of an aliphatic dicarboxylic acid (salt), saccharide, amino acid, protein, inorganic salt, etc., and a solvent.
- an aliphatic dicarboxylic acid salt
- saccharide amino acid
- protein protein
- inorganic salt etc.
- phase separation operation may be particularly difficult.
- Patent Document 7 does not provide a method for recovering the target organic acid as crystals.
- the present invention eliminates the inhibition of phase separation due to the generation of the above-mentioned solids and the obstacles to the production operation associated therewith, and is effective and stable from an aqueous solution containing an aliphatic dicarboxylic acid obtained by microbial conversion from a biological material. It is another object of the present invention to provide a method for producing an aliphatic dicarboxylic acid.
- a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism comprising the steps of (1) to (4) below: Production method.
- Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms
- Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step PH adjustment step (3) for obtaining a solution having a pH of 1.0 or more and 5.0 or less by mixing a solution containing a solution and an acid, a microorganism separation step (4) for separating microorganisms from the solution obtained in the pH adjustment step
- a contact step in which an aqueous solution obtained by separating microorganisms in the microorganism separation step is brought into contact with a solvent.
- the solvent is an organic solvent having an inorganic value / organic value ratio (I / O value) of 0.2 or more and 2.3 or less and a normal pressure (1 atm) and a boiling point of 40 ° C. or more. 4.
- a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material comprising the following steps (1) to (3).
- a contact step in which an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material is brought into contact with a solvent that can be phase-separated with the aqueous solution.
- a phase separation step for separating the solvent by phase separation. 6.
- the extracted solid content is subjected to solid-liquid separation, and the obtained liquid is returned to at least one of the step before the contact step between the aqueous solution and the solvent and the step after the phase separation step.
- Production method. 10 10.
- 11. 10 The method for producing an aliphatic dicarboxylic acid according to item 9, wherein the solid-liquid separation is performed using a filter.
- a contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is brought into contact with a solvent that is phase-separated with the aqueous solution.
- Extraction phase concentration step in which water concentration in phase increases (3) Crystallization step for precipitation of aliphatic dicarboxylic acid from liquid after extraction phase concentration step (4) Solid recovery of aliphatic dicarboxylic acid precipitated in crystallization step Liquid separation step (5) Crystallization mother liquor recycling step for returning at least part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step prior to the crystallization step. 15.
- the method for producing an aliphatic dicarboxylic acid according to item 14 further comprising a protonation step of adding an acid to the aqueous solution containing the aliphatic dicarboxylic acid obtained from the biological material before the contacting step. 16. 16.
- An acid having a pKa of less than 4 is used as an acid to be added in the protonation step, and the pH of the aqueous solution containing the aliphatic dicarboxylic acid obtained from the biological material in the protonation step is maintained in the range of more than 1 and less than 4.
- 20. The method for producing an aliphatic dicarboxylic acid according to any one of items 15 to 19, wherein: 21.
- the solvent used in the contacting step has a ratio of an inorganic value to an organic value of 0.2 or more and 2.3 or less, and a boiling point of 40 ° C. or more, any one of the items 14 to 20 above 2.
- a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism comprising the steps of (1) to (6) below: How to manufacture.
- Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms
- Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step A pH adjusting step for obtaining a liquid having a pH of 1.0 or more and 5.0 or less by mixing a solution containing the microbial cells and an acid
- Body separation step (4)
- Contact step of contacting the aqueous solution obtained by separating the cells in the cell separation step with a solvent capable of phase separation with the aqueous solution (5) Generated by contact between the aqueous solution and the solvent Solid content removing step for removing solid content (6)
- Phase separation step for separating the solvent by phase separation 30.
- a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism comprising the steps of (1) to (8) below: How to manufacture.
- Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms
- Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step A pH adjusting step for obtaining a liquid having a pH of 1.0 or more and 5.0 or less by mixing a solution containing the microbial cells and an acid
- Bacteria for separating microbial cells from the liquid obtained in the pH adjusting step Body separation step (4) Contact step in which the aqueous solution obtained by separating the cells in the cell separation step is brought into contact with a solvent capable of phase separation with the aqueous solution (5)
- Aliphatic dicarboxylic acid recovered in the contact step In which the concentration of water in the extraction phase increases due to the concentration (6) Crystallization step (7) Crystallization of depositing aliphatic dicarboxylic acid from the liquid after the extraction phase concentration step
- an aliphatic dicarboxylic acid can be efficiently and stably produced from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological material.
- microbial cells can be efficiently separated even by low-speed centrifugation, so that the equipment load can be reduced, and the separation time in solvent extraction and the intermediate insoluble matter can be reduced.
- Aliphatic dicarboxylic acid can be purified efficiently.
- FIG. 6 shows one form of the manufacturing method of aliphatic dicarboxylic acid which has the contact process of the aqueous solution and solvent containing aliphatic dicarboxylic acid, a phase-separation process, and a solid content removal process.
- the underlined number indicates a primer consisting of the sequence of SEQ ID NO. 6 is a drawing showing composition changes in dilution / distillation operations of the extract in Example 2-1 and Example 3-1.
- 4 is a drawing showing a contact step of an aqueous solution containing succinic acid and a solvent, a phase separation step, and a solid content removal step in Example 2-4.
- 6 is a drawing showing a contact step of an aqueous solution containing succinic acid and a solvent, a phase separation step, and a solid content removal step in Example 2-8.
- 1st this invention is a method of manufacturing aliphatic dicarboxylic acid from the aqueous solution containing aliphatic dicarboxylic acid obtained by making a biological raw material and microorganisms react, Comprising: The following processes (1 ) To (4).
- Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms
- a contact process in which an aqueous solution obtained by separating microorganisms in the microorganism separation process is contacted with a solvent.
- the steps other than the above (1) to (3) may include any conventionally known steps. Hereafter, it explains in detail according to each process.
- Aliphatic dicarboxylic acid production process in which aliphatic dicarboxylic acid is accumulated in the reaction liquid by reacting biological raw material with microorganisms
- biological materials include wood, rice straw, rice husk, rice bran, old rice, corn, sugar cane, cassava, sago palm, okara, corn cob, tapioca cass, bagasse, vegetable oil cass, persimmon, buckwheat, soybean, fat, waste paper, paper Residue, marine product residue, livestock excrement, sewage sludge, food waste and the like.
- plant resources such as wood, rice straw, rice husk, rice bran, old rice, corn, sugar cane, cassava, sago palm, okara, corn cob, tapioca cass, bagasse, vegetable oil residue, persimmon, buckwheat, soybean, oil, waste paper and paper residue More preferred are wood, rice straw, rice husk, old rice, corn, sugar cane, cassava, sago palm, straw, oil, waste paper and papermaking residue, and most preferred are corn, sugar cane, cassava and sago palm.
- the biological raw material generally contains a lot of alkali metals and alkaline earth metals such as nitrogen element, Na, K, Mg, and Ca. *
- the biological raw material is not particularly limited.
- the biological raw material is converted into a carbon source through a known pretreatment such as a chemical treatment such as acid and alkali, a biological treatment using a microorganism, a physical treatment, and a saccharification step. Be guided.
- the process is not particularly limited, but includes, for example, a miniaturization process by pretreatment such as chipping, shaving, and crushing of a biological material. If necessary, a grinding step by a grinder or a mill is further included.
- the biological raw material thus refined is further guided to a carbon source through pretreatment and saccharification processes.
- Specific methods thereof include, for example, chemical methods such as acid treatment with strong acids such as sulfuric acid, nitric acid, hydrochloric acid and phosphoric acid, alkali treatment, ammonia freeze steaming explosion method, solvent extraction, supercritical fluid treatment and oxidant treatment, Examples include physical methods such as fine pulverization, steam explosion, microwave treatment, and electron beam irradiation, and biological treatment such as hydrolysis by microorganisms or enzyme treatment.
- microorganism The type of microorganism is not particularly limited, but examples include intestinal bacteria such as Escherichia coli, Bacillus bacteria, coryneform bacteria, etc., and it is preferable to use aerobic microorganisms, facultative anaerobic microorganisms, or microaerobic microorganisms. .
- the aerobic microorganism include, for example, Coryneform Bacterium, Bacillus genus bacteria, Rhizobium genus bacteria, Escherichia genus bacteria, Lactobacillus genus bacteria, and Lactobacillus genus bacteria.
- coryneform bacteria are preferable.
- Escherichia bacteria examples include, for example, Escherichia coli.
- Lactobacillus examples include Lactobacillus helveticas (J Appl Microbiol, 2001, 91, p846-852).
- Bacillus bacteria examples include Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus pumilus, Bacillus stearothermophilus, and the like.
- Rhizobium bacteria examples include Rhizobium etli.
- the coryneform bacterium is not particularly limited as long as it is classified as such.
- bacteria belonging to the genus Corynebacterium, bacteria belonging to the genus Brevibacterium, bacteria belonging to the genus Arthrobacter, myco Examples include bacteria belonging to the genus Mycobacterium, the genus Microbacterium, the bacteria belonging to the genus Micrococcus, and the like.
- bacteria belonging to the genus Corynebacterium and bacteria belonging to the genus Brevibacterium are preferable, and bacteria classified as Corynebacterium glutamicum are classified as Brevibacterium flavum. More preferred are bacteria, bacteria classified as Brevibacterium ammoniagenes, and bacteria classified as Brevibacterium lactofermentum.
- Microorganisms that convert carbon sources into aliphatic dicarboxylic acids include wild strains of microorganisms as described above, mutant strains obtained by normal mutation treatment such as UV irradiation and NTG treatment, and genetics such as cell fusion and genetic recombination methods. And recombinants derived by a conventional method.
- genetic recombinants those obtained by known methods such as enhancing the expression of biosynthetic enzyme genes and decreasing the expression of degrading enzyme genes are used for each aliphatic dicarboxylic acid.
- a microorganism that has been genetically modified so that pyruvate carboxylase activity is enhanced compared to an unmodified form, and a lactodehydrogenase activity that is unmodified form examples include microorganisms that have been genetically modified so as to be reduced.
- parent strain of coryneform bacterium used for recombinant production include Brevibacterium flavum MJ-233 (FERM BP-1497), MJ-233 AB-41 (FERM BP-1498), Brevibacterium ammoniagenes ATCC6872, Corynebacterium glutamicum ATCC31831, Brevibacterium lactofermentum ATCC 13869, and the like.
- Brevibacterium flavum is currently sometimes classified as Corynebacterium glutamicum (Lielbl, W., Ehrmann, M., Ludwig, W. and Schleiffer, K.H., International Journal of Systematic). Bacteriology, 1991, vol. 41, p255-260).
- Brevibacterium flavum MJ-233 strain and its mutant MJ-233 AB-41 strain are the same as Corynebacterium glutamicum MJ-233 strain and MJ-233 AB-41 strain, respectively. Stocks.
- Brevibacterium flavum MJ-233 was established on April 28, 1975, by the Ministry of International Trade and Industry, Institute of Industrial Science, Microbial Industrial Technology Research Institute (currently an independent administrative agency, National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center) (Japan 305-8666, Japan) Deposited as FERM P-3068 at Tsukuba City, Ibaraki Prefecture, 1-chome, 1st-chome, 1st-century 6th), transferred to an international deposit based on the Budapest Treaty on May 1, 1981, and deposited under the FERM BP-1497 deposit number. Has been.
- PC pyruvate carboxylase
- Transformation can be performed by, for example, an electric pulse method (Res. Microbiol., Vol. 144, p. 181-185, 1993).
- Enhancement of PC activity can be achieved by a known method such as J. Org. Bacteriol. , 158, 55-62, (1984), and can be confirmed by measuring PC activity.
- LDH lactodehydrogenase
- the LDH activity may completely disappear.
- the decrease or disappearance of the LDH activity can be confirmed by measuring the LDH activity by a known method (L. Kanarek, et al., J. Biol. Chem. 239, 4202 (1964), etc.).
- the microorganism used in the method for producing succinic acid is preferably a microorganism in which both PC activity enhancement and LDH activity reduction have been made, as described in the Examples below.
- Examples of such microorganisms include the Brevibacterium flavum MJ233 / PC-4 / ⁇ LDH strain described in Japanese Patent Application Laid-Open No. 2008-259451.
- microorganism that converts the carbon source into the aliphatic dicarboxylic acid is not limited to the above microorganism, and any known microorganism can be used.
- Succinic acid can be accumulated in the reaction solution by reacting a microorganism that converts the carbon source to succinic acid with a carbon source in a reaction solution such as a medium containing the carbon source.
- a slant cultured in a solid medium such as an agar medium may be directly used for the reaction.
- a microorganism that has been grown in advance (seed culture) in a liquid medium it is preferable to use a microorganism that has been grown in advance (seed culture) in a liquid medium.
- a normal medium used for culturing the above microorganisms can be used.
- a general medium in which natural nutrient sources such as meat extract, yeast extract and peptone are added to a composition comprising inorganic salts such as ammonium sulfate, potassium phosphate and magnesium sulfate can be used.
- Species culture is usually performed for coryneform bacteria while aeration and agitation are performed within a temperature range of 25 ° C. to 35 ° C., which is the optimum temperature for growth, and oxygen is supplied.
- the culture time may be a time for obtaining a certain amount of cells, but is usually 6 to 96 hours.
- aliphatic dicarboxylic acid examples include lactic acid, succinic acid, malic acid, fumaric acid, oxaloacetic acid, citric acid, isocitric acid, 2-oxoglutaric acid, cis-aconitic acid, pyruvic acid, and acetic acid.
- dicarboxylic acids such as succinic acid, malic acid and fumaric acid are preferable, and succinic acid is particularly preferable.
- a plurality of types of aliphatic dicarboxylic acids may be included.
- reaction solution In a reaction solution containing a carbon source derived from the biological material, a reaction between the carbon source and a microorganism that converts the carbon source to an aliphatic dicarboxylic acid is carried out to react the aliphatic dicarboxylic acid in the reaction solution. Can be accumulated.
- hexoses such as glucose, mannose, galactose, fructose, sorbose and tagatose
- pentoses such as arabinose, xylose, ribose, xylulose and ribulose
- maltose sucrose, lactose, trehalose
- Disaccharides or polysaccharides such as starch and cellulose, butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, monoctinic acid
- Fatty acids such as arachidic acid, eicosenoic acid, arachidonic acid, behenic acid, erucic acid, docosapentaenoic acid, docosahexaenoic
- the concentration of the carbon source used in the reaction solution is not particularly limited, but it is advantageous to make it as high as possible within the range that does not inhibit the production of succinic acid. It is preferably 30% (W / V), more preferably 10 to 20% (W / V).
- the reaction solution containing the carbon source is not particularly limited, and may be, for example, a medium for culturing microorganisms or a buffer solution such as a phosphate buffer solution.
- the reaction solution is preferably an aqueous solution further containing a nitrogen source and an inorganic salt.
- the nitrogen source is not particularly limited as long as it is a nitrogen source that can be assimilated by microorganisms to generate succinic acid.
- a nitrogen source that can be assimilated by microorganisms to generate succinic acid.
- organic or inorganic nitrogen compounds such as food, peptone, yeast extract, meat extract and corn steep liquor.
- the inorganic salt for example, metal salts such as various phosphates, sulfates, magnesium, potassium, manganese, iron and zinc are used. Moreover, it is preferable to add factors that promote growth such as vitamins such as biotin, pantothenic acid, inositol, and nicotinic acid, nucleotides, and amino acids as necessary. In order to suppress foaming during the reaction, it is preferable to add an appropriate amount of a commercially available antifoaming agent to the culture solution.
- the reaction liquid preferably contains, for example, carbonate ion, bicarbonate ion or carbon dioxide gas (carbon dioxide gas) in addition to the above-described carbon source, nitrogen source and inorganic salt.
- Carbonate ion or bicarbonate ion is supplied from magnesium carbonate, sodium carbonate, sodium bicarbonate, potassium carbonate and potassium bicarbonate, which can also be used as a neutralizing agent. Or it can also supply from these salts or carbon dioxide gas.
- carbonate or bicarbonate salt examples include magnesium carbonate, ammonium carbonate, sodium carbonate, potassium carbonate, ammonium bicarbonate, sodium bicarbonate, and potassium bicarbonate.
- the amount of carbonate ion or bicarbonate ion added to the reaction solution is preferably 1 to 500 mM, more preferably 2 to 300 mM, and even more preferably 3 to 200 mM.
- the content of carbon dioxide gas per liter of the solution is preferably 50 mg to 25 g, more preferably 100 mg to 15 g, and further preferably 150 mg to 10 g.
- the pH in the reaction system is measured by a pH sensor, and the pH is adjusted by adding a neutralizing agent so that it falls within a predetermined pH range.
- the pH value is adjusted to a range in which the activity is most effectively exhibited according to the type of microorganisms such as bacteria and mold used.
- a neutralizing agent Continuous addition or intermittent addition may be sufficient.
- Examples of the neutralizing agent include ammonia, ammonium carbonate, urea, alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal carbonate, and alkaline earth metal carbonate. Among these, ammonia, ammonium carbonate, and urea are preferable.
- alkali (earth) metal hydroxide examples include NaOH, KOH, Ca (OH) 2 and Mg (OH) 2 , and mixtures thereof.
- alkali (earth) metal carbonates e.g., Na 2 CO 3, K 2 CO 3, CaCO 3, etc. MgCO 3 and NaKCO 3, etc. and mixtures thereof.
- the pH in the production reaction is usually preferably 5 to 10, more preferably 6 to 9.5.
- the temperature of the production reaction is preferably 37 to 45 ° C, more preferably 39 ° C to 45 ° C, further preferably 39 ° C to 43 ° C, and particularly preferably 39 to 41 ° C. .
- the reaction time is preferably 1 hour to 168 hours, more preferably 3 hours to 72 hours.
- the amount of the microorganism used for the reaction is not particularly limited, but is preferably 1 to 700 g / L, more preferably 10 to 500 g / L, and further preferably 20 to 400 g / L. .
- the production reaction may be carried out with aeration and stirring, but it may be carried out in an anaerobic atmosphere in which oxygen is not supplied without aeration.
- anaerobic atmosphere here, for example, the container is sealed and reacted without aeration, supplied with an inert gas such as nitrogen gas and reacted, or the inert gas containing carbon dioxide gas is vented.
- the microorganism that converts the carbon source into succinic acid and the method for obtaining the liquid containing the succinic acid and the microorganism using the microorganism have been described as examples.
- the microorganism that converts the carbon source into an aliphatic dicarboxylic acid other than succinic acid There are also many known methods for producing aliphatic dicarboxylic acids using them.
- reaction conditions such as reaction temperature and pressure in producing aliphatic dicarboxylic acid by microorganisms depend on the activity of microorganisms such as selected cells and molds, but suitable conditions for obtaining aliphatic dicarboxylic acid May be selected according to each case.
- the fermentation broth after microbial conversion may be appropriately concentrated in consideration of operability and efficiency in the subsequent purification step.
- concentration method For example, the method of distribute
- An aliphatic dicarboxylic acid can be produced by applying the following purification steps (2) to (4) to the “liquid containing an aliphatic dicarboxylic acid and a microorganism” obtained as described above.
- I- (2) pH adjusting step for obtaining a solution having a pH of 1.0 or more and 5.0 or less by mixing a solution containing an aliphatic dicarboxylic acid obtained in the step of producing an aliphatic dicarboxylic acid and a microorganism and an acid.
- the pH of the fermentation suspension when separating cells from a liquid (fermentation suspension) containing an aliphatic dicarboxylic acid and a microorganism is usually preferably 1.0 or more and 5.0 or less, and preferably 1.5 or more and 4. 0 or less is more preferable, and 2.0 or more and 3.5 or less is more preferable.
- the pH of the fermentation suspension it is not particularly limited as long as it is an acid, but the pH can be adjusted using an acid such as hydrochloric acid, sulfuric acid, phosphoric acid and nitric acid.
- the aliphatic dicarboxylic acid When the aliphatic dicarboxylic acid is recovered as a salt aqueous solution in the aqueous solution containing the aliphatic dicarboxylic acid when the aliphatic dicarboxylic acid is recovered in the solvent in the contact step described later, the aliphatic dicarboxylic acid and / or the aliphatic dicarboxylic acid In some cases, the amount of the salt of the aliphatic dicarboxylic acid extracted in the solvent that is phase-separated from the aqueous solution containing the aliphatic dicarboxylic acid is small. Therefore, it is preferable to protonate by adjusting the pH by adding an acid to the aqueous solution.
- the hydrogen ion concentration (pH) of the fermentation liquid may be adjusted in order to advance the fermentation efficiently.
- the aliphatic dicarboxylic acid When alkali neutralization is performed, the aliphatic dicarboxylic acid is present as an aqueous salt solution, and thus it is particularly preferable to perform protonation.
- the aliphatic dicarboxylic acid when ammonia is used as a neutralizing agent in the fermentation operation, the aliphatic dicarboxylic acid is present as an ammonium salt, so that protonation with an acid is preferably performed.
- Protonation may be carried out at any stage as long as it is before the contact step described later.
- the acid used for protonation needs to be salt-exchanged with an aliphatic dicarboxylate, it is usually stronger than an aliphatic dicarboxylic acid, that is, an acid having an acid dissociation constant pKa smaller than that of an aliphatic dicarboxylic acid, usually a pKa of 4 It is preferable to use less acid.
- the acid used for protonation may be an organic acid, an inorganic acid, a monovalent acid or a polyvalent acid, but an inorganic acid is preferred. .
- the amount of acid to be added depends on the strength of the acid to be used, but it is usually preferable to add an acid in an amount of 0.1 to 5 times the amount of the cation constituting the aliphatic dicarboxylate. Usually, acid addition is adjusted by pH.
- the pH of the acid used for protonation depends on the acid strength pKa of the aliphatic dicarboxylic acid, it is preferably at least pKa or less. Further, the pH is preferably in the range of more than 1 and less than 4.
- the protonation step may be included in the pH adjustment step, or the protonation step may be performed in the pH adjustment step.
- Microorganism separation step for separating microorganisms from the liquid obtained in the pH adjustment step
- the methods for separating and removing microorganisms from the fermentation suspension include sedimentation separation, centrifugal separation and filtration separation, and a combination thereof. Is used. Industrially, it is carried out by methods such as centrifugation and membrane filtration separation. In centrifugation, centrifugal sedimentation, centrifugal filtration, or the like can be used.
- the centrifugal separation method is not limited to the type of separator and the operating conditions as long as it can remove microorganisms of usually 70% or more, preferably 80% or more, more preferably 90% or more. It is preferable to separate with a centrifugal force of 100,000 G.
- the separation operation can be performed either batchwise or continuously.
- the membrane used for membrane filtration is not particularly limited as long as it is a pore size membrane through which aliphatic dicarboxylic acid can pass and microorganisms and aggregates thereof cannot pass through, but microfiltration membranes and ultrafiltration membranes are preferred, and microfiltration membranes are preferred. More preferred.
- the material of the film is not particularly limited, and for example, an organic film such as polyolefin, polysulfine, polyacrylonitrile and polyvinylidene fluoride, or an inorganic film such as ceramic can be used.
- either a dead end type or a cross flow type can be used.
- a method of performing membrane filtration after roughly removing microorganisms by centrifugation or the like is also used.
- I- (4) Contacting step in which an aqueous solution obtained by separating microorganisms in the microorganism separation step is contacted with a solvent Contacting an aqueous solution containing an aliphatic dicarboxylic acid after separation of microorganisms in the microorganism separation step and the solvent To extract the aliphatic dicarboxylic acid. You may make it contact with a solvent, after concentrating the said aqueous solution.
- the aliphatic dicarboxylic acid can be selectively extracted into the solvent, and saccharides, amino acids and inorganic salts having high water solubility are mainly distributed in the aqueous phase.
- By-product salt generated by protonation of the aliphatic dicarboxylate is distributed to the aqueous phase and can be easily separated from the aliphatic dicarboxylic acid.
- the by-product salt is ammonium sulfate
- ammonium sulfate can be recovered as ammonium sulfate containing organic components of amino acid and saccharide together with the amino acid and saccharide recovered in the aqueous phase by a treatment such as concentration, crystallization, and drying.
- This ammonium sulfate is useful as a fertilizer because it contains an organic substance moderately.
- the solvent used in the contacting step is not particularly limited as long as it is an organic solvent that is phase-separated from an aqueous solution containing an aliphatic dicarboxylic acid, but the ratio of inorganic value / organic value (hereinafter abbreviated as I / O value). Is preferably 0.2 or more and 2.3 or less, and more preferably 0.3 or more and 2.0 or less.
- the aliphatic dicarboxylic acid can be selectively extracted and efficiently separated from the impurities.
- the solvent used in the contacting step is preferably atmospheric pressure (1 atm) and a boiling point of 40 ° C. or higher, and more preferably 60 ° C. or higher.
- the boiling point at normal pressure is preferably 120 ° C. or lower, more preferably 100 ° C. or lower, and preferably 90 ° C. or lower.
- the solvent By using the solvent, it is possible to avoid the risk that the solvent evaporates and ignites, the problem that the solvent evaporates and the extraction efficiency of the aliphatic dicarboxylic acid decreases, and the problem that the solvent is difficult to recycle. .
- Inorganic values and organic values have been proposed by organic conceptual diagrams ("systematic organic qualitative analysis", Satoshi Fujita, Kazama Shobo (1974)) and have been set in advance for functional groups constituting organic compounds. The organic value and the inorganic value are calculated based on the numerical values, and the ratio is obtained.
- Examples of the organic solvent having an I / O value of 0.2 or more and 2.3 or less and a boiling point of 40 ° C. or more at normal pressure include ketone solvents such as methyl ethyl ketone, methyl isobutyl ketone and acetone, tetrahydrofuran, dioxane and the like.
- Examples include ether solvents, ester solvents such as ethyl acetate, nitrile solvents such as acetonitrile, and alcohols having 3 or more carbon chains such as propanol, butanol and octanol.
- Table 1 shows the I / O value and boiling point of each organic solvent.
- the solvent as described above is brought into contact with the aqueous solution obtained by separating the microorganisms in the microorganism separation step to extract the aliphatic dicarboxylic acid.
- the solvent is preferably added in a volume of 0.5 to 5 with respect to the volume 1 of the “aqueous solution obtained by separating microorganisms in the microorganism separation step”, and more preferably in a volume of 1 to 3. preferable.
- the temperature in the contacting step is not particularly limited as long as the aliphatic dicarboxylic acid is extracted, but is preferably 10 to 50 ° C, more preferably 20 to 40 ° C.
- the aliphatic dicarboxylic acid is recovered in the solvent.
- the aqueous solution contains an amino acid or a saccharide
- both the amino acid and the saccharide are distributed in the aqueous solution phase, so that the aliphatic dicarboxylic acid can be separated from the saccharide and the amino acid.
- the operation of bringing the aqueous solution containing the aliphatic dicarboxylic acid into contact with the aqueous solution and the solvent may be performed in one step or in multiple steps, but is preferably performed in multiple steps. Further, the solvent may be flowed in parallel or countercurrent with respect to the aqueous solution containing the aliphatic dicarboxylic acid.
- the aliphatic dicarboxylic acid obtained in the solvent may be subjected to further purification operations such as column treatment and crystallization treatment. Further, when a plurality of aliphatic dicarboxylic acids are contained, an operation of separating only the target aliphatic dicarboxylic acid may be further performed.
- the time for contacting is not particularly limited as long as the aliphatic dicarboxylic acid is sufficiently extracted, and is usually preferably 1 second to 5 hours, although it depends on the contact device and the contact conditions.
- the pressure at the time of contact is not particularly limited as long as the aliphatic dicarboxylic acid is sufficiently extracted, but when it is continuously used, it is usually operated at atmospheric pressure.
- a succinic acid derivative can be produced using the obtained succinic acid after producing succinic acid by the method of the present invention.
- succinic acid derivative for example, succinate, succinic anhydride, maleic anhydride, succinic acid ester, succinimide, 1,4-diaminobutane, succinic acid nitrile, 2-pyrrolidone, N-methylpyrrolidone (NMP), N-ethylpyrrolidone (NEP), 1,4-butanediol (1,4-BG), tetrahydrofuran (THF), ⁇ -butyrolactone (GBL), polytetramethine ether glycol (PTMG), and the like.
- NMP N-methylpyrrolidone
- NEP N-ethylpyrrolidone
- 1,4-butanediol 1,4-butanediol
- THF tetrahydrofuran
- GBL ⁇ -butyrolactone
- PTMG polytetramethine ether glycol
- a succinic acid-containing polymer can be produced by conducting a polymerization reaction using succinic acid produced by the production method of the present invention as a raw material.
- succinic acid produced in the present invention is processed into polymers such as polyester and polyamide. Can be used.
- succinic acid-containing polymer examples include a succinic acid polyester obtained by polymerizing a diol such as butanediol and ethylene glycol and succinic acid, and a diamine such as hexamethylenediamine and succinic acid. And succinic acid polyamide obtained.
- the second invention is a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, the fat comprising the following steps (1) to (3): It is a manufacturing method of a group dicarboxylic acid.
- a contact step in which an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material is brought into contact with a solvent that can be phase-separated with the aqueous solution.
- the steps other than the above (1) to (3) may include any conventionally known steps. Hereafter, it explains in detail according to each process.
- the solvent is not particularly limited as long as it can be phase-separated from an aqueous solution containing an aliphatic dicarboxylic acid, and the solvents described above in I- (4) can be used.
- the contact operation between the aqueous solution containing the aliphatic dicarboxylic acid and the solvent is performed as described above in I- (4).
- the temperature at the time of contact is not particularly limited as long as the aliphatic dicarboxylic acid is extracted, but it is preferably 30 to 60 ° C.
- the contact temperature By setting the contact temperature to 30 ° C. or higher, it is possible to suppress the increase in the viscosity of the solvent, to shorten the time required for sedimentation of solids described later, and to prevent the solids from floating in the solvent phase. It is possible to prevent solid content from being mixed into the phase. Moreover, by making a contact temperature 60 degrees C or less, the extraction rate of aliphatic dicarboxylic acid can be improved and efficiency is good.
- the aqueous solution and the solvent are phase-separated, but in some cases, a phase containing a solid content at the phase interface (hereinafter sometimes referred to as an intermediate phase) may be formed.
- the intermediate phase makes it difficult to separate the solvent phase (hereinafter sometimes referred to as the extraction phase) and the aqueous phase (hereinafter sometimes referred to as the extraction residual phase), or impurities to the extraction phase There is a risk of increasing the amount of contamination. Therefore, it is preferable to remove the intermediate phase.
- a particularly preferable form of the contacting step is to separate an extraction phase liquid, an intermediate phase, and an extracted residual phase by mixing an aqueous solution containing an aliphatic dicarboxylic acid and a solvent after mixing with a mixer settler.
- the intermediate phase is recovered and separated into solid and liquid, and the separated and recovered liquid is phase-separated as necessary, and then returned to at least one of the step before the contact step and the step after the phase separation step.
- the contact device may be any device as long as it can contact the aqueous solution containing the aliphatic dicarboxylic acid and the solvent and the solvent phase, recover the aqueous solution phase, and remove the solid content in the solid content removal step described later.
- the above-mentioned mixer-settler type extraction device is preferable because the device is simple and easy to operate.
- the mixer may be of any type as long as an aqueous solution containing an aliphatic dicarboxylic acid and a solvent capable of phase separation are sufficiently mixed, and examples thereof include a container having a stirring device and a static mixer.
- the mixer is preferably a static mixer from the viewpoint of wide operating tolerance and equipment cost.
- the method for producing an aliphatic dicarboxylic acid according to the second aspect of the present invention includes a contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is contacted with a solvent capable of phase separation with the aqueous solution, and the liquid is phase-separated after the contact step.
- a phase separation step described later is included.
- a solvent capable of phase separation from the aqueous solution is added to an aqueous solution containing an aliphatic dicarboxylic acid, and after sufficiently mixing, in the phase separation step, an extraction phase, an intermediate phase, and a residual extraction phase are added.
- These can be separated and recovered by a method of taking out from the vicinity of each phase through a discharge port, a method of taking out sequentially from the bottom of the contacted container, and the like.
- the intermediate phase containing a large amount of solids can be taken out together with the extraction phase, or can be taken out together with the extracted residual phase.
- the contact step when carried out by continuous operation, it is obtained by contacting and mixing with an aqueous solution containing an aliphatic dicarboxylic acid and a mixer part having a mixer for contacting and mixing the aqueous solution and a solvent capable of phase separation.
- a contact device hereinafter referred to as a mixer-settler type extractor
- a phase separation step of phase separation by allowing the mixed liquid to stand.
- the extractor phase, the intermediate phase and the extracted residual phase can also be recovered in the settler part.
- Solid content removal step for removing solid content generated by contact of the aqueous solution and the solvent in the contact step
- an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material The solid content generated by contact with the aqueous solution and the phase-separable solvent is removed.
- an aqueous solution containing an aliphatic dicarboxylic acid often contains higher molecular structures such as biological proteins. Proteins and the like are usually highly water-soluble, and most of them are distributed to the aqueous solution phase in the extraction operation. However, when they come into contact with the solvent in the contact process, the higher-order structure is destroyed and denatured, and both water and solvent Some of them do not dissolve and become solids.
- the solid content removal step according to the present invention, the solid content that is not dissolved in the water or the solvent generated in the contact step is removed.
- the solid content generated in the contact process tends to gather mainly in the vicinity of the liquid-liquid interface, which makes it difficult to separate the phases. Even if solid content is generated near the liquid-liquid interface, if it is contacted by a batch method, it will not cause a major problem in operation if the generated solid content is removed each time and the extracted phase and the extracted residual phase are recovered. There is a case.
- the liquid containing solids may have an adverse effect in the subsequent process.
- the extract containing aliphatic dicarboxylic acid recovered in the contact process may be concentrated due to the low concentration of aliphatic dicarboxylic acid, but if there is a solid content, the solid content adheres to the heating surface such as a reboiler and is burnt. , Deteriorate the heat transfer efficiency.
- the solid content removal method is not particularly limited, and for example, it may be performed by a method of filtering the solid content or a method of separating with a device having a function of phase-separating each phase after bringing incompatible liquids into contact with each other. it can.
- a solvent is added to a fermentation broth containing an aliphatic dicarboxylic acid and mixed well, and then an extraction phase, an intermediate phase containing a large amount of solids, and a residue phase can be separated and recovered.
- a mixer-settler type extractor comprising a mixer part for mixing the fermentation liquor and the solvent and a settler part for separating the liquid mixture into liquids, the extractor, the intermediate phase containing a large amount of solids, and the residue Each phase can be recovered.
- the mixer may be of any type as long as the fermented liquid and the solvent are sufficiently mixed, and examples thereof include a stirring tank and a static mixer.
- examples thereof include a stirring tank and a static mixer.
- the bubbles that have been entrained in the stirrer by stirring to the generated solid matter adhere to the solid matter, and the settling of the solid matter in the subsequent settler is significantly inhibited. Be careful.
- the mixer is preferably a static mixer.
- the type of settler is not particularly limited.
- a type in which the extraction phase, the intermediate phase, and the extracted residual phase are respectively collected in a single tank type, and a type in which the extraction phase, the intermediate phase, and the extracted residual phase are respectively recovered in a multi-tank type, and the like.
- the intermediate phase rich in solids also includes an extraction phase and a residual phase
- solid-liquid separation is performed to separate the solid and the extracted phase and / or the residual phase, and the extracted phase and / or the residual phase is recovered. be able to.
- the solid-liquid separation method is not particularly limited, but any method of sedimentation separation and filtration separation can be used.
- the solid content may be separated by sedimentation in a gravitational field, or the solid content may be separated by sedimentation in a centrifugal field.
- centrifugal sedimentation is preferred because of its sedimentation speed.
- the method may be batch operation or continuous operation.
- Examples of the continuous centrifugal sedimentator include a screw decanter and a separation plate centrifugal sedimentator.
- the method is classified according to filter medium, filtration pressure, continuous operation / batch operation, etc., but any method is not particularly limited as long as the solid content can be separated from the extraction phase and / or the extraction residual phase.
- the aperture of the filter medium is preferably 0.1 ⁇ m or more and 10 ⁇ m or less.
- the permeation flux becomes sufficient, and the filtration time can be shortened.
- the thickness is 10 ⁇ m or less, the solid content is sufficiently separated.
- the material of the filter medium needs to be insoluble in the solvent, it is preferable to use Teflon (registered trademark) or the like.
- Teflon registered trademark
- any of a vacuum type, a pressure type, and a centrifugal type can be used.
- the system may be a continuous system or a batch system.
- the solid content may be removed by a method of selectively separating the solid content, or may be removed together with the liquid used in the contact step. However, in order to increase the recovery efficiency of the aliphatic dicarboxylic acid, only the solid content is removed. It is preferable to remove it selectively. When it is removed together with the liquid used in the contacting step, it is preferable to selectively remove only the solid content by solid-liquid separation in the subsequent step.
- the solid content is taken out and removed together with the liquid used in the contact step, and the solid content is further separated from the mixture of the taken out liquid and the solid matter, and then only the liquid is again the step before the contact step and the phase separation. It is preferable to return to at least one of the steps after the step.
- phase separation step for separating the solvent by phase separation can be performed by standing in a tank for a certain period of time or can be performed by a centrifugal separator.
- the mixer-settler type extractor as described above has a settler part that causes phase separation by allowing the mixed liquid obtained by contact mixing to stand, and by allowing the liquid to stand for a certain time in the settler part. Can be phase separated.
- the phase separation step may be performed continuously or batchwise.
- the temperature at the time of phase separation is not particularly limited as long as each phase can be separated, but it is preferably 30 to 60 ° C. and is preferably treated at the same temperature as the contact operation.
- phase separation temperature By setting the phase separation temperature to 30 ° C. or higher, the liquid viscosity is prevented from becoming high, the solid content is easily separated, and the solid content in the solvent phase and the increase in the amount of the solvent mixed in the solid content are suppressed. Can do.
- phase separation temperature By setting the phase separation temperature to 60 ° C. or lower, it is possible to prevent succinic acid from being back-extracted into the aqueous solution during the phase separation process.
- the time for phase separation is not particularly limited as long as each phase is phase-separated, and it is usually 1 minute to 5 hours, although it depends on the contact apparatus, contact conditions and phase separation method.
- phase separation time By setting the phase separation time to 1 minute or longer, phase separation is sufficient, and mixing of the aqueous solution and solid content into the solvent phase and the mixing of the solvent and solid content into the aqueous solution phase can be prevented. Further, by setting the phase separation time to 5 hours or less, it is possible to prevent the phase separation apparatus from becoming unnecessarily large and efficient.
- the pressure at the time of phase separation is not particularly limited as long as the aliphatic dicarboxylic acid is sufficiently extracted, but it is usually preferable to operate at atmospheric pressure when continuously performed.
- the settler used in the phase separation step may be any system as long as it can phase-separate the liquid after bringing the aqueous solution containing the aliphatic dicarboxylic acid into contact with the solvent capable of phase separation. May be.
- one that collects the extraction phase, the intermediate phase, and the residual phase in a single tank one that collects the extraction phase, the intermediate phase, and the residual phase in a multi-tank system, and centrifugal separation using a rotating device And the like for recovering each phase.
- the intermediate phase containing a large amount of solid usually contains at least one liquid selected from the liquid of the extraction phase and the liquid of the extraction residual phase
- the intermediate phase is obtained by the same method as described above in II- (2). Can be separated into solid and liquid, and at least one liquid selected from the liquid of the extraction phase and the liquid of the extraction residual phase can be separated and recovered.
- the aliphatic dicarboxylic acid of the second aspect of the present invention is not particularly limited as long as it is produced from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, and is an aliphatic hydrocarbon having two carboxy groups. Any one can be adopted.
- the carbon number of the aliphatic dicarboxylic acid is preferably 2 or more and 40 or less, more preferably 3 or more and 20 or less, and particularly preferably 3 or more and 10 or less.
- the carbon number of the hydrocarbon group 40 or less it becomes easy to exist as a stable aqueous solution. Moreover, it becomes easy to exist stably in the solvent which can be phase-separated with aqueous solution by making C2 or more into carbon number.
- the aliphatic hydrocarbon group portion of the aliphatic dicarboxylic acid may be linear or cyclic.
- oxalic acid succinic acid, glutaric acid, adipic acid, sebacic acid, malic acid, fumaric acid, oxaloacetic acid, 2-oxoglutaric acid, cis-aconitic acid, dodecanedioic acid, dimer acid, etc.
- adipic acid, succinic acid and dimer acid are preferable, and succinic acid and adipic acid are particularly preferable.
- an aliphatic dicarboxylic acid is derived from a carbon source derived from a biological material.
- a carbon source derived from a biological material.
- fermentation methods by microbial conversion chemical conversion methods including reaction steps such as hydrolysis, dehydration reaction, hydration reaction, oxidation reaction, reduction reaction, and combinations thereof
- the dicarboxylic acid is preferably synthesized by the above.
- a fermentation method by microbial conversion using a microorganism having an ability to produce an aliphatic dicarboxylic acid is preferable. Fermentation by microbial conversion using microorganisms capable of producing aliphatic dicarboxylic acids is carried out as described above in I- (1).
- the fermented liquid when using a fermented liquid in the method of this invention, it is preferable to use the fermented liquid after removing microorganisms.
- the method for removing microorganisms is performed as described above in I- (3).
- the aliphatic dicarboxylic acid obtained by microbial conversion from the carbon source derived from the biological material is usually dissolved in the aqueous solution, but before the subsequent contact step, the dicarboxylic acid is present.
- a part of may be present as a solid, for example, by precipitation.
- the aqueous solution containing an aliphatic dicarboxylic acid obtained by microbial conversion usually contains saccharides, amino acids, etc. used in the microbial conversion, and this aqueous solution and solvent are used in a subsequent contact step.
- this aqueous solution and solvent are used in a subsequent contact step.
- solids are easily generated, the problems of the present invention become remarkable, and the effect of the solution according to the present invention is also increased.
- extraction phase concentration process The extraction phase concentration step is performed as described later in III- (2).
- Crystallization process The crystallization step is performed as described later in III- (3).
- the aliphatic dicarboxylic acid slurry obtained in the crystallization step separates the aliphatic dicarboxylic acid crystal and the mother liquor by solid-liquid separation operation, as described later in III- (4).
- the third invention is a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, and comprises the following steps (1) to (5): It is a manufacturing method of the aliphatic dicarboxylic acid containing this.
- a contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is brought into contact with a solvent that is phase-separated with the aqueous solution.
- Extraction phase concentration step in which water concentration in phase increases (3) Crystallization step for precipitation of aliphatic dicarboxylic acid from liquid after extraction phase concentration step (4) Solid recovery of aliphatic dicarboxylic acid precipitated in crystallization step Liquid separation step (5) Crystallization mother liquor recycling step for returning at least part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step prior to the crystallization step
- the steps other than the above (1) to (5) may include any conventionally known steps. Hereafter, it explains in detail according to each process.
- the phase containing the extraction phase, the extracted residual phase, and the solid content is recovered from the settler portion of the mixer settler using the mixer settler, and the solid content recovered from the settler portion. More preferably, the phase containing is separated into solid and liquid and the liquid phase is recovered.
- concentration of the aliphatic dicarboxylic acid in the extraction phase is Since it is dilute, a concentration operation is required.
- the concentration is not particularly limited, but it is preferable that the solubility of the aliphatic dicarboxylic acid in the final concentrated solution is not more than the saturation solubility and is as close to the saturation solubility as possible.
- the solvent used for extraction of the aliphatic dicarboxylic acid often forms the lowest azeotropic composition with water, and the azeotropic composition is often a solvent-rich composition. Therefore, with the concentration operation, a large amount of the solvent is distilled off, and the solvent concentration in the concentrated solution is lower than that before the concentration.
- Aliphatic dicarboxylic acids obtained from biological raw materials generally contain many water-soluble impurities, and therefore, in the subsequent crystallization process, a system in which water is present has a higher purification effect than a system in which a solvent coexists. I can expect.
- the concentration of the solvent contained in the solution containing the aliphatic dicarboxylic acid after the extraction phase concentration step is preferably 10% by weight or less. It is more preferable to set the weight% or less.
- water is added in at least one of the steps before concentration of the extraction phase and the concentration operation. It is preferable.
- Crystallization process for precipitating aliphatic dicarboxylic acid from the liquid after the extraction phase concentration process usually involves converting the aliphatic dicarboxylic acid in the solution containing the extraction phase to the solubility difference of the dicarboxylic acid, etc. Is a step of precipitating solid aliphatic dicarboxylic acid using
- the crystallization method may be any method as long as the aliphatic dicarboxylic acid is precipitated as a solid from the solution containing the extraction phase.
- a cooling crystallization method in which crystallization is performed by utilizing the temperature dependence of solubility by changing the solution temperature, and a solvent in the solution is volatilized by evaporating the solvent from the solution by operations such as heating and decompression.
- cooling methods include, for example, a method in which a solution containing an extraction phase is circulated through an external heat exchanger and the like, and a pipe through which a refrigerant flows is placed in the solution containing the extraction phase. There are ways to do it.
- the crystallization operation may be a batch operation or a continuous operation.
- the continuous operation is preferable because the variation in the particle size of the solid aliphatic dicarboxylic acid obtained can be reduced, and the energy required for crystallization can be reduced efficiently during mass production.
- the crystallizer need not be a special crystallization tank, and a known stirring tank can be used.
- Solid-liquid separation step for recovering the aliphatic dicarboxylic acid precipitated in the crystallization step
- the aliphatic dicarboxylic acid slurry obtained by the crystallization separates the aliphatic dicarboxylic acid crystal and the mother liquor by solid-liquid separation operation.
- the separation method is not particularly limited, and examples thereof include filtration separation and sedimentation separation.
- the solid-liquid separation operation may be batch or continuous.
- Examples of the efficient solid-liquid separator include a continuous centrifugal filter and a centrifugal sedimentator such as a decanter.
- the wet cake recovered by the solid-liquid separation operation can be rinsed with cold water or the like.
- Crystallization mother liquor recycling step in which at least a part of the crystallization mother liquor after recovery of the aliphatic dicarboxylic acid obtained in the solid-liquid separation step is returned to any step prior to the crystallization step. At least a part of the mother liquor and / or cleaning liquid obtained in the process can be recycled to a process prior to the crystallization process.
- the process of recycling the crystallization mother liquor is not particularly limited, but can be recycled to the contact process and the concentration process.
- the extraction tower becomes larger, but impurities that are easily distributed to the aqueous phase (having a small distribution coefficient) can be selectively removed from within the recycling system.
- the extraction tower may be small, but all non-volatile impurities will accumulate in the recycling system.
- Purge water is usually discharged after treating organic substances such as activated sludge, but purge water contains aliphatic dicarboxylic acid and has a low pH. It is effective as
- the amount and location of recycling can be determined according to conditions such as the purity, yield and yield of the desired aliphatic dicarboxylic acid.
- the aliphatic dicarboxylic acid produced by the production method of the third invention is the same as the aliphatic dicarboxylic acid of the second invention.
- an aliphatic dicarboxylic acid is derived from a carbon source derived from a biological material.
- the protonation step of adding an acid to an aqueous solution containing an aliphatic dicarboxylic acid may be a step performed at any stage as long as it is a step prior to the contact step.
- a decolorization process using an adsorbent such as activated carbon an ion exchange process for removing coexisting ions with an ion exchange resin, a process for hydrogenating coexisting unsaturated dicarboxylic acid, and a crystallization process for further purification.
- an adsorbent such as activated carbon
- an ion exchange process for removing coexisting ions with an ion exchange resin a process for hydrogenating coexisting unsaturated dicarboxylic acid
- a crystallization process for further purification can be considered.
- Reference Example 1 Preparation of a succinic acid fermenting strain (A) Extraction of genomic DNA of Brevibacterium flavum MJ233 Brevibacterium flavum MJ233 was introduced on April 28, 1975 at the Institute of Microbial Industrial Technology, Ministry of International Trade and Industry. (Currently the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center) (deposited as FERM P-3068, 1st East, 1st Street, Tsukuba City, Ibaraki Prefecture, Japan 305-8586), May 1981 It was transferred to an international deposit based on the Budapest Treaty on the 1st and deposited under the deposit number of FERM BP-1497.
- a medium [urea 2 g, (NH 4 ) 2 SO 4 7 g, KH 2 PO 4 0.5 g, K 2 HPO 4 0.5 g, MgSO 4 .7H 2 O 0.5 g, FeSO 4 ⁇ 7H 2 O 6 mg, MnSO 4. 4-5H 2 O 6 mg, biotin 200 ⁇ g, thiamine 200 ⁇ g, yeast extract 1 g, casamino acid 1 g, glucose 20 g, dissolved in 1 L of distilled water]
- the Brevibacterium flavum MJ233 strain is cultured until the late logarithmic growth phase The cells were collected by centrifugation (10000 g, 5 minutes).
- the obtained bacterial cells were suspended in 0.15 mL of a 10 mM NaCl / 20 mM Tris buffer solution (pH 8.0) / 1 mM EDTA ⁇ 2Na solution containing lysozyme at a concentration of 10 mg / mL.
- proteinase K was added to the suspension so that the final concentration was 100 ⁇ g / mL, and the mixture was incubated at 37 ° C. for 1 hour.
- sodium dodecyl sulfate was added to a final concentration of 0.5%, and the mixture was incubated at 50 ° C. for 6 hours for lysis.
- (B) Construction of a plasmid for PC promoter substitution The DNA fragment of the N-terminal region of the pyruvate carboxylase gene derived from Brevibacterium flavum MJ233 strain was obtained using the DNA prepared in (A) above as a template and the entire genome sequence was reported. This was performed by PCR using synthetic DNA (SEQ ID NO: 1 and SEQ ID NO: 2) designed based on the sequence of the gene of Corynebacterium glutamicum ATCC13032 strain (Cgl0689 of GenBank Database Accession No. BA00000036). The DNA of SEQ ID NO: 1 used was phosphorylated at the 5 ′ end.
- composition of reaction solution Template DNA 1 ⁇ L, Pfx DNA polymerase (manufactured by Invitrogen) 0.2 ⁇ L, 1 ⁇ concentration attached buffer, 0.3 ⁇ M each primer, 1 mM MgSO 4 , 0.25 ⁇ M dNTPs were mixed to make a total volume of 20 ⁇ L.
- DNA thermal cycler PTC-200 manufactured by MJ Research was used, and a cycle of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds, and 72 ° C. for 1 minute was repeated 35 times. However, the heat retention at 94 ° C. in the first cycle was 1 minute 20 seconds, and the heat retention at 72 ° C. in the final cycle was 4 minutes.
- the TZ4 promoter fragment derived from Brevibacterium flavum MJ233 strain is constitutively highly expressed using plasmid pMJPC1 (Japanese Patent Application Laid-Open No. 2005-95169) as a template, and the synthetic DNAs described in SEQ ID NO: 3 and SEQ ID NO: 4 It was prepared by PCR using The DNA of SEQ ID NO: 4 used was phosphorylated at the 5 'end.
- composition of reaction solution 1 ⁇ L of template DNA, 0.2 ⁇ L of Pfx DNA polymerase (manufactured by Invitrogen), 1 ⁇ concentrated buffer, 0.3 ⁇ M each primer, 1 mM MgSO 4 , 0.25 ⁇ M dNTPs were mixed to make a total volume of 20 ⁇ L.
- DNA thermal cycler PTC-200 manufactured by MJ Research
- a cycle of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds and 72 ° C. for 30 seconds was repeated 25 times.
- the heat retention at 94 ° C. in the first cycle was 1 minute 20 seconds
- the heat retention at 72 ° C. in the final cycle was 3 minutes.
- the amplification product was confirmed by separation by 1.0% agarose (SeaKem GTG agarose: manufactured by FMC BioProducts) gel electrophoresis and visualization by ethidium bromide staining to detect a fragment of about 0.5 kb.
- the PC gene N-terminal fragment prepared above and the TZ4 promoter fragment were mixed, and ligation kit ver. 2 (manufactured by Takara Shuzo), cleaved with the restriction enzyme PstI, separated by 1.0% agarose (SeaKem GTG agarose: manufactured by FMCBioProducts) gel electrophoresis, and a DNA fragment of about 1.0 kb was extracted with QIAQuick Gel Extraction Kit (Manufactured by QIAGEN) was used as a TZ4 promoter: PC gene N-terminal fragment.
- DNA fragment was mixed with DNA prepared by cutting E. coli plasmid pHSG299 (Takara Shuzo) with PstI, and ligation kit ver. 2 (Takara Shuzo) were used for connection.
- Escherichia coli (DH5 ⁇ strain) was transformed with the obtained plasmid DNA.
- the recombinant Escherichia coli thus obtained was smeared on an LB agar medium containing 50 ⁇ g / mL kana machine and 50 ⁇ g / mL X-Gal.
- a clone that formed white colonies on the medium was subjected to liquid culture by a conventional method, and then the plasmid DNA was purified.
- the obtained plasmid DNA was cleaved with the restriction enzyme PstI, whereby an inserted fragment of about 1.0 kb was recognized, and this was named pMJPC17.1.
- the DNA fragment of the 5 ′ upstream region of the pyruvate carboxylase gene derived from Brevibacterium flavum MJ233 strain was obtained by using the DNA prepared in Reference Example 1 (A) as a template, and Corynebacterium It was carried out by PCR using a synthetic DNA (SEQ ID NO: 5 and SEQ ID NO: 6) designed based on the sequence of the gene of Glutamicum ATCC13032 strain (GenBank Database Accession No. BA00000036).
- composition of reaction solution 1 ⁇ L of template DNA, 0.2 ⁇ L of Pfx DNA polymerase (manufactured by Invitrogen), 1 ⁇ concentration buffer, 0.3 ⁇ M each primer, 1 mM MgSO 4 , 0.25 ⁇ M dNTPs were mixed to make a total volume of 20 ⁇ L.
- DNA thermal cycler PTC-200 manufactured by MJ Research
- a cycle consisting of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds and 72 ° C. for 30 seconds was repeated 35 times.
- the heat retention at 94 ° C. in the first cycle was 1 minute 20 seconds
- the heat retention at 72 ° C. in the final cycle was 5 minutes.
- the amplification product was confirmed by separation by 1.0% agarose (SeaKem GTG agarose: manufactured by FMC BioProducts) gel electrophoresis and visualization by ethidium bromide staining, and a fragment of about 0.7 kb was detected.
- the target DNA fragment was collected from the gel using a QIAQuick Gel Extraction Kit (manufactured by QIAGEN).
- the recovered DNA fragment was phosphorylated at the 5 ′ end with T4 polynucleotide kinase (T4 Polynucleotide Kinase: manufactured by Takara Shuzo), and then ligated kit ver. 2 (Takara Shuzo) was used to bind to the SmaI site of E. coli vector pUC119 (Takara Shuzo), and Escherichia coli (DH5 ⁇ strain) was transformed with the resulting plasmid DNA.
- the recombinant Escherichia coli thus obtained was smeared on an LB agar medium containing 50 ⁇ g / mL ampicillin and 50 ⁇ g / mL X-Gal. Clones that formed white colonies on this medium were subjected to liquid culture by a conventional method, and then plasmid DNA was purified. The obtained plasmid DNA was subjected to a PCR reaction using the synthetic DNAs shown in SEQ ID NO: 7 and SEQ ID NO: 6 as primers.
- composition of reaction solution 1 ng of the above plasmid, Ex-Taq DNA polymerase (Takara Shuzo) 0.2 ⁇ L, 1-fold concentration buffer, 0.2 ⁇ M each primer, 0.25 ⁇ M dNTPs were mixed to make a total volume of 20 ⁇ L.
- DNA thermal cycler PTC-200 manufactured by MJ Research
- a cycle consisting of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds and 72 ° C. for 50 seconds was repeated 20 times.
- the heat retention at 94 ° C. in the first cycle was 1 minute 20 seconds
- the heat retention at 72 ° C. in the final cycle was 5 minutes.
- pMJPC5.1 a plasmid that recognizes an amplification product of about 0.7 kb was selected and named pMJPC5.1.
- pMJPC17.1 and pMJPC5.1 were cut with restriction enzyme XbaI and mixed, and ligation kit ver. 2 (Takara Shuzo) were used for connection.
- the DNA fragment cleaved with restriction enzyme SacI and restriction enzyme SphI was separated by 0.75% agarose (SeaKem GTG agarose: manufactured by FMCBioProducts) gel electrophoresis, and a DNA fragment of about 1.75 kb was manufactured by QIAQUICK Gel Extraction Kit (QIAGEN Kit) ).
- Escherichia coli (DH5 ⁇ strain) was transformed with the obtained plasmid DNA.
- the recombinant Escherichia coli thus obtained was smeared on an LB agar medium containing 50 ⁇ g / mL kana machine and 50 ⁇ g / mLX-Gal. Clones that formed white colonies on this medium were subjected to liquid culture by a conventional method, and then plasmid DNA was purified.
- Transformation of Brevibacterium flavum MJ233 / ⁇ LDH strain was performed by the electric pulse method (Res. Microbiol., Vol. 144, p.181-185, 1993), and the obtained transformant contained 25 ⁇ g / mL of kanamycin.
- LBG agar medium [tryptone 10 g, yeast extract 5 g, NaCl 5 g, glucose 20 g, and agar 15 g dissolved in 1 L of distilled water] was smeared.
- the strain grown on the medium is a plasmid in which pMJPC17.2 cannot replicate in the Brevibacterium flavum MJ233 strain
- the PC gene of the plasmid and the same gene on the genome of Brevibacterium flavum MJ233 strain As a result of homologous recombination with the kanamycin, the kanamycin resistance gene and sacB gene derived from the plasmid should be inserted on the genome.
- the above homologous recombinant strain was subjected to liquid culture in an LBG medium containing 25 ⁇ g / mL kanamycin. A portion equivalent to about 1 million cells of this culture was smeared on a 10% sucrose-containing LBG medium. As a result, sacB gene was eliminated by the second homologous recombination, and several tens of strains considered to be insensitive to sucrose were obtained.
- the strains thus obtained include those in which the TZ4 promoter derived from pMJPC17.2 is inserted upstream of the PC gene and those that have returned to the wild type. Confirmation of whether the PC gene is promoter-substituted type or wild type can be easily confirmed by subjecting the cells obtained by liquid culture in LBG medium to direct PCR reaction and detecting the PC gene. .
- the pH was kept at 7.2% or lower using 28% aqueous ammonia, the main culture was started at aeration of 3.0 L / min, back pressure of 0.05 MPa, and stirring at 750 rpm. After the dissolved oxygen concentration dropped to almost 0, when it started to rise again and reached 1 ppm, about 5.3 g of a 72% sucrose aqueous solution sterilized in advance was added. Each time the dissolved oxygen concentration increased again, the addition of aqueous sucrose solution was repeated by the above method, and continued for 19 hours after the start of culture.
- Reaction temperature is 40 ° C.
- stirring speed is 150 per minute
- pH is adjusted to 7.35 by sequential addition of neutralizing agent (ammonium bicarbonate: 171 g, 28% ammonia water: 354 g, distilled water: 529 g).
- neutralizing agent ammonium bicarbonate: 171 g, 28% ammonia water: 354 g, distilled water: 529 g. The process was terminated when the residual sucrose in the reaction solution became 0.1 g / L or less.
- the succinic acid fermentation broth prepared in this way (containing the bacterial cells) was used in the following examples and comparative examples.
- the succinic acid fermentation broth was subjected to centrifugal separation (15,000 G, 5 minutes), and the resulting supernatant was subjected to composition analysis.
- the results shown in Table 2 were obtained.
- Example 1 [Example 1-1] ⁇ Cell separation> (Example 1-1-1) After adjusting the pH of the succinic acid fermentation liquor obtained in Reference Example 2 (C) above to 2.5 with 95% sulfuric acid, 10 mL was placed in a centrifuge tube, centrifuged at 500 G for 8 minutes, and then centrifuged. The turbidity (OD660) of the supernatant was measured. The turbidity (OD660) before cell separation was 24.
- Example 1-1-2 The cells were separated in the same manner as in Example 1-1-1 except that the pH was adjusted to 3.0, and the turbidity (OD660) of the supernatant after centrifugation was measured.
- Example 1-1-3 The cells were separated in the same manner as in Example 1-1-1 except that the pH was adjusted to 4.0, and the turbidity (OD660) of the supernatant after centrifugation was measured.
- Example 1-1-4 The cells were separated in the same manner as in Example 1-1-1 except that the pH was adjusted to 5.0, and the turbidity (OD660) of the supernatant after centrifugation was measured.
- Example 1-1-1-1 Bacterial cells were separated from the succinic acid fermentation broth (pH 7.6) in the same manner as in Example 1-1-1, and the turbidity (OD660) of the supernatant after centrifugation was measured.
- centrifugation was performed at 4200G for 8 minutes, and the same experiment as in Examples 1-1-1 to 1-1-4 and Comparative Example 1-1-1 was performed.
- Example 1-2 ⁇ Cell separation + MEK extraction> (Example 1-2-1) 50 mL of the succinic acid fermentation broth obtained in Reference Example 2 (C) above was placed in a centrifuge tube, adjusted to pH 2.5 with 95% sulfuric acid, and then centrifuged at 9500 G for 10 minutes. About the obtained supernatant liquid, turbidity (OD660) was measured using a part, and MEK extraction was performed by the following procedure using the rest.
- turbidity OD660
- the area (organic phase clear rate) in the organic phase where there is no suspended matter is a substantial proportion of the organic phase that can be recovered when extracting an organic acid such as succinic acid.
- light scattering measurement was performed using a part of the supernatant.
- Light scattering measurement was performed using an ELS-800 type apparatus manufactured by Otsuka Electronics Co., Ltd., ELS-8000 ver. Performed with 4.0 software.
- the incident slit was set to ⁇ 0.2 and ⁇ 0.1.
- the light source used was a He-Ne laser 10 mW, and the detector used was a photomultiplier tube for photocounting.
- the measurement was carried out using a 1 cm square cell at room temperature and using each treatment solution stock solution. The measurement angle was 90 ° C. and the solvent was water.
- Example 1-2-2 Cell separation was carried out in the same manner as in Example 1-2-1, except that the pH of the succinic acid fermentation broth obtained in Reference Example 2 (C) was adjusted to 3.0, and the resulting supernatant was After adjusting the pH to 2.5 with 95% sulfuric acid, measurement of OD660 and extraction of MEK were performed in the same manner as in Example 1-2-1, and the phase separation time, the amount of intermediate insoluble matter, the organic phase clear rate, the average The particle size and light scattering intensity were measured.
- Example 1-2-3 Except that the pH before cell separation was adjusted to 4.0, cell separation and MEK extraction were performed in the same manner as in Example 1-2-2, and OD660, phase separation time, amount of intermediate insoluble matter, and organic phase clear The rate, average particle size, and light scattering intensity were measured.
- Example 1-2-4 Cell separation and MEK extraction were carried out in the same manner as in Example 1-2-2 except that the pH before cell separation was adjusted to 5.0, OD660, phase separation time, amount of intermediate insoluble matter, organic phase clear The rate, average particle size, and light scattering intensity were measured.
- Example 1-2-1 (Comparative Example 1-2-1) Except that the pH before cell separation was adjusted to 7.6, cell separation and MEK extraction were performed in the same manner as in Example 1-2-2, and OD660, phase separation time, amount of intermediate insoluble matter, and organic phase clear The rate, average particle size, and light scattering intensity were measured.
- Example 1-3 Bacterial cell separation and MEK extraction were performed in the same manner as in Example 1-2-1 except that centrifugation was performed at 1000 G for 8 minutes, and OD660, phase separation time, and amount of intermediate insoluble matter were measured. As a result, OD660 was 0.191, phase separation time was 26 seconds, and the amount of intermediate insoluble matter was small. From this, it was found that MEK extraction can be performed efficiently even at low speed centrifugation.
- Example 1-4 (A) Preparation of succinic acid fermentation broth by jar fermenter A succinic acid fermentation broth was prepared according to Reference Example 2 using the strain prepared in Reference Example 1. However, the temperature during the succinic acid production reaction was 39 ° C. and the pH was 7.6. The obtained succinic acid fermentation broth was subjected to centrifugal separation (15,000 G, 5 minutes), and the resulting supernatant was subjected to composition analysis. The results are shown in Table 5.
- each pH sample was centrifuged at 500 G for 8 minutes. Of the obtained centrifugal supernatant, the pH 2.0-0.5 sample was readjusted to pH 2.5 using 28% aqueous ammonia, and the pH 7.6-3.0 sample was 95. Each was readjusted to pH 2.5 using% sulfuric acid.
- Turbidity (OD660) was measured using a part of the supernatant adjusted to pH 2.5. The measurement results are shown in Table 6 (centrifugation conditions: 500 G, 8 minutes). In addition, the turbidity (OD660) of the succinic acid fermentation liquid before cell separation was 25.
- Example 1-5 (A) Preparation of succinic acid fermentation broth by jar fermenter (A-1) Seed culture and main culture Using the strain prepared in Reference Example 1, according to the method described in Reference Example 2 (A) and (B) Culture and main culture were performed.
- reaction temperature is 40 ° C.
- stirring rotation speed is 200 rotations per minute
- pH is adjusted to 7.35 by sequential addition of a neutralizing agent (ammonium bicarbonate: 171 g, 28% aqueous ammonia: 354 g, distilled water: 529 g).
- a neutralizing agent ammonium bicarbonate: 171 g, 28% aqueous ammonia: 354 g, distilled water: 529 g.
- the succinic acid fermentation broth was subjected to centrifugal separation (15,000 G, 5 minutes), and the resulting supernatant was subjected to composition analysis.
- the results shown in Table 7 were obtained.
- the result of measuring the turbidity (OD660) of the succinic acid fermentation broth was 30.
- Example 1-6 (A) pH adjustment, centrifugation, and membrane separation
- the succinic acid fermentation broth obtained in Example 1-5 (A-2) was adjusted to pH 2.5 with 95% sulfuric acid. 35 mL of this was dispensed into a centrifuge tube, centrifuged at 4200 G for 8 minutes, and the resulting supernatant was subjected to membrane filtration (membrane pore size 0.22, 5 ⁇ m or no membrane filtration).
- Example 1-5 35 mL of the succinic acid fermentation broth obtained in Example 1-5 (A-2) was dispensed into a centrifuge tube and centrifuged at 4200 G for 8 minutes. Filtration (membrane pore size 0.22, 5 ⁇ m or no membrane filtration). Thereafter, the filtrate was adjusted to pH 2.5 with 95% sulfuric acid. Moreover, turbidity (OD660) measurement was performed on a portion of the membrane filtrate whose pH was adjusted to 2.5. Table 9 shows the measurement results.
- Example 2 high-performance liquid chromatography (LC) was used for quantitative analysis of acids and saccharides, and amino acid quantitative analysis was performed using an amino acid analyzer under the following conditions.
- LC liquid chromatography
- amino acid quantitative analysis was performed using an amino acid analyzer under the following conditions.
- protein quantification the sample was hydrolyzed with hydrochloric acid, and the increment of the total amino acid content before and after hydrolysis was regarded as the protein amount.
- ⁇ Hydrolysis for protein quantification A sample of 10 mg or 100 mg was precisely weighed and a 1 mL constant volume with pure water was dispensed to 200 ⁇ L, dried and heated in a hydrochloric acid atmosphere at 150 ° C. for 1 hour to hydrolyze the protein. After drying this, 200 ⁇ L of pure water was added and redissolved, and after filtration through a 0.45 ⁇ m filter, the filtrate was subjected to amino acid analysis.
- the respective weights were 2503 g of the extraction phase, 1490 g of the extraction residual phase, and 132 g of the intermediate phase.
- the composition of the extracted phase is shown in Table 10 below.
- the recovered extract substantially removes MEK by continuous distillation.
- the distilled distillate is recovered as an azeotropic composition of MEK and water, that is, 11 wt% water-containing MEK, but there is a concern that succinic acid may precipitate depending on the degree of concentration of the kettle residue. Therefore, 103 g of water was added to 2503 g of the extract so that the distilled distillate was 11 wt% hydrous MEK and the kettle residue was 30 wt% succinic acid solution.
- the amount of water added was calculated according to the following calculation.
- FIG. 3 shows a phase diagram when the MEK and succinic acid concentrations of the extract were C MEK, 0 and C SA, 0 , respectively, and the diluted MEK and succinic acid concentrations were C MEK, 1 and C SA, 1 respectively. .
- FIG. 3 shows the composition change of MEK and succinic acid in the dilution of the extract with water and subsequent distillation.
- the diluent composition (C MEK, 1 , C SA, 1 ) is obtained. Furthermore, the amount of dilution water added can be calculated as (C MEK, 0 -C MEK, 1 ) / C MEK, 1 with respect to the extract.
- the composition of the extract was (0.8220, 0.0353)
- the composition after dilution was (0.7895, 0.0339)
- Distillation was carried out using a 500 ml round bottom flask equipped with a packed column with a diameter of 40 mm and packed with a coil pack with a diameter of 5 mm and a height of 30 cm, and a normal pressure continuous distillation apparatus equipped with a distillation apparatus. About 300 ml of the diluted solution was added to the round bottom flask, and the flask was cooked while being heated in an oil bath, and the inside of the system was stabilized while applying total return.
- the temperature of the oil bath was 120 ° C.
- the column top temperature was 74 ° C. (azeotropic temperature), and the column bottom temperature was 101 ° C.
- set the return ratio to 1, continuously extract the distillate and continuously supply the feed liquid to the middle stage of the packed tower, and continuously extract the residual liquid from the kettle. Started.
- the feed liquid was heated to 60 ° C. in advance with a preheater and then fed to the packed tower.
- the supply liquid started to be supplied at 100 ml / hour so as not to disturb the tower top temperature and tower bottom temperature, and increased to 400 ml / hour over about 1 hour.
- the temperature of the oil bath was gradually increased as the supply liquid increased, and finally it was 140 ° C. It took 7 hours to distill 2606 g of the diluted solution.
- the bottom flask after distillation was relatively clean.
- ⁇ Crystal> The liquid from which MEK was distilled off was transferred to a jacketed 500 ml separable flask, and kept warm at 80 ° C. by passing warm water through the jacket while stirring. Thereafter, using a circulating thermostatic bath with a program, the warm water passing through the jacket was cooled to 20 ° C. over 1 hour, succinic acid was cooled and crystallized, and after reaching 20 ° C., the mixture was further aged at 20 ° C. for 1 hour.
- the obtained slurry was vacuum filtered and further washed with 150 g of cold water to collect a wet cake. Further, the obtained wet cake was dried at 80 ° C. under a maximum pressure using a vacuum dryer, and finally 78 g of succinic acid was recovered.
- the composition analysis results of the obtained succinic acid are shown in Table 12 below.
- Example 2-2 In Example 2-1, the treatment was performed in the same manner except that the extraction temperature was 20 ° C. The number of extracted intermediate phases was clearly larger than that of Example 2-1, and the recovered extracted phase, intermediate phase, and extracted residual phase were 2322 g, 316 g, and 1485 g, respectively. The results of composition analysis of the extracted phase are shown in Table 13 below.
- Example 2-3 In Example 2-1, the treatment was performed in the same manner except that the extraction temperature was 60 ° C.
- the extracted intermediate phase was clearly less than in Example 2-2, and the recovered extracted phase, intermediate phase, and extracted residual phase were 2498 g, 131 g, and 1495 g, respectively.
- the results of composition analysis of the extracted phase are shown in Table 14 below.
- the suspension was continuously extracted at a rate of 50 g / min while being vigorously stirred so that the inside of the vessel became uniform while being supplied to the 500 ml jacketed agitation vessel at a rate of 20 g / min and 30 g / min, respectively.
- the extracted suspension is supplied to the first tank of a three-tank settler whose temperature is controlled by flowing hot water of 30 ° C. through the jacket, and is separated into liquids, and the extracted residual phase is continuously extracted from the bottom of the first tank. It was.
- the extract overflowed the weir between the first tank and the second tank and was supplied to the second tank. A considerable amount of insoluble components were suspended in the extract supplied to the second tank.
- the extract remaining in each tank, the extracted residual phase, and the insoluble components remaining in the first tank intermediate phase and the second tank bottom were recovered.
- the recovered extracted phase, extracted residual phase, and intermediate phase were 2370 g, extracted residual phase 1490 g, and intermediate phase 260 g, respectively.
- the results of composition analysis of the extracted phase are shown in Table 15 below.
- aqueous succinic acid solution after the addition of sulfuric acid is a jacketed static mixer (Noritake 1/4 (1) -N40-174-0 (inner diameter 5 mm, number of elements 24)) and three tanks with jackets of 600 ml, 400 ml and 300 ml respectively.
- the succinic acid was continuously extracted by mixing with the MEK solution and separating the liquid using a settling formula.
- the extract liquid overflowed the weir between the first tank and the second tank and was supplied to the second tank.
- insoluble components that could not be separated in the first tank were allowed to settle to the bottom, and only the clear extract was allowed to overflow the weir between the second and third tanks and supplied to the third tank.
- the clear extract was overflowed from the liquid interface vicinity, and the extract was collect
- the extract remaining in each tank, the extracted residual phase, and the insoluble components remaining in the first tank intermediate phase and the second tank bottom were recovered.
- the recovered extracted phase, extracted residual phase, and intermediate phase were 2435 g, extracted residual phase 1490 g, and intermediate phase 197 g, respectively.
- the results of composition analysis of the extracted phase are shown in Table 18 below.
- Example 2-6 In accordance with the method of Example 2-1, protonation and extraction operations were performed to recover 2503 g of extraction phase, 1490 g of extraction residual phase, and 132 g of intermediate phase.
- the intermediate phase was centrifuged at 2000 G for 10 minutes with a centrifugal settling machine to recover 130 g of the clarified liquid.
- Table 20 below shows the composition analysis results of the liquid obtained by mixing the clarified liquid and the extract.
- Example 2-7 In accordance with the method of Example 2-1, protonation and extraction operations were performed to recover 2503 g of extraction phase, 1490 g of extraction residual phase, and 132 g of intermediate phase.
- the intermediate phase was subjected to pressure filtration with a PTFE membrane filter having an opening of 0.5 ⁇ m to recover 130 g of the clarified liquid.
- Table 22 shows the composition analysis results of the liquid obtained by mixing the clarified liquid and the extract.
- the recovered extract was 2637 g in total, including the extract recovered by the mixer settler and the clarified liquid from which the solid content was removed by pressure filtration of the intermediate phase.
- Table 24 shows the results of analysis of the composition of the recovered extract, the extract recovered by the mixer settler, and the mixture of the clarified liquid from which the solid content was removed by pressure filtration.
- ⁇ Distillation> 190 g of water was added to 2637 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 432 g of the residue from the kettle. The distillation took 7 hours, but the bottom flask after the distillation was relatively clean.
- Example 2-1 According to the method of Example 2-1, except that the intermediate phase was recovered as the extraction phase, 2633 g of the extraction phase and 1490 g of the extracted residual phase were recovered by the same method.
- ⁇ Distillation> 109 g of water was added to 2633 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 300 g of the residue from the kettle.
- the oil bath temperature was 140 ° C, but the bottom temperature decreased several hours after the start of continuous distillation (MEK detection), so the supply amount was lowered and the bottom temperature> 100 ° C was maintained at the oil bath temperature of 140 ° C. The operation was continued under conditions that allowed it. Results Distillation took 9 hours. In addition, a large amount of adhesive was adhered to the inner surface of the bottom flask after distillation.
- ⁇ Crystal> The liquid from which MEK was distilled off was transferred to a jacketed 500 ml separable flask, and kept warm at 80 ° C. by passing warm water through the jacket while stirring. Thereafter, using a circulating thermostatic bath with a program, the warm water passing through the jacket was cooled to 20 ° C. over 1 hour, succinic acid was cooled and crystallized, and after reaching 20 ° C., the mixture was further aged at 20 ° C. for 1 hour.
- the obtained slurry was vacuum filtered and further washed with 150 g of cold water to collect a wet cake. Further, the obtained wet cake was dried with a vacuum drier at 80 ° C. and a maximum reduced pressure, and finally 83 g of succinic acid was recovered.
- the results of composition analysis of the obtained succinic acid are shown in Table 26 below.
- Example 2-2 In accordance with the method of Example 2-1, a protonated succinic acid aqueous solution was supplied from the top of the continuous extraction tower used in Example 2-9 at 200 g / hour, and 10% hydrous MEK was supplied from the bottom at 300 g / hour. As a result, a large amount of insoluble components were generated in the tower, and liquid-liquid separation was inhibited, and continuous operation was not possible.
- Example 3 In this example, the quantitative analysis of acids, saccharides, quantitative analysis of amino acids, and protein were performed in the same manner as in Example 2.
- Example 3-1 ⁇ Protonation process> To 1500 g of the succinic acid fermentation broth having the composition shown in Table 2, 98% sulfuric acid was added to adjust the pH to 2.5. Here, the addition amount of 98% sulfuric acid was 150 g.
- the succinic acid aqueous solution after the addition of sulfuric acid consists of a jacketed static mixer (Noritake 1/4 (1) -N40-174-0 (inner diameter ⁇ 5 mm, number of elements 24)) and three tanks with jackets of 600 ml, 400 ml and 300 ml, respectively.
- the succinic acid was continuously extracted by mixing with the MEK solution and separating the liquids using a settling formula.
- the extraction phase overflowed the weir between the first tank and the second tank and was supplied to the second tank.
- insoluble components that could not be separated in the first tank were allowed to settle to the bottom, and only the clear extraction phase was allowed to overflow the weir between the second tank and the third tank and supplied to the third tank.
- the clear extraction phase was overflowed from the vicinity of the liquid interface to recover the extraction phase, and finally the extraction phase 688 g, the extracted residual phase 1613 g, and the intermediate phase 173 g were recovered.
- the intermediate phase was subjected to pressure filtration with a PTFE membrane filter having an opening of 0.5 ⁇ m to recover 172 g of the clarified liquid.
- the temperature of the extraction tower was controlled at 30 ° C. by passing warm water through the jacket. Finally, 1777 g of the extracted phase was recovered. The recovered extracted phase was combined with the clarified liquid recovered by the mixer settler, and the total amount of the liquid containing the aliphatic dicarboxylic acid was 2637 g. When the composition was analyzed, the results were as shown in Table 28 below.
- ⁇ Crystal> The liquid from which MEK was distilled off was transferred to a jacketed 500 ml separable flask, and kept warm at 80 ° C. by passing warm water through the jacket while stirring. Thereafter, using a circulating thermostatic bath with a program, the warm water passing through the jacket was cooled to 20 ° C. over 1 hour, succinic acid was cooled and crystallized, and after reaching 20 ° C., the mixture was further aged at 20 ° C. for 1 hour.
- the resulting slurry was vacuum filtered to separate the crystallization mother liquor. Further, the obtained wet cake of the crystallized product was washed with 250 g of cold water to recover the washing liquid, and a wet cake containing succinic acid as a main component was obtained. Further, the obtained wet cake was dried with a vacuum dryer at a maximum pressure of 80 ° C., and finally 114 g of succinic acid was recovered.
- Table 30 The results of composition analysis of the obtained succinic acid are shown in Table 30 below.
- the crystallization mother liquor and the cleaning liquid were mixed to obtain a recovered liquid of 562 g, and the composition thereof was as shown in Table 31 below.
- Example 3-2 ⁇ Protonation> 98% sulfuric acid was added to 1384 g of the succinic acid fermentation broth to adjust the pH to 2.5. Here, the addition amount of 98% sulfuric acid was 138 g. To this protonated liquid, 281 g corresponding to half of the recovered liquid recovered in Example 3-1 was added to prepare 1803 g of an aqueous succinic acid solution.
- Example 3-1 A succinic acid aqueous solution was extracted with 10% aqueous MEK in the same manner as in Example 3-1.
- the mixer settler supplies 0.5% by weight 10% water-containing MEK to the aqueous succinic acid solution at 20 g / min and 10 g / min, respectively, and the recovered intermediate phase is subjected to pressure filtration.
- Countercurrent multistage continuous extraction was performed with 1.0% by weight 10% water-containing MEK. As a result, 2882 g of extracted phase and 1593 g of extracted residual phase were recovered. Its composition was as follows.
- Example 3-3 The steps from protonation to crystallization were carried out in the same procedure as in Example 3-2 to obtain 573 g of a recovered liquid which was a mixture of 114 g of succinic acid, the crystallization mother liquor and the washing liquid.
- Table 35 below shows the composition of succinic acid obtained
- Table 36 below shows the composition of the collected liquid.
- Example 3-4 The steps from protonation to crystallization were carried out in the same procedure as in Example 2 to obtain 114 g of recovered liquid, which was a mixture of succinic acid 114 g, crystallization mother liquor and washing liquid.
- the composition of succinic acid obtained is shown in Table 37 below.
- the composition of the recovered liquid obtained is shown in Table 38 below.
- non-amino organic acids such as succinic acid, fumaric acid and malic acid can be obtained according to the present invention.
- the succinic acid or the composition containing the succinic acid can be used for food additives, pharmaceuticals, cosmetics and the like.
- fumaric acid or a composition containing fumaric acid can be used for food additives, unsaturated polyester resins, paper sizing agents, and the like.
- malic acid or a composition containing malic acid can be used for food additives, cosmetics, deodorants, detergents and dyes.
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Abstract
Description
1.生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(4)を含む脂肪族ジカルボン酸の製造方法。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物を分離する微生物分離工程
(4)微生物分離工程で微生物を分離して得られた水溶液と、溶剤とを接触させる接触工程
2.微生物がコリネ型細菌である、前項1に記載の方法。
3.脂肪族ジカルボン酸がコハク酸である、前項1又は2に記載の方法。
4.溶剤が無機性値/有機性値の比(I/O値)が0.2以上2.3以下であり、常圧(1気圧)で沸点が40℃以上の有機溶媒である、前項1~3のいずれか1項に記載の方法。
5.生物由来原料から得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(3)を含む脂肪族ジカルボン酸の製造方法。
(1)生物由来原料から得られる脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(2)接触工程で前記水溶液と前記溶剤の接触により発生した固形分を除去する固形分除去工程
(3)前記溶剤を相分離により分離する相分離工程
6.前記固形分除去工程において、前記固形分の除去を、セトラーを用いて行う前項5に記載の脂肪族ジカルボン酸の製造方法。
7.前記接触工程において、前記水溶液と前記溶剤との接触を、スタティックミキサーを用いて行う前項5または前項6に記載の脂肪族ジカルボン酸の製造方法。
8.前記固形分除去工程において、前記固形分の除去を、前記水溶液および前記溶剤から選ばれる少なくとも1種の液体と共に抜き出すことにより行う前項5から前項7のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
9.抜き出した固形分を固液分離し、得られた液体を前記水溶液と前記溶剤との接触工程以前の工程および相分離工程以降の工程の少なくとも一方に戻す、前項8に記載の脂肪族ジカルボン酸の製造方法。
10.前記固液分離を遠心沈降法により行う、前項9に記載の脂肪族ジカルボン酸の製造方法。
11.前記固液分離をフィルターを用いて行う、前項9に記載の脂肪族ジカルボン酸の製造方法。
12.前記溶剤を相分離により分離した後に残った液体と、該液体と相分離する溶剤とを、向流多段抽出塔で接触させ、後に該溶剤を相分離により分離する工程を含む、前項5から前項11のいずれか1項に記載の脂肪族ジカルボン酸の製造方法
13.前記接触工程を30~60℃で行う、前項5から前項12のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
14.生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(5)を含む脂肪族ジカルボン酸の製造方法。
(1)脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離する溶剤とを接触させる接触工程
(2)接触工程で回収された脂肪族ジカルボン酸を濃縮する工程であって、該濃縮により抽出相中の水濃度が上昇する抽出相濃縮工程
(3)抽出相濃縮工程後の液から脂肪族ジカルボン酸を析出させる晶析工程
(4)晶析工程で析出した脂肪族ジカルボン酸を回収する固液分離工程
(5)固液分離工程で得られた脂肪族ジカルボン酸を回収した後の晶析母液の少なくとも一部を晶析工程より前のいずれかの工程に戻す晶析母液リサイクル工程
15.接触工程より前に、前記生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液に酸を加えるプロトン化工程を含む、前項14に記載の脂肪族ジカルボン酸の製造方法。
16.晶析工程に供給される、抽出相濃縮工程後の脂肪族ジカルボン酸を含む溶液中に含まれる溶剤の濃度が10重量%以下である、前項14または15に記載の脂肪族ジカルボン酸の製造方法。
17.晶析工程に供給される、前記脂肪族ジカルボン酸を含む溶液中に含まれる溶剤の濃度が1重量%以下である、前項16に記載の脂肪族ジカルボン酸の製造方法。
18.固液分離工程で脂肪族ジカルボン酸を回収した後の母液から、脂肪族ジカルボン酸以外の成分を除去した後、当該液の少なくとも一部を接触工程から晶析工程までの間の少なくともいずれか1の工程にリサイクルする、前項14から前項17のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
19.抽出相濃縮工程以前のいずれかの工程において、水を加えることを特徴とする前項15から前項18のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
20.プロトン化工程において加える酸としてpKaが4未満の酸を用い、且つ該プロトン化工程における前記生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液のpHを、1より大きく4未満の範囲に維持することを特徴とする、前項15から前項19のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
21.接触工程で用いる溶剤が、有機性値に対する無機性値の比率が0.2以上2.3以下であり、且つ沸点が40℃以上であることを特徴とする、前項14から前項20のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
22.接触工程より前に、微生物を除去する工程を含むことを特徴とする、前項14から前項21のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
23.接触工程より前に、タンパク質を除去する工程を含むことを特徴とする、前項14から前項22のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
24.接触工程でミキサーセトラーを用い、該ミキサーセトラーのセトラー部から、抽出相、抽残相および固形分を含む相を回収することを特徴とする、前項14から前項23のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
25.前記セトラー部から回収した固形分を含む相を固液分離し、液相を回収することを特徴とする前項24に記載の脂肪族ジカルボン酸の製造方法。
26.接触工程で更に向流多段抽出塔を用い、セトラー部で得られた抽残相から脂肪族ジカルボン酸を回収することを特徴とする、前項24または前項25に記載の脂肪族ジカルボン酸の製造方法。
27.晶析工程において冷却を行うことを特徴とする、前項14から前項26のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
28.晶析工程が減圧操作を含むことを特徴とする、前項14から前項27のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。
29.生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(6)を含む脂肪族ジカルボン酸を製造する方法。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物菌体とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物菌体を分離する菌体分離工程
(4)菌体分離工程で菌体を分離して得られた水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(5)前記水溶液と前記溶剤の接触により発生した固形分を除去する固形分除去工程
(6)前記溶剤を相分離により分離する相分離工程
30.生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(8)を含む脂肪族ジカルボン酸を製造する方法。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物菌体とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物菌体を分離する菌体分離工程
(4)菌体分離工程で菌体を分離して得られた水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(5)接触工程で回収された脂肪族ジカルボン酸を濃縮する工程であって、該濃縮により抽出相中の水濃度が上昇する抽出相濃縮工程
(6)抽出相濃縮工程後の液から脂肪族ジカルボン酸を析出させる晶析工程
(7)晶析工程で析出した脂肪族ジカルボン酸を回収する固液分離工程
(8)固液分離工程で得られた脂肪族ジカルボン酸を回収した後の晶析母液の少なくとも一部を、晶析工程より前のいずれかの工程に戻す晶析母液リサイクル工程 That is, according to the present invention, the following inventions are provided.
1. A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism, comprising the steps of (1) to (4) below: Production method.
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step PH adjustment step (3) for obtaining a solution having a pH of 1.0 or more and 5.0 or less by mixing a solution containing a solution and an acid, a microorganism separation step (4) for separating microorganisms from the solution obtained in the pH adjustment step ) A contact step in which an aqueous solution obtained by separating microorganisms in the microorganism separation step is brought into contact with a solvent. 2. The method according to
3. 3. The method according to
4). The solvent is an organic solvent having an inorganic value / organic value ratio (I / O value) of 0.2 or more and 2.3 or less and a normal pressure (1 atm) and a boiling point of 40 ° C. or more. 4. The method according to any one of items 3.
5. A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, comprising the following steps (1) to (3).
(1) A contact step in which an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material is brought into contact with a solvent that can be phase-separated with the aqueous solution. (2) A solid content generated by the contact between the aqueous solution and the solvent in the contact step. (3) A phase separation step for separating the solvent by phase separation. 6. The method for producing an aliphatic dicarboxylic acid according to 5 above, wherein in the solid content removing step, the solid content is removed using a settler.
7). 7. The method for producing an aliphatic dicarboxylic acid according to 5 or 6 above, wherein in the contacting step, the aqueous solution and the solvent are contacted using a static mixer.
8). The aliphatic dicarboxylic acid according to any one of the preceding
9. The extracted solid content is subjected to solid-liquid separation, and the obtained liquid is returned to at least one of the step before the contact step between the aqueous solution and the solvent and the step after the phase separation step. Production method.
10. 10. The method for producing an aliphatic dicarboxylic acid according to
11. 10. The method for producing an aliphatic dicarboxylic acid according to
12 Including the step of bringing the liquid remaining after the solvent is separated by phase separation into contact with the solvent to be phase-separated with the liquid in a countercurrent multi-stage extraction tower, and then separating the solvent by phase separation. 12. Manufacturing method of aliphatic dicarboxylic acid of any one of 11 13. The method for producing an aliphatic dicarboxylic acid according to any one of
14 A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, comprising the following steps (1) to (5).
(1) A contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is brought into contact with a solvent that is phase-separated with the aqueous solution. (2) A step of concentrating the aliphatic dicarboxylic acid recovered in the contact step, which is extracted by the concentration. Extraction phase concentration step in which water concentration in phase increases (3) Crystallization step for precipitation of aliphatic dicarboxylic acid from liquid after extraction phase concentration step (4) Solid recovery of aliphatic dicarboxylic acid precipitated in crystallization step Liquid separation step (5) Crystallization mother liquor recycling step for returning at least part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step prior to the crystallization step. 15. The method for producing an aliphatic dicarboxylic acid according to item 14, further comprising a protonation step of adding an acid to the aqueous solution containing the aliphatic dicarboxylic acid obtained from the biological material before the contacting step.
16. 16. The method for producing an aliphatic dicarboxylic acid according to item 14 or 15, wherein the concentration of the solvent contained in the solution containing the aliphatic dicarboxylic acid after the extraction phase concentration step supplied to the crystallization step is 10% by weight or less. .
17. The method for producing an aliphatic dicarboxylic acid according to item 16 above, wherein the concentration of the solvent contained in the solution containing the aliphatic dicarboxylic acid supplied to the crystallization step is 1% by weight or less.
18. After removing components other than the aliphatic dicarboxylic acid from the mother liquor after collecting the aliphatic dicarboxylic acid in the solid-liquid separation step, at least one of the liquid from the contact step to the crystallization step is removed. 18. The method for producing an aliphatic dicarboxylic acid according to any one of items 14 to 17, which is recycled to the step.
19. 19. The method for producing an aliphatic dicarboxylic acid according to any one of items 15 to 18 above, wherein water is added in any step before the extraction phase concentration step.
20. An acid having a pKa of less than 4 is used as an acid to be added in the protonation step, and the pH of the aqueous solution containing the aliphatic dicarboxylic acid obtained from the biological material in the protonation step is maintained in the range of more than 1 and less than 4. 20. The method for producing an aliphatic dicarboxylic acid according to any one of items 15 to 19, wherein:
21. The solvent used in the contacting step has a ratio of an inorganic value to an organic value of 0.2 or more and 2.3 or less, and a boiling point of 40 ° C. or more, any one of the items 14 to 20 above 2. A process for producing an aliphatic dicarboxylic acid according to
22. [22] The method for producing an aliphatic dicarboxylic acid according to any one of [14] to [21] above, which comprises a step of removing microorganisms before the contacting step.
23. 23. The method for producing an aliphatic dicarboxylic acid according to any one of items 14 to 22, further comprising a step of removing the protein before the contacting step.
24. 24. The method according to any one of items 14 to 23, wherein a mixer settler is used in the contacting step, and an extraction phase, an extraction residual phase, and a phase containing a solid content are recovered from a setter portion of the mixer setter. A method for producing an aliphatic dicarboxylic acid.
25. 25. The method for producing an aliphatic dicarboxylic acid according to the item 24, wherein the phase containing the solid content recovered from the settler part is subjected to solid-liquid separation, and the liquid phase is recovered.
26. The method for producing an aliphatic dicarboxylic acid according to item 24 or 25, wherein an aliphatic dicarboxylic acid is further recovered from the extracted residual phase obtained in the settler part using a countercurrent multistage extraction tower in the contacting step. .
27. 27. The method for producing an aliphatic dicarboxylic acid according to any one of items 14 to 26, wherein cooling is performed in a crystallization step.
28. 28. The method for producing an aliphatic dicarboxylic acid according to any one of items 14 to 27, wherein the crystallization step includes a pressure reduction operation.
29. A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism, comprising the steps of (1) to (6) below: How to manufacture.
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step A pH adjusting step for obtaining a liquid having a pH of 1.0 or more and 5.0 or less by mixing a solution containing the microbial cells and an acid (3) Bacteria for separating microbial cells from the liquid obtained in the pH adjusting step Body separation step (4) Contact step of contacting the aqueous solution obtained by separating the cells in the cell separation step with a solvent capable of phase separation with the aqueous solution (5) Generated by contact between the aqueous solution and the solvent Solid content removing step for removing solid content (6) Phase separation step for separating the solvent by phase separation 30. A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism, comprising the steps of (1) to (8) below: How to manufacture.
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step A pH adjusting step for obtaining a liquid having a pH of 1.0 or more and 5.0 or less by mixing a solution containing the microbial cells and an acid (3) Bacteria for separating microbial cells from the liquid obtained in the pH adjusting step Body separation step (4) Contact step in which the aqueous solution obtained by separating the cells in the cell separation step is brought into contact with a solvent capable of phase separation with the aqueous solution (5) Aliphatic dicarboxylic acid recovered in the contact step In which the concentration of water in the extraction phase increases due to the concentration (6) Crystallization step (7) Crystallization of depositing aliphatic dicarboxylic acid from the liquid after the extraction phase concentration step Aliphatic dicarbo precipitated in the process Solid-liquid separation step for recovering acid (8) Crystal for returning at least a part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step before the crystallization step Analysis mother liquor recycling process
I.第1の本発明
第1の本発明は、生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(4)を含む脂肪族ジカルボン酸の製造方法である。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物を分離する微生物分離工程
(4)微生物分離工程で微生物を分離して得られた水溶液と、溶剤とを接触させる接触工程 Hereinafter, embodiments of the present invention will be described in detail.
I. 1st this invention 1st this invention is a method of manufacturing aliphatic dicarboxylic acid from the aqueous solution containing aliphatic dicarboxylic acid obtained by making a biological raw material and microorganisms react, Comprising: The following processes (1 ) To (4).
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step PH adjustment step (3) for obtaining a solution having a pH of 1.0 or more and 5.0 or less by mixing a solution containing a solution and an acid, a microorganism separation step (4) for separating microorganisms from the solution obtained in the pH adjustment step ) A contact process in which an aqueous solution obtained by separating microorganisms in the microorganism separation process is contacted with a solvent.
(生物由来原料)
生物由来原料としては、例えば、木材、稲わら、籾殻、米ぬか、古米、とうもろこし、サトウキビ、キャッサバ、サゴヤシ、おから、コーンコブ、タピオカカス、バガス、植物油カス、芋、そば、大豆、油脂、古紙、製紙残渣、水産物残渣、家畜排泄物、下水汚泥および食品廃棄物等が挙げられる。 I- (1) Aliphatic dicarboxylic acid production process (biological raw material) in which aliphatic dicarboxylic acid is accumulated in the reaction liquid by reacting biological raw material with microorganisms
Examples of biological materials include wood, rice straw, rice husk, rice bran, old rice, corn, sugar cane, cassava, sago palm, okara, corn cob, tapioca cass, bagasse, vegetable oil cass, persimmon, buckwheat, soybean, fat, waste paper, paper Residue, marine product residue, livestock excrement, sewage sludge, food waste and the like.
微生物の種類は特に制限されないが、エシェリヒア・コリ等の腸内細菌、バチルス属細菌、コリネ型細菌などが挙げられ、好気性微生物、通性嫌気性微生物または微好気性微生物を使用することが好ましい。 (Microorganism)
The type of microorganism is not particularly limited, but examples include intestinal bacteria such as Escherichia coli, Bacillus bacteria, coryneform bacteria, etc., and it is preferable to use aerobic microorganisms, facultative anaerobic microorganisms, or microaerobic microorganisms. .
脂肪族ジカルボン酸としては、例えば、乳酸、コハク酸、リンゴ酸、フマル酸、オキザロ酢酸、クエン酸、イソクエン酸、2-オキソグルタル酸、シス-アコニット酸、ピルビン酸および酢酸などが挙げられる。この中では、コハク酸、リンゴ酸およびフマル酸などのジカルボン酸が好ましく、コハク酸が特に好ましい。脂肪族ジカルボン酸は複数種類含まれてもよい。 (Aliphatic dicarboxylic acid)
Examples of the aliphatic dicarboxylic acid include lactic acid, succinic acid, malic acid, fumaric acid, oxaloacetic acid, citric acid, isocitric acid, 2-oxoglutaric acid, cis-aconitic acid, pyruvic acid, and acetic acid. Among these, dicarboxylic acids such as succinic acid, malic acid and fumaric acid are preferable, and succinic acid is particularly preferable. A plurality of types of aliphatic dicarboxylic acids may be included.
前記生物由来原料から誘導される炭素源を含む反応液中で、該炭素源を脂肪族ジカルボン酸に変換する微生物と、該炭素源とを反応させることにより、脂肪族ジカルボン酸を反応液中に蓄積させることができる。 (Reaction solution)
In a reaction solution containing a carbon source derived from the biological material, a reaction between the carbon source and a microorganism that converts the carbon source to an aliphatic dicarboxylic acid is carried out to react the aliphatic dicarboxylic acid in the reaction solution. Can be accumulated.
脂肪族ジカルボン酸と微生物とを含む液(発酵懸濁液)から菌体を分離する際の発酵懸濁液のpHは、通常1.0以上5.0以下が好ましく、1.5以上4.0以下がより好ましく、2.0以上3.5以下がさらに好ましい。 I- (2) pH adjusting step for obtaining a solution having a pH of 1.0 or more and 5.0 or less by mixing a solution containing an aliphatic dicarboxylic acid obtained in the step of producing an aliphatic dicarboxylic acid and a microorganism and an acid. The pH of the fermentation suspension when separating cells from a liquid (fermentation suspension) containing an aliphatic dicarboxylic acid and a microorganism is usually preferably 1.0 or more and 5.0 or less, and preferably 1.5 or more and 4. 0 or less is more preferable, and 2.0 or more and 3.5 or less is more preferable.
微生物を発酵懸濁液より分離除去する方法としては、沈降分離、遠心分離およびろ過分離並びにそれらを組み合わせた方法などが用いられる。工業的には、遠心分離および膜ろ過分離などの方法で行われる。遠心分離においては、遠心沈降および遠心ろ過などを用いることができる。 I- (3) Microorganism separation step for separating microorganisms from the liquid obtained in the pH adjustment step The methods for separating and removing microorganisms from the fermentation suspension include sedimentation separation, centrifugal separation and filtration separation, and a combination thereof. Is used. Industrially, it is carried out by methods such as centrifugation and membrane filtration separation. In centrifugation, centrifugal sedimentation, centrifugal filtration, or the like can be used.
微生物分離工程で微生物を分離した後の脂肪族ジカルボン酸を含む水溶液と、溶剤とを接触させて、脂肪族ジカルボン酸を抽出する。当該水溶液を濃縮した後に溶剤と接触させてもよい。接触工程により、脂肪族ジカルボン酸を選択的に溶剤中に抽出することができ、水溶性の高い糖類、アミノ酸類および無機塩類は主に水溶液相に分配される。 I- (4) Contacting step in which an aqueous solution obtained by separating microorganisms in the microorganism separation step is contacted with a solvent Contacting an aqueous solution containing an aliphatic dicarboxylic acid after separation of microorganisms in the microorganism separation step and the solvent To extract the aliphatic dicarboxylic acid. You may make it contact with a solvent, after concentrating the said aqueous solution. By the contacting step, the aliphatic dicarboxylic acid can be selectively extracted into the solvent, and saccharides, amino acids and inorganic salts having high water solubility are mainly distributed in the aqueous phase.
接触工程において使用する溶剤としては、脂肪族ジカルボン酸を含有する水溶液と相分離する有機溶媒であれば特に制限は無いが、無機性値/有機性値の比(以下、I/O値と略記することがある)が0.2以上2.3以下であることが好ましく、0.3以上2.0以下であることがより好ましい。 (solvent)
The solvent used in the contacting step is not particularly limited as long as it is an organic solvent that is phase-separated from an aqueous solution containing an aliphatic dicarboxylic acid, but the ratio of inorganic value / organic value (hereinafter abbreviated as I / O value). Is preferably 0.2 or more and 2.3 or less, and more preferably 0.3 or more and 2.0 or less.
接触工程の温度は、脂肪族ジカルボン酸が抽出される温度であれば特に限定されないが、10~50℃が好ましく、20~40℃がより好ましい。 (Contact operation)
The temperature in the contacting step is not particularly limited as long as the aliphatic dicarboxylic acid is extracted, but is preferably 10 to 50 ° C, more preferably 20 to 40 ° C.
第2の発明は、生物由来原料から得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(3)を含む脂肪族ジカルボン酸の製造方法である。
(1)生物由来原料から得られる脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(2)接触工程で前記水溶液と前記溶剤の接触により発生した固形分を除去する固形分除去工程
(3)前記溶剤を相分離により分離する相分離工程 II. Second invention The second invention is a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, the fat comprising the following steps (1) to (3): It is a manufacturing method of a group dicarboxylic acid.
(1) A contact step in which an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material is brought into contact with a solvent that can be phase-separated with the aqueous solution. (3) Phase separation step of separating the solvent by phase separation
接触工程により、脂肪族ジカルボン酸を選択的に溶剤中に抽出することができ、水溶性の高い糖類、アミノ酸類、無機塩類は主に水溶液相に分配される。 II- (1) Contacting process in which an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material is brought into contact with a solvent that can be phase-separated from the aqueous solution The aliphatic dicarboxylic acid is selectively extracted into the solvent by the contacting process Highly water-soluble saccharides, amino acids and inorganic salts are mainly distributed in the aqueous phase.
接触装置は、脂肪族ジカルボン酸を含む水溶液と溶剤との接触および溶剤相、水溶液相の回収、および後述する固形分除去工程において固形分の除去ができればどのような装置であってもよいが、装置が簡単で操作も容易な、上記のミキサーセトラー型抽出装置が好ましい。 (Contact device)
The contact device may be any device as long as it can contact the aqueous solution containing the aliphatic dicarboxylic acid and the solvent and the solvent phase, recover the aqueous solution phase, and remove the solid content in the solid content removal step described later. The above-mentioned mixer-settler type extraction device is preferable because the device is simple and easy to operate.
第2の本発明の脂肪族ジカルボン酸の製造方法は、脂肪族ジカルボン酸を含有する水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程と、接触工程後に液を相分離させる、後述する相分離工程を含む。 (Contact process and phase separation process)
The method for producing an aliphatic dicarboxylic acid according to the second aspect of the present invention includes a contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is contacted with a solvent capable of phase separation with the aqueous solution, and the liquid is phase-separated after the contact step. A phase separation step described later is included.
II-(2)接触工程で前記水溶液と前記溶剤の接触により発生した固形分を除去する固形分除去工程
固形分除去工程では、生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離可能な溶剤との接触により発生した固形分を除去する。
II- (2) Solid content removal step for removing solid content generated by contact of the aqueous solution and the solvent in the contact step In the solid content removal step, an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, The solid content generated by contact with the aqueous solution and the phase-separable solvent is removed.
(相分離操作)
相分離工程における相分離操作は、槽で一定時間静置することでも可能であるし、遠心分離装置により行うこともできる。上記のようなミキサーセトラー型抽出機では、接触混合することにより得られた混合液を静置することで相分離させるセトラー部を有しており、セトラー部において液を一定時間静置することにより、相分離することができる。 II- (3) Phase separation step for separating the solvent by phase separation (phase separation operation)
The phase separation operation in the phase separation step can be performed by standing in a tank for a certain period of time or can be performed by a centrifugal separator. The mixer-settler type extractor as described above has a settler part that causes phase separation by allowing the mixed liquid obtained by contact mixing to stand, and by allowing the liquid to stand for a certain time in the settler part. Can be phase separated.
相分離工程に用いるセトラーは、脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離可能な溶剤とを接触させた後の液を相分離することが可能なものであれば、如何なる方式であってもよい。 (Phase separator)
The settler used in the phase separation step may be any system as long as it can phase-separate the liquid after bringing the aqueous solution containing the aliphatic dicarboxylic acid into contact with the solvent capable of phase separation. May be.
第2の本発明の脂肪族ジカルボン酸としては、生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液から製造されるものであれば特に限定されず、カルボキシ基を2つ有する脂肪族炭化水素であれば如何なるものも採用することができる。 <Aliphatic dicarboxylic acid>
The aliphatic dicarboxylic acid of the second aspect of the present invention is not particularly limited as long as it is produced from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, and is an aliphatic hydrocarbon having two carboxy groups. Any one can be adopted.
抽出相中に選択的に回収された脂肪族ジカルボン酸を最終的に脂肪族ジカルボン酸結晶として回収するには、通常、濃縮工程および晶析工程が必要となる。ここで抽出相から脂肪族ジカルボン酸結晶を回収する一般的な方法を説明するが、本発明に採用可能な後続工程はこれらに制限されない。 <Subsequent process>
In order to finally collect the aliphatic dicarboxylic acid selectively recovered in the extraction phase as an aliphatic dicarboxylic acid crystal, a concentration step and a crystallization step are usually required. Here, a general method for recovering the aliphatic dicarboxylic acid crystals from the extraction phase will be described, but the subsequent steps that can be employed in the present invention are not limited thereto.
抽出相濃縮工程は、III-(2)において後述するように行う。 (Extraction phase concentration process)
The extraction phase concentration step is performed as described later in III- (2).
晶析工程は、III-(3)において後述するように行う。 (Crystallization process)
The crystallization step is performed as described later in III- (3).
晶析工程で得られた脂肪族ジカルボン酸スラリーは、III-(4)において後述するように、固液分離操作により脂肪族ジカルボン酸結晶および母液を分離する。 (Solid-liquid separation process)
The aliphatic dicarboxylic acid slurry obtained in the crystallization step separates the aliphatic dicarboxylic acid crystal and the mother liquor by solid-liquid separation operation, as described later in III- (4).
第3の本発明は、生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(5)を含む脂肪族ジカルボン酸の製造方法である。
(1)脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離する溶剤とを接触させる接触工程
(2)接触工程で回収された脂肪族ジカルボン酸を濃縮する工程であって、該濃縮により抽出相中の水濃度が上昇する抽出相濃縮工程
(3)抽出相濃縮工程後の液から脂肪族ジカルボン酸を析出させる晶析工程
(4)晶析工程で析出した脂肪族ジカルボン酸を回収する固液分離工程
(5)固液分離工程で得られた脂肪族ジカルボン酸を回収した後の晶析母液の少なくとも一部を晶析工程より前のいずれかの工程に戻す晶析母液リサイクル工程 III. Third invention The third invention is a method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, and comprises the following steps (1) to (5): It is a manufacturing method of the aliphatic dicarboxylic acid containing this.
(1) A contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is brought into contact with a solvent that is phase-separated with the aqueous solution. (2) A step of concentrating the aliphatic dicarboxylic acid recovered in the contact step, which is extracted by the concentration. Extraction phase concentration step in which water concentration in phase increases (3) Crystallization step for precipitation of aliphatic dicarboxylic acid from liquid after extraction phase concentration step (4) Solid recovery of aliphatic dicarboxylic acid precipitated in crystallization step Liquid separation step (5) Crystallization mother liquor recycling step for returning at least part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step prior to the crystallization step
接触工程は、II-(1)で上述したように行う。また、接触工程より前に、上記した方法により、微生物またはタンパク質を除去する工程を含むことが好ましい。 III- (1) Contacting step of contacting an aqueous solution containing an aliphatic dicarboxylic acid with a solvent that is phase-separated from the aqueous solution The contacting step is performed as described above in II- (1). Moreover, it is preferable to include the process of removing microorganisms or protein by an above-described method before a contact process.
一般に、抽出相における脂肪族ジカルボン酸濃度は希薄であるため濃縮操作が必要となる。濃縮度は特に限定されるものではないが、最終濃縮液中の脂肪族ジカルボン酸の溶解度が飽和溶解度以下であり、かつ極力飽和溶解度に近いほうが好ましい。 III- (2) A step of concentrating the aliphatic dicarboxylic acid recovered in the contacting step, wherein the concentration of water in the extraction phase increases due to the concentration. Generally, the concentration of the aliphatic dicarboxylic acid in the extraction phase is Since it is dilute, a concentration operation is required. The concentration is not particularly limited, but it is preferable that the solubility of the aliphatic dicarboxylic acid in the final concentrated solution is not more than the saturation solubility and is as close to the saturation solubility as possible.
晶析工程は、通常、抽出相を含有する溶液中の脂肪族ジカルボン酸を、ジカルボン酸の溶解度差などを利用して固体の脂肪族ジカルボン酸を析出させる工程である。 III- (3) Crystallization process for precipitating aliphatic dicarboxylic acid from the liquid after the extraction phase concentration process The crystallization process usually involves converting the aliphatic dicarboxylic acid in the solution containing the extraction phase to the solubility difference of the dicarboxylic acid, etc. Is a step of precipitating solid aliphatic dicarboxylic acid using
晶析で得られた脂肪族ジカルボン酸スラリーは固液分離操作により脂肪族ジカルボン酸結晶および母液を分離する。分離方法は、特に限定されるものではなく、例えば、ろ過分離および沈降分離などが挙げられる。 III- (4) Solid-liquid separation step for recovering the aliphatic dicarboxylic acid precipitated in the crystallization step The aliphatic dicarboxylic acid slurry obtained by the crystallization separates the aliphatic dicarboxylic acid crystal and the mother liquor by solid-liquid separation operation. The separation method is not particularly limited, and examples thereof include filtration separation and sedimentation separation.
固液分離工程で得られた母液および/または洗浄液の少なくとも一部は、晶析工程より前の工程にリサイクルすることができる。 III- (5) Crystallization mother liquor recycling step in which at least a part of the crystallization mother liquor after recovery of the aliphatic dicarboxylic acid obtained in the solid-liquid separation step is returned to any step prior to the crystallization step. At least a part of the mother liquor and / or cleaning liquid obtained in the process can be recycled to a process prior to the crystallization process.
また、接触工程で脂肪族ジカルボン酸を溶剤中に回収する際に、該脂肪族ジカルボン酸を含有する水溶液において、脂肪族ジカルボン酸が塩の水溶液として存在する場合、I-(2)において上述したプロトン化と同様の方法により塩をプロトン化することが好ましい。 <Protonation process>
Further, when the aliphatic dicarboxylic acid is recovered as a salt aqueous solution in the aqueous solution containing the aliphatic dicarboxylic acid when the aliphatic dicarboxylic acid is recovered in the solvent in the contacting step, it is described above in I- (2). It is preferable to protonate the salt by a method similar to protonation.
さらに本発明の方法により得られた脂肪族ジカルボン酸に、その用途に応じて、乾燥処理および精製処理に代表されるような、他の工程を適用することもできる。例えば、活性炭等の吸着剤による脱色工程、イオン交換樹脂により共存イオン類を除去するイオン交換工程、共存する不飽和ジカルボン酸を水添処理する工程およびさらに高度精製するための晶析工程などの処理が考えられる。 <Other processes>
Furthermore, other steps, such as drying and purification, can be applied to the aliphatic dicarboxylic acid obtained by the method of the present invention, depending on its use. For example, a decolorization process using an adsorbent such as activated carbon, an ion exchange process for removing coexisting ions with an ion exchange resin, a process for hydrogenating coexisting unsaturated dicarboxylic acid, and a crystallization process for further purification. Can be considered.
り制限されるものではない。 Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples.
(A)ブレビバクテリウム・フラバムMJ233株ゲノムDNAの抽出
ブレビバクテリウム・フラバムMJ233は、1975年4月28日に通商産業省工業技術院微生物工業技術研究所(現独立行政法人 産業技術総合研究所 特許生物寄託センター)(〒305-8566 日本国茨城県つくば市東1丁目1番地1 中央第6)に受託番号FERM P-3068として寄託され、1981年5月1日にブダペスト条約に基づく国際寄託に移管され、FERM BP-1497の受託番号で寄託されている。 Reference Example 1: Preparation of a succinic acid fermenting strain (A) Extraction of genomic DNA of Brevibacterium flavum MJ233 Brevibacterium flavum MJ233 was introduced on April 28, 1975 at the Institute of Microbial Industrial Technology, Ministry of International Trade and Industry. (Currently the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center) (deposited as FERM P-3068, 1st East, 1st Street, Tsukuba City, Ibaraki Prefecture, Japan 305-8586), May 1981 It was transferred to an international deposit based on the Budapest Treaty on the 1st and deposited under the deposit number of FERM BP-1497.
ブレビバクテリウム・フラバムMJ233株由来ピルベートカルボキシラーゼ遺伝子のN末端領域のDNA断片の取得は、上記(A)で調製したDNAを鋳型とし、全ゲノム配列が報告されているコリネバクテリウム・グルタミカム ATCC13032株の該遺伝子の配列(GenBank Database Accession No.BA000036のCgl0689)を基に設計した合成DNA(配列番号1および配列番号2)を用いたPCRによって行った。尚、配列番号1のDNAは5’末端がリン酸化されたものを用いた。 (B) Construction of a plasmid for PC promoter substitution The DNA fragment of the N-terminal region of the pyruvate carboxylase gene derived from Brevibacterium flavum MJ233 strain was obtained using the DNA prepared in (A) above as a template and the entire genome sequence was reported. This was performed by PCR using synthetic DNA (SEQ ID NO: 1 and SEQ ID NO: 2) designed based on the sequence of the gene of Corynebacterium glutamicum ATCC13032 strain (Cgl0689 of GenBank Database Accession No. BA00000036). The DNA of SEQ ID NO: 1 used was phosphorylated at the 5 ′ end.
ブレビバクテリウム・フラバムMJ233/ΔLDH(LDH活性が低下した株:特開2005-95169)の形質転換に用いるプラスミドDNAは、pMJPC17.2のプラスミドDNA用いて塩化カルシウム法(Journal of Molecular Biology,53,159,1970)により形質転換した大腸菌JM110株から再調製した。 (C) Production of PC enhanced strain
Plasmid DNA used for transformation of Brevibacterium flavum MJ233 / ΔLDH (strain with reduced LDH activity: Japanese Patent Application Laid-Open No. 2005-95169) was obtained using the plasmid DNA of pMJPC17.2 using the calcium chloride method (Journal of Molecular Biology, 53, 159, 1970) and re-prepared from E. coli strain JM110.
(A)種培養
尿素:4g、硫酸アンモニウム:14g、リン酸1カリウム:0.5g、リン酸2カリウム0.5g、硫酸マグネシウム・7水和物:0.5g、硫酸第一鉄・7水和物:20mg、硫酸マンガン・水和物:20mg、D-ビオチン:200μg、塩酸チアミン:200μg、酵母エキス:1g、カザミノ酸:1gを蒸留水に溶解、1000mLに調整した培地100mLを500mLの三角フラスコに入れ、121℃、20分加熱滅菌した。これを室温まで冷やし、あらかじめ滅菌した50%グルコース水溶液4mLを添加し、上記で構築したブレビバクテリウム・フラバムMJ233/PC-5/ΔLDH株を接種して16時間30℃にて振とう(160rpm)培養した。 Reference Example 2: Preparation of succinic acid fermentation broth by jar fermenter (A) Seed culture Urea: 4 g, ammonium sulfate: 14 g, 1 potassium phosphate: 0.5 g, 2 potassium phosphate 0.5 g, magnesium sulfate 7 hydrate Product: 0.5 g, ferrous sulfate heptahydrate: 20 mg, manganese sulfate hydrate: 20 mg, D-biotin: 200 μg, thiamine hydrochloride: 200 μg, yeast extract: 1 g, casamino acid: 1 g Into a 500 mL Erlenmeyer flask was added 100 mL of the medium dissolved in 1000 mL and sterilized by heating at 121 ° C. for 20 minutes. This was cooled to room temperature, 4 mL of a 50% glucose aqueous solution sterilized in advance was added, and the Brevibacterium flavum MJ233 / PC-5 / ΔLDH strain constructed above was inoculated and shaken at 30 ° C. for 16 hours (160 rpm). Cultured.
硫酸アンモニウム:3.0g、85%リン酸:6.7g、塩化カリウム:4.9g、硫酸マグネシウム・7水和物:1.5g、硫酸第一鉄・7水和物:120mg、硫酸マンガン・水和物:120mg、コーンスティープリカー(王子コーンスターチ社製)30.0g、10N水酸化カリウム水溶液:11.0g、消泡剤(CE457:日本油脂製):2.5gを蒸留水に溶解して調整した培地2.0Lを5Lの発酵糟に入れ、121℃、20分加熱滅菌した。これを室温まで冷やしてから28% アンモニア水を加えてpHを7.0に調整した後、予めフィルター滅菌したD-ビオチン、塩酸チアミン各0.2g/L水溶液:15mL、及びあらかじめ滅菌した720g/Lショ糖水溶液:110mLを添加し、これに前述の種培養液を100mL加えて、30℃に保温した。 (B) Main culture Ammonium sulfate: 3.0 g, 85% phosphoric acid: 6.7 g, potassium chloride: 4.9 g, magnesium sulfate heptahydrate: 1.5 g, ferrous sulfate heptahydrate: 120 mg , Manganese sulfate hydrate: 120 mg, corn steep liquor (manufactured by Oji Cornstarch) 30.0 g, 10N aqueous potassium hydroxide solution: 11.0 g, antifoaming agent (CE457: manufactured by NOF Corporation): 2.5 g of distilled water 2.0 L of the medium prepared by dissolving in the solution was placed in a 5 L fermenter and sterilized by heating at 121 ° C. for 20 minutes. This was cooled to room temperature and then adjusted to pH 7.0 by adding 28% aqueous ammonia, then D-biotin, thiamine hydrochloride 0.2 g / L aqueous solution: 15 mL each in advance, and 720 g / L previously sterilized. L Sucrose aqueous solution: 110 mL was added, and 100 mL of the above seed culture solution was added thereto, and the mixture was kept at 30 ° C.
85% リン酸:1.6g、硫酸マグネシウム・7水和物:1.1g、硫酸第一鉄・7水和物:43mg、硫酸マンガン・水和物:43mg、10N水酸化カリウム水溶液:2.86gを蒸留水に溶解、42mLに調整後、121℃、20分加熱滅菌処理し、反応濃縮培地を作製した。 (C) Succinic acid production reaction 85% Phosphoric acid: 1.6 g, Magnesium sulfate heptahydrate: 1.1 g, Ferrous sulfate heptahydrate: 43 mg, Manganese sulfate / hydrate: 43 mg, 10N Aqueous potassium hydroxide solution: 2.86 g was dissolved in distilled water, adjusted to 42 mL, and then heat sterilized at 121 ° C. for 20 minutes to prepare a reaction concentrated medium.
[実施例1-1]
<菌体分離>
(実施例1-1-1)
上記参考例2(C)で得られたコハク酸発酵液22mLを、95%硫酸でpHを2.5に調整したのち10mLを遠心チューブに入れ、500Gで8分遠心分離を行い、遠心分離後の上澄液の濁度(OD660)を測定した。尚、菌体分離前の濁度(OD660)は24であった。 [Example 1]
[Example 1-1]
<Cell separation>
(Example 1-1-1)
After adjusting the pH of the succinic acid fermentation liquor obtained in Reference Example 2 (C) above to 2.5 with 95% sulfuric acid, 10 mL was placed in a centrifuge tube, centrifuged at 500 G for 8 minutes, and then centrifuged. The turbidity (OD660) of the supernatant was measured. The turbidity (OD660) before cell separation was 24.
pHを3.0に調整した以外は実施例1-1-1と同様にして菌体分離を行い、遠心分離後の上澄液の濁度(OD660)を測定した。 (Example 1-1-2)
The cells were separated in the same manner as in Example 1-1-1 except that the pH was adjusted to 3.0, and the turbidity (OD660) of the supernatant after centrifugation was measured.
pHを4.0に調整した以外は実施例1-1-1と同様にして菌体分離を行い、遠心分離後の上澄液の濁度(OD660)を測定した。 (Example 1-1-3)
The cells were separated in the same manner as in Example 1-1-1 except that the pH was adjusted to 4.0, and the turbidity (OD660) of the supernatant after centrifugation was measured.
pHを5.0に調整した以外は実施例1-1-1と同様にして菌体分離を行い、遠心分離後の上澄液の濁度(OD660)を測定した。 (Example 1-1-4)
The cells were separated in the same manner as in Example 1-1-1 except that the pH was adjusted to 5.0, and the turbidity (OD660) of the supernatant after centrifugation was measured.
コハク酸発酵液(pH7.6)を実施例1-1-1と同様にして菌体分離を行い、遠心分離後の上澄液の濁度(OD660)を測定した。 (Comparative Example 1-1-1)
Bacterial cells were separated from the succinic acid fermentation broth (pH 7.6) in the same manner as in Example 1-1-1, and the turbidity (OD660) of the supernatant after centrifugation was measured.
<菌体分離+MEK抽出>
(実施例1-2-1)
上記参考例2(C)で得られたコハク酸発酵液50mLを遠心チューブに入れ、95%硫酸でpHを2.5に調整したのち、9500Gで10分遠心分離を行った。得られた上澄液について、一部を用いて濁度(OD660)の測定を行い、残りを用いて以下の手順でMEK抽出を行った。 [Example 1-2]
<Cell separation + MEK extraction>
(Example 1-2-1)
50 mL of the succinic acid fermentation broth obtained in Reference Example 2 (C) above was placed in a centrifuge tube, adjusted to pH 2.5 with 95% sulfuric acid, and then centrifuged at 9500 G for 10 minutes. About the obtained supernatant liquid, turbidity (OD660) was measured using a part, and MEK extraction was performed by the following procedure using the rest.
上記参考例2(C)で得られたコハク酸発酵液のpHを3.0に調整した以外は実施例1-2-1と同様にして菌体分離を行い、得られた上澄液を95%硫酸でpHを2.5に調整したのち、実施例1-2-1と同様にしてOD660の測定、MEK抽出を行い、分相時間、中間不溶分の量、有機相クリア率、平均粒径、光散乱強度を測定した。 (Example 1-2-2)
Cell separation was carried out in the same manner as in Example 1-2-1, except that the pH of the succinic acid fermentation broth obtained in Reference Example 2 (C) was adjusted to 3.0, and the resulting supernatant was After adjusting the pH to 2.5 with 95% sulfuric acid, measurement of OD660 and extraction of MEK were performed in the same manner as in Example 1-2-1, and the phase separation time, the amount of intermediate insoluble matter, the organic phase clear rate, the average The particle size and light scattering intensity were measured.
菌体分離前のpHを4.0に調整した以外は実施例1-2-2と同様にして菌体分離・MEK抽出を行い、OD660、分相時間、中間不溶分の量、有機相クリア率、平均粒径、光散乱強度を測定した。 (Example 1-2-3)
Except that the pH before cell separation was adjusted to 4.0, cell separation and MEK extraction were performed in the same manner as in Example 1-2-2, and OD660, phase separation time, amount of intermediate insoluble matter, and organic phase clear The rate, average particle size, and light scattering intensity were measured.
菌体分離前のpHを5.0に調整した以外は実施例1-2-2と同様にして菌体分離・MEK抽出を行い、OD660、分相時間、中間不溶分の量、有機相クリア率、平均粒径、光散乱強度を測定した。 (Example 1-2-4)
Cell separation and MEK extraction were carried out in the same manner as in Example 1-2-2 except that the pH before cell separation was adjusted to 5.0, OD660, phase separation time, amount of intermediate insoluble matter, organic phase clear The rate, average particle size, and light scattering intensity were measured.
菌体分離前のpHを7.6に調整した以外は実施例1-2-2と同様にして菌体分離・MEK抽出を行い、OD660、分相時間、中間不溶分の量、有機相クリア率、平均粒径、光散乱強度を測定した。 (Comparative Example 1-2-1)
Except that the pH before cell separation was adjusted to 7.6, cell separation and MEK extraction were performed in the same manner as in Example 1-2-2, and OD660, phase separation time, amount of intermediate insoluble matter, and organic phase clear The rate, average particle size, and light scattering intensity were measured.
遠心分離を1000Gで8分間行った以外は実施例1-2-1と同様にして菌体分離・MEK抽出を行い、OD660、分相時間、中間不溶分の量を測定した。その結果、OD660は0.191であり、分相時間は26秒で、中間不溶分の量は少なかった。このことから、低速遠心でも効率よくMEK抽出が行えることがわかった。 [Example 1-3]
Bacterial cell separation and MEK extraction were performed in the same manner as in Example 1-2-1 except that centrifugation was performed at 1000 G for 8 minutes, and OD660, phase separation time, and amount of intermediate insoluble matter were measured. As a result, OD660 was 0.191, phase separation time was 26 seconds, and the amount of intermediate insoluble matter was small. From this, it was found that MEK extraction can be performed efficiently even at low speed centrifugation.
(A)ジャーファーメンターによるコハク酸発酵液の調整
参考例1で作製した菌株を用いて、参考例2に従ってコハク酸発酵液を作製した。ただし、コハク酸生成反応時の温度は39℃、pHは7.6で行った。なお、得られたコハク酸発酵液を遠心分離(15,000G,5分)処理し、得られた上澄液について組成分析を行った。結果を表5に示す。 [Example 1-4]
(A) Preparation of succinic acid fermentation broth by jar fermenter A succinic acid fermentation broth was prepared according to Reference Example 2 using the strain prepared in Reference Example 1. However, the temperature during the succinic acid production reaction was 39 ° C. and the pH was 7.6. The obtained succinic acid fermentation broth was subjected to centrifugal separation (15,000 G, 5 minutes), and the resulting supernatant was subjected to composition analysis. The results are shown in Table 5.
作製したコハク酸発酵液を95%硫酸で各々pHを7.6、7.0、6.5、6.0、5.5、5.0、4.5、4.0、3.5、3.0、2.5、2.0、1.5、1.0および0.5に調整した後、35mLずつ遠心チューブに分注し、40℃で10~30分間保温した。 (B) pH adjustment and centrifugal separation of the succinic acid fermentation broth prepared with 95% sulfuric acid were adjusted to pH 7.6, 7.0, 6.5, 6.0, 5.5, 5.0, 4.5, After adjusting to 4.0, 3.5, 3.0, 2.5, 2.0, 1.5, 1.0 and 0.5, dispense 35 mL each into a centrifuge tube, Incubated for 30 minutes.
pH2.5に調整済みの発酵液上澄液の一部を用いてMEK抽出を行った。発酵液上澄液20gにMEK10gを加え40℃で5分間懸濁した後、静置し、分相時間(水相と有機相が分離するまでの時間)を測定し、水相と有機相の間に存在する中間不溶分の存在量を確認し、有機相における浮遊物の存在しない領域(有機相クリア率)を目視にて確認した。 (C) MEK extraction MEK extraction was performed using a part of the fermentation broth supernatant adjusted to pH 2.5. 10 g of MEK was added to 20 g of the supernatant of the fermentation broth, suspended at 40 ° C. for 5 minutes, allowed to stand, and the phase separation time (time until the aqueous phase and the organic phase were separated) was measured. The amount of intermediate insolubles present between them was confirmed, and the region in the organic phase where there was no suspended matter (organic phase clear rate) was visually confirmed.
(A)ジャーファーメンターによるコハク酸発酵液の調整
(A-1)種培養および本培養
参考例1で作製した菌株を用いて、参考例2(A)および(B)記載の方法に従って、種培養、本培養を行った。 [Example 1-5]
(A) Preparation of succinic acid fermentation broth by jar fermenter (A-1) Seed culture and main culture Using the strain prepared in Reference Example 1, according to the method described in Reference Example 2 (A) and (B) Culture and main culture were performed.
85%リン酸:0.3g、硫酸マグネシウム・7水和物:0.2g、硫酸第一鉄・7水和物:9mg、硫酸マンガン・水和物:9mg、10N水酸化カリウム水溶液:0.57gを蒸留水に溶解、8.4mLに調整後、121℃、20分加熱滅菌処理し、反応濃縮培地を作製した。 (A-2) Succinic acid production reaction 85% phosphoric acid: 0.3 g, magnesium sulfate heptahydrate: 0.2 g, ferrous sulfate heptahydrate: 9 mg, manganese sulfate hydrate: 9 mg 10N aqueous potassium hydroxide solution: 0.57 g was dissolved in distilled water, adjusted to 8.4 mL, and then heat-sterilized at 121 ° C. for 20 minutes to prepare a reaction concentrated medium.
得られたコハク酸発酵液を95%硫酸でpH2.5に調整した後、40mLずつを膜濾過(膜孔径0.22μm、5μm)にて菌体分離した。一方、比較例として得られたコハク酸発酵液を、40mLずつを各々膜濾過(膜孔径0.22μm、5μm)した後、濾液は95%硫酸を用いてpHを2.5に調整した。得られたpHを2.5に調整済みの膜濾過液の一部を用いて濁度(OD660)測定を行った。測定結果を表8に示す。 (B) Membrane filtration and pH adjustment The obtained succinic acid fermentation broth was adjusted to pH 2.5 with 95% sulfuric acid, and then 40 mL of cells were separated by membrane filtration (membrane pore diameter 0.22 μm, 5 μm). On the other hand, 40 mL each of succinic acid fermentation broth obtained as a comparative example was subjected to membrane filtration (membrane pore diameter 0.22 μm, 5 μm), and then the filtrate was adjusted to pH 2.5 using 95% sulfuric acid. Turbidity (OD660) measurement was performed using a part of the obtained membrane filtrate adjusted to pH 2.5. Table 8 shows the measurement results.
上記膜濾過液の一部を用いてMEK抽出を行った。膜濾過液2gにMEK1gを加え、室温で5分間懸濁した後、静置し、分相時間(水相と有機相が分離するまでの時間)を測定し、水相と有機相の間に存在する中間不溶分の存在量を確認し、有機相における浮遊物の存在しない領域(有機相クリア率)を目視にて確認した。MEK抽出結果を表8に示す。 (C) MEK extraction MEK extraction was performed using a part of the membrane filtrate. Add 1 g of MEK to 2 g of membrane filtrate, suspend at room temperature for 5 minutes, let stand, measure the phase separation time (time until the water phase and the organic phase separate), and between the water phase and the organic phase The amount of the intermediate insoluble component present was confirmed, and the region in the organic phase where there was no suspended matter (organic phase clear rate) was visually confirmed. The MEK extraction results are shown in Table 8.
(A)pH調整および遠心分離、膜分離
実施例1-5(A-2)で得られたコハク酸発酵液を95%硫酸でpHを2.5に調整した。これを35mLずつ遠心チューブに分注し、4200Gで8分間遠心し、得られた上澄液を各々膜濾過(膜孔径0.22、5μmまたは、膜濾過なし)した。 [Example 1-6]
(A) pH adjustment, centrifugation, and membrane separation The succinic acid fermentation broth obtained in Example 1-5 (A-2) was adjusted to pH 2.5 with 95% sulfuric acid. 35 mL of this was dispensed into a centrifuge tube, centrifuged at 4200 G for 8 minutes, and the resulting supernatant was subjected to membrane filtration (membrane pore size 0.22, 5 μm or no membrane filtration).
上記、膜濾過液を用いてMEK抽出を行った。上記濾過液20gにMEK10gを加え40℃で5分間懸濁した後、静置し、分相時間(水相と有機相が分離するまでの時間)を測定し、水相と有機相の間に存在する中間不溶分の存在量を確認し、有機相における浮遊物の存在しない領域(有機相クリア率)を目視にて確認した。また、濾過液の一部を用いて光散乱測定を行い、平均粒径、光散乱強度を測定した。MEK抽出および光散乱測定結果を表9に示す。なお、光散乱測定は、実施例1-2-1記載の測定方法に従って行った。 (B) MEK extraction MEK extraction was performed using the above membrane filtrate. 10 g of MEK was added to 20 g of the above filtrate and suspended at 40 ° C. for 5 minutes, then allowed to stand, and the phase separation time (time until the aqueous phase and the organic phase were separated) was measured, and between the aqueous phase and the organic phase. The amount of the intermediate insoluble component present was confirmed, and the region in the organic phase where there was no suspended matter (organic phase clear rate) was visually confirmed. Moreover, light scattering measurement was performed using a part of the filtrate, and the average particle diameter and light scattering intensity were measured. The results of MEK extraction and light scattering measurement are shown in Table 9. The light scattering measurement was performed according to the measurement method described in Example 1-2-1.
本実施例における酸類、糖類の定量分析は、高速液体クロマトグラフィー(LC)を用い、アミノ酸の定量分析はアミノ酸分析計を用い以下の条件で測定を行った。またタンパク質の定量は試料を塩酸により加水分解処理し、加水分解前後の総アミノ酸量の増分をタンパク質量と見なした。 [Example 2]
In this example, high-performance liquid chromatography (LC) was used for quantitative analysis of acids and saccharides, and amino acid quantitative analysis was performed using an amino acid analyzer under the following conditions. For protein quantification, the sample was hydrolyzed with hydrochloric acid, and the increment of the total amino acid content before and after hydrolysis was regarded as the protein amount.
カラム;信和化工(株)製 ULTRON PS-80H 8.0mmI.D.× 30cm
溶離液:水(過塩素酸)(過塩素酸60%水溶液1.8ml/1L-H2O)
温度:60℃ <Analysis of acids and sugars>
Column: ULTRON PS-80H 8.0 mmI. D. × 30cm
Eluent: water (perchloric acid) (perchloric acid 60% aqueous solution 1.8 ml / 1 L-H 2 O)
Temperature: 60 ° C
装置:日立アミノ酸分析計 L-8900
分析条件:生体アミノ酸分離条件-ニンヒドリン発色法(570nm,440nm)
標準品:PF(和光アミノ酸混合液ANII型0.8ml+B型0.8ml→10ml)
注入量:10μl <Amino acid analysis>
Equipment: Hitachi Amino Acid Analyzer L-8900
Analysis conditions: biogenic amino acid separation conditions-ninhydrin coloring method (570 nm, 440 nm)
Standard product: PF (Wako amino acid mixed solution ANII type 0.8 ml + B type 0.8 ml → 10 ml)
Injection volume: 10 μl
試料10mgまたは100mgを精秤し、純水で1mL定容としたものを、200μL分注、乾固し、塩酸雰囲気下150℃、1時間加熱し、タンパク質を加水分解処理した。これを乾固させた後純水200μLを加えて再溶解させ、0.45μmフィルターでろ過後、ろ液をアミノ酸の分析に供した。 <Hydrolysis for protein quantification>
A sample of 10 mg or 100 mg was precisely weighed and a 1 mL constant volume with pure water was dispensed to 200 μL, dried and heated in a hydrochloric acid atmosphere at 150 ° C. for 1 hour to hydrolyze the protein. After drying this, 200 μL of pure water was added and redissolved, and after filtration through a 0.45 μm filter, the filtrate was subjected to amino acid analysis.
<プロトン化>
表2に示す組成のコハク酸発酵液1500gに98%硫酸を加え、pHを2.5に調整した(以下、コハク酸水溶液と記載することがある)。ここで98%硫酸の添加量は、150gであった。 [Example 2-1]
<Protonation>
98% sulfuric acid was added to 1500 g of the succinic acid fermentation broth having the composition shown in Table 2 to adjust the pH to 2.5 (hereinafter sometimes referred to as an aqueous succinic acid solution). Here, the addition amount of 98% sulfuric acid was 150 g.
硫酸添加後のコハク酸水溶液を、ボトム弁付の容量5Lのジャケット付攪拌槽に入れ、さらにメチルエチルケトン(以下、MEKと記載することがある)に予め水を添加した10%含水メチルエチルケトン溶液2475gを加え(MEK溶液/コハク酸水溶液=1.5(w/w))、ジャケットに温水を通液することで内温を30℃に制御しながら30分間攪拌した。その後攪拌を止め、内温を30℃に制御しながら約1時間静置した。静置後液々界面近傍のMEK相には中間相が存在していた。 <Contact process (batch operation)>
The aqueous succinic acid solution after addition of sulfuric acid is placed in a 5 L jacketed stirring tank with a bottom valve, and 2475 g of a 10% hydrous methyl ethyl ketone solution in which water has been added in advance to methyl ethyl ketone (hereinafter sometimes referred to as MEK) is added. (MEK solution / succinic acid aqueous solution = 1.5 (w / w)) The mixture was stirred for 30 minutes while controlling the internal temperature to 30 ° C. by passing warm water through the jacket. Thereafter, stirring was stopped, and the mixture was allowed to stand for about 1 hour while controlling the internal temperature at 30 ° C. An intermediate phase was present in the MEK phase near the liquid-liquid interface after standing.
回収した抽出液は連続蒸留によりMEKを実質的に除去する。ここで蒸留留出液はMEKと水の共沸組成物、すなわち11wt%含水MEKとして回収されるが、釜残液の濃縮度合い次第ではコハク酸が析出する懸念がある。そこで蒸留留出液が11wt%含水MEK、釜残液が30wt%コハク酸溶液となるよう抽出液2503gに対し103gの水を添加した。ここで水の添加量は下記計算に従い算出した。 <Distillation>
The recovered extract substantially removes MEK by continuous distillation. Here, the distilled distillate is recovered as an azeotropic composition of MEK and water, that is, 11 wt% water-containing MEK, but there is a concern that succinic acid may precipitate depending on the degree of concentration of the kettle residue. Therefore, 103 g of water was added to 2503 g of the extract so that the distilled distillate was 11 wt% hydrous MEK and the kettle residue was 30 wt% succinic acid solution. Here, the amount of water added was calculated according to the following calculation.
抽出液のMEKおよびコハク酸濃度をそれぞれCMEK,0、CSA,0、希釈後のMEKおよびコハク酸濃度をそれぞれCMEK,1、CSA,1とした際の相図を図3に示す。図3では、抽出液の水による希釈、その後の蒸留におけるMEKおよびコハク酸の組成変化が示されている。 (Dilution of extract)
FIG. 3 shows a phase diagram when the MEK and succinic acid concentrations of the extract were C MEK, 0 and C SA, 0 , respectively, and the diluted MEK and succinic acid concentrations were C MEK, 1 and C SA, 1 respectively. . FIG. 3 shows the composition change of MEK and succinic acid in the dilution of the extract with water and subsequent distillation.
CSA=(CSA,0/CMEK,0)CMEK
また蒸留操作線は釜残液(0,0.3)と留出液(0.89,0)結ぶ線、
CSA=0.3-(0.3/0.89)CMEK
で表現され、希釈後の液はそれぞれの線の交点にあることから、
CSA,1=(CSA,0/CMEK,0)CMEK,1 ・・・・eq.(1)
CSA,1=0.3-(0.3/0.89)CMEK,1 ・・・・eq.(2) In FIG. 3, the dilution operation line is a line connecting the extract composition (C MEK, 0 , C SA, 0 ) and the origin (0,0),
C SA = (C SA, 0 / C MEK, 0 ) C MEK
In addition, the distillation operation line is a line connecting the kettle residue (0,0.3) and the distillate (0.89,0)
C SA = 0.3− (0.3 / 0.89) C MEK
Since the diluted liquid is at the intersection of each line,
C SA, 1 = (C SA, 0 / C MEK, 0 ) C MEK, 1 ... Eq. (1)
C SA, 1 = 0.3− (0.3 / 0.89) C MEK, 1 ... Eq. (2)
MEKを留去した液はジャケット付500mlセパラブルフラスコへ移し、攪拌下ジャケットに温水を通水することで80℃に保温した。その後プログラム付循環恒温槽を用い、ジャケットへ通水する温水を1時間かけて20℃まで冷却し、コハク酸を冷却晶析し、20℃到達後さらに20℃で1時間熟成した。 <Crystal>
The liquid from which MEK was distilled off was transferred to a jacketed 500 ml separable flask, and kept warm at 80 ° C. by passing warm water through the jacket while stirring. Thereafter, using a circulating thermostatic bath with a program, the warm water passing through the jacket was cooled to 20 ° C. over 1 hour, succinic acid was cooled and crystallized, and after reaching 20 ° C., the mixture was further aged at 20 ° C. for 1 hour.
実施例2-1において、抽出温度を20℃にする以外同じ方法で処理を行った。抽出後の中間相は実施例2-1に比べて明らかに多く、回収された抽出相、中間相、抽残相はそれぞれ2322g、316g、1485gであった。抽出相を組成分析した結果を下記表13に示す。 [Example 2-2]
In Example 2-1, the treatment was performed in the same manner except that the extraction temperature was 20 ° C. The number of extracted intermediate phases was clearly larger than that of Example 2-1, and the recovered extracted phase, intermediate phase, and extracted residual phase were 2322 g, 316 g, and 1485 g, respectively. The results of composition analysis of the extracted phase are shown in Table 13 below.
実施例2-1において、抽出温度を60℃にする以外同じ方法で処理を行った。 [Example 2-3]
In Example 2-1, the treatment was performed in the same manner except that the extraction temperature was 60 ° C.
<プロトン化>
上述のコハク酸発酵液1500gに98%硫酸を加えpHを2.5に調整した。ここで98%硫酸添加量は約150gであった。 [Example 2-4]
<Protonation>
To 1500 g of the above succinic acid fermentation broth, 98% sulfuric acid was added to adjust the pH to 2.5. Here, the addition amount of 98% sulfuric acid was about 150 g.
硫酸添加後のコハク酸水溶液を、図4に示すような、500mlジャケット付攪拌槽および各槽が600ml、400ml、300mlのジャケット付3槽式のセトラーを用い、MEK溶液と混合、液々分離することでコハク酸を連続抽出した。すなわちコハク酸水溶液1650gおよび予め水を添加した10%含水メチルエチルケトン(MEK)溶液2475g(MEK溶液/コハク酸水溶液=1.5(w/w))を、ジャケットに30℃の温水を流し温度制御された500mlジャケット付攪拌槽にそれぞれ20g/分、30g/分の速度で供給するとともに、槽内が均一となるよう激しく攪拌しながら、懸濁液を50g/分の速度で連続的に抜き出した。 <Extraction (mixer (stirring tank) / settler)>
The aqueous solution of succinic acid after the addition of sulfuric acid is mixed and separated from the MEK solution using a 500 ml jacketed stirring tank and a 600 ml, 400 ml, and 300 ml jacketed 3-settler settling tank as shown in FIG. Thus, succinic acid was continuously extracted. That is, 1650 g of succinic acid aqueous solution and 2475 g of 10% hydrous methyl ethyl ketone (MEK) solution (MEK solution / succinic acid aqueous solution = 1.5 (w / w)) to which water has been added in advance, and hot water at 30 ° C. are passed through the jacket to control the temperature In addition, the suspension was continuously extracted at a rate of 50 g / min while being vigorously stirred so that the inside of the vessel became uniform while being supplied to the 500 ml jacketed agitation vessel at a rate of 20 g / min and 30 g / min, respectively.
回収した抽出液は2370gに対し97gの水を添加し、実施例2-1と同様の方法で連続蒸留を行い、釜残液275gを回収した。蒸留には7時間を要したが、蒸留後のボトムフラスコは比較的きれいであった。得られた釜残液を組成分析した結果を下記表16に示す。 <Distillation>
97 g of water was added to 2370 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 275 g of the residue from the kettle. The distillation took 7 hours, but the bottom flask after the distillation was relatively clean. Table 16 below shows the results of composition analysis of the obtained kettle residue.
回収した釜残液は実施例2-1と同様の方法により晶析し、最終的に74gのコハク酸を回収した。得られたコハク酸を組成分析した結果を下記表17に示す。 <Crystal>
The recovered pot residue was crystallized in the same manner as in Example 2-1, and finally 74 g of succinic acid was recovered. The results of composition analysis of the obtained succinic acid are shown in Table 17 below.
上述のコハク酸発酵液1500gに98%硫酸を加えpHを2.5に調整した。ここで98%硫酸添加量は約150gであった。 <Protonation>
To 1500 g of the above succinic acid fermentation broth, 98% sulfuric acid was added to adjust the pH to 2.5. Here, the addition amount of 98% sulfuric acid was about 150 g.
硫酸添加後のコハク酸水溶液は、ジャケット付スタティックミキサー(ノリタケ1/4(1)-N40-174-0(内径5mm、エレメント数24))および各槽が600ml、400ml、300mlのジャケット付3槽式のセトラーを用い、MEK溶液と混合、液々分離することでコハク酸を連続抽出した。 <Extraction (mixer (static mixer) / settler)>
The aqueous succinic acid solution after the addition of sulfuric acid is a jacketed static mixer (
回収した抽出液は2435gに対し100gの水を添加し、実施例2-1と同様の方法で連続蒸留を行い、釜残液286gを回収した。蒸留には7時間を要したが、蒸留後のボトムフラスコは比較的きれいであった。 <Distillation>
100 g of water was added to 2435 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 286 g of the residue from the kettle. The distillation took 7 hours, but the bottom flask after the distillation was relatively clean.
回収した釜残液は実施例2-1と同様の方法により晶析し、最終的に76gのコハク酸を回収した。得られたコハク酸を組成分析した結果を下記表19に示す。 <Crystal>
The recovered kettle residue was crystallized in the same manner as in Example 2-1, and finally 76 g of succinic acid was recovered. The results of composition analysis of the obtained succinic acid are shown in Table 19 below.
実施例2-1の方法に従い、プロトン化、抽出操作を行い、抽出相2503g、抽残相1490g、中間相132gを回収した。中間相は遠心沈降機で2000Gで10分間振切り清澄液130gを回収した。清澄液と抽出液とを混合して得られた液の組成分析結果を、下記表20に示す。 [Example 2-6]
In accordance with the method of Example 2-1, protonation and extraction operations were performed to recover 2503 g of extraction phase, 1490 g of extraction residual phase, and 132 g of intermediate phase. The intermediate phase was centrifuged at 2000 G for 10 minutes with a centrifugal settling machine to recover 130 g of the clarified liquid. Table 20 below shows the composition analysis results of the liquid obtained by mixing the clarified liquid and the extract.
回収した抽出液は2633gに対し108gの水を添加し、実施例2-1と同様の方法で連続蒸留を行い、釜残液310gを回収した。蒸留には7時間を要したが、蒸留後のボトムフラスコは比較的きれいであった。 <Distillation>
108 g of water was added to 2633 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 310 g of the residue from the kettle. The distillation took 7 hours, but the bottom flask after the distillation was relatively clean.
回収した釜残液は実施例2-1と同様の方法により晶析し、最終的に82gのコハク酸を回収した。得られたコハク酸を組成分析した結果を、下記表21に示す。 <Crystal>
The recovered pot residue was crystallized in the same manner as in Example 2-1, and finally 82 g of succinic acid was recovered. The results of composition analysis of the obtained succinic acid are shown in Table 21 below.
実施例2-1の方法に従い、プロトン化、抽出操作を行い、抽出相2503g、抽残相1490g、中間相132gを回収した。中間相は目開き0.5μmのPTFE製メンブランフィルターで加圧ろ過することで清澄液130gを回収した。清澄液と抽出液とを混合して得られた液の組成分析結果を、下記表22に示す。 [Example 2-7]
In accordance with the method of Example 2-1, protonation and extraction operations were performed to recover 2503 g of extraction phase, 1490 g of extraction residual phase, and 132 g of intermediate phase. The intermediate phase was subjected to pressure filtration with a PTFE membrane filter having an opening of 0.5 μm to recover 130 g of the clarified liquid. Table 22 shows the composition analysis results of the liquid obtained by mixing the clarified liquid and the extract.
回収した抽出液は2503gに対し108gの水を添加し、実施例2-1と同様の方法で連続蒸留を行い、釜残液310gを回収した。蒸留には7時間を要したが、蒸留後のボトムフラスコは比較的きれいであった。 <Distillation>
108 g of water was added to 2503 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 310 g of the residue from the kettle. The distillation took 7 hours, but the bottom flask after the distillation was relatively clean.
回収した釜残液は実施例2-1と同様の方法により晶析し、最終的に82gのコハク酸を回収した。得られたコハク酸を組成分析した結果を、下記表23に示す。 <Crystal>
The recovered pot residue was crystallized in the same manner as in Example 2-1, and finally 82 g of succinic acid was recovered. The results of composition analysis of the obtained succinic acid are shown in Table 23 below.
実施例5のプロトン化、抽出操作において、コハク酸水溶液1650gに対して含水MEK供給量を825g((MEK溶液/コハク酸水溶液=0.5(w/w))をそれぞれ20g/分、含水MEK供給量を10g/分でスタティックミキサーにする供給する以外は同じ方法に従い、抽出相688g、抽残相1613g、中間相173gを回収した。中間相は目開き0.5μmのPTFE製メンブランフィルターで加圧ろ過することで清澄液172gを回収した。 [Example 2-8]
In the protonation and extraction operations of Example 5, 825 g of hydrous MEK supply ((MEK solution / succinic aqueous solution = 0.5 (w / w)) was 20 g / min and hydrous MEK to 1650 g of succinic acid aqueous solution. Except for feeding to a static mixer at a feed rate of 10 g / min, an extraction phase of 688 g, an extraction residual phase of 1613 g, and an intermediate phase of 173 g were recovered, and the intermediate phase was added with a PTFE membrane filter having an opening of 0.5 μm. 172 g of the clarified liquid was recovered by pressure filtration.
図5に示すようなプロセスフローにて、連続抽出操作により溶剤相にコハク酸を回収した。抽残相1613gは、内径φ20mm、高さ2mのジャケット付攪拌型連続抽出塔(理論段10段)を用い連続抽出しコハク酸を回収した。ここで抽残相は200g/時で塔頂部から供給し、ボトムから予め10wt%含水に調整したMEK溶液200g/時で連続抽出した。連続相は抽残相、分散相はMEK相とした(軽液分散)。また抽出塔の温度はジャケットに温水を通液することで30℃に制御した。最終的に抽出液を1777g回収した。 <Continuous extraction>
In the process flow as shown in FIG. 5, succinic acid was recovered in the solvent phase by a continuous extraction operation. The extracted residual phase 1613 g was continuously extracted by using a jacketed continuous extraction tower (10 theoretical plates) with an inner diameter of 20 mm and a height of 2 m to recover succinic acid. Here, the extracted residual phase was supplied from the top of the column at 200 g / hour, and continuously extracted from the bottom at 200 g / hour of MEK solution adjusted in advance to a water content of 10 wt%. The continuous phase was the extracted residual phase, and the dispersed phase was the MEK phase (light liquid dispersion). The temperature of the extraction tower was controlled at 30 ° C. by passing warm water through the jacket. Finally, 1777 g of the extract was recovered.
回収した抽出液は2637gに対し190gの水を添加し、実施例2-1と同様の方法で連続蒸留を行い、釜残液432gを回収した。蒸留には7時間を要したが、蒸留後のボトムフラスコは比較的きれいであった。 <Distillation>
190 g of water was added to 2637 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 432 g of the residue from the kettle. The distillation took 7 hours, but the bottom flask after the distillation was relatively clean.
回収した釜残液は実施例2-1と同様の方法により晶析し、最終的に114gのコハク酸を回収した。得られたコハク酸を組成分析した結果を、下記表25に示す。 <Crystal>
The recovered pot residue was crystallized in the same manner as in Example 2-1, and finally 114 g of succinic acid was recovered. The results of composition analysis of the obtained succinic acid are shown in Table 25 below.
実施例2-1の方法に従い、中間相を抽出相として回収する以外同じ方法では抽出相2633g、抽残相1490gを回収した。 [Comparative Example 2-1]
According to the method of Example 2-1, except that the intermediate phase was recovered as the extraction phase, 2633 g of the extraction phase and 1490 g of the extracted residual phase were recovered by the same method.
回収した抽出液は2633gに対し109gの水を添加し、実施例2-1と同様の方法で連続蒸留を行い、釜残液300gを回収した。連続蒸留ではオイルバス温度を140℃としたが、連続蒸留開始後数時間後にはボトム温度が低下したので(MEK検出)、供給量を下げ、オイルバス温度140℃でボトム温度>100℃が維持できる条件で運転を継続した。結果蒸留には9時間を要した。また蒸留後のボトムフラスコ内面には粘着物が多量に付着していた。 <Distillation>
109 g of water was added to 2633 g of the recovered extract, and continuous distillation was performed in the same manner as in Example 2-1, to recover 300 g of the residue from the kettle. In continuous distillation, the oil bath temperature was 140 ° C, but the bottom temperature decreased several hours after the start of continuous distillation (MEK detection), so the supply amount was lowered and the bottom temperature> 100 ° C was maintained at the oil bath temperature of 140 ° C. The operation was continued under conditions that allowed it. Results Distillation took 9 hours. In addition, a large amount of adhesive was adhered to the inner surface of the bottom flask after distillation.
MEKを留去した液はジャケット付500mlセパラブルフラスコへ移し、攪拌下ジャケットに温水を通水することで80℃に保温した。その後プログラム付循環恒温槽を用い、ジャケットへ通水する温水を1時間かけて20℃まで冷却し、コハク酸を冷却晶析し、20℃到達後さらに20℃で1時間熟成した。 <Crystal>
The liquid from which MEK was distilled off was transferred to a jacketed 500 ml separable flask, and kept warm at 80 ° C. by passing warm water through the jacket while stirring. Thereafter, using a circulating thermostatic bath with a program, the warm water passing through the jacket was cooled to 20 ° C. over 1 hour, succinic acid was cooled and crystallized, and after reaching 20 ° C., the mixture was further aged at 20 ° C. for 1 hour.
実施例2-1の方法に従い、プロトン化したコハク酸水溶液を実施例2-9で用いた連続抽出塔塔頂部から200g/時で供給するとともに、ボトムから10%含水MEKを300g/時で供給したところ、塔内で多量の不溶成分が発生し、液々分離を阻害し、連続運転はできなかった。 [Comparative Example 2-2]
In accordance with the method of Example 2-1, a protonated succinic acid aqueous solution was supplied from the top of the continuous extraction tower used in Example 2-9 at 200 g / hour, and 10% hydrous MEK was supplied from the bottom at 300 g / hour. As a result, a large amount of insoluble components were generated in the tower, and liquid-liquid separation was inhibited, and continuous operation was not possible.
本実施例における酸類、糖類の定量分析、アミノ酸の定量分析、タンパク質の定量は実施例2と同様に行った。 [Example 3]
In this example, the quantitative analysis of acids, saccharides, quantitative analysis of amino acids, and protein were performed in the same manner as in Example 2.
<プロトン化工程>
表2に示す組成のコハク酸発酵液1500gに、98%硫酸を加えpHを2.5に調整した。ここで98%硫酸添加量は150gであった。 [Example 3-1]
<Protonation process>
To 1500 g of the succinic acid fermentation broth having the composition shown in Table 2, 98% sulfuric acid was added to adjust the pH to 2.5. Here, the addition amount of 98% sulfuric acid was 150 g.
硫酸添加後のコハク酸水溶液は、ジャケット付スタティックミキサー(ノリタケ1/4(1)-N40-174-0(内径Φ5mm、エレメント数24))および各槽が600ml、400ml、300mlのジャケット付3槽式のセトラーを用い、MEK溶液と混合、液々分離することでコハク酸を連続抽出した。 <Contact process>
The succinic acid aqueous solution after the addition of sulfuric acid consists of a jacketed static mixer (
回収した抽残相1613gは、内径Φ20mm,高さ2mのジャケット付攪拌型連続抽出塔(理論段10段)を用い、10%含水MEK1613g(MEK溶液(重量)/コハク酸水溶液(重量)=1.0(重量/重量))で連続抽出した。ここで抽残相は200g/時間の速度で塔頂部から供給し、塔底から予め含水量10重量%に調整したMEK溶液を200g/時間の速度で流した。連続相は抽残相、分散相はMEK相とした(軽液分散)。 <Continuous extraction>
1613 g of the recovered residual phase was obtained by using a jacketed continuous extraction tower (10 theoretical plates) with an inner diameter of Φ20 mm and a height of 2 m (10 stages of theoretical plate), 10% hydrous MEK 1613 g (MEK solution (weight) / succinic acid aqueous solution (weight) = 1. 0.0 (weight / weight)). Here, the extracted residual phase was supplied from the top of the tower at a rate of 200 g / hour, and a MEK solution adjusted in advance to a water content of 10% by weight was passed from the bottom of the tower at a rate of 200 g / hour. The continuous phase was the extracted residual phase, and the dispersed phase was the MEK phase (light liquid dispersion).
回収した抽出相は連続蒸留によりMEKを実質的に除去する。ここで蒸留留出液への水の添加量は、実施例2-1と同様に算出した。蒸留後の釜残液は432gであった。またその組成を分析した結果を、下記表29に示す。 <Distillation>
The recovered extracted phase substantially removes MEK by continuous distillation. Here, the amount of water added to the distillate was calculated in the same manner as in Example 2-1. The residue in the kettle after distillation was 432 g. The results of analyzing the composition are shown in Table 29 below.
MEKを留去した液はジャケット付500mlセパラブルフラスコへ移し、攪拌下ジャケットに温水を通水することで80℃に保温した。その後プログラム付循環恒温槽を用い、ジャケットへ通水する温水を1時間かけて20℃まで冷却し、コハク酸を冷却晶析し、20℃到達後さらに20℃で1時間熟成した。 <Crystal>
The liquid from which MEK was distilled off was transferred to a jacketed 500 ml separable flask, and kept warm at 80 ° C. by passing warm water through the jacket while stirring. Thereafter, using a circulating thermostatic bath with a program, the warm water passing through the jacket was cooled to 20 ° C. over 1 hour, succinic acid was cooled and crystallized, and after reaching 20 ° C., the mixture was further aged at 20 ° C. for 1 hour.
<プロトン化>
前記コハク酸発酵液1384gに、98%硫酸を加えpH2.5とした。ここで98%硫酸添加量は138gであった。このプロトン化した液に、実施例3-1で回収された回収液の半分に当たる281gを加え、コハク酸水溶液1803gを調整した。 [Example 3-2]
<Protonation>
98% sulfuric acid was added to 1384 g of the succinic acid fermentation broth to adjust the pH to 2.5. Here, the addition amount of 98% sulfuric acid was 138 g. To this protonated liquid, 281 g corresponding to half of the recovered liquid recovered in Example 3-1 was added to prepare 1803 g of an aqueous succinic acid solution.
実施例3-1と同様の方法で、コハク酸水溶液を10%含水MEKで抽出した。ミキサーセトラーではコハク酸水溶液に対し0.5重量倍の10%含水MEKをそれぞれ20g/分、10g/分で供給し、回収した中間相は加圧ろ過、抽残相はさらに抽残相に対し1.0重量倍の10%含水MEKで向流多段連続抽出を行った。結果、抽出相2882g、抽残相1593gを回収した。その組成は以下の通りであった。 <Extraction>
A succinic acid aqueous solution was extracted with 10% aqueous MEK in the same manner as in Example 3-1. The mixer settler supplies 0.5% by weight 10% water-containing MEK to the aqueous succinic acid solution at 20 g / min and 10 g / min, respectively, and the recovered intermediate phase is subjected to pressure filtration. Countercurrent multistage continuous extraction was performed with 1.0% by weight 10% water-containing MEK. As a result, 2882 g of extracted phase and 1593 g of extracted residual phase were recovered. Its composition was as follows.
実施例3-1と同様の方法で蒸留を行い、コハク酸濃縮液438gを回収した。その組成は下記表32の通りであった。 <Distillation>
Distillation was carried out in the same manner as in Example 3-1, and 438 g of a succinic acid concentrate was recovered. The composition was as shown in Table 32 below.
実施例3-1と同様の方法で晶析を行い、コハク酸113gと晶析母液および洗浄液の混合液である回収液568gを回収した。下記表33に得られたコハク酸の組成を、下記表34に得られた回収液の組成を示す。 <Crystal>
Crystallization was carried out in the same manner as in Example 3-1, and 568 g of a recovery liquid, which was a mixed liquid of 113 g of succinic acid, a crystallization mother liquid, and a cleaning liquid, was recovered. Table 33 below shows the composition of succinic acid obtained, and Table 34 below shows the composition of the collected liquid.
実施例3-2と同じ手順でプロトン化から晶析までの工程を実施して、コハク酸114gと晶析母液および洗浄液の混合物である回収液573gを得た。下記表35に得られたコハク酸の組成を、下記表36に得られた回収液の組成を示す。 [Example 3-3]
The steps from protonation to crystallization were carried out in the same procedure as in Example 3-2 to obtain 573 g of a recovered liquid which was a mixture of 114 g of succinic acid, the crystallization mother liquor and the washing liquid. Table 35 below shows the composition of succinic acid obtained, and Table 36 below shows the composition of the collected liquid.
実施例2と同じ手順でプロトン化から晶析までの工程を実施して、コハク酸114gと、晶析母液および洗浄液の混合物である回収液564gを得た。下記表37に得られたコハク酸の組成を示す。下記表38に得られた回収液の組成を示す。 [Example 3-4]
The steps from protonation to crystallization were carried out in the same procedure as in Example 2 to obtain 114 g of recovered liquid, which was a mixture of succinic acid 114 g, crystallization mother liquor and washing liquid. The composition of succinic acid obtained is shown in Table 37 below. The composition of the recovered liquid obtained is shown in Table 38 below.
Claims (30)
- 生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(4)を含む脂肪族ジカルボン酸の製造方法。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物を分離する微生物分離工程
(4)微生物分離工程で微生物を分離して得られた水溶液と、溶剤とを接触させる接触工程 A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism, comprising the steps of (1) to (4) below: Production method.
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step PH adjustment step (3) for obtaining a solution having a pH of 1.0 or more and 5.0 or less by mixing a solution containing a solution and an acid, a microorganism separation step (4) for separating microorganisms from the solution obtained in the pH adjustment step ) A contact process in which an aqueous solution obtained by separating microorganisms in the microorganism separation process is contacted with a solvent. - 微生物がコリネ型細菌である、請求項1に記載の方法。 The method according to claim 1, wherein the microorganism is a coryneform bacterium.
- 脂肪族ジカルボン酸がコハク酸である、請求項1又は2に記載の方法。 The method according to claim 1 or 2, wherein the aliphatic dicarboxylic acid is succinic acid.
- 溶剤が無機性値/有機性値の比(I/O値)が0.2以上2.3以下であり、常圧(1気圧)で沸点が40℃以上の有機溶媒である、請求項1~3のいずれか1項に記載の方法。 The solvent is an organic solvent having an inorganic value / organic value ratio (I / O value) of 0.2 or more and 2.3 or less and a normal pressure (1 atm) and a boiling point of 40 ° C. or more. 4. The method according to any one of items 1 to 3.
- 生物由来原料から得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(3)を含む脂肪族ジカルボン酸の製造方法。
(1)生物由来原料から得られる脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(2)接触工程で前記水溶液と前記溶剤の接触により発生した固形分を除去する固形分除去工程
(3)前記溶剤を相分離により分離する相分離工程 A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, comprising the following steps (1) to (3).
(1) A contact step in which an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material is brought into contact with a solvent that can be phase-separated with the aqueous solution. (3) Phase separation step of separating the solvent by phase separation - 前記固形分除去工程において、前記固形分の除去を、セトラーを用いて行う請求項5に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 5, wherein in the solid content removal step, the solid content is removed using a settler.
- 前記接触工程において、前記水溶液と前記溶剤との接触を、スタティックミキサーを用いて行う請求項5または請求項6に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 5 or 6, wherein in the contacting step, the aqueous solution and the solvent are contacted using a static mixer.
- 前記固形分除去工程において、前記固形分の除去を、前記水溶液および前記溶剤から選ばれる少なくとも1種の液体と共に抜き出すことにより行う請求項5から請求項7のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The aliphatic dicarboxylic acid according to any one of claims 5 to 7, wherein in the solid content removing step, the solid content is removed together with at least one liquid selected from the aqueous solution and the solvent. Acid production method.
- 抜き出した固形分を固液分離し、得られた液体を前記水溶液と前記溶剤との接触工程以前の工程および相分離工程以降の工程の少なくとも一方に戻す、請求項8に記載の脂肪族ジカルボン酸の製造方法。 The aliphatic dicarboxylic acid according to claim 8, wherein the extracted solid content is subjected to solid-liquid separation, and the obtained liquid is returned to at least one of a step before the contact step between the aqueous solution and the solvent and a step after the phase separation step. Manufacturing method.
- 前記固液分離を遠心沈降法により行う、請求項9に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 9, wherein the solid-liquid separation is performed by a centrifugal sedimentation method.
- 前記固液分離をフィルターを用いて行う、請求項9に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 9, wherein the solid-liquid separation is performed using a filter.
- 前記溶剤を相分離により分離した後に残った液体と、該液体と相分離する溶剤とを、向流多段抽出塔で接触させ、後に該溶剤を相分離により分離する工程を含む、請求項5から請求項11のいずれか1項に記載の脂肪族ジカルボン酸の製造方法 The method comprises the steps of bringing a liquid remaining after the solvent is separated by phase separation and a solvent to be phase-separated with the liquid in a countercurrent multistage extraction tower, and then separating the solvent by phase separation. The manufacturing method of the aliphatic dicarboxylic acid of any one of Claim 11
- 前記接触工程を30~60℃で行う、請求項5から請求項12のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to any one of claims 5 to 12, wherein the contacting step is performed at 30 to 60 ° C.
- 生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(5)を含む脂肪族ジカルボン酸の製造方法。
(1)脂肪族ジカルボン酸を含む水溶液と、該水溶液と相分離する溶剤とを接触させる接触工程
(2)接触工程で回収された脂肪族ジカルボン酸を濃縮する工程であって、該濃縮により抽出相中の水濃度が上昇する抽出相濃縮工程
(3)抽出相濃縮工程後の液から脂肪族ジカルボン酸を析出させる晶析工程
(4)晶析工程で析出した脂肪族ジカルボン酸を回収する固液分離工程
(5)固液分離工程で得られた脂肪族ジカルボン酸を回収した後の晶析母液の少なくとも一部を晶析工程より前のいずれかの工程に戻す晶析母液リサイクル工程 A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained from a biological raw material, comprising the following steps (1) to (5).
(1) A contact step in which an aqueous solution containing an aliphatic dicarboxylic acid is brought into contact with a solvent that is phase-separated with the aqueous solution. (2) A step of concentrating the aliphatic dicarboxylic acid recovered in the contact step, which is extracted by the concentration. Extraction phase concentration step in which water concentration in phase increases (3) Crystallization step for precipitation of aliphatic dicarboxylic acid from liquid after extraction phase concentration step (4) Solid recovery of aliphatic dicarboxylic acid precipitated in crystallization step Liquid separation step (5) Crystallization mother liquor recycling step for returning at least part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step prior to the crystallization step - 接触工程より前に、前記生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液に酸を加えるプロトン化工程を含む、請求項14に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 14, further comprising a protonation step of adding an acid to an aqueous solution containing the aliphatic dicarboxylic acid obtained from the biological material before the contacting step.
- 晶析工程に供給される、抽出相濃縮工程後の脂肪族ジカルボン酸を含む溶液中に含まれる溶剤の濃度が10重量%以下である、請求項14または15に記載の脂肪族ジカルボン酸の製造方法。 The production of an aliphatic dicarboxylic acid according to claim 14 or 15, wherein the concentration of the solvent contained in the solution containing the aliphatic dicarboxylic acid after the extraction phase concentration step supplied to the crystallization step is 10% by weight or less. Method.
- 晶析工程に供給される、前記脂肪族ジカルボン酸を含む溶液中に含まれる溶剤の濃度が1重量%以下である、請求項16に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 16, wherein the concentration of the solvent contained in the solution containing the aliphatic dicarboxylic acid supplied to the crystallization step is 1% by weight or less.
- 固液分離工程で脂肪族ジカルボン酸を回収した後の母液から、脂肪族ジカルボン酸以外の成分を除去した後、当該液の少なくとも一部を接触工程から晶析工程までの間の少なくともいずれか1の工程にリサイクルする、請求項14から請求項17のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 After removing components other than the aliphatic dicarboxylic acid from the mother liquor after collecting the aliphatic dicarboxylic acid in the solid-liquid separation step, at least one of the liquid from the contact step to the crystallization step is removed. The method for producing an aliphatic dicarboxylic acid according to any one of claims 14 to 17, wherein the aliphatic dicarboxylic acid is recycled to the step.
- 抽出相濃縮工程以前のいずれかの工程において、水を加えることを特徴とする請求項15から請求項18のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to any one of claims 15 to 18, wherein water is added in any step before the extraction phase concentration step.
- プロトン化工程において加える酸としてpKaが4未満の酸を用い、且つ該プロトン化工程における前記生物由来原料から得られた脂肪族ジカルボン酸を含む水溶液のpHを、1より大きく4未満の範囲に維持することを特徴とする、請求項15から請求項19のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 An acid having a pKa of less than 4 is used as an acid to be added in the protonation step, and the pH of the aqueous solution containing the aliphatic dicarboxylic acid obtained from the biological material in the protonation step is maintained in the range of more than 1 and less than 4. The method for producing an aliphatic dicarboxylic acid according to any one of claims 15 to 19, wherein:
- 接触工程で用いる溶剤が、有機性値に対する無機性値の比率が0.2以上2.3以下であり、且つ沸点が40℃以上であることを特徴とする、請求項14から請求項20のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The solvent used in the contacting step has a ratio of an inorganic value to an organic value of 0.2 or more and 2.3 or less, and a boiling point of 40 ° C or more. The manufacturing method of the aliphatic dicarboxylic acid of any one.
- 接触工程より前に、微生物を除去する工程を含むことを特徴とする、請求項14から請求項21のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to any one of claims 14 to 21, further comprising a step of removing microorganisms before the contacting step. *
- 接触工程より前に、タンパク質を除去する工程を含むことを特徴とする、請求項14から請求項22のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to any one of claims 14 to 22, further comprising a step of removing the protein before the contacting step. *
- 接触工程でミキサーセトラーを用い、該ミキサーセトラーのセトラー部から、抽出相、抽残相および固形分を含む相を回収することを特徴とする、請求項14から請求項23のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 24. The method according to any one of claims 14 to 23, wherein a mixer settler is used in the contact step, and an extraction phase, a residual phase and a phase containing solid content are recovered from a settling portion of the mixer setler. The manufacturing method of aliphatic dicarboxylic acid of description.
- 前記セトラー部から回収した固形分を含む相を固液分離し、液相を回収することを特徴とする請求項24に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to claim 24, wherein the phase containing the solid content recovered from the settler part is subjected to solid-liquid separation, and the liquid phase is recovered.
- 接触工程で更に向流多段抽出塔を用い、セトラー部で得られた抽残相から脂肪族ジカルボン酸を回収することを特徴とする、請求項24または請求項25に記載の脂肪族ジカルボン酸の製造方法。 The aliphatic dicarboxylic acid according to claim 24 or 25, wherein an aliphatic dicarboxylic acid is recovered from the extracted residual phase obtained in the settler section using a countercurrent multistage extraction column in the contacting step. Production method.
- 晶析工程において冷却を行うことを特徴とする、請求項14から請求項26のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to any one of claims 14 to 26, wherein cooling is performed in the crystallization step.
- 晶析工程が減圧操作を含むことを特徴とする、請求項14から請求項27のいずれか1項に記載の脂肪族ジカルボン酸の製造方法。 The method for producing an aliphatic dicarboxylic acid according to any one of claims 14 to 27, wherein the crystallization step includes a decompression operation.
- 生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(6)を含む脂肪族ジカルボン酸を製造する方法。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物菌体とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物菌体を分離する菌体分離工程
(4)菌体分離工程で菌体を分離して得られた水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(5)前記水溶液と前記溶剤の接触により発生した固形分を除去する固形分除去工程
(6)前記溶剤を相分離により分離する相分離工程 A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism, comprising the steps of (1) to (6) below: How to manufacture.
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction liquid by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step A pH adjusting step for obtaining a liquid having a pH of 1.0 or more and 5.0 or less by mixing a solution containing the microbial cells and an acid (3) Bacteria for separating microbial cells from the liquid obtained in the pH adjusting step Body separation step (4) Contact step of contacting the aqueous solution obtained by separating the cells in the cell separation step with a solvent capable of phase separation with the aqueous solution (5) Generated by contact between the aqueous solution and the solvent Solid content removal step for removing solid content (6) Phase separation step for separating the solvent by phase separation - 生物由来原料と微生物とを反応させて得られる脂肪族ジカルボン酸を含む水溶液から、脂肪族ジカルボン酸を製造する方法であって、以下の工程(1)~(8)を含む脂肪族ジカルボン酸を製造する方法。
(1)生物由来原料と微生物とを反応させることにより脂肪族ジカルボン酸を反応液中に蓄積させる脂肪族ジカルボン酸生成工程
(2)脂肪族ジカルボン酸生成工程で得られた脂肪族ジカルボン酸と微生物菌体とを含む液と、酸とを混合することにより、pH1.0以上5.0以下の液を得るpH調整工程
(3)pH調整工程で得られた液から微生物菌体を分離する菌体分離工程
(4)菌体分離工程で菌体を分離して得られた水溶液と、該水溶液と相分離可能な溶剤とを接触させる接触工程
(5)接触工程で回収された脂肪族ジカルボン酸を濃縮する工程であって、該濃縮により抽出相中の水濃度が上昇する抽出相濃縮工程
(6)抽出相濃縮工程後の液から脂肪族ジカルボン酸を析出させる晶析工程
(7)晶析工程で析出した脂肪族ジカルボン酸を回収する固液分離工程
(8)固液分離工程で得られた脂肪族ジカルボン酸を回収した後の晶析母液の少なくとも一部を、晶析工程より前のいずれかの工程に戻す晶析母液リサイクル工程 A method for producing an aliphatic dicarboxylic acid from an aqueous solution containing an aliphatic dicarboxylic acid obtained by reacting a biological material with a microorganism, comprising the steps of (1) to (8) below: How to manufacture.
(1) Aliphatic dicarboxylic acid production step for accumulating aliphatic dicarboxylic acid in the reaction solution by reacting biological material and microorganisms (2) Aliphatic dicarboxylic acid and microorganism obtained in the aliphatic dicarboxylic acid production step A pH adjusting step for obtaining a liquid having a pH of 1.0 or more and 5.0 or less by mixing a solution containing the microbial cells and an acid (3) Bacteria for separating microbial cells from the liquid obtained in the pH adjusting step Body separation step (4) Contact step in which the aqueous solution obtained by separating the cells in the cell separation step is brought into contact with a solvent capable of phase separation with the aqueous solution (5) Aliphatic dicarboxylic acid recovered in the contact step In which the concentration of water in the extraction phase increases due to the concentration (6) Crystallization step (7) Crystallization of depositing aliphatic dicarboxylic acid from the liquid after the extraction phase concentration step Aliphatic dicarbo precipitated in the process Solid-liquid separation step for recovering acid (8) Crystal for returning at least a part of the crystallization mother liquor after recovering the aliphatic dicarboxylic acid obtained in the solid-liquid separation step to any step before the crystallization step Analysis mother liquor recycling process
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2015118111A2 (en) | 2014-02-07 | 2015-08-13 | Basf Se | Modified microorganism with improved biomass separation behaviour |
WO2015117916A1 (en) | 2014-02-07 | 2015-08-13 | Basf Se | Improved microorganisms for succinic acid production |
WO2018211093A1 (en) | 2017-05-19 | 2018-11-22 | Basf Se | Process for producing an organic compound |
EP3502241A1 (en) | 2017-12-21 | 2019-06-26 | Basf Se | Modified microorganism for improved production of succinate |
Families Citing this family (2)
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EP3950680A4 (en) | 2019-03-25 | 2022-11-30 | Toray Industries, Inc. | Method for producing 3-hydroxyadipic acid-3,6-lactone |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0330685A (en) * | 1989-06-28 | 1991-02-08 | Michigan Biotechnol Inst | Fermentation and purification of succinic acid |
JPH03180187A (en) * | 1989-12-08 | 1991-08-06 | Kyowa Hakko Kogyo Co Ltd | Production of l-malic acid |
JPH04271787A (en) * | 1991-02-28 | 1992-09-28 | Mitsubishi Petrochem Co Ltd | Production of d-lactic acid and genus pseudomonas bacterium |
JPH053780A (en) * | 1991-06-26 | 1993-01-14 | Mitsui Toatsu Chem Inc | Method for separating microorganism |
JPH0568582A (en) * | 1991-09-09 | 1993-03-23 | Nikko Kyodo Co Ltd | Method for treating culture solution for secreting and producing hirudins |
JP2005027533A (en) * | 2003-07-09 | 2005-02-03 | Mitsubishi Chemicals Corp | Method for producing organic acid |
JP2006328374A (en) * | 2005-04-26 | 2006-12-07 | Mitsubishi Chemicals Corp | Manufacturing method of polyester |
JP2008094884A (en) * | 2006-10-06 | 2008-04-24 | Mitsubishi Chemicals Corp | Stretched film made of polyester derived from biomass resource and method for producing the same |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS50117714A (en) * | 1974-03-02 | 1975-09-16 | ||
US4544477A (en) * | 1983-10-12 | 1985-10-01 | Standard Oil Company | Polar solvent extraction and dedusting process |
GB8828695D0 (en) * | 1988-12-08 | 1989-01-11 | Exxon Chemical Patents Inc | Minimising catalyst loss in production of alcohols |
US5426219A (en) * | 1993-07-26 | 1995-06-20 | A.E. Staley Manufacturing Co. | Process for recovering organic acids |
JP2008035732A (en) * | 2006-08-02 | 2008-02-21 | Toyota Central Res & Dev Lab Inc | Method for producing organic acid |
-
2010
- 2010-10-07 BR BR112012007968A patent/BR112012007968A2/en not_active Application Discontinuation
- 2010-10-07 JP JP2011535468A patent/JP5724876B2/en active Active
- 2010-10-07 WO PCT/JP2010/067694 patent/WO2011043443A1/en active Application Filing
-
2015
- 2015-04-02 JP JP2015075950A patent/JP5991400B2/en active Active
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0330685A (en) * | 1989-06-28 | 1991-02-08 | Michigan Biotechnol Inst | Fermentation and purification of succinic acid |
JPH03180187A (en) * | 1989-12-08 | 1991-08-06 | Kyowa Hakko Kogyo Co Ltd | Production of l-malic acid |
JPH04271787A (en) * | 1991-02-28 | 1992-09-28 | Mitsubishi Petrochem Co Ltd | Production of d-lactic acid and genus pseudomonas bacterium |
JPH053780A (en) * | 1991-06-26 | 1993-01-14 | Mitsui Toatsu Chem Inc | Method for separating microorganism |
JPH0568582A (en) * | 1991-09-09 | 1993-03-23 | Nikko Kyodo Co Ltd | Method for treating culture solution for secreting and producing hirudins |
JP2005027533A (en) * | 2003-07-09 | 2005-02-03 | Mitsubishi Chemicals Corp | Method for producing organic acid |
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JP2008094884A (en) * | 2006-10-06 | 2008-04-24 | Mitsubishi Chemicals Corp | Stretched film made of polyester derived from biomass resource and method for producing the same |
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JP5991400B2 (en) | 2016-09-14 |
JPWO2011043443A1 (en) | 2013-03-04 |
JP5724876B2 (en) | 2015-05-27 |
JP2015119738A (en) | 2015-07-02 |
BR112012007968A2 (en) | 2015-09-08 |
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